Insecticidal proteins and methods of use thereof

Abstract

The present invention relates to insecticidal proteins and methods of use thereof. The present disclosure provides compositions and methods for controlling pests. The methods involve transforming organisms with nucleic acid sequences that encode insecticidal proteins. In particular, the nucleic acid sequences can be used to make plants and microorganisms that have insecticidal activity. Thus, the present disclosure provides transformed bacteria, plants, plant cells, plant tissues, and seeds. Compositions are insecticidal nucleic acids and proteins of bacterial species. The sequences can be used to construct expression vectors for subsequent transformation into organisms of interest, including plants, as probes to isolate other homologous (or partially homologous) genes. The pesticidal proteins can be used to control, kill, or inhibit the growth of populations of Lepidopteran, Coleopteran, Dipteran, fungal, Hemipteran, and nematode pests, and to produce compositions having insecticidal activity.

Description

[0001] This application is a divisional application of Chinese patent application 201480061497.6, "Insecticidal Protein and Method of Using Thereof," filed on September 11, 2014.

[0002] References to sequence lists submitted electronically A sequence list named "5345PCT_sequence_listing.txt", created on August 28, 2014, and measuring 576 kilobytes in size, is submitted with this specification in a computer-readable form. This sequence list is part of this specification and is incorporated herein by reference in its entirety. Technical Field

[0003] This disclosure relates to the field of molecular biology. This disclosure provides novel genes encoding insecticidal proteins. These insecticidal proteins, along with the nucleic acid sequences encoding them, can be used to prepare insecticides and to produce transgenic insect-resistant plants. Background Technology

[0004] Biological control of agriculturally significant insect pests using microbial agents such as fungi, bacteria, or other insects offers an environmentally friendly and commercially attractive alternative to synthetic chemical pesticides. Generally, the use of biopesticides poses a lower risk of pollution and environmental harm, and they provide higher target specificity than traditional broad-spectrum chemical insecticides. Furthermore, biopesticides are often cheaper to produce, thus increasing the economic yield of many crops.

[0005] Known Bacillus genus ( Bacillus Certain species of microorganisms possess insecticidal activity against a range of insect pests, including Lepidoptera ( Lepidoptera Diptera ( Diptera ), Coleoptera ( Coleoptera ), Hemiptera ( Hemiptera Pests and other pests. Bacillus thuringiensis (Bt) Bacillus thuringiensis (Bt) and Bacillus juvenilee ( Bacillus popilliae (This is) the most successful biological control agent discovered to date. It has long been believed that insect pathogenicity is caused by Bacillus larvae (…). B. larv ae), Bacillus subtilis ( B. lentimorbus ), Bacillus spheroidae ( B. sphaericus ) and Bacillus cereus ( B. cereus These bacteria are caused by strains of bacteria. Microbial insecticides, especially those derived from Bacillus strains, have played an important role in agriculture as an alternative to chemical pest control.

[0006] In this field, crops have been genetically engineered to produce insecticidal proteins secreted by Bacillus subtilis, thereby developing crops with enhanced insect resistance. For example, corn and cotton plants have been genetically engineered to produce insecticidal proteins isolated from Bt strains. These genetically engineered crops are now widely used in agriculture, providing farmers with an environmentally friendly alternative to traditional insect control methods. While these genetically engineered insect-resistant crops have proven commercially successful, they are resistant only to a narrow range of economically important insect pests. In some cases, insects can develop resistance to different insecticidal compounds, necessitating the search for alternative biological control agents to manage pests.

[0007] Therefore, there is still a need for new insecticidal proteins with different ranges of insecticidal activity against insect pests, such as insecticidal proteins that are active against a variety of insects in the Lepidoptera and Coleoptera orders, including but not limited to insect pests that have developed resistance to existing insecticides. Summary of the Invention

[0008] This disclosure provides compositions and methods for conferring insecticidal activity to bacteria, plants, plant cells, tissues, and seeds. The compositions comprise nucleic acid molecules encoding sequences of insecticidal and insecticidal polypeptides, a vector containing those nucleic acid molecules, and a host cell containing said vector. The compositions also comprise insecticidal polypeptide sequences and antibodies against those polypeptides. The nucleic acid sequences can be used in DNA constructs or expression cassettes for transformation and expression in organisms, including microorganisms and plants. The nucleotide or amino acid sequences can be synthetic sequences designed for expression in organisms, including but not limited to microorganisms or plants. The compositions also comprise transformed bacteria, plants, plant cells, tissues, and seeds.

[0009] Specifically, this disclosure provides encoding Pseudomonas ( PseudomonasThis disclosure provides isolated or recombinant nucleic acid molecules of the insecticidal protein-72 (PIP-72) polypeptide, comprising amino acid substitutions, amino acid deletions, amino acid insertions, amino acid fragments, and combinations thereof. Furthermore, this disclosure also covers amino acid sequences corresponding to the PIP-72 polypeptide. This disclosure provides isolated or recombinant nucleic acid molecules capable of encoding the PIP-72 polypeptide shown in SEQ ID NO: 849, and amino acid substitutions, amino acid deletions, amino acid insertions, amino acid fragments, and combinations thereof. This disclosure also covers nucleic acid sequences complementary to the nucleic acid sequences in the embodiments herein, or nucleic acid sequences hybridized to the sequences in the embodiments herein. This disclosure also provides isolated or recombinant PIP-72 polypeptide shown in SEQ ID NO: 849, and amino acid substitutions, amino acid deletions, amino acid insertions, amino acid fragments, and combinations thereof.

[0010] This disclosure provides methods for producing the aforementioned polypeptides, and methods for using these polypeptides to control or kill lepidopteran, coleopteran, nematode, fungal, and / or dipteran pests. The transgenic plants of the embodiments express one or more of the insecticidal sequences disclosed herein. In various embodiments, the transgenic plants also contain one or more additional genes for insect resistance, for example, one or more additional genes for controlling coleopteran, lepidopteran, hemiptera, or nematode pests. Those skilled in the art will understand that the transgenic plants may contain any genes conferring agronomic traits of interest.

[0011] The disclosure also includes methods for detecting nucleic acids and peptides of the embodiments described above in a sample. A kit is also provided for detecting the presence of the PIP-72 peptide, or the presence of a nucleotide sequence encoding the PIP-72 peptide, in a sample. The kit is provided together with all reagents and control samples required to perform the method for detecting the expected factor, as well as instructions for use.

[0012] The compositions and methods described herein can be used to produce organisms with enhanced resistance or tolerance to pests. These organisms and compositions containing them are of great value to agricultural production. The compositions described herein can also be used to produce modified or improved proteins with insecticidal activity, or to detect the presence of PIP-72 peptides or nucleic acids in products or organisms.

[0013] This invention relates to the following technical solutions: 1. A DNA construct comprising a heterologous nucleic acid molecule, said heterologous nucleic acid molecule encoding a target for the western maize rootworm (Maize Rootworm). Diabrotica virgifera virgifera PIP-72 polypeptide with insecticidal activity.

[0014] 2. The DNA construct according to Scheme 1, wherein the encoded PIP-72 polypeptide comprises an amino acid sequence having at least 50% identity with SEQ ID NO:2.

[0015] 3. The DNA construct according to Scheme 1 or 2, wherein the encoded PIP-72 polypeptide, compared with the corresponding amino acid shown in SEQ ID NO: 2, is present at amino acid positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 42, 44, 45. Positions 46, 47, 48, 49, 50, 51, 52, 53, 54, 56, 57, 58, 59, 60, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, or 86 contain 1 to 45 amino acid substitutions.

[0016] 4. The DNA construct according to Scheme 1 or 2, wherein the encoded PIP-72 polypeptide is identical to SEQ ID NO: Compared to the corresponding amino acids shown in Figure 2, the amino acids include 1 to 45 amino acid substitutions at positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 17, 18, 19, 20, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 42, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 56, 58, 60, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, or 86.

[0017] 5. The DNA construct according to Scheme 1, 2, or 4, wherein the encoded PIP-72 polypeptide comprises the amino acid sequence shown below: in The second Xaa is Gly, Ala, Cys, Asp, Glu, Ile, Lys, Leu, Asn, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 3 is either Ile or Trp; The fourth Xaa is Thr, Ala, Asp, Glu, His, Ile, Lys, Leu, Arg, Ser, Val, Trp, or Tyr; The Xaa in the 5th position is Val, Ala, Cys, Gly, His, Ile, or Tyr; The Xaa at position 6 is Thr, Ala, Cys, Phe, Gly, His, Ile, Lys, Met, Pro, Gln, Arg, Ser, Trp, or Tyr; Xaa at position 7 is either Asn, Ala, or Val; The Xaa at position 8 is Asn, Ala, Cys, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Gln, Arg, Ser, Thr, or Val; The Xaa at position 9 is either Ser, Ala, Cys, Gly, or Thr; The Xaa at the 10th position is Ser, Ala, Glu, Phe, Gly, His, Ile, Lys, Leu, Asn, Pro, Gln, Arg, Thr, or Trp; The Xaa at position 11 is Asn, Ala, Cys, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Gln, Ser, Thr, Val, or Tyr; The Xaa at position 12 is Pro, Ala, Cys, Asp, Glu, Gly, His, Lys, Leu, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 13 is Ile, Asn, Gln, or Val; Xaa at position 14 is Glu, Ala, Cys, Phe, His, Lys, or Gln; Xaa at position 15 is Val, Ala, Cys, Ile, Met, or Arg; Xaa at position 17 is Ile, Glu, or Val; Xaa at position 18 is either Asn or Ser; The Xaa at position 19 is His, Ala, Glu, Lys, Leu, Pro, Arg, Ser, or Tyr; Xaa at position 20 is Trp, Ala, or Thr; Xaa at position 22 is Ser, Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Thr, Val, or Tyr; Xaa at position 23 is Asp, Ala, Gly, His, Lys, Met, Asn, Gln, Ser, Thr, or Val; Xaa at position 24 is Gly, Asp, or Phe; Xaa at position 25 is Asp, Ala, Glu, Phe, Asn, or Gln; Xaa at position 26 is Thr, Glu, or Pro; Xaa at position 27 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Asn, Gln, Arg, or Thr; Xaa at position 28 is Phe, Pro, Trp, or Tyr; Xaa at position 29 is Phe, Ala, Cys, Ile, Leu, Gln, Arg, Trp, or Tyr; The Xaa at position 30 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Lys, Leu, Met, Asn, Pro, Gln, Arg, Thr, Val, Trp, or Tyr; Xaa at position 31 is Val, Ile, or Leu; Xaa at position 32 is Gly, Ala, Asp, Glu, Phe, His, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 33 is Asn, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Pro, Gln, Arg, Ser, Thr, Val, or Tyr; Xaa at position 34 is Gly, Glu, Phe, His, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Tyr; Xaa at position 35 is Lys, Ala, Cys, Asp, Gly, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Val; Xaa at position 36 is Gln, Ala, Cys, Glu, Gly, His, Ile, Lys, Leu, Asn, Pro, Arg, Ser, Thr, or Val; Xaa at position 37 is Glu, Ala, Cys, Asp, Phe, Gly, Ile, Lys, Leu, Met, Asn, Ser, Thr, or Val; Xaa at position 38 is Thr, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Val, Trp, or Tyr; Xaa at position 39 is either Trp or Phe; The Xaa at position 40 is Asp, Ala, Cys, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 42 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, Ile, Lys, Leu, Met, Asn, Gln, Arg, Thr, Val, Trp, or Tyr; Xaa at position 44 is Ser, Ala, Asp, Glu, Gly, Leu, Met, Asn, Pro, Gln, Thr, Val, or Tyr; Xaa at position 45 is Arg, Lys, or Ser; Xaa at position 46 is Gly, Ala, or Gln; Xaa at position 47 is Phe, Cys, Val, or Tyr; Xaa at position 48 is Val, Ile, or Leu; Xaa at position 49 is Leu, Cys, Phe, Met, Arg, or Tyr; Xaa at position 50 is Ser, Ala, Cys, Asp, Ile, Met, Pro, Gln, Thr, or Val; Xaa at position 51 is Leu, Ala, Cys, Met, or Val; Xaa at position 52 is Lys, Cys, Phe, His, Ile, Leu, Met, Asn, Arg, Ser, Thr, Trp, or Tyr; Xaa at position 53 is Lys, Ala, Cys, Asp, Glu, Phe, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, or Tyr; Xaa at position 54 is Asn, Cys, Asp, Glu, Phe, Gly, Lys, Met, Gln, Arg, Ser, or Trp; Xaa at position 56 is Ala, Gly, Leu, Asn, Pro, Gln, Arg, Ser, or Thr; Xaa at position 57 is Gln, Glu, Leu, Met, Ser, or Thr; Xaa at position 58 is His, Ala, Asp, Phe, Leu, Met, Asn, Arg, Trp, or Tyr; Xaa at position 60 is Tyr, Glu, or Phe; Xaa at position 63 is Gln, Cys, Gly, Ile, Leu, Met, Asn, Thr, Val, or Tyr; Xaa at position 64 is Ala, Phe, Gly, His, Arg, Ser, or Tyr; Xaa at position 65 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Leu, Asn, Thr, or Val; Xaa at position 66 is Ser, Ala, or Gly; Xaa at position 67 is Lys, Ala, Cys, Asp, Phe, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 68 is Ile, Asp, Leu, or Val; Xaa at position 69 is Glu, Ala, Cys, Asp, Phe, His, Ile, Leu, Met, Gln, Arg, Ser, Thr, Val, or Tyr; Xaa at position 70 is Val, Cys, or Ile; Xaa at position 71 is Asp, Ala, Cys, Gly, His, Ile, Leu, Met, Asn, Ser, Thr, Val, or Tyr; Xaa at position 72 is Asn, Ala, Cys, Asp, Glu, Gly, Lys, Met, Pro, Gln, Arg, Ser, Thr, Val, or Trp; Xaa at position 73 is Asn, Ala, Cys, Asp, Phe, Gly, His, Ile, Leu, Ser, Thr, Val, or Tyr; Xaa at position 74 is Ala, Cys, Asp, Phe, Gly, His, Ile, Leu, Asn, Gln, Arg, Ser, Thr, Val, or Tyr; Xaa at position 75 is Val, Cys, Ile, or Leu; Xaa at position 76 is Lys, Ala, Cys, Phe, His, Ile, Leu, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 77 is Asp or Tyr; Xaa at position 78 is Gln, Ala, Cys, Asp, Phe, Gly, His, Ile, Leu, Met, Asn, Arg, Ser, Thr, Val, or Tyr; Xaa at position 79 is Gly, Arg, Ala, Cys, Asp, Glu, Phe, His, Lys, Leu, Asn, Gln, Arg, Ser, Thr, Trp, or Tyr; Xaa at position 80 is Arg, Ala, Cys, Asp, Phe, Gly, His, Ile, Leu, Asn, Ser, Thr, Val, or Tyr; Xaa at position 81 is Leu, Ala, Cys, Asp, Phe, Gly, His, Ile, Asn, Pro, Arg, Ser, Thr, or Val; Xaa at position 82 is Ile, Ala, Leu, Met, Arg, or Val; Xaa at position 83 is Glu, Ala, Cys, Asp, Phe, Gly, His, Ile, Lys, Leu, Asn, Pro, Arg, Ser, Thr, Val, or Tyr; Xaa at position 84 is Pro, Ala, Cys, Glu, Ile, Ser, Val, Trp, or Tyr; Xaa at position 85 is Leu, Cys, Gly, or Val; and Xaa at position 86 is either Ser, Ala, Ile, Thr, or Val.

[0018] 6. The DNA construct according to Scheme 1, 2 or 3, wherein the encoded PIP-72 polypeptide comprises the amino acid sequence shown below: in The second Xaa is Gly, Ala, Cys, Asp, Glu, Ile, Lys, Leu, Asn, Arg, Ser, Thr, Val, Trp, or Tyr; The Xaa in the third position is Ile, Leu, Val, or Trp; The fourth Xaa is Thr, Ala, Asp, Glu, His, Ile, Lys, Leu, Arg, Ser, Val, Trp, or Tyr; The Xaa in the 5th position is Val, Ala, Cys, Gly, His, Ile, Leu, or Tyr; The Xaa at position 6 is Thr, Ala, Cys, Phe, Gly, His, Ile, Lys, Met, Pro, Gln, Arg, Ser, Trp, or Tyr; Xaa at position 7 is either Asn, Ala, or Val; The Xaa at position 8 is Asn, Lys, Gly, Ser, Gln, Arg, Thr, Ala, Cys, Asp, Glu, His, Ile, Leu, Met, or Val; The Xaa at position 9 is either Ser, Ala, Cys, Gly, or Thr; The Xaa at position 11 is Asn, Lys, Thr, Gln, Arg, Ser, Ala, Cys, Asp, Glu, Gly, His, Ile, Leu, Met, Val, or Tyr; The Xaa at position 12 is Pro, Thr, Lys, Ser, Arg, Ala, Cys, Asp, Glu, Gly, His, Leu, Asn, Gln, Arg, Val, Trp, or Tyr; Xaa at position 13 is Ile, Asn, Gln, Leu, or Val; Xaa at position 14 is Glu, Ala, Cys, Phe, His, Lys, Asp, or Gln; The Xaa at position 15 is Val, Ala, Ile, Leu, Cys, Met, or Arg; Xaa at position 16 is either Ala or Ser; Xaa at position 17 is Ile, Glu, Leu, or Val; Xaa at position 18 is Asn, Gln, Thr, or Ser; The Xaa at position 19 is His, Lys, Ala, Arg, Glu, Leu, Pro, Ser, or Tyr; Xaa at position 20 is Trp, Ala, or Thr; Xaa at position 21 is Gly, Arg, or Lys; Xaa at position 22 is Ser, Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Thr, Val, or Tyr; Xaa at position 23 is Asp, Ala, Gly, His, Lys, Met, Asn, Gln, Ser, Thr, or Val; Xaa at position 24 is Gly, Asp, or Phe; Xaa at position 25 is Asp, Ala, Glu, Phe, Asn, or Gln; Xaa at position 26 is Thr, Glu, Asp, Ser, or Pro; Xaa at position 27 is Ser, Thr, Lys, Arg, Ala, Cys, Asp, Glu, Phe, Gly, His, Asn, or Gln; Xaa at position 28 is Phe, Tyr, Pro, or Trp; Xaa at position 29 is Phe, Ala, Cys, Ile, Leu, Gln, Arg, Trp, or Tyr; The Xaa at position 30 is Ser, Gly, Lys, Thr, Arg, Ala, Cys, Asp, Glu, Phe, His, Leu, Met, Asn, Pro, Gln, Val, Trp, or Tyr; Xaa at position 31 is Val, Ile, Met, or Leu; Xaa at position 32 is Gly, Ala, Asp, Glu, Phe, His, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 33 is Asn, Ser, Gln, Pro, Thr, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Arg, Val, or Tyr; Xaa at position 34 is Gly, Glu, Phe, His, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Tyr; Xaa at position 35 is Lys, Glu, Ala, Cys, Asp, Gly, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Val; Xaa at position 36 is Gln, Ala, Cys, Glu, Gly, His, Ile, Lys, Leu, Asn, Pro, Arg, Ser, Thr, or Val; Xaa at position 37 is Glu, Asp, Ala, Cys, Phe, Gly, Ile, Lys, Leu, Met, Asn, Ser, Thr, or Val; Xaa at position 38 is Thr, Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Gln, Arg, Val, Trp, or Tyr; Xaa at position 39 is either Trp or Phe; The Xaa at position 40 is Asp, Ala, Cys, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 42 is Ser, Asn, Thr, Ala, Cys, Asp, Glu, Phe, Gly, Ile, Lys, Leu, Met, Arg, Val, Trp, Tyr, or Gln; Xaa at position 44 is Ser, Asp, Ala, Leu, Thr, Glu, Ile, Ala, Gly, Leu, Met, Asn, Pro, Gln, Val, Tyr, or Val; Xaa at position 45 is Arg, Lys, or Ser; Xaa at position 46 is Gly, Ala, or Gln; Xaa at position 47 is Phe, Tyr, Cys, Val, or Trp; Xaa at position 48 is Leu, Met, Ile, Cys, Phe, Met, Arg, Tyr, or Val; Xaa at position 49 is Leu, Met, Ile, or Val; Xaa at position 50 is Ser, Ala, Tyr, Cys, Asp, Ile, Met, Pro, Gln, Val, or Thr; Xaa at position 51 is Leu, Val, Ala, Cys, Met, or Ile; Xaa at position 52 is Lys, Cys, Phe, His, Ile, Leu, Met, Asn, Arg, Ser, Thr, Gln, Trp, or Tyr; Xaa at position 53 is Lys, Arg, Met, Leu, Ile, Ala, Cys, Asp, Glu, Phe, His, Asn, Gln, Ser, Thr, Tyr, or Val; Xaa at position 54 is Asn, Cys, Asp, Glu, Phe, Gly, Lys, Met, Gln, Arg, Ser, or Trp; Xaa at position 55 is Gly, Ser, or Thr; Xaa at position 56 is Ala, Thr, Gln, Ser, Gly, Leu, Pro, Arg, or Asn; Xaa at position 57 is Gln, Glu, Leu, Met, Ser, Val, Ala, Asn, Ile, or Thr; Xaa at position 58 is His, Ala, Lys, Asp, Phe, Leu, Met, Asn, Arg, Trp, Tyr, or Thr; Xaa at position 59 is Pro, Thr, or Ser; Xaa at position 60 is Tyr, Glu, or Phe; Xaa at position 62 is Val, Ile, or Leu; Xaa at position 63 is Gln, Ser, Cys, Gly, Ile, Leu, Met, Asn, Thr, Val, or Tyr; Xaa at position 64 is Ala, Gln, Asn, Phe, Gly, His, Arg, Ser, or Tyr; Xaa at position 65 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Leu, Asn, Val, or Thr; Xaa at position 66 is Ser, Ala, or Gly; Xaa at position 67 is Lys, Gln, Asn, or Arg; Xaa at position 67 is Lys, Ala, Cys, Asp, Phe, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 68 is Ile, Asp, Leu, or Val; Xaa at position 69 is Glu, Ala, Cys, Asp, Phe, His, Ile, Leu, Met, Gln, Arg, Ser, Thr, Val, or Tyr; Xaa at position 70 is Val, Ile, Cys, or Leu; Xaa at position 71 is Asp, Glu, Tyr, Ala, Cys, Gly, His, Ile, Leu, Met, Asn, Ser, Thr, Val, or Trp; Xaa at position 72 is Asn, Ala, Cys, Asp, Glu, Gly, Lys, Met, Pro, Gln, Arg, Ser, Thr, Val, His, or Trp; Xaa at position 73 is Asn, Ser, Asp, Gln, Thr, Ala, Cys, Phe, Gly, His, Ile, Leu, Val, Tyr, or Glu; Xaa at position 74 is Ala, Thr, Met, Ile, Lys, Ser, Leu, Val, Cys, Asp, Phe, Gly, His, Asn, Gln, Tyr, or Arg; Xaa at position 75 is Val, Cys, Ile, or Leu; Xaa at position 76 is Lys, Ala, Cys, Phe, His, Ile, Leu, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 77 is Asp or Tyr; Xaa at position 78 is Gln, His, Ser, Asn, Ala, Cys, Asp, Phe, Gly, Ile, Leu, Met, Asn, Arg, Val, Tyr, or Thr; Xaa at position 79 is Gly, Arg, Ala, Cys, Asp, Glu, Phe, His, Lys, Leu, Asn, Gln, Arg, Ser, Thr, Trp, or Tyr; Xaa at position 80 is Arg, Glu, Gln, Lys, Asp, Ala, Cys, Phe, Gly, His, Ile, Leu, Ser, Thr, Val, Tyr, or Asn; Xaa at position 81 is Leu, Pro, Thr, Ile, Val, Ala, Cys, Asp, Phe, Gly, His, or Ser; Xaa at position 82 is Ile, Ala, Leu, Met, Arg, and Val; Xaa at position 83 is Glu, His, Asn, Leu, Gln, Ile, Ala, Cys, Asp, Phe, Gly, Lys, Pro, Arg, Ser, Thr, Tyr, or Val; Xaa at position 84 is Pro, Ala, Cys, Glu, Ile, Ser, Val, Trp, or Tyr; Xaa at position 85 is Leu, Val, Cys, Gly, or Ala; and Xaa at position 86 is either Ser, Ala, Tyr, Asn, Ile, Val, or Thr.

[0019] 7. The DNA construct according to any one of schemes 1, 2, 3 and 4, wherein the encoded PIP-72 polypeptide comprises an amino acid motif represented by positions 37 to 51 of the following sequences: SEQ ID NO: 846, SEQ ID NO: 847, SEQ ID NO: 848 or SEQ ID NO: 849.

[0020] 8. A DNA construct comprising: A polynucleotide encoding a polypeptide having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher sequence identity with the following amino acid sequences: SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 30, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 929, SEQ ID NO: 930, SEQ ID NO: 931, SEQ ID NO: 937, SEQ ID NO: 938, SEQ ID NO: 942, SEQ ID NO: 947 or SEQ ID NO: 948; and A heterologous regulatory sequence, wherein the polynucleotide is operatively linked to the heterologous regulatory sequence.

[0021] 9. A DNA construct comprising a chimeric nucleic acid molecule encoding a chimeric PIP-72 polypeptide, the chimeric PIP-72 polypeptide comprising at least a first portion and a second portion, the first portion comprising a first PIP-72 polypeptide moiety, the second portion comprising a complementary second PIP-72 polypeptide moiety, wherein the first PIP-72 polypeptide and the second PIP-72 polypeptide have different amino acid sequences in the respective portions.

[0022] 10. An isolated polynucleotide comprising a nucleic acid molecule, said nucleic acid molecule encoding a receptor for the western maize rootworm (…). Diabrotica virgifera virgifera PIP-72 polypeptide with insecticidal activity.

[0023] 11. The isolated polynucleotide according to Scheme 10, wherein the encoded PIP-72 polypeptide comprises an amino acid sequence having at least 50% identity with SEQ ID NO: 2.

[0024] 12. The isolated polynucleotide according to Scheme 10 or 11, wherein the encoded PIP-72 polypeptide, compared with the corresponding amino acid shown in SEQ ID NO: 2, is present at amino acid positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 42, 44, 45. Positions 46, 47, 48, 49, 50, 51, 52, 53, 54, 56, 57, 58, 59, 60, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, or 86 contain 1 to 45 amino acid substitutions.

[0025] 13. The isolated polynucleotide according to scheme 10 or 11, wherein the encoded PIP-72 polypeptide is associated with SEQ ID NO: Compared to the corresponding amino acids shown in Figure 2, the amino acids include 1 to 45 amino acid substitutions at positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 17, 18, 19, 20, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 42, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 56, 58, 60, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, and 86.

[0026] 14. The isolated polynucleotide according to any one of schemes 10, 11 and 13, wherein the encoded PIP-72 polypeptide comprises the amino acid sequence shown in the following formula: in The second Xaa is Gly, Ala, Cys, Asp, Glu, Ile, Lys, Leu, Asn, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 3 is either Ile or Trp; The fourth Xaa is Thr, Ala, Asp, Glu, His, Ile, Lys, Leu, Arg, Ser, Val, Trp, or Tyr; The Xaa in the 5th position is Val, Ala, Cys, Gly, His, Ile, or Tyr; The Xaa at position 6 is Thr, Ala, Cys, Phe, Gly, His, Ile, Lys, Met, Pro, Gln, Arg, Ser, Trp, or Tyr; Xaa at position 7 is either Asn, Ala, or Val; The Xaa at position 8 is Asn, Ala, Cys, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Gln, Arg, Ser, Thr, or Val; The Xaa at position 9 is either Ser, Ala, Cys, Gly, or Thr; The Xaa at the 10th position is Ser, Ala, Glu, Phe, Gly, His, Ile, Lys, Leu, Asn, Pro, Gln, Arg, Thr, or Trp; The Xaa at position 11 is Asn, Ala, Cys, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Gln, Ser, Thr, Val, or Tyr; The Xaa at position 12 is Pro, Ala, Cys, Asp, Glu, Gly, His, Lys, Leu, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 13 is Ile, Asn, Gln, or Val; Xaa at position 14 is Glu, Ala, Cys, Phe, His, Lys, or Gln; Xaa at position 15 is Val, Ala, Cys, Ile, Met, or Arg; Xaa at position 17 is Ile, Glu, or Val; Xaa at position 18 is either Asn or Ser; The Xaa at position 19 is His, Ala, Glu, Lys, Leu, Pro, Arg, Ser, or Tyr; Xaa at position 20 is Trp, Ala, or Thr; Xaa at position 22 is Ser, Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Thr, Val, or Tyr; Xaa at position 23 is Asp, Ala, Gly, His, Lys, Met, Asn, Gln, Ser, Thr, or Val; Xaa at position 24 is Gly, Asp, or Phe; Xaa at position 25 is Asp, Ala, Glu, Phe, Asn, or Gln; Xaa at position 26 is Thr, Glu, or Pro; Xaa at position 27 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Asn, Gln, Arg, or Thr; Xaa at position 28 is Phe, Pro, Trp, or Tyr; Xaa at position 29 is Phe, Ala, Cys, Ile, Leu, Gln, Arg, Trp, or Tyr; The Xaa at position 30 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Lys, Leu, Met, Asn, Pro, Gln, Arg, Thr, Val, Trp, or Tyr; Xaa at position 31 is Val, Ile, or Leu; Xaa at position 32 is Gly, Ala, Asp, Glu, Phe, His, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 33 is Asn, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Pro, Gln, Arg, Ser, Thr, Val, or Tyr; Xaa at position 34 is Gly, Glu, Phe, His, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Tyr; Xaa at position 35 is Lys, Ala, Cys, Asp, Gly, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Val; Xaa at position 36 is Gln, Ala, Cys, Glu, Gly, His, Ile, Lys, Leu, Asn, Pro, Arg, Ser, Thr, or Val; Xaa at position 37 is Glu, Ala, Cys, Asp, Phe, Gly, Ile, Lys, Leu, Met, Asn, Ser, Thr, or Val; Xaa at position 38 is Thr, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Val, Trp, or Tyr; Xaa at position 39 is either Trp or Phe; The Xaa at position 40 is Asp, Ala, Cys, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 42 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, Ile, Lys, Leu, Met, Asn, Gln, Arg, Thr, Val, Trp, or Tyr; Xaa at position 44 is Ser, Ala, Asp, Glu, Gly, Leu, Met, Asn, Pro, Gln, Thr, Val, or Tyr; Xaa at position 45 is Arg, Lys, or Ser; Xaa at position 46 is Gly, Ala, or Gln; Xaa at position 47 is Phe, Cys, Val, or Tyr; Xaa at position 48 is Val, Ile, or Leu; Xaa at position 49 is Leu, Cys, Phe, Met, Arg, or Tyr; Xaa at position 50 is Ser, Ala, Cys, Asp, Ile, Met, Pro, Gln, Thr, or Val; Xaa at position 51 is Leu, Ala, Cys, Met, or Val; Xaa at position 52 is Lys, Cys, Phe, His, Ile, Leu, Met, Asn, Arg, Ser, Thr, Trp, or Tyr; Xaa at position 53 is Lys, Ala, Cys, Asp, Glu, Phe, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, or Tyr; Xaa at position 54 is Asn, Cys, Asp, Glu, Phe, Gly, Lys, Met, Gln, Arg, Ser, or Trp; Xaa at position 56 is Ala, Gly, Leu, Asn, Pro, Gln, Arg, Ser, or Thr; Xaa at position 57 is Gln, Glu, Leu, Met, Ser, or Thr; Xaa at position 58 is His, Ala, Asp, Phe, Leu, Met, Asn, Arg, Trp, or Tyr; Xaa at position 60 is Tyr, Glu, or Phe; Xaa at position 63 is Gln, Cys, Gly, Ile, Leu, Met, Asn, Thr, Val, or Tyr; Xaa at position 64 is Ala, Phe, Gly, His, Arg, Ser, or Tyr; Xaa at position 65 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Leu, Asn, Thr, or Val; Xaa at position 66 is Ser, Ala, or Gly; Xaa at position 67 is Lys, Ala, Cys, Asp, Phe, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 68 is Ile, Asp, Leu, or Val; Xaa at position 69 is Glu, Ala, Cys, Asp, Phe, His, Ile, Leu, Met, Gln, Arg, Ser, Thr, Val, or Tyr; Xaa at position 70 is Val, Cys, or Ile; Xaa at position 71 is Asp, Ala, Cys, Gly, His, Ile, Leu, Met, Asn, Ser, Thr, Val, or Tyr; Xaa at position 72 is Asn, Ala, Cys, Asp, Glu, Gly, Lys, Met, Pro, Gln, Arg, Ser, Thr, Val, or Trp; Xaa at position 73 is Asn, Ala, Cys, Asp, Phe, Gly, His, Ile, Leu, Ser, Thr, Val, or Tyr; Xaa at position 74 is Ala, Cys, Asp, Phe, Gly, His, Ile, Leu, Asn, Gln, Arg, Ser, Thr, Val, or Tyr; Xaa at position 75 is Val, Cys, Ile, or Leu; Xaa at position 76 is Lys, Ala, Cys, Phe, His, Ile, Leu, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 77 is Asp or Tyr; Xaa at position 78 is Gln, Ala, Cys, Asp, Phe, Gly, His, Ile, Leu, Met, Asn, Arg, Ser, Thr, Val, or Tyr; Xaa at position 79 is Gly, Arg, Ala, Cys, Asp, Glu, Phe, His, Lys, Leu, Asn, Gln, Arg, Ser, Thr, Trp, or Tyr; Xaa at position 80 is Arg, Ala, Cys, Asp, Phe, Gly, His, Ile, Leu, Asn, Ser, Thr, Val, or Tyr; Xaa at position 81 is Leu, Ala, Cys, Asp, Phe, Gly, His, Ile, Asn, Pro, Arg, Ser, Thr, or Val; Xaa at position 82 is Ile, Ala, Leu, Met, Arg, or Val; Xaa at position 83 is Glu, Ala, Cys, Asp, Phe, Gly, His, Ile, Lys, Leu, Asn, Pro, Arg, Ser, Thr, Val, or Tyr; Xaa at position 84 is Pro, Ala, Cys, Glu, Ile, Ser, Val, Trp, or Tyr; Xaa at position 85 is Leu, Cys, Gly, or Val; and Xaa at position 86 is either Ser, Ala, Ile, Thr, or Val.

[0027] 15. The isolated polynucleotide according to any one of schemes 10, 11 and 13, wherein the encoded PIP-72 polypeptide comprises the amino acid sequence shown in the following formula: in The second Xaa is Gly, Lys, or Ala; Xaa in the 3rd position is either Ile or Leu; Xaa at position 4 is Thr or Ser; Xaa at position 5 is Val or Ile; Xaa at position 6 is Thr or Lys; Xaa at position 8 is either Asn, Lys, Gly, or Ser; Xaa at position 9 is either Ser or Ala; Xaa at position 11 is either Asn, Lys, His, or Thr; Xaa at the 12th position is Pro, Thr, Lys, or Ser; Xaa at position 13 is either Ile or Val; Xaa at position 14 is either Glu or Asp; Xaa at position 15 is Val, Ala, or Ile; Xaa at position 16 is either Ala or Ser; Xaa at position 17 is either Ile or Val; Xaa at position 18 is either Asn or Ser; Xaa at position 19 is His, Lys, Arg, Gln, or Ala; Xaa at position 21 is either Gly or Arg; Xaa at position 22 is Ser, Lys, Asn, Asp, or Thr; Xaa at position 25 is either Asp or Asn; Xaa at position 26 is Thr or Asp; Xaa at position 27 is Ser, Thr, Asn, or Lys; Xaa at position 28 is Phe, Tyr, or Pro; Xaa at position 29 is either Phe or Tyr; Xaa at position 30 is Ser, Gly, or Lys; Xaa at position 31 is Val, Ile, or Met; Xaa at position 32 is Gly, Ala, or Asp; Xaa at position 33 is Asn, Ser, Gln, or Pro; Xaa at position 35 is Lys, Glu, or Ser; Xaa at position 36 is Gln, Asn, or Ser; Xaa at position 37 is either Glu or Asp; Xaa at position 38 is Thr or Ser; Xaa at position 42 is either Ser or Asn; Xaa at position 44 is Ser, Asp, Ala, or Leu; Xaa at position 47 is either Phe or Tyr; Xaa at position 48 is either Leu or Met; Xaa at position 49 is either Leu or Met; Xaa at position 50 is Ser, Ala, or Tyr; Xaa at position 51 is either Leu or Val; Xaa at position 52 is either Lys or Gln; Xaa at position 53 is Lys, Arg, Met, or Leu; Xaa at position 54 is either Asn, Lys, or Gly; Xaa at position 55 is either Gly or Ser; Xaa at position 56 is Ala, Thr, Gln, or Ser; Xaa at position 57 is Gln, Val, or Ala; Xaa at position 58 is His, Ala, Lys, Tyr, or Thr; Xaa at position 59 is either Pro or Thr; Xaa at position 62 is either Val or Ile; Xaa at position 63 is Gln, Ser, or Leu; Xaa at position 64 is Ala, Gln, or Ser; Xaa at position 65 is either Ser or Thr; Xaa at position 67 is Lys, Gln, Arg, or Asn; Xaa at position 69 is Glu, Lys, or Val; Xaa at position 70 is either Val or Ile; Xaa at position 71 is Asp, Glu, or Tyr; Xaa at position 72 is either Asn, His, Ser, or Asp; Xaa at position 73 is Asn, Ser, or Asp; Xaa at position 74 is Ala, Thr, Met, Ile, or Lys; Xaa at position 76 is either Lys or Thr; Xaa at position 78 is Gln, His, or Ser; Xaa at position 80 is Arg, Glu, or Gln; Xaa at position 81 is Leu, Pro, Ala, or Thr; Xaa at position 82 is either Ile or Leu; Xaa at position 83 is Glu, His, Asn, Gln, or Leu; Xaa at position 85 is Leu, Val, or Ala; and Xaa at position 86 is Ser, Ala, Tyr, or Asn.

[0028] 16. The isolated polynucleotide according to any one of schemes 10, 11 and 13, wherein the encoded PIP-72 polypeptide comprises the amino acid sequence shown in the following formula: in The second Xaa is Gly, Lys, Ala, or Arg; The Xaa in the third position is Ile, Leu, or Val; Xaa at position 4 is Thr or Ser; The Xaa at the 5th position is Val, Ile, or Leu; Xaa at position 6 is Thr, Lys, Ser, or Arg; The Xaa at position 8 is either Asn, Lys, Gly, Ser, Gln, Arg, Thr, or Ala; Xaa at position 9 is Ser, Ala, or Thr; Xaa at position 11 is Asn, Lys, Thr, Gln, Arg, His, or Ser; Xaa at position 12 is Pro, Thr, Lys, Ser, or Arg; Xaa at position 13 is Ile, Val, or Leu; Xaa at position 14 is either Glu or Asp; Xaa at position 15 is Val, Ala, Ile, or Leu; Xaa at position 16 is either Ala or Ser; Xaa at position 17 is Ile, Val, or Leu; Xaa at position 18 is Asn, Ser, Gln, or Thr; Xaa at position 19 is His, Lys, Ala, Gln, Asn, or Arg; Xaa at position 21 is Gly, Arg, or Lys; Xaa at position 22 is Ser, Lys, Asn, Thr, Arg, Asp, Glu, or Gln; Xaa at position 25 is Asp, Asn, Glu, or Gln; Xaa at position 26 is Thr, Asp, Ser, or Glu; Xaa at position 27 is Ser, Thr, Lys, Asn, Gln, or Arg; Xaa at position 28 is Phe, Tyr, Pro, or Trp; Xaa at position 29 is Phe, Tyr, or Trp; Xaa at position 30 is Ser, Gly, Lys, Thr, or Arg; Xaa at position 31 is Val, Ile, Met, or Leu; Xaa at position 32 is Gly, Ala, Asp, or Glu; Xaa at position 33 is Asn, Ser, Gln, Pro, or Thr; Xaa at position 35 is Lys, Glu, Ser, Arg, or Thr; Xaa at position 36 is Gln, Ser, Asn, or Thr; Xaa at position 37 is either Glu or Asp; Xaa at position 38 is Thr or Ser; Xaa at position 42 is Ser, Asn, Thr, or Gln; Xaa at position 44 is Ser, Asp, Ala, Leu, Thr, Glu, Ile, or Val; Xaa at position 47 is Phe, Tyr, or Trp; Xaa at position 48 is Leu, Met, Ile, or Val; Xaa at position 49 is Leu, Met, Ile, or Val; Xaa at position 50 is Ser, Ala, Tyr, or Thr; Xaa at position 51 is Leu, Val, or Ile; Xaa at position 52 is Lys, Gln, Arg, or Asn; Xaa at position 53 is Lys, Arg, Met, Leu, Ile, or Val; Xaa at position 54 is either Asn, Lys, Gly, Gln, or Arg; Xaa at position 55 is Gly, Ser, or Thr; Xaa at position 56 is Ala, Thr, Gln, Ser, or Asn; Xaa at position 57 is Gln, Val, Ala, Asn, Leu, or Ile; Xaa at position 58 is His, Ala, Lys, Tyr, or Thr; Xaa at position 59 is Pro, Thr, or Ser; Xaa at position 62 is Val, Ile, or Leu; Xaa at position 63 is Gln, Ser, Leu, Asn, Thr, Ile, or Val; Xaa at position 64 is Ala, Gln, Ser, Asn, or Thr; Xaa at position 65 is either Ser or Thr; Xaa at position 67 is Lys, Gln, Asn, or Arg; Xaa at position 69 is Glu, Val, Asp, Lys, Arg, Ile, or Leu; Xaa at position 70 is Val, Ile, or Leu; Xaa at position 71 is Asp, Glu, Tyr, or Trp; Xaa at position 72 is Asn, His, Ser, Asp, Gln, Thr, or Glu; Xaa at position 73 is Asn, Ser, Asp, Gln, Thr, or Glu; Xaa at position 74 is Ala, Thr, Met, Ile, Lys, Ser, Leu, Val, or Arg; Xaa at position 76 is Lys, Thr, Arg, or Ser; Xaa at position 78 is Gln, His, Ser, Asn, or Thr; Xaa at position 80 is Arg, Glu, Gln, Lys, Asp, or Asn; Xaa at position 81 is Leu, Pro, Thr, Ile, Val, Ala, or Ser; Xaa at position 82 is Ile, Leu, or Val; Xaa at position 83 is Glu, His, Asn, Leu, Gln, Ile, or Val; Xaa at position 85 is Leu, Val, or Ala; and Xaa at position 86 is either Ser, Ala, Tyr, Asn, or Thr.

[0029] 17. The isolated polynucleotide according to schemes 21, 22 or 23, wherein the encoded PIP-72 polypeptide comprises the amino acid sequence shown in the following formula: in The second Xaa is Gly, Ala, Cys, Asp, Glu, Ile, Lys, Leu, Asn, Arg, Ser, Thr, Val, Trp, or Tyr; The Xaa in the third position is Ile, Leu, Val, or Trp; The fourth Xaa is Thr, Ala, Asp, Glu, His, Ile, Lys, Leu, Arg, Ser, Val, Trp, or Tyr; The Xaa in the 5th position is Val, Ala, Cys, Gly, His, Ile, Leu, or Tyr; The Xaa at position 6 is Thr, Ala, Cys, Phe, Gly, His, Ile, Lys, Met, Pro, Gln, Arg, Ser, Trp, or Tyr; Xaa at position 7 is either Asn, Ala, or Val; The Xaa at position 8 is Asn, Lys, Gly, Ser, Gln, Arg, Thr, Ala, Cys, Asp, Glu, His, Ile, Leu, Met, or Val; The Xaa at position 9 is either Ser, Ala, Cys, Gly, or Thr; The Xaa at the 10th position is Ser, Ala, Glu, Phe, Gly, His, Ile, Lys, Leu, Asn, Pro, Gln, Arg, Thr, or Trp; The Xaa at position 11 is Asn, Lys, Thr, Gln, Arg, Ser, Ala, Cys, Asp, Glu, Gly, His, Ile, Leu, Met, Val, or Tyr; The Xaa at position 12 is Pro, Thr, Lys, Ser, Arg, Ala, Cys, Asp, Glu, Gly, His, Leu, Asn, Gln, Arg, Val, Trp, or Tyr; Xaa at position 13 is Ile, Asn, Gln, Leu, or Val; Xaa at position 14 is Glu, Ala, Cys, Phe, His, Lys, Asp, or Gln; The Xaa at position 15 is Val, Ala, Ile, Leu, Cys, Met, or Arg; Xaa at position 16 is either Ala or Ser; Xaa at position 17 is Ile, Glu, Leu, or Val; Xaa at position 18 is Asn, Gln, Thr, or Ser; The Xaa at position 19 is His, Lys, Ala, Arg, Glu, Leu, Pro, Ser, or Tyr; Xaa at position 20 is Trp, Ala, or Thr; Xaa at position 21 is Gly, Arg, or Lys; Xaa at position 22 is Ser, Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Thr, Val, or Tyr; Xaa at position 23 is Asp, Ala, Gly, His, Lys, Met, Asn, Gln, Ser, Thr, or Val; Xaa at position 24 is Gly, Asp, or Phe; Xaa at position 25 is Asp, Ala, Glu, Phe, Asn, or Gln; Xaa at position 26 is Thr, Glu, Asp, Ser, or Pro; Xaa at position 27 is Ser, Thr, Lys, Arg, Ala, Cys, Asp, Glu, Phe, Gly, His, Asn, or Gln; Xaa at position 28 is Phe, Tyr, Pro, or Trp; Xaa at position 29 is Phe, Ala, Cys, Ile, Leu, Gln, Arg, Trp, or Tyr; The Xaa at position 30 is Ser, Gly, Lys, Thr, Arg, Ala, Cys, Asp, Glu, Phe, His, Leu, Met, Asn, Pro, Gln, Val, Trp, or Tyr; Xaa at position 31 is Val, Ile, Met, or Leu; Xaa at position 32 is Gly, Ala, Asp, Glu, Phe, His, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 33 is Asn, Ser, Gln, Pro, Thr, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Arg, Val, or Tyr; Xaa at position 34 is Gly, Glu, Phe, His, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Tyr; Xaa at position 35 is Lys, Glu, Ala, Cys, Asp, Gly, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Val; Xaa at position 36 is Gln, Ala, Cys, Glu, Gly, His, Ile, Lys, Leu, Asn, Pro, Arg, Ser, Thr, or Val; Xaa at position 37 is Glu, Asp, Ala, Cys, Phe, Gly, Ile, Lys, Leu, Met, Asn, Ser, Thr, or Val; Xaa at position 38 is Thr, Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Gln, Arg, Val, Trp, or Tyr; Xaa at position 39 is either Trp or Phe; The Xaa at position 40 is Asp, Ala, Cys, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 42 is Ser, Asn, Thr, Ala, Cys, Asp, Glu, Phe, Gly, Ile, Lys, Leu, Met, Arg, Val, Trp, Tyr, or Gln; Xaa at position 44 is Ser, Asp, Ala, Leu, Thr, Glu, Ile, Ala, Gly, Leu, Met, Asn, Pro, Gln, Val, Tyr, or Val; Xaa at position 45 is Arg, Lys, or Ser; Xaa at position 46 is Gly, Ala, or Gln; Xaa at position 47 is Phe, Tyr, Cys, Val, or Trp; Xaa at position 48 is Leu, Met, Ile, Cys, Phe, Met, Arg, Tyr, or Val; Xaa at position 49 is Leu, Met, Ile, or Val; Xaa at position 50 is Ser, Ala, Tyr, Cys, Asp, Ile, Met, Pro, Gln, Val, or Thr; Xaa at position 51 is Leu, Val, Ala, Cys, Met, or Ile; Xaa at position 52 is Lys, Cys, Phe, His, Ile, Leu, Met, Asn, Arg, Ser, Thr, Gln, Trp, or Tyr; Xaa at position 53 is Lys, Arg, Met, Leu, Ile, Ala, Cys, Asp, Glu, Phe, His, Asn, Gln, Ser, Thr, Tyr, or Val; Xaa at position 54 is Asn, Cys, Asp, Glu, Phe, Gly, Lys, Met, Gln, Arg, Ser, or Trp; Xaa at position 55 is Gly, Ser, or Thr; Xaa at position 56 is Ala, Thr, Gln, Ser, Gly, Leu, Pro, Arg, or Asn; Xaa at position 57 is Gln, Glu, Leu, Met, Ser, Val, Ala, Asn, Ile, or Thr; Xaa at position 58 is His, Ala, Lys, Asp, Phe, Leu, Met, Asn, Arg, Trp, Tyr, or Thr; Xaa at position 59 is Pro, Thr, or Ser; Xaa at position 60 is Tyr, Glu, or Phe; Xaa at position 62 is Val, Ile, or Leu; Xaa at position 63 is Gln, Ser, Cys, Gly, Ile, Leu, Met, Asn, Thr, Val, or Tyr; Xaa at position 64 is Ala, Gln, Asn, Phe, Gly, His, Arg, Ser, or Tyr; Xaa at position 65 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Leu, Asn, Val, or Thr; Xaa at position 66 is Ser, Ala, or Gly; Xaa at position 67 is Lys, Gln, Asn, or Arg; Xaa at position 67 is Lys, Ala, Cys, Asp, Phe, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 68 is Ile, Asp, Leu, or Val; Xaa at position 69 is Glu, Ala, Cys, Asp, Phe, His, Ile, Leu, Met, Gln, Arg, Ser, Thr, Val, or Tyr; Xaa at position 70 is Val, Ile, Cys, or Leu; Xaa at position 71 is Asp, Glu, Tyr, Ala, Cys, Gly, His, Ile, Leu, Met, Asn, Ser, Thr, Val, or Trp; Xaa at position 72 is Asn, Ala, Cys, Asp, Glu, Gly, Lys, Met, Pro, Gln, Arg, Ser, Thr, Val, His, or Trp; Xaa at position 73 is Asn, Ser, Asp, Gln, Thr, Ala, Cys, Phe, Gly, His, Ile, Leu, Val, Tyr, or Glu; Xaa at position 74 is Ala, Thr, Met, Ile, Lys, Ser, Leu, Val, Cys, Asp, Phe, Gly, His, Asn, Gln, Tyr, or Arg; Xaa at position 75 is Val, Cys, Ile, or Leu; Xaa at position 76 is Lys, Ala, Cys, Phe, His, Ile, Leu, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 77 is Asp or Tyr; Xaa at position 78 is Gln, His, Ser, Asn, Ala, Cys, Asp, Phe, Gly, Ile, Leu, Met, Asn, Arg, Val, Tyr, or Thr; Xaa at position 79 is Gly, Arg, Ala, Cys, Asp, Glu, Phe, His, Lys, Leu, Asn, Gln, Arg, Ser, Thr, Trp, or Tyr; Xaa at position 80 is Arg, Glu, Gln, Lys, Asp, Ala, Cys, Phe, Gly, His, Ile, Leu, Ser, Thr, Val, Tyr, or Asn; Xaa at position 81 is Leu, Pro, Thr, Ile, Val, Ala, Cys, Asp, Phe, Gly, His, or Ser; Xaa at position 82 is Ile, Ala, Leu, Met, Arg, and Val; Xaa at position 83 is Glu, His, Asn, Leu, Gln, Ile, Ala, Cys, Asp, Phe, Gly, Lys, Pro, Arg, Ser, Thr, Tyr, or Val; Xaa at position 84 is Pro, Ala, Cys, Glu, Ile, Ser, Val, Trp, or Tyr; Xaa at position 85 is Leu, Val, Cys, Gly, or Ala; and Xaa at position 86 is either Ser, Ala, Tyr, Asn, Ile, Val, or Thr.

[0030] 18. The isolated polynucleotide according to any one of schemes 10 to 17, wherein the encoded PIP-72 polypeptide comprises an amino acid motif represented by positions 37 to 51 of the following sequences: SEQ ID NO: 846, SEQ ID NO: 847, SEQ ID NO: 848 or SEQ ID NO: 849.

[0031] 19. An isolated polynucleotide comprising a nucleic acid molecule encoding a chimeric PIP-72 polypeptide, the chimeric PIP-72 polypeptide comprising at least a first portion and a second portion, the first portion comprising a first PIP-72 polypeptide moiety, the second portion comprising a complementary second PIP-72 polypeptide moiety, wherein the first PIP-72 polypeptide and the second PIP-72 polypeptide have different amino acid sequences in the respective portions.

[0032] 20. An expression cassette comprising isolated polynucleotides operatively linked to a heterologous regulatory element as described in any one of schemes 11 to 19.

[0033] 21. The expression cassette according to claim 20, wherein the regulatory element is a promoter capable of expressing a protein in a plant.

[0034] 22. A transgenic plant comprising a DNA construct as described in any one of schemes 1 to 9.

[0035] 23. A transgenic plant stably transformed using a DNA construct as described in any one of schemes 1 to 9.

[0036] 24. A seed produced by the plant described in Scheme 22 or 23, wherein the seed contains the nucleic acid molecule.

[0037] 25. A progeny plant produced from the seeds described in Scheme 24.

[0038] 26. A host cell transformed using a DNA construct according to any one of schemes 1 to 9.

[0039] 27. The host cell according to Scheme 26, wherein the host cell is a bacterial cell or a plant cell.

[0040] 28. The host cell according to Scheme 27, wherein the plant cell is a monocotyledonous plant cell or a dicotyledonous plant cell.

[0041] 29. A recombinant PIP-72 polypeptide, said recombinant PIP-72 polypeptide being effective against the western maize rootworm (… Diabrotica virgifera virgifera It has insecticidal activity.

[0042] 30. The recombinant PIP-72 polypeptide according to Scheme 29, wherein the PIP-72 polypeptide comprises an amino acid sequence having at least 50% identity with SEQ ID NO: 2.

[0043] 31. The recombinant PIP-72 polypeptide according to Scheme 29 or 30, wherein the PIP-72 polypeptide, compared with the corresponding amino acid shown in SEQ ID NO: 2, is present at amino acid concentrations 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 42, 44, 45. Positions 46, 47, 48, 49, 50, 51, 52, 53, 54, 56, 57, 58, 59, 60, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, or 86 contain 1 to 45 amino acid substitutions.

[0044] 32. The recombinant PIP-72 polypeptide according to Scheme 29 or 30, wherein the PIP-72 polypeptide is related to SEQ ID NO: Compared to the corresponding amino acids shown in Figure 2, the amino acids include 1 to 45 amino acid substitutions at positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 17, 18, 19, 20, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 42, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 56, 58, 60, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, or 86.

[0045] 33. The recombinant PIP-72 polypeptide according to schemes 29, 30 or 32, wherein the PIP-72 polypeptide comprises the amino acid sequence shown in the following formula: in The second Xaa is Gly, Ala, Cys, Asp, Glu, Ile, Lys, Leu, Asn, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 3 is either Ile or Trp; The fourth Xaa is Thr, Ala, Asp, Glu, His, Ile, Lys, Leu, Arg, Ser, Val, Trp, or Tyr; The Xaa in the 5th position is Val, Ala, Cys, Gly, His, Ile, or Tyr; The Xaa at position 6 is Thr, Ala, Cys, Phe, Gly, His, Ile, Lys, Met, Pro, Gln, Arg, Ser, Trp, or Tyr; Xaa at position 7 is either Asn, Ala, or Val; The Xaa at position 8 is Asn, Ala, Cys, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Gln, Arg, Ser, Thr, or Val; The Xaa at position 9 is either Ser, Ala, Cys, Gly, or Thr; The Xaa at the 10th position is Ser, Ala, Glu, Phe, Gly, His, Ile, Lys, Leu, Asn, Pro, Gln, Arg, Thr, or Trp; The Xaa at position 11 is Asn, Ala, Cys, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Gln, Ser, Thr, Val, or Tyr; The Xaa at position 12 is Pro, Ala, Cys, Asp, Glu, Gly, His, Lys, Leu, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 13 is Ile, Asn, Gln, or Val; Xaa at position 14 is Glu, Ala, Cys, Phe, His, Lys, or Gln; Xaa at position 15 is Val, Ala, Cys, Ile, Met, or Arg; Xaa at position 17 is Ile, Glu, or Val; Xaa at position 18 is either Asn or Ser; The Xaa at position 19 is His, Ala, Glu, Lys, Leu, Pro, Arg, Ser, or Tyr; Xaa at position 20 is Trp, Ala, or Thr; Xaa at position 22 is Ser, Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Thr, Val, or Tyr; Xaa at position 23 is Asp, Ala, Gly, His, Lys, Met, Asn, Gln, Ser, Thr, or Val; Xaa at position 24 is Gly, Asp, or Phe; Xaa at position 25 is Asp, Ala, Glu, Phe, Asn, or Gln; Xaa at position 26 is Thr, Glu, or Pro; Xaa at position 27 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Asn, Gln, Arg, or Thr; Xaa at position 28 is Phe, Pro, Trp, or Tyr; Xaa at position 29 is Phe, Ala, Cys, Ile, Leu, Gln, Arg, Trp, or Tyr; The Xaa at position 30 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Lys, Leu, Met, Asn, Pro, Gln, Arg, Thr, Val, Trp, or Tyr; Xaa at position 31 is Val, Ile, or Leu; Xaa at position 32 is Gly, Ala, Asp, Glu, Phe, His, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 33 is Asn, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Pro, Gln, Arg, Ser, Thr, Val, or Tyr; Xaa at position 34 is Gly, Glu, Phe, His, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Tyr; Xaa at position 35 is Lys, Ala, Cys, Asp, Gly, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Val; Xaa at position 36 is Gln, Ala, Cys, Glu, Gly, His, Ile, Lys, Leu, Asn, Pro, Arg, Ser, Thr, or Val; Xaa at position 37 is Glu, Ala, Cys, Asp, Phe, Gly, Ile, Lys, Leu, Met, Asn, Ser, Thr, or Val; Xaa at position 38 is Thr, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Val, Trp, or Tyr; Xaa at position 39 is either Trp or Phe; The Xaa at position 40 is Asp, Ala, Cys, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 42 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, Ile, Lys, Leu, Met, Asn, Gln, Arg, Thr, Val, Trp, or Tyr; Xaa at position 44 is Ser, Ala, Asp, Glu, Gly, Leu, Met, Asn, Pro, Gln, Thr, Val, or Tyr; Xaa at position 45 is Arg, Lys, or Ser; Xaa at position 46 is Gly, Ala, or Gln; Xaa at position 47 is Phe, Cys, Val, or Tyr; Xaa at position 48 is Val, Ile, or Leu; Xaa at position 49 is Leu, Cys, Phe, Met, Arg, or Tyr; Xaa at position 50 is Ser, Ala, Cys, Asp, Ile, Met, Pro, Gln, Thr, or Val; Xaa at position 51 is Leu, Ala, Cys, Met, or Val; Xaa at position 52 is Lys, Cys, Phe, His, Ile, Leu, Met, Asn, Arg, Ser, Thr, Trp, or Tyr; Xaa at position 53 is Lys, Ala, Cys, Asp, Glu, Phe, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, or Tyr; Xaa at position 54 is Asn, Cys, Asp, Glu, Phe, Gly, Lys, Met, Gln, Arg, Ser, or Trp; Xaa at position 56 is Ala, Gly, Leu, Asn, Pro, Gln, Arg, Ser, or Thr; Xaa at position 57 is Gln, Glu, Leu, Met, Ser, or Thr; Xaa at position 58 is His, Ala, Asp, Phe, Leu, Met, Asn, Arg, Trp, or Tyr; Xaa at position 60 is Tyr, Glu, or Phe; Xaa at position 63 is Gln, Cys, Gly, Ile, Leu, Met, Asn, Thr, Val, or Tyr; Xaa at position 64 is Ala, Phe, Gly, His, Arg, Ser, or Tyr; Xaa at position 65 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Leu, Asn, Thr, or Val; Xaa at position 66 is Ser, Ala, or Gly; Xaa at position 67 is Lys, Ala, Cys, Asp, Phe, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 68 is Ile, Asp, Leu, or Val; Xaa at position 69 is Glu, Ala, Cys, Asp, Phe, His, Ile, Leu, Met, Gln, Arg, Ser, Thr, Val, or Tyr; Xaa at position 70 is Val, Cys, or Ile; Xaa at position 71 is Asp, Ala, Cys, Gly, His, Ile, Leu, Met, Asn, Ser, Thr, Val, or Tyr; Xaa at position 72 is Asn, Ala, Cys, Asp, Glu, Gly, Lys, Met, Pro, Gln, Arg, Ser, Thr, Val, or Trp; Xaa at position 73 is Asn, Ala, Cys, Asp, Phe, Gly, His, Ile, Leu, Ser, Thr, Val, or Tyr; Xaa at position 74 is Ala, Cys, Asp, Phe, Gly, His, Ile, Leu, Asn, Gln, Arg, Ser, Thr, Val, or Tyr; Xaa at position 75 is Val, Cys, Ile, or Leu; Xaa at position 76 is Lys, Ala, Cys, Phe, His, Ile, Leu, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 77 is Asp or Tyr; Xaa at position 78 is Gln, Ala, Cys, Asp, Phe, Gly, His, Ile, Leu, Met, Asn, Arg, Ser, Thr, Val, or Tyr; Xaa at position 79 is Gly, Arg, Ala, Cys, Asp, Glu, Phe, His, Lys, Leu, Asn, Gln, Arg, Ser, Thr, Trp, or Tyr; Xaa at position 80 is Arg, Ala, Cys, Asp, Phe, Gly, His, Ile, Leu, Asn, Ser, Thr, Val, or Tyr; Xaa at position 81 is Leu, Ala, Cys, Asp, Phe, Gly, His, Ile, Asn, Pro, Arg, Ser, Thr, or Val; Xaa at position 82 is Ile, Ala, Leu, Met, Arg, or Val; Xaa at position 83 is Glu, Ala, Cys, Asp, Phe, Gly, His, Ile, Lys, Leu, Asn, Pro, Arg, Ser, Thr, Val, or Tyr; Xaa at position 84 is Pro, Ala, Cys, Glu, Ile, Ser, Val, Trp, or Tyr; Xaa at position 85 is Leu, Cys, Gly, or Val; and Xaa at position 86 is either Ser, Ala, Ile, Thr, or Val.

[0046] 34. The recombinant PIP-72 polypeptide according to schemes 29, 30 or 31, wherein the PIP-72 polypeptide comprises the amino acid sequence shown in the following formula: in The second Xaa is Gly, Ala, Cys, Asp, Glu, Ile, Lys, Leu, Asn, Arg, Ser, Thr, Val, Trp, or Tyr; The Xaa in the third position is Ile, Leu, Val, or Trp; The fourth Xaa is Thr, Ala, Asp, Glu, His, Ile, Lys, Leu, Arg, Ser, Val, Trp, or Tyr; The Xaa in the 5th position is Val, Ala, Cys, Gly, His, Ile, Leu, or Tyr; The Xaa at position 6 is Thr, Ala, Cys, Phe, Gly, His, Ile, Lys, Met, Pro, Gln, Arg, Ser, Trp, or Tyr; Xaa at position 7 is either Asn, Ala, or Val; The Xaa at position 8 is Asn, Lys, Gly, Ser, Gln, Arg, Thr, Ala, Cys, Asp, Glu, His, Ile, Leu, Met, or Val; The Xaa at position 9 is either Ser, Ala, Cys, Gly, or Thr; The Xaa at position 11 is Asn, Lys, Thr, Gln, Arg, Ser, Ala, Cys, Asp, Glu, Gly, His, Ile, Leu, Met, Val, or Tyr; The Xaa at position 12 is Pro, Thr, Lys, Ser, Arg, Ala, Cys, Asp, Glu, Gly, His, Leu, Asn, Gln, Arg, Val, Trp, or Tyr; Xaa at position 13 is Ile, Asn, Gln, Leu, or Val; Xaa at position 14 is Glu, Ala, Cys, Phe, His, Lys, Asp, or Gln; The Xaa at position 15 is Val, Ala, Ile, Leu, Cys, Met, or Arg; Xaa at position 16 is either Ala or Ser; Xaa at position 17 is Ile, Glu, Leu, or Val; Xaa at position 18 is Asn, Gln, Thr, or Ser; The Xaa at position 19 is His, Lys, Ala, Arg, Glu, Leu, Pro, Ser, or Tyr; Xaa at position 20 is Trp, Ala, or Thr; Xaa at position 21 is Gly, Arg, or Lys; Xaa at position 22 is Ser, Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Thr, Val, or Tyr; Xaa at position 23 is Asp, Ala, Gly, His, Lys, Met, Asn, Gln, Ser, Thr, or Val; Xaa at position 24 is Gly, Asp, or Phe; Xaa at position 25 is Asp, Ala, Glu, Phe, Asn, or Gln; Xaa at position 26 is Thr, Glu, Asp, Ser, or Pro; Xaa at position 27 is Ser, Thr, Lys, Arg, Ala, Cys, Asp, Glu, Phe, Gly, His, Asn, or Gln; Xaa at position 28 is Phe, Tyr, Pro, or Trp; Xaa at position 29 is Phe, Ala, Cys, Ile, Leu, Gln, Arg, Trp, or Tyr; The Xaa at position 30 is Ser, Gly, Lys, Thr, Arg, Ala, Cys, Asp, Glu, Phe, His, Leu, Met, Asn, Pro, Gln, Val, Trp, or Tyr; Xaa at position 31 is Val, Ile, Met, or Leu; Xaa at position 32 is Gly, Ala, Asp, Glu, Phe, His, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 33 is Asn, Ser, Gln, Pro, Thr, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Arg, Val, or Tyr; Xaa at position 34 is Gly, Glu, Phe, His, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Tyr; Xaa at position 35 is Lys, Glu, Ala, Cys, Asp, Gly, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Val; Xaa at position 36 is Gln, Ala, Cys, Glu, Gly, His, Ile, Lys, Leu, Asn, Pro, Arg, Ser, Thr, or Val; Xaa at position 37 is Glu, Asp, Ala, Cys, Phe, Gly, Ile, Lys, Leu, Met, Asn, Ser, Thr, or Val; Xaa at position 38 is Thr, Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Gln, Arg, Val, Trp, or Tyr; Xaa at position 39 is either Trp or Phe; The Xaa at position 40 is Asp, Ala, Cys, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 42 is Ser, Asn, Thr, Ala, Cys, Asp, Glu, Phe, Gly, Ile, Lys, Leu, Met, Arg, Val, Trp, Tyr, or Gln; Xaa at position 44 is Ser, Asp, Ala, Leu, Thr, Glu, Ile, Ala, Gly, Leu, Met, Asn, Pro, Gln, Val, Tyr, or Val; Xaa at position 45 is Arg, Lys, or Ser; Xaa at position 46 is Gly, Ala, or Gln; Xaa at position 47 is Phe, Tyr, Cys, Val, or Trp; Xaa at position 48 is Leu, Met, Ile, Cys, Phe, Met, Arg, Tyr, or Val; Xaa at position 49 is Leu, Met, Ile, or Val; Xaa at position 50 is Ser, Ala, Tyr, Cys, Asp, Ile, Met, Pro, Gln, Val, or Thr; Xaa at position 51 is Leu, Val, Ala, Cys, Met, or Ile; Xaa at position 52 is Lys, Cys, Phe, His, Ile, Leu, Met, Asn, Arg, Ser, Thr, Gln, Trp, or Tyr; Xaa at position 53 is Lys, Arg, Met, Leu, Ile, Ala, Cys, Asp, Glu, Phe, His, Asn, Gln, Ser, Thr, Tyr, or Val; Xaa at position 54 is Asn, Cys, Asp, Glu, Phe, Gly, Lys, Met, Gln, Arg, Ser, or Trp; Xaa at position 55 is Gly, Ser, or Thr; Xaa at position 56 is Ala, Thr, Gln, Ser, Gly, Leu, Pro, Arg, or Asn; Xaa at position 57 is Gln, Glu, Leu, Met, Ser, Val, Ala, Asn, Ile, or Thr; Xaa at position 58 is His, Ala, Lys, Asp, Phe, Leu, Met, Asn, Arg, Trp, Tyr, or Thr; Xaa at position 59 is Pro, Thr, or Ser; Xaa at position 60 is Tyr, Glu, or Phe; Xaa at position 62 is Val, Ile, or Leu; Xaa at position 63 is Gln, Ser, Cys, Gly, Ile, Leu, Met, Asn, Thr, Val, or Tyr; Xaa at position 64 is Ala, Gln, Asn, Phe, Gly, His, Arg, Ser, or Tyr; Xaa at position 65 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Leu, Asn, Val, or Thr; Xaa at position 66 is Ser, Ala, or Gly; Xaa at position 67 is Lys, Gln, Asn, or Arg; Xaa at position 67 is Lys, Ala, Cys, Asp, Phe, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 68 is Ile, Asp, Leu, or Val; Xaa at position 69 is Glu, Ala, Cys, Asp, Phe, His, Ile, Leu, Met, Gln, Arg, Ser, Thr, Val, or Tyr; Xaa at position 70 is Val, Ile, Cys, or Leu; Xaa at position 71 is Asp, Glu, Tyr, Ala, Cys, Gly, His, Ile, Leu, Met, Asn, Ser, Thr, Val, or Trp; Xaa at position 72 is Asn, Ala, Cys, Asp, Glu, Gly, Lys, Met, Pro, Gln, Arg, Ser, Thr, Val, His, or Trp; Xaa at position 73 is Asn, Ser, Asp, Gln, Thr, Ala, Cys, Phe, Gly, His, Ile, Leu, Val, Tyr, or Glu; Xaa at position 74 is Ala, Thr, Met, Ile, Lys, Ser, Leu, Val, Cys, Asp, Phe, Gly, His, Asn, Gln, Tyr, or Arg; Xaa at position 75 is Val, Cys, Ile, or Leu; Xaa at position 76 is Lys, Ala, Cys, Phe, His, Ile, Leu, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 77 is Asp or Tyr; Xaa at position 78 is Gln, His, Ser, Asn, Ala, Cys, Asp, Phe, Gly, Ile, Leu, Met, Asn, Arg, Val, Tyr, or Thr; Xaa at position 79 is Gly, Arg, Ala, Cys, Asp, Glu, Phe, His, Lys, Leu, Asn, Gln, Arg, Ser, Thr, Trp, or Tyr; Xaa at position 80 is Arg, Glu, Gln, Lys, Asp, Ala, Cys, Phe, Gly, His, Ile, Leu, Ser, Thr, Val, Tyr, or Asn; Xaa at position 81 is Leu, Pro, Thr, Ile, Val, Ala, Cys, Asp, Phe, Gly, His, or Ser; Xaa at position 82 is Ile, Ala, Leu, Met, Arg, and Val; Xaa at position 83 is Glu, His, Asn, Leu, Gln, Ile, Ala, Cys, Asp, Phe, Gly, Lys, Pro, Arg, Ser, Thr, Tyr, or Val; Xaa at position 84 is Pro, Ala, Cys, Glu, Ile, Ser, Val, Trp, or Tyr; Xaa at position 85 is Leu, Val, Cys, Gly, or Ala; and Xaa at position 86 is either Ser, Ala, Tyr, Asn, Ile, Val, or Thr.

[0047] 35. The recombinant PIP-72 polypeptide according to any one of claims 29, 30 and 31, wherein the PIP-72 polypeptide comprises an amino acid motif represented by positions 37 to 51 of the following sequences: SEQ ID NO: 846, SEQ ID NO: 847, SEQ ID NO: 848 or SEQ ID NO: 849.

[0048] 36. A recombinant polypeptide, said recombinant polypeptide being selected from polypeptides having at least 95% identity with the following amino acid sequences: SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 30, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 929, SEQ ID NO: 930, SEQ ID NO: 931, SEQ ID NO: 937, SEQ ID NO: 938, SEQ ID NO: 942, SEQ ID NO: 947 or SEQ ID NO: 948.

[0049] 37. A chimeric PIP-72 polypeptide comprising at least a first portion and a second portion, the first portion comprising a first PIP-72 polypeptide moiety, the second portion comprising a complementary second PIP-72 polypeptide moiety, wherein the first PIP-72 polypeptide and the second PIP-72 polypeptide have different amino acid sequences in the respective portions.

[0050] 38. A composition comprising any one of embodiments 29 to 35, the polypeptide of embodiment 36, or the chimeric PIP-72 polypeptide of embodiment 37.

[0051] 39. A fusion protein comprising any one of embodiments 29 to 35, the polypeptide of embodiment 36, or the chimeric PIP-72 polypeptide of embodiment 37.

[0052] 40. A method for controlling insect pest populations, the method comprising contacting the insect pest population with an insecticidal effective amount of any one of schemes 29 to 35, the polypeptide of scheme 36, or the chimeric PIP-72 polypeptide of scheme 37.

[0053] 41. A method for inhibiting or killing insect pests, the method comprising contacting the insect pests with a composition comprising an insecticidal effective amount of any one of schemes 29 to 35, the polypeptide of scheme 36, or the chimeric PIP-72 polypeptide of scheme 37.

[0054] 42. A method for controlling insect pest populations resistant to insecticidal proteins, the method comprising contacting the insect pest population with an insecticidal effective amount of any one of schemes 29 to 35, the polypeptide of scheme 36, or the chimeric PIP-72 polypeptide of scheme 37.

[0055] 43. A method for preventing insect infection of transgenic plants and providing insect resistance management, the method comprising expressing in the plant any one of schemes 29 to 35, the polypeptide of scheme 36, or the chimeric PIP-72 polypeptide of scheme 37.

[0056] 44. A transgenic plant comprising the DNA construct described in Scheme 8.

[0057] 45. A transgenic plant stably transformed using the DNA construct described in the protocol.

[0058] 46. ​​A seed produced by the plant described in Scheme 44 or 45, wherein the seed contains the nucleic acid molecule.

[0059] 47. A progeny plant produced from the seeds described in Scheme 46.

[0060] 48. A host cell transformed using the DNA construct described in Scheme 8.

[0061] 49. The host cell according to claim 48, wherein the host cell is a bacterial cell or a plant cell.

[0062] 50. The host cell according to Scheme 49, wherein the plant cell is a monocotyledonous plant cell or a dicotyledonous plant cell.

[0063] 51. A method for identifying the nucleotide sequence of a PIP-72 polypeptide, a polypeptide of scheme 36, or a chimeric PIP-72 polypeptide of scheme 37, in a biological sample, the method comprising contacting the sample with a polynucleotide hybridized to the nucleotide sequence under strict hybridization conditions, and detecting the binding of the polynucleotide to the nucleotide sequence, wherein the binding is diagnosed against the nucleotide sequence in the sample.

[0064] 52. A method for identifying, in a sample, any one of schemes 29 to 35, the polypeptide of scheme 36, or the chimeric PIP-72 polypeptide of scheme 37, the method comprising contacting the sample with an antibody specifically bound to the polypeptide and detecting the binding, wherein the binding is diagnosed for the polypeptide present in the sample. Attached Figure Description

[0065] Figure 1 The amino acid sequence alignments of the following substances are shown: PIP-72Aa (SEQ ID NO: 2), PIP-72Ba (SEQ ID NO: 4), PIP-72Ca (SEQ ID NO: 6), PIP-72Cb (SEQ ID NO: 8), PIP-72Da (SEQ ID NO: 10), PIP-72Db (SEQ ID NO: 12), PIP-72Dc (SEQ ID NO: 14), PIP-72Ea (SEQ ID NO: 16), PIP-72Fa (SEQ ID NO: 18), GBP_A3175 (SEQ ID NO: 20), SRBS_294080 (SEQ ID NO: 22), JG43047 (SEQ ID NO: 24), SwiRh_4910 (SEQ ID NO: 26), PIP-72Ff (SEQ ID NO: 28), PFL_6283 (SEQ ID NO: 294080). 30), PIP-72Gb (SEQ ID NO: 32), XBJ1_1078 (SEQ ID NO: 34), plu2373 (SEQ ID NO: 36), and PIP-72Ge (SEQ ID NO: 38). Sequence differences are highlighted in the figure. Amino acids 37 to 51 (motif 1) in PIP-72Aa (SEQ ID NO: 2) are underlined.

[0066] Figure 2The amino acid sequence alignments of the following substances are shown: PIP-72Aa (SEQ ID NO: 2), PIP-72Ab (SEQ ID NO: 927), PIP-72Ba (SEQ ID NO: 4), PIP-72Bb (SEQ ID NO: 928), PIP-72Ca (SEQ ID NO: 6), PIP-72Cb (SEQ ID NO: 8), WP_030131237 (SEQ ID NO: 929), PIP-72Da (SEQ ID NO: 10), PIP-72Db (SEQ ID NO: 12), PIP-72Dc (SEQ ID NO: 14), PIP-72Fa (SEQ ID NO: 18), and GBP_A3175 (SEQ ID NO: 20). The amino acid differences between PIP-72Aa (SEQ ID NO: 2) and its homologs are highlighted with shading.

[0067] Figure 3 The amino acid sequence alignments of the following substances are shown: PIP-72Aa (SEQ ID NO: 2), PIP-72Ba (SEQ ID NO: 4), PIP-72Ca (SEQ ID NO: 6), PIP-72Cb (SEQ ID NO: 8), PIP-72Da (SEQ ID NO: 10), PIP-72Db (SEQ ID NO: 12), and PIP-72Dc (SEQ ID NO: 14). Amino acid differences between PIP-72Aa (SEQ ID NO: 2) and its homologs are highlighted with shading.

[0068] Figure 4 The amino acid sequence alignments of the following substances are shown: WP_030131237 (SEQ ID NO: 929), PIP-72Ca (SEQ ID NO: 6), PIP-72Cb (SEQ ID NO: 8), PIP-72Da (SEQ ID NO: 10), PIP-72Db (SEQ ID NO: 12), and PIP-72Dc (SEQ ID NO: 14). The amino acid differences between PIP-72Da (SEQ ID NO: 10) and its homologs are highlighted with shading.

[0069] Figure 5The amino acid sequence alignments of the following substances are shown: PIP-72Fh (SEQ ID NO: 932), PIP-72Gi (SEQ ID NO: 941), PIP-72Fi (SEQ ID NO: 933), PIP-72Gl (SEQ ID NO: 944), and PIP-72Fa (SEQ ID NO: 14). The amino acid differences between PIP-72Ca (SEQ ID NO: 2) and its homologs are highlighted with shading.

[0070] Figure 6 The T0 GH efficacy results for events generated by the constructs PHP61664, PHP61666, PHP61668, PHP64465, PHP64468, PHP64471, and PHP69828 are shown. The efficacy of events derived from these constructs compared to the negative control events was observed based on measurements of root protection against Western maize rootworm damage. The method developed by Oleson et al. in 2005 was employed. J. Econ Entomol [98(1):1-8], the degree of root protection is measured by the number of damaged root nodes (CRWNIS = maize rootworm damage score). The root damage score is measured from "0" to "3", where "0" indicates no visible damage to the root, "1" indicates one damaged root node, "2" indicates two damaged root nodes, and "3" indicates three damaged root nodes (the highest score). Each symbol (triangle, square, or circle) in the figure represents a single event. Detailed Implementation

[0071] It should be understood that this disclosure is not limited to the specific methods, protocols, cell lines, genera, and reagents described, as these can vary. It should also be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of this disclosure.

[0072] Unless the context clearly specifies otherwise, the singular forms “a,” “an,” and “the” used herein include multiple referents. Thus, for example, reference to “a cell” includes multiple such cells, and reference to “the protein” includes reference to one or more proteins, as well as their equivalents known to those skilled in the art, etc. Unless otherwise expressly stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains.

[0073] This disclosure relates to compositions and methods for controlling pests. These methods involve transforming organisms with a nucleic acid sequence encoding a PIP-72 polypeptide. Specifically, the nucleic acid sequence of the embodiments can be used to prepare plants and microorganisms with insecticidal activity. Therefore, this disclosure provides transformed bacteria, plants, plant cells, plant tissues, and seeds. The compositions are insecticidal nucleic acids and proteins of bacterial strains. The nucleic acid sequence can be used to construct expression vectors for subsequent transformation into organisms of interest, can be used as probes for isolating other homologous (or partially homologous) genes, and can be used to produce altered PIP-72 polypeptides by methods known in the art (e.g., site-directed mutagenesis, domain exchange, or DNA shuffling). The PIP-72 polypeptide can be used to control or kill populations of lepidopteran, coleopteran, dipteran, fungal, hemiptera, and nematode pests, and can be used to produce compositions with insecticidal activity. Insect pests of interest include, but are not limited to, lepidopteran species, including, but not limited to, diamondback moths, such as the grain borer (Prunella vulgaris). Helicoverpa zea Boddie; soybean looper moth, such as the soybean looper moth ( Pseudoplusia includens Walker; bean caterpillars, such as the bean noctuid moth ( Anticarsia gemmatalis Hübner); and Coleoptera species, including but not limited to: western maize rootworm ( Diabrotica virgifera - WCRW, Southern Corn Rootworm ( Diabrotica undecimpunctata howardi SCRW, Northern Corn Rootworm Diabrotica barberi )- NCRW.

[0074] The term "insecticide toxin" or "insecticide protein" as used herein refers to toxins or proteins homologous to one or more pests (including, but not limited to, members of the orders Lepidoptera, Diptera, Hemiptera, and Coleoptera or Nematoda). Insecticide proteins have been isolated from organisms including, for example, species of Bacillus, species of Pseudomonas, and species of Luminobacterium. Photorhabdus ) species, pathogenic bacilli ( Xenorhabdus ) species, Clostridium bifermentans, and Japanese beetle-like spores ( Paenibacillus popilliae Insecticidal proteins include, but are not limited to: insecticidal proteins from Pseudomonas species, such as PSEEN3174 (Monalysin; (2011)). PLoS Pathogens 7:1-13); from biocontrol Pseudomonas ( Pseudomonas protegens Insecticidal proteins of strains CHA0 and Pf-5 (formerly known as Pseudomonas fluorescens) (Pechy-Tarr, (2008)). Environmental Microbiology 10:2368-2386; GenBank accession number EU400157); from *Pseudomonas taiwanensis* (Pseudomonas Taiwanensis Insecticidal proteins (Liu et al., (2010)). J. Agric . Food Chem ., 58:12343-12349); from Alcaligenes pseudomonas ( Pseudomonas pseudoalcligenes Insecticidal proteins (Zhang et al., (2009)). Annals of Microbiology 59:45-50; and Li et al., (2007) Plant Cell Tiss. Organ Cult .89:159-168); Insecticidal proteins from species of the genera *Proteobacterium* and *Pathogenic Bacteria* (Hinchliffe et al., (2010)). The Open Toxicology Journal , 3:101-118; and Morgan et al., (2001) Applied and Envir. Micro . 67:2062-2069); US Patent 6,048,838, US Patent 6,379,946; PIP-1 peptide in US Patent Serial No. 13 / 792861; AfIP-1A and / or AfIP-1B peptides in US Patent Serial No. 13 / 800233; PHI-4 peptide in US Patent Serial No. 13 / 839702; PIP-47 peptide in US Patent Serial No. 61 / 866747 Peptides; insecticidal proteins as described in U.S. Patent Serial Nos. 61 / 863761 and 61 / 863763; and δ-endotoxins, including but not limited to Cry1, Cry2, Cry3, Cry4, Cry5, Cry6, Cry7, Cry8, Cry9, Cry10, Cry11, Cry12, Cry13, Cry14, Cry15, Cry16, Cry17, Cry18, Cry19, and Cry 20. Cry21, Cry22, Cry23, Cry24, Cry25, Cry26, Cry27, Cry28, Cry29, Cry30, Cry31, Cry32, Cry 33. Cry34, Cry35, Cry36, Cry37, Cry38, Cry39, Cry40, Cry41, Cry42, Cry43, Cry44, Cry45, Cry4 6. Cry47, Cry49, Cry51, Cry52, Cry53, Cry54, Cry55, Cry56, Cry57, Cry58, Cry59, Cry60, Cry61, Cry62, Cry63, Cry64, Cry65, Cry66, Cry67, Cry68, Cry69, Cry70, and Cry71 type δ-endotoxin genes, as well as Bacillus thuringiensis (Bt). B. thuringiensisThe cytolytic genes cyt1 and cyt2. Members of these classes of Bacillus thuringiensis insecticidal proteins include, but are not limited to, Cry1Aa1 (accession number AAA22353); Cry1Aa2 (accession number AAA22552); Cry1Aa3 (accession number BAA00257); Cry1Aa4 (accession number CAA31886); Cry1Aa5 (accession number BAA04468); Cry1Aa6 (accession number AAA86265); Cry1Aa7 (accession number AAD46139); Cry1Aa8 (accession number I26149); Cry1Aa9 (accession number BAA77213); Cry1Aa10 (accession number AAD55382); and Cry1Aa1. Cry1Aa12 (Login ID AAP80146); Cry1Aa13 (Login ID AAM44305); Cry1Aa14 (Login ID AAP40639); Cry1Aa15 (Login ID AAY66993); Cry1Aa16 (Login ID HQ439776); Cry1Aa17 (Login ID HQ439788); Cry1Aa18 (Login ID HQ439790); Cry1Aa19 (Login ID HQ685121); Cry1Aa20 (Login ID JF340156); Cry1Aa21 (Login ID JN651496); Cry1Aa22 (Login ID KC158223); Cry1Ab1 (Login ID AAA22330); Cry1Ab2 (Login ID AAA22613); Cry1Ab3 (Login ID AAA22561); Cry1Ab4 (Login ID BAA00071); Cry1Ab5 (Login ID CAA28405); Cry1Ab6 (Login ID AAA22420); Cry1Ab7 (Login ID CAA31620); Cry1Ab8 (Login ID AAA22551); Cry1Ab9 (Login ID CAA38701); Cry1Ab10 (Login ID A29125); Cry1Ab11 (Login ID I12419) Cry1Ab12 (Logo AAC64003); Cry1Ab13 (Logo AAN76494); Cry1Ab14 (Logo AAG16877); Cry1Ab15 (Logo AAO13302); Cry1Ab16 (Logo AAK55546); Cry1Ab17 (Logo AAT46415); Cry1Ab18 (Logo AAQ88259); Cry1Ab19 (Logo AAW31761); Cry1Ab20 (Logo ABB72460); Cry1Ab21 (Logo ABS18384); Cry1Ab22 (Logo ABW87320);Cry1Ab23 (Login ID HQ439777); Cry1Ab24 (Login ID HQ439778); Cry1Ab25 (Login ID HQ685122); Cry1Ab26 (Login ID HQ847729); Cry1Ab27 (Login ID JN135249); Cry1Ab28 (Login ID JN135250); Cry1Ab29 (Login ID JN135251); Cry1Ab30 (Login ID JN135252); Cry1Ab31 (Login ID JN135253); Cry1Ab32 (Login ID JN135254); Cry1Ab33 (Login ID AAS93798); Cry1Ab3 4 (Grant No. KC156668); Cry1Ab sample (Grant No. AAK14336); Cry1Ab sample (Grant No. AAK14337); Cry1Ab sample (Grant No. AAK14338); Cry1Ab sample (Grant No. ABG88858); Cry1Ac1 (Grant No. AAA22331); Cry1Ac2 (Grant No. AAA22338); Cry1Ac3 (Grant No. CAA38098); Cry1Ac4 (Grant No. AAA73077); Cry1Ac5 (Grant No. AAA22339); Cry1Ac6 (Grant No. AAA86266); Cry1Ac7 (Grant No. AAB46989); Cry Cry1Ac8 (Grant No. AAC44841); Cry1Ac9 (Grant No. AAB49768); Cry1Ac10 (Grant No. CAA05505); Cry1Ac11 (Grant No. CAA10270); Cry1Ac12 (Grant No. I12418); Cry1Ac13 (Grant No. AAD38701); Cry1Ac14 (Grant No. AAQ06607); Cry1Ac15 (Grant No. AAN07788); Cry1Ac16 (Grant No. AAU87037); Cry1Ac17 (Grant No. AAX18704); Cry1Ac18 (Grant No. AAY88347); Cry1Ac19 (Grant No. AB) D37053); Cry1Ac20 (accession number ABB89046); Cry1Ac21 (accession number AAY66992); Cry1Ac22 (accession number ABZ01836); Cry1Ac23 (accession number CAQ30431); Cry1Ac24 (accession number ABL01535); Cry1Ac25 (accession number FJ513324); Cry1Ac26 (accession number FJ617446); Cry1Ac27 (accession number FJ617447); Cry1Ac28 (accession number ACM90319); Cry1Ac29 (accession number DQ438941); Cry1Ac30 (accession number GQ227507);Cry1Ac31 (Accession number: GU446674); Cry1Ac32 (Accession number: HM061081); Cry1Ac33 (Accession number: GQ866913); Cry1Ac34 (Accession number: HQ230364); Cry1Ac35 (Accession number: JF340157); Cry1Ac36 (Accession number: JN387137); Cry1Ac37 (Accession number: JQ317685); Cry1Ad1 (Accession number: AAA22340); Cry1Ad2 (Accession number: CAA01880); Cry1Ae1 (Accession number: AAA22410); Cry1Af1 (Accession number: AAB82749); Cry1Ag1 (Accession number: AAD46137); Cry1Ah1 (Accession number: AAQ14326); Cry1Ah2 (Accession number: ABB76664); Cry1Ah3 (Accession number: HQ439779); Cry1Ai1 (Accession number: AAO39719); Cry1Ai2 (Accession number: HQ439780); Cry1A-like (Accession number: AAK14339); Cry1Ba1 (Accession number: CAA29898); Cry1Ba2 (Accession number: CAA65003); Cry1Ba3 (Accession number: AAK63251); Cry1Ba4 (Accession number: AAK51084); Cry1Ba5 (Accession number: ABO20894); Cry1Ba6 (Accession number: ABL60921); Cry1Ba7 (Accession number: HQ439781); Cry1Bb1 (Accession number: AAA22344); Cry1Bb2 (Accession number: HQ439782); Cry1Bc1 (Accession number: CAA86568); Cry1Bd1 (Accession number: AAD10292); Cry1Bd2 (Accession number: AAM93496); Cry1Be1 (Accession number: AAC32850); Cry1Be2 (Accession number: AAQ52387); Cry1BeCry1Ca9 (Grant No. AAL79362); Cry1Ca10 (Grant No. AAN16462); Cry1Ca11 (Grant No. AAX53094); Cry1Ca12 (Grant No. HM070027); Cry1Ca13 (Grant No. HQ412621); Cry1Ca14 (Grant No. JN651493); Cry1Cb1 (Grant No. M97880); Cry1Cb2 (Grant No. AAG35409); Cry1Cb3 (Grant No. ACD50894); Cry1Cb (Grant No. AAX63901); Cry1Da1 (Grant No. CAA38099); Cry1Da2 (Grant No. I7) 6415); Cry1Da3 (Grant number HQ439784); Cry1Db1 (Grant number CAA80234); Cry1Db2 (Grant number AAK48937); Cry1Dc1 (Grant number ABK35074); Cry1Ea1 (Grant number CAA37933); Cry1Ea2 (Grant number CAA39609); Cry1Ea3 (Grant number AAA22345); Cry1Ea4 (Grant number AAD04732); Cry1Ea5 (Grant number A15535); Cry1Ea6 (Grant number AAL50330); Cry1Ea7 (Grant number AAW72936); Cry1Ea8 (Grant number A BX11258); Cry1Ea9 (Grant number HQ439785); Cry1Ea10 (Grant number ADR00398); Cry1Ea11 (Grant number JQ652456); Cry1Eb1 (Grant number AAA22346); Cry1Fa1 (Grant number AAA22348); Cry1Fa2 (Grant number AAA22347); Cry1Fa3 (Grant number HM070028); Cry1Fa4 (Grant number HM439638); Cry1Fb1 (Grant number CAA80235); Cry1Fb2 (Grant number BAA25298); Cry1Fb3 (Grant number AAF21767); Cry1F b4 (Grant No. AAC10641); Cry1Fb5 (Grant No. AAO13295); Cry1Fb6 (Grant No. ACD50892); Cry1Fb7 (Grant No. ACD50893); Cry1Ga1 (Grant No. CAA80233); Cry1Ga2 (Grant No. CAA70506); Cry1Gb1 (Grant No. AAD10291); Cry1Gb2 (Grant No. AAO13756); Cry1Gc1 (Grant No. AAQ52381); Cry1Ha1 (Grant No. CAA80236); Cry1Hb1 (Grant No. AAA79694); Cry1Hb2 (Grant No. HQ439786);Cry1H (Grant No. AAF01213); Cry1Ia1 (Grant No. CAA44633); Cry1Ia2 (Grant No. AAA22354); Cry1Ia3 (Grant No. AAC36999); Cry1Ia4 (Grant No. AAB00958); Cry1Ia5 (Grant No. CAA70124); Cry1Ia6 (Grant No. AAC26910); Cry1Ia7 (Grant No. AAM73516); Cry1Ia8 (Grant No. AAK66742); Cry1Ia9 (Grant No. AAQ08616); Cry1Ia10 (Grant No. AAP86782); Cry1Ia11 (Grant No. CAC8 5964); Cry1Ia12 (Grant No. AAV53390); Cry1Ia13 (Grant No. ABF83202); Cry1Ia14 (Grant No. ACG63871); Cry1Ia15 (Grant No. FJ617445); Cry1Ia16 (Grant No. FJ617448); Cry1Ia17 (Grant No. GU989199); Cry1Ia18 (Grant No. ADK23801); Cry1Ia19 (Grant No. HQ439787); Cry1Ia20 (Grant No. JQ228426); Cry1Ia21 (Grant No. JQ228424); Cry1Ia22 (Grant No. JQ228427); Cry1Ia23 (Login ID JQ228428); Cry1Ia24 (Login ID JQ228429); Cry1Ia25 (Login ID JQ228430); Cry1Ia26 (Login ID JQ228431); Cry1Ia27 (Login ID JQ228432); Cry1Ia28 (Login ID JQ228433); Cry1Ia29 (Login ID JQ228434); Cry1Ia30 (Login ID JQ317686); Cry1Ia31 (Login ID JX944038); Cry1Ia32 (Login ID JX944039); Cry1Ia33 (Login ID JX944040); Cry1Ib Cry1Ib1 (Login ID AAA82114); Cry1Ib2 (Login ID ABW88019); Cry1Ib3 (Login ID ACD75515); Cry1Ib4 (Login ID HM051227); Cry1Ib5 (Login ID HM070028); Cry1Ib6 (Login ID ADK38579); Cry1Ib7 (Login ID JN571740); Cry1Ib8 (Login ID JN675714); Cry1Ib9 (Login ID JN675715); Cry1Ib10 (Login ID JN675716); Cry1Ib11 (Login ID JQ228423); Cry1Ic1 (Login ID AAC62933);Cry1Ic2 (Accession number AAE71691); Cry1Id1 (Accession number AAD44366); Cry1Id2 (Accession number JQ228422); Cry1Ie1 (Accession number AAG43526); Cry1Ie2 (Accession number HM439636); Cry1Ie3 (Accession number KC156647); Cry1Ie4 (Accession number KC156681); Cry1If1 (Accession number AAQ52382); Cry1Ig1 (Accession number KC156701); Cry1I-like (Accession number AAC31094); Cry1I-like (Accession number ABG88859); Cry1Ja1 (Accession number AAA22341); Cry1Ja2 (Accession number HM070030); Cry1Ja3 (Accession number JQ228425); Cry1Jb1 (Accession number AAA98959); Cry1Jc1 (Accession number AAC31092); Cry1Jc2 (Accession number AAQ52372); Cry1Jd1 (Accession number CAC50779); Cry1Ka1 (Accession number AAB00376); Cry1Ka2 (Accession number HQ439783); Cry1La1 (Accession number AAS60191); Cry1La2 (Accession number HM070031); Cry1Ma1 (Accession number FJ884067); Cry1Ma2 (Accession number KC156659); Cry1Na1 (Accession number KC,156648); Cry1Nb1 (Accession number KC156678); Cry1-like (Accession number AAC31091); Cry2Aa1 (Accession number AAA22335); Cry2Aa2 (Accession number AAA83516); Cry2Aa3 (Accession number D86064); Cry2Aa4 (Accession number AAC04867); Cry2Aa5 (Accession number CAA10671); Cry2Aa6 (Accession number CAA10672); Cry2Aa7 (Accession number CAA10670); Cry2Aa8 (Accession number AAO13734); Cry2Aa9 (Accession number AAO13750); Cry2Aa10 (Accession number AAQ04263); Cry2Aa11 (Accession number AAQ52384); Cry2Aa12 (Accession number ABI83671); Cry2Aa13 (Accession number ABL01536); Cry2Aa14 (Accession number ACF04939); Cry2Aa15 (Accession number JN426947); Cry2Ab1 (Accession number AAA22342); Cry2Ab2 (Accession number CAA39075); Cry2Ab3 (Accession number AAG36762); Cry2Ab4 (Accession number AAO13296); Cry2Ab5 (Accession number AAQ04609);Cry2Ab6 (Grant No. AAP59457); Cry2Ab7 (Grant No. AAZ66347); Cry2Ab8 (Grant No. ABC95996); Cry2Ab9 (Grant No. ABC74968); Cry2Ab10 (Grant No. EF157306); Cry2Ab11 (Grant No. CAM84575); Cry2Ab12 (Grant No. ABM21764); Cry2Ab13 (Grant No. ACG76120); Cry2Ab14 (Grant No. ACG76121); Cry2Ab15 (Grant No. HM037126); Cry2Ab16 (Grant No. GQ866914); Cry2Ab17 ( Login ID HQ439789); Cry2Ab18 (Login ID JN135255); Cry2Ab19 (Login ID JN135256); Cry2Ab20 (Login ID JN135257); Cry2Ab21 (Login ID JN135258); Cry2Ab22 (Login ID JN135259); Cry2Ab23 (Login ID JN135260); Cry2Ab24 (Login ID JN135261); Cry2Ab25 (Login ID JN415485); Cry2Ab26 (Login ID JN426946); Cry2Ab27 (Login ID JN415764); Cry2Ab28 (Login ID JN 651494); Cry2Ac1 (Grant number CAA40536); Cry2Ac2 (Grant number AAG35410); Cry2Ac3 (Grant number AAQ52385); Cry2Ac4 (Grant number ABC95997); Cry2Ac5 (Grant number ABC74969); Cry2Ac6 (Grant number ABC74793); Cry2Ac7 (Grant number CAL18690); Cry2Ac8 (Grant number CAM09325); Cry2Ac9 (Grant number CAM09326); Cry2Ac10 (Grant number ABN15104); Cry2Ac11 (Grant number CAM83895); Cry2Ac 12 (Login ID CAM83896); Cry2Ad1 (Login ID AAF09583); Cry2Ad2 (Login ID ABC86927); Cry2Ad3 (Login ID CAK29504); Cry2Ad4 (Login ID CAM32331); Cry2Ad5 (Login ID CAO78739); Cry2Ae1 (Login ID AAQ52362); Cry2Af1 (Login ID ABO30519); Cry2Af2 (Login ID GQ866915); Cry2Ag1 (Login ID ACH91610); Cry2Ah1 (Login ID EU939453); Cry2Ah2 (Login ID ACL80665);Cry2Ah3 (GU073380); Cry2Ah4 (KC156702); Cry2Ai1 (FJ788388); Cry2Aj (GROUP ID); Cry2Ak1 (KC156660); Cry2Ba1 (KC156658); Cry3Aa1 (AAA22336); Cry3Aa2 (AAA22541); Cry3Aa3 (CAA68482); Cry3Aa4 (AAA22542); Cry3Aa5 (AAA50255); Cry3Aa6 (AAC43266); Cry3Aa 7 (Login ID CAB41411); Cry3Aa8 (Login ID AAS79487); Cry3Aa9 (Login ID AAW05659); Cry3Aa10 (Login ID AAU29411); Cry3Aa11 (Login ID AAW82872); Cry3Aa12 (Login ID ABY49136); Cry3Ba1 (Login ID CAA34983); Cry3Ba2 (Login ID CAA00645); Cry3Ba3 (Login ID JQ397327); Cry3Bb1 (Login ID AAA22334); Cry3Bb2 (Login ID AAA74198); Cry3Bb3 (Login ID I15475); Cr y3Ca1 (Registration No. CAA42469); Cry4Aa1 (Registration No. CAA68485); Cry4Aa2 (Registration No. BAA00179); Cry4Aa3 (Registration No. CAD30148); Cry4Aa4 (Registration No. AFB18317); Cry4A sample (Registration No. AAY96321); Cry4Ba1 (Registration No. CAA30312); Cry4Ba2 (Registration No. CAA30114); Cry4Ba3 (Registration No. AAA22337); Cry4Ba4 (Registration No. BAA00178); Cry4Ba5 (Registration No. CAD30095); Cry4Ba sample (Registration No. ABC47686); Cry4Ca1 (Grant No. EU646202); Cry4Cb1 (Grant No. FJ403208); Cry4Cb2 (Grant No. FJ597622); Cry4Cc1 (Grant No. FJ403207); Cry5Aa1 (Grant No. AAA67694); Cry5Ab1 (Grant No. AAA67693); Cry5Ac1 (Grant No. I34543); Cry5Ad1 (Grant No. ABQ82087); Cry5Ba1 (Grant No. AAA68598); Cry5Ba2 (Grant No. ABW88931); Cry5Ba3 (Grant No. AFJ04417); Cry5Ca1 (Grant No. HM461869);Cry5Ca2 (Login ID ZP_04123426); Cry5Da1 (Login ID HM461870); Cry5Da2 (Login ID ZP_04123980); Cry5Ea1 (Login ID HM485580); Cry5Ea2 (Login ID ZP_04124038); Cry6Aa1 (Login ID AAA22357); Cry6Aa2 (Login ID AAM46849); Cry6Aa3 (Login ID ABH03377); Cry6Ba1 (Login ID AAA22358); Cry7Aa1 (Login ID AAA22351); Cry7Ab1 (Login ID AAA21120); Cry7Ab ...Da2 (Login ID ZP_04123980); Cry5Da2 (Login ID ZP_04 Accession number AAA21121); Cry7Ab3 (accession number ABX24522); Cry7Ab4 (accession number EU380678); Cry7Ab5 (accession number ABX79555); Cry7Ab6 (accession number ACI44005); Cry7Ab7 (accession number ADB89216); Cry7Ab8 (accession number GU145299); Cry7Ab9 (accession number ADD92572); Cry7Ba1 (accession number ABB70817); Cry7Bb1 (accession number KC156653); Cry7Ca1 (accession number ABR67863); Cry7Cb1 (accession number KC156698); Cry7Da 1 (Login ID ACQ99547); Cry7Da2 (Login ID HM572236); Cry7Da3 (Login ID KC156679); Cry7Ea1 (Login ID HM035086); Cry7Ea2 (Login ID HM132124); Cry7Ea3 (Login ID EEM19403); Cry7Fa1 (Login ID HM035088); Cry7Fa2 (Login ID EEM19090); Cry7Fb1 (Login ID HM572235); Cry7Fb2 (Login ID KC156682); Cry7Ga1 (Login ID HM572237); Cry7Ga2 (Login ID KC156669); Cry 7Gb1 (GROUP BY KC156650); Cry7Gc1 (GROUP BY KC156654); Cry7Gd1 (GROUP BY KC156697); Cry7Ha1 (GROUP BY KC156651); Cry7Ia1 (GROUP BY KC156665); Cry7Ja1 (GROUP BY KC156671); Cry7Ka1 (GROUP BY KC156680); Cry7Kb1 (GROUP BY BAM99306); Cry7La1 (GROUP BY BAM99307); Cry8Aa1 (GROUP BY AAA21117); Cry8Ab1 (GROUP BY EU044830); Cry8Ac1 (GROUP BY KC156662);Cry8Ad1 (Accession number KC156684); Cry8Ba1 (Accession number AAA21118); Cry8Bb1 (Accession number CAD57542); Cry8Bc1 (Accession number CAD57543); Cry8Ca1 (Accession number AAA21119); Cry8Ca2 (Accession number AAR98783); Cry8Ca3 (Accession number EU625349); Cry8Ca4 (Accession number ADB54826); Cry8Da1 (Accession number BAC07226); Cry8Da2 (Accession number BD133574); Cry8Da3 (Accession number BD133575); Cry8Db1 (Accession number BAF93483); Cry8Ea1 (Accession number AAQ73470); Cry8Ea2 (Accession number EU047597); Cry8Ea3 (Accession number KC855216); Cry8Fa1 (Accession number AAT48690); Cry8Fa2 (Accession number HQ174208); Cry8Fa3 (Accession number AFH78109); Cry8Ga1 (Accession number AAT46073); Cry8Ga2 (Accession number ABC42043); Cry8Ga3 (Accession number FJ198072); Cry8Ha1 (Accession number AAW81032); Cry8Ia1 (Accession number EU381044); Cry8Ia2 (Accession number GU073381); Cry8Ia3 (Accession number HM044664); Cry8Ia4 (Accession number KC156674); Cry8Ib1 (Accession number GU325772); Cry8Ib2 (Accession number KC156677); Cry8Ja1 (Accession number EU625348); Cry8Ka1 (Accession number FJ422558); Cry8Ka2 (Accession number ACN87262); Cry8Kb1 (Accession number HM123758); Cry8Kb2 (Accession number KC156675); Cry8La1 (Accession number GU325771); Cry8Ma1 (Accession number HM044665); Cry8Ma2 (Accession number EEM86551); Cry8Ma3 (Accession number HM210574); Cry8Na1 (Accession number HM640939); Cry8Pa1 (Accession number HQ388415); Cry8Qa1 (Accession number HQ441166); Cry8Qa2 (Accession number KC152468); Cry8Ra1 (Accession number AFP87548); Cry8Sa1 (Accession number JQ740599); Cry8Ta1 (Accession number KC156673); Cry8-like (Accession number FJ770571); Cry8-like (Accession number ABS53003); Cry9Aa1 (Accession number CAA41122);Cry9Aa2 (Accession number: CAA41425); Cry9Aa3 (Accession number: GQ249293); Cry9Aa4 (Accession number: GQ249294); Cry9Aa5 (Accession number: JX174110); Cry9Aa-like (Accession number: AAQ52376); Cry9Ba1 (Accession number: CAA52927); Cry9Ba2 (Accession number: GU299522); Cry9Bb1 (Accession number: AAV28716); Cry9Ca1 (Accession number: CAA85764); Cry9Ca2 (Accession number: AAQ52375); Cry9Da1 (Accession number: BAA19948); Cry9Da2 (Accession number: AAB97923); Cry9Da3 (Accession number: GQ249293); Cry9Da4 (Accession number: GQ249297); Cry9Db1 (Accession number: AAX78439); Cry9Dc1 (Accession number: KC156683); Cry9Ea1 (Accession number: BAA34908); Cry9Ea2 (Accession number: AAO12908); Cry9Ea3 (Accession number: ABM21765); Cry9Ea4 (Accession number: ACE88267); Cry9Ea5 (Accession number: ACF04743); Cry9Ea6 (Accession number: ACG63872); Cry9Ea7 (Accession number: FJ380927); Cry9Ea8 (Accession number: GQ249292); Cry9Ea9 (Accession number: JN651495); Cry9Eb1 (Accession number: CAC50780); Cry9Eb2 (Accession number: GQ249298); Cry9Eb3 (Accession number: KC156646); Cry9Ec1 (Accession number: AAC63366); Cry9Ed1 (Accession number: AAX78440); Cry9Ee1 (Accession number: GQ249296); Cry9Ee2 (Accession number: KC156664); Cry9Fa1 (Accession number: KC156692); Cry9Ga1 (Accession number: KC156699); Cry9-like (Accession number: AAC63366); Cry10Aa1 (Accession number: AAA22614); Cry10Aa2 (Accession number: E00614); Cry10Aa3 (Accession number: CAD30098); Cry10Aa4 (Accession number: AFB18318); Cry10A-like (Accession number: DQ167578); Cry11Aa1 (Accession number: AAA22352); Cry11Aa2 (Accession number: AAA22611); Cry11Aa3 (Accession number: CAD30081); Cry11Aa4 (Accession number: AFB18319); Cry11Aa-like (Accession number: DQ166531); Cry11Ba1 (Accession number: CAA60504); Cry11Bb1 (Accession number: AAC97162);Cry11Bb2 (Grant number HM068615); Cry12Aa1 (Grant number AAA22355); Cry13Aa1 (Grant number AAA22356); Cry14Aa1 (Grant number AAA21516); Cry14Ab1 (Grant number KC156652); Cry15Aa1 (Grant number AAA22333); Cry16Aa1 (Grant number CAA63860); Cry17Aa1 (Grant number CAA67841); Cry18Aa1 (Grant number CAA67506); Cry18Ba1 (Grant number AAF89667); Cry18Ca1 (Grant number AAF89668); Cry 19Aa1 (Grant No. CAA68875); Cry19Ba1 (Grant No. BAA32397); Cry19Ca1 (Grant No. AFM37572); Cry20Aa1 (Grant No. AAB93476); Cry20Ba1 (Grant No. ACS93601); Cry20Ba2 (Grant No. KC156694); Cry20sample (Grant No. GQ144333); Cry21Aa1 (Grant No. I32932); Cry21Aa2 (Grant No. I66477); Cry21Ba1 (Grant No. BAC06484); Cry21Ca1 (Grant No. JF521577); Cry21Ca2 (Grant No. K C156687); Cry21Da1 (Grant number JF521578); Cry22Aa1 (Grant number I34547); Cry22Aa2 (Grant number CAD43579); Cry22Aa3 (Grant number ACD93211); Cry22Ab1 (Grant number AAK50456); Cry22Ab2 (Grant number CAD43577); Cry22Ba1 (Grant number CAD43578); Cry22Bb1 (Grant number KC156672); Cry23Aa1 (Grant number AAF76375); Cry24Aa1 (Grant number AAC61891); Cry24Ba1 (Grant number BAD326) 57); Cry24Ca1 (Grant No. CAJ43600); Cry25Aa1 (Grant No. AAC61892); Cry26Aa1 (Grant No. AAD25075); Cry27Aa1 (Grant No. BAA82796); Cry28Aa1 (Grant No. AAD24189); Cry28Aa2 (Grant No. AAG00235); Cry29Aa1 (Grant No. CAC80985); Cry30Aa1 (Grant No. CAC80986); Cry30Ba1 (Grant No. BAD00052); Cry30Ca1 (Grant No. BAD67157); Cry30Ca2 (Grant No. ACU24781);Cry30Da1 (Grant No. EF095955); Cry30Db1 (Grant No. BAE80088); Cry30Ea1 (Grant No. ACC95445); Cry30Ea2 (Grant No. FJ499389); Cry30Fa1 (Grant No. ACI22625); Cry30Ga1 (Grant No. ACG60020); Cry30Ga2 (Grant No. HQ638217); Cry31Aa1 (Grant No. BAB11757); Cry31Aa2 (Grant No. AAL87458); Cry31Aa3 (Grant No. BAE79808); Cry31Aa4 (Grant No. BAF32571); Cry31 Aa5 (Grant No. BAF32572); Cry31Aa6 (Grant No. BAI44026); Cry31Ab1 (Grant No. BAE79809); Cry31Ab2 (Grant No. BAF32570); Cry31Ac1 (Grant No. BAF34368); Cry31Ac2 (Grant No. AB731600); Cry31Ad1 (Grant No. BAI44022); Cry32Aa1 (Grant No. AAG36711); Cry32Aa2 (Grant No. GU063849); Cry32Ab1 (Grant No. GU063850); Cry32Ba1 (Grant No. BAB78601); Cry32Ca1 (Grant No. BAF32572); Cry32 (GROUP NO BAB78602); Cry32Cb1 (GROUP NO KC156708); Cry32Da1 (GROUP NO BAB78603); Cry32Ea1 (GROUP NO GU324274); Cry32Ea2 (GROUP NO KC156686); Cry32Eb1 (GROUP NO KC156663); Cry32Fa1 (GROUP NO KC156656); Cry32Ga1 (GROUP NO KC156657); Cry32Ha1 (GROUP NO KC156661); Cry32Hb1 (GROUP NO KC156666); Cry32Ia1 (GROUP NO KC156667); Cry32Ja1 (GROUP NO KC156608); Cry32Da1 (GROUP NO BAB78603); Cry32Ea1 (GROUP NO KC156608 ... Cry32Ka1 (KC156688); Cry32La1 (KC156689); Cry32Ma1 (KC156690); Cry32Mb1 (KC156704); Cry32Na1 (KC156691); Cry32Oa1 (KC156703); Cry32Pa1 (KC156705); Cry32Qa1 (KC156706); Cry32Ra1 (KC156707); Cry32Sa1 (KC156709); Cry32Ta1 (KC156710);Cry32Ua1 (Accession number KC156655); Cry33Aa1 (Accession number AAL26871); Cry34Aa1 (Accession number AAG50341); Cry34Aa2 (Accession number AAK64560); Cry34Aa3 (Accession number AAT29032); Cry34Aa4 (Accession number AAT29030); Cry34Ab1 (Accession number AAG41671); Cry34Ac1 (Accession number AAG50118); Cry34Ac2 (Accession number AAK64562); Cry34Ac3 (Accession number AAT29029); Cry34Ba1 (Accession number AAK64565); Cry34Ba2 (Accession number AAT29033); Cry34Ba3 (Accession number AAT29031); Cry35Aa1 (Accession number AAG50342); Cry35Aa2 (Accession number AAK64561); Cry35Aa3 (Accession number AAT29028); Cry35Aa4 (Accession number AAT29025); Cry35Ab1 (Accession number AAG41672); Cry35Ab2 (Accession number AAK64563); Cry35Ab3 (Accession number AY536891); Cry35Ac1 (Accession number AAG50117); Cry35Ba1 (Accession number AAK64566); Cry35Ba2 (Accession number AAT29027); Cry35Ba3 (Accession number AAT29026); Cry36Aa1 (Accession number AAK64558); Cry37Aa1 (Accession number AAF76376); Cry38Aa1 (Accession number AAK6455Cry45Aa (login ID BAD22577); Cry46Aa (login ID BAC79010); Cry46Aa2 (login ID BAG68906); Cry46Ab (login ID BAD35170); Cry47Aa (login ID AAY24695); Cry48Aa (login ID CAJ18351); Cry48Aa2 (login ID CAJ86545); Cry48Aa3 (login ID CAJ86546); Cry48Ab (login ID CAJ86548); Cry48Ab2 (login ID CAJ86549); Cry49Aa (login ID CAH56541); Cry49Aa2 (login ID CAH56541); Cry49Aa2 (login ID BAD22577); Cry46Aa ... Accession number CAJ86541); Cry49Aa3 (accession number CAJ86543); Cry49Aa4 (accession number CAJ86544); Cry49Ab1 (accession number CAJ86542); Cry50Aa1 (accession number BAE86999); Cry50Ba1 (accession number GU446675); Cry50Ba2 (accession number GU446676); Cry51Aa1 (accession number ABI14444); Cry51Aa2 (accession number GU570697); Cry52Aa1 (accession number EF613489); Cry52Ba1 (accession number FJ361760); Cry53Aa1 (accession number EF613489); 633476); Cry53Ab1 (GROUP BY FJ361759); Cry54Aa1 (GROUP BY ACA52194); Cry54Aa2 (GROUP BY GQ140349); Cry54Ba1 (GROUP BY GU446677); Cry55Aa1 (GROUP BY ABW88932); Cry54Ab1 (GROUP BY JQ916908); Cry55Aa2 (GROUP BY AAE33526); Cry56Aa1 (GROUP BY ACU57499); Cry56Aa2 (GROUP BY GQ483512); Cry56Aa3 (GROUP BY JX025567); Cry57Aa1 (GROUP BY ANC872) 61); Cry58Aa1 (Grant No. ANC87260); Cry59Ba1 (Grant No. JN790647); Cry59Aa1 (Grant No. ACR43758); Cry60Aa1 (Grant No. ACU24782); Cry60Aa2 (Grant No. EAO57254); Cry60Aa3 (Grant No. EEM99278); Cry60Ba1 (Grant No. GU810818); Cry60Ba2 (Grant No. EAO57253); Cry60Ba3 (Grant No. EEM99279); Cry61Aa1 (Grant No. HM035087); Cry61Aa2 (Grant No. HM132125);Cry61Aa3 (Grant No. EEM19308); Cry62Aa1 (Grant No. HM054509); Cry63Aa1 (Grant No. BAI44028); Cry64Aa1 (Grant No. BAJ05397); Cry65Aa1 (Grant No. HM461868); Cry65Aa2 (Grant No. ZP_04123838); Cry66Aa1 (Grant No. HM485581); Cry66Aa2 (Grant No. ZP_04099945); Cry67Aa1 (Grant No. HM485582); Cry67Aa2 (Grant No. ZP_04148882); Cry68Aa1 (Grant No. HQ113114); Cry69Aa1 (Grant No. HQ401006); Cr y69Aa2 (Login ID JQ821388); Cry69Ab1 (Login ID JN209957); Cry70Aa1 (Login ID JN646781); Cry70Ba1 (Login ID ADO51070); Cry70Bb1 (Login ID EEL67276); Cry71Aa1 (Login ID JX025568); Cry72Aa1 (Login ID JX025569); Cyt1Aa (GenBank Login ID X03182); Cyt1Ab (GenBank Login ID X98793); Cyt1B (GenBank Login ID U37196); Cyt2A (GenBank Login ID Z14147); and Cyt2B (GenBank Login ID U52043).

[0075] Examples of delta-endotoxins include, but are not limited to: the Cry1A protein in U.S. Patents 5,880,275, 7,858,849, 8,530,411, 8,575,433, and 8,686,233; and DIG-3 or DIG-11 toxins (α-helical 1 and / or α-spiral of cry proteins such as Cry1A, Cry3A) in U.S. Patents 8,304,604, 8,304,605, and 8,476,226. (N-terminal deletion of the Cry2 variant); Cry1B in U.S. Patent Application Serial No. 10 / 525,318; Cry1C in U.S. Patent 6,033,874; Cry1F in U.S. Patents 5,188,960 and 6,218,188; Cry1A / F chimeras in U.S. Patents 7,070,982, 6,962,705 and 6,713,063; Cry2 proteins, such as those in U.S. Patent 7,064,249. Cry2Ab protein; Cry3A protein, including but not limited to engineered hybrid insecticidal protein (eHIP) formed by a unique combination of variable and conserved blocks of at least two different Cry proteins (US Patent Application Publication 2010 / 0017914); Cry4 protein; Cry5 protein; Cry6 protein; Cry8 protein as described in US Patents 7,329,736, 7,449,552, 7,803,943, 7,476,781, 7,105,332, 7,378,499, and 7,462,760; Cry9 protein, such as members of the Cry9A, Cry9B, Cry9C, Cry9D, Cry9E, and Cry9F families, including but not limited to Cry9D protein as described in US Patent 8,802,933 and Cry9B protein as described in US Patent 8,802,934; Naimov et al., (2008) Applied and Environmental MicrobiologyCry15 protein in 74:7145-7151; Cry22 and Cry34Ab1 proteins in U.S. Patents 6,127,180, 6,624,145, and 6,340,593; CryET33 and CryET34 proteins in U.S. Patents 6,248,535, 6,326,351, 6,399,330, 6,949,626, 7,385,107, and 7,504,229; U.S. Patent Publications 2006 / 0191034, 2012 / 0278954, and PCT Publication WO CryET33 and CryET34 homologs in 2012 / 139004; Cry35Ab1 protein in U.S. Patents 6,083,499, 6,548,291, and 6,340,593; Cry46 protein, Cry51 protein, and Cry binary toxin; TIC901 or related toxins; TIC807 in U.S. Patent Application Publication 2008 / 0295207; PCT US ET29, ET37, TIC809, TIC810, TIC812, TIC127, and TIC128 in WO 2006 / 033867; TIC853 toxin in US Patent 8,513,494; AXMI-027, AXMI-036, and AXMI-038 in US Patent 8,236,757; AXMI-031, AXMI-039, AXMI-040, and AXMI-049 in US Patent 7,923,602; AXMI-018, AXMI-020, and AXMI-021 in WO 2006 / 083891; WO AXMI-010 in WO 2005 / 038032; AXMI-003 in WO 2005 / 021585; AXMI-008 in U.S. Patent Application Publication 2004 / 0250311; AXMI-006 in U.S. Patent Application Publication 2004 / 0216186; AXMI-007 in U.S. Patent Application Publication 2004 / 0210965; AXMI-009 in U.S. Patent Application Publication 2004 / 0210964; AXMI-014 in U.S. Patent Application Publication 2004 / 0197917; AXMI-004 in U.S. Patent Application Publication 2004 / 0197916; WO AXMI-028 and AXMI-029 in 2006 / 119457; AXMI-007, AXMI-008, AXMI-0080rf2, AXMI-009, AXMI-014 and AXMI-004 in WO2004 / 074462; AXMI-150 in U.S. Patent 8,084,416; AXMI-205 in U.S. Patent Application Publication 2011 / 0023184;AXMI-011, AXMI-012, AXMI-013, AXMI-015, AXMI-019, AXMI-044, AXMI-037, AXMI-043, AXMI-033, AXMI-034, AXMI-022, AXMI-023, AXMI-041, AXMI-063, and AXMI-064 in U.S. Patent Application Publication 2010 / 0197592; AXMI-R1 and related proteins in WO 2011 / 103248; AXMI221Z, AXMI222z, AXMI223z, AXMI224z, and AXMI225z in WO 2011 / 103248; WO AXMI218, AXMI219, AXMI220, AXMI226, AXMI227, AXMI228, AXMI229, AXMI230, and AXMI231 in U.S. Patent 2011 / 103247 and U.S. Patent 8,759,619; AXMI-115, AXMI-113, AXMI-005, AXMI-163, and AXMI-184 in U.S. Patent 8,334,431; AXMI-001, AXMI-002, AXMI-030, AXMI-035, and AXMI-045 in U.S. Patent Application Publication 2010 / 0298211; AXMI-066 and AXMI-076 in U.S. Patent Application Publication 2009 / 0144852; AXMI128, AXMI130, AXMI131, AXMI133, and A in U.S. Patent 8,318,900. XMI140, AXMI141, AXMI142, AXMI143, AXMI144, AXMI146, AXMI148, AXMI149, AXMI152, AXMI153, AXMI154, AXMI155, AXMI156, AXMI157, AXMI158, AXMI162, AXMI165, AXMI166, AXMI167, AXMI 168, AXMI169, AXMI170, AXMI171, AXMI172, AXMI173, AXMI174, AXMI175, AXMI176, AXMI177, AX MI178, AXMI179, AXMI180, AXMI181, AXMI182, AXMI185, AXMI186, AXMI187, AXMI188, AXMI189;AXMI079, AXMI080, AXMI081, AXMI082, AXMI091, AXMI092, AXMI096, AXMI097, AXMI098, AXMI099, AXMI100, AXMI101, AXMI102, AXMI103, AXMI104, AXMI107, AXMI108, AXMI109, AXMI110, AXMI111, AXMI112, AXMI114, AXMI116, AXMI117, AXMI118, AXMI119, AXMI120, AXMI121, and AXMI122 from U.S. Patent Application Publication 2010 / 0005543 AXMI123, AXMI124, AXMI1257, AXMI1268, AXMI127, AXMI129, AXMI164, AXMI151, AXMI161, AXMI183, AXMI132, AXMI138, AXMI137; AXMI270 in US Patent Application Publication US20140223598; AXMI279 in US Patent Application Publication US20140223599; cry proteins, such as Cry1A and Cry3A with modified proteolytic sites in US Patent 8,319,019; and Cry1Ac, Cry2Aa, and Cry1Ca toxin proteins from Bacillus thuringiensis strain VBTS 2528 in US Patent Application Publication 2011 / 0064710. Other Cry proteins are also well known to those skilled in the art (see Crickmore et al., “Bacillus thuringiensis toxin nomenclature” (2011) at lifesci.sussex.ac.uk / home / Neil_Crickmore / Bt / (accessible via the “www” prefix on the World Wide Web)). The insecticidal activity of Cry proteins is well known to those skilled in the art (for a review, see van Frannkenhuyzen, (2009) J. Invert. Path. 101:1-16). The use of Cry proteins as traits in transgenic plants is well known to those skilled in the art, and Cry transgenic plants (including, but not limited to, plants expressing Cry1Ac, Cry1Ac+Cry2Ab, Cry1Ab, Cry1A.105, Cry1F, Cry1Fa2, Cry1F+Cry1Ac, Cry2Ab, Cry3A, mCry3A, Cry3Bb1, Cry34Ab1, Cry35Ab1, Vip3A, mCry3A, Cry9c, and CBI-Bt) have been subject to regulatory approval (see Sanahuja, (2011)). Plant Biotech Journal9:283-300; and CERA (2010) GM Crop Database Center for Environmental Risk Assessment (CERA), ILSI Research Foundation, Washington DC, at cera-gmc.org / index.php?action=gm_crop_database (accessible via the World Wide Web with the prefix "www"). More than one insecticidal protein well known to those skilled in the art can also be expressed in plants, such as: Vip3Ab and Cry1Fa (US2012 / 0317682); Cry1BE and Cry1F (US2012 / 0311746); Cry1CA and Cry1AB (US2012 / 0311745); Cry1F and CryCa (US2012 / 0317681); Cry1DA and Cry1BE (US2012 / 0331590); Cry1DA and Cry1Fa (US2012 / 0331589); C Cry1AB and Cry1BE (US2012 / 0324606); Cry1Fa and Cry2Aa, Cry1I and Cry1E (US2012 / 0324605); Cry34Ab / 35Ab and Cry6Aa (US20130167269); Cry34Ab / VCry35Ab and Cry3Aa (US20130167268); Cry1Ab and Cry1F (US20140182018); Cry3A and Cry1Ab or Vip3Aa (US20130116170). Insecticidal proteins also include insecticidal lipases (including acyl hydrolases in US Patent 7,491,869) and cholesterol oxidases, such as those from the genus Streptomyces (…). Streptomyces Cholesterol oxidase (Purcell et al. (1993)) Biochem Biophys Res Commun 15:1406-1413). Insecticidal proteins also include VIP (plant-derived insecticidal protein) toxins, etc., of U.S. patents 5,877,012, 6,107,279, 6,137,033, 7,244,820, 7,615,686, and 8,237,020. Other VIP proteins are well known to those skilled in the art (see lifesci.sussex.ac.uk / home / Neil_Crickmore / Bt / vip.html (accessible via the World Wide Web with the prefix "www")). Insecticidal proteins also include toxin complex (TC) proteins, which can be produced by pathogenic bacteria, luminescent bacteria, and spore-forming bacteria (Bacillus). PaenibacillusThese proteins are obtained from organisms such as (see U.S. Patents 7,491,698 and 8,084,418). Some TC proteins have “independent” insecticidal activity, while others enhance the activity of independent toxins produced by the same given organism. The toxicity of “independent” TC proteins (from, for example, *Luminobacterium*, *Pathobacterium*, or *Bacillus* species) can be enhanced by one or more TC protein “synergists” from different genera of source organisms. There are three main types of TC proteins. As mentioned herein, type A proteins (“protein A”) are independent toxins. Type B proteins (“protein B”) and type C proteins (“protein C”) enhance the toxicity of type A proteins. Examples of type A proteins are TcbA, TcdA, XptA1, and XptA2. Examples of type B proteins are TcaC, TcdB, XptB1Xb, and XptC1Wi. Examples of type C proteins are TccC, XptC1Xb, and XptB1Wi. Insecticidal proteins also include spider, snake, and scorpion venom proteins. Examples of spider venom peptides include, but are not limited to, lycotoxin-1 peptide and its mutants (US Patent 8,334,366).

[0076] In some embodiments, the PIP-72 polypeptide comprises an amino acid sequence deduced from the full-length nucleic acid sequence disclosed herein, as well as an amino acid sequence shorter than the full-length sequence due to the use of an alternative downstream starting site or due to processing to produce a shorter protein with insecticidal activity. Processing can occur in the organism expressing the protein or in the pest after protein ingestion.

[0077] Therefore, this paper provides novel isolated or recombinant nucleic acid sequences conferring insecticidal activity. The amino acid sequence of the PIP-72 polypeptide is also provided. Cells containing proteins translated from these PIP-72 polypeptide genes can control or kill pests that feed on these cells.

[0078] bacterial strains This disclosure relates, in one aspect, to bacterial strains expressing the PIP-72 polypeptide. In some embodiments, the bacterial strain is a member of the genus *Halomonas* (…). Halomonas ) species, species of the genus *Luminobacterium*, species of the genus *Pathogenic Bacillus*, species of the genus *Burkholderia* ( Burkholderia ) species, Paludibacterium Species or species of the genus *Pseudomonas*. In some embodiments, the bacterial strain is *Haloxylon anticlocatemus* (…). Halomonas anticariensis ), luminescent bacteria ( Photorhabdus luminescens ), Burkholderia nematode ( Xenorhabdus bovienii Burkholderia melioides ( Burkholderia pseudomallei Burkholderia polyphaga ( Burkholderia multivorans Burkholderia tsukia ( Burkholderia thailandensis ), Paludibacterium yongneupense Pseudomonas rhodopseudomonas (Pseudomonas rhodesiae ); Pseudomonas tataricus ( Pseudomonas entomophila ), Pseudomonas aeruginosa ( Pseudomonas chlororaphis ); Pseudomonas mongolica ( Pseudomonas mandelii ); Ice-forming Pseudomonas ( Pseudomonas congelans ); Pseudomonas mongolica; Pseudomonas proteus ( Pseudomonasplecoglossicida ), biocontrol Pseudomonas, Pseudomonas tianxiong ( Pseudomonasficuserectae ); Pseudomonas molluscum ( Pseudomonas mosselii ) or Pseudomonas brassicae ( Pseudomonas brassicacearum The bacterial strain is described in some embodiments as a pure biological culture of *Pseudomonas aeruginosa* strain SS143D5, which was deposited on February 7, 2013, with accession number NRRL B-50810 at the Agricultural Research Culture Collection (NRRL) (1815 North University Street, Peoria, Illinois 616040) (nrrl.ncaur.usda.gov, accessible via the World Wide Web using the "www" prefix). This deposit will be preserved in accordance with the terms of the Budapest Treaty on the International Recognition of Microbial Deposits for Patent Proceedings. These deposits are held solely for the convenience of those skilled in the art and do not constitute an admission that the deposits are required under 35 USC §112. During the pending period of this application, the Director of the United States Patent and Trademark Office and authorized personnel may access the deposits upon his request. Upon grant of any claim of this application, the applicant will make samples of the deposits at the Agricultural Research Culture Collection (NRRL), 1815 North University Street, Peoria, Illinois 61604, publicly available in accordance with 37 CFR § 1.808. The deposits will be held at the NRRL, a public depository, for 30 years, or 5 years after the most recent request, or until the expiry of this patent, whichever is longer, and will be replaced if inactivated during this period. Upon patent grant, the deposits will be irrevocably and unrestrictedly or unconditionally available to the public. Furthermore, the applicant has satisfied all the requirements of 37 CFR §§ 1,801-1,809, including providing indications of the viability of the samples at the time of deposit. The applicant has no right to exempt any legal restrictions on the transfer or commercial transport of biological material. The applicant does not exempt any infringement of the rights granted to them under this patent. However, it should be understood that the availability of the deposits should not be construed as a license to practice the invention in violation of the patent rights granted by government statute.

[0079] Nucleic acid molecules and their variants and fragments This disclosure relates, in one aspect, to isolated or recombinant nucleic acid molecules comprising a nucleic acid sequence encoding a PIP-72 polypeptide or its biologically active portion; and to nucleic acid molecules sufficient to be used as hybridization probes for identifying nucleic acid molecules encoding proteins having regions of sequence homology. As used herein, the term “nucleic acid molecule” refers to DNA molecules (e.g., recombinant DNA, cDNA, genomic DNA, plastid DNA, mitochondrial DNA) and RNA molecules (e.g., mRNA), as well as analogs of DNA or RNA produced using nucleotide analogs. Nucleic acid molecules can be single-stranded or double-stranded, but double-stranded DNA is preferred.

[0080] In this document, "isolated" nucleic acid molecule (or DNA) refers to a nucleic acid sequence (or DNA) that is no longer in its natural environment, such as in vitro. "Recombinant" nucleic acid molecule (or DNA) refers to a nucleic acid sequence (or DNA) in a recombinant bacterial or plant host cell. In some embodiments, the "isolated" or "recombinant" nucleic acid does not contain a sequence naturally located flanking the nucleic acid in the genomic DNA of the source organism (i.e., the sequence located at the 5' and 3' ends of the nucleic acid) (preferably a protein-coding sequence). For the purposes of this disclosure, "isolated" or "recombinant" excludes isolated chromosomes when used to refer to nucleic acid molecules. For example, in various embodiments, a recombinant nucleic acid molecule encoding the PIP-72 polypeptide may contain a nucleic acid sequence of less than about 5 kb, 4 kb, 3 kb, 2 kb, 1 kb, 0.5 kb, or 0.1 kb, which is naturally located flanking the nucleic acid molecule in the genomic DNA of the source cell.

[0081] In some embodiments, the isolated nucleic acid molecule encoding the PIP-72 polypeptide has one or more alterations in its nucleic acid sequence compared to the natural or genomic nucleic acid sequence. In some embodiments, the alterations in the natural or genomic nucleic acid sequence include, but are not limited to: alterations in the nucleic acid sequence due to genetic code degeneracy; alterations in the nucleic acid sequence due to amino acid substitutions, insertions, deletions, and / or additions compared to the natural or genomic sequence; removal of one or more introns; deletion of one or more upstream or downstream regulatory regions; and deletion of 5' and / or 3' untranslated regions associated with the genomic nucleic acid sequence. In some embodiments, the nucleic acid molecule encoding the PIP-72 polypeptide is a non-genomic sequence.

[0082] This disclosure envisions various polynucleotides encoding the PIP-72 polypeptide or related proteins. These polynucleotides, when operably linked to suitable promoters, transcription termination sequences, and / or polyadenylation sequences, can be used to generate the PIP-72 polypeptide in host cells. These polynucleotides can also be used as probes to isolate homologous or substantially homologous polynucleotides encoding the PIP-72 polypeptide or related proteins.

[0083] Sources of polynucleotides encoding the PIP-72 polypeptide or related proteins include, but are not limited to, *Haloxylon anticlocatemus*, *Luminobacterium luminiferum*, *Burkholderia burgdorferi*, *Burkholderia melioides*, *Burkholderia polyphaga*, and *Burkholderia thamnoides*. Paludibacterium yongneupense *Pseudomonas rhodopseudomonas*; *Pseudomonas taeniophilus*, *Pseudomonas aeruginosa*; *Pseudomonas monnieri*; *Pseudomonas icarius*; *Pseudomonas monnieri*; *Pseudomonas proteus*, *Pseudomonas biocontrolus*, *Pseudomonas spp.*; *Pseudomonas moss* or *Pseudomonas brassicae* strains. Sources of polynucleotides encoding the PIP-72 polypeptide or related proteins include, but are not limited to: *Pseudomonas aeruginosa* strains containing the PIP-72Aa polypeptide encoded by SEQ ID NO: 2, or the PIP-72Aa polynucleotide shown in SEQ ID NO: 1; *Pseudomonas repens* strains containing the PIP-72Ba polypeptide encoded by SEQ ID NO: 4, or the PIP-72Ba polynucleotide shown in SEQ ID NO: 3; *Pseudomonas aeruginosa* strains containing the PIP-72Ca polypeptide encoded by SEQ ID NO: 6, or the PIP-72Ca polynucleotide shown in SEQ ID NO: 5; *Pseudomonas meningitidis* strains containing the PIP-72Cb polypeptide encoded by SEQ ID NO: 8, or the PIP-72Cb polynucleotide shown in SEQ ID NO: 7; and *Pseudomonas icariina* strains containing the PIP-72Da polypeptide encoded by SEQ ID NO: 10, or the PIP-72Cb polynucleotide shown in SEQ ID NO: 10. 9. The following strains are listed: PIP-72Da polynucleotide; *Pseudomonas meningitidis* strain containing the PIP-72Db polypeptide encoded by SEQ ID NO: 12, and the PIP-72Db polynucleotide encoded by SEQ ID NO: 11; *Pseudomonas serrata* strain containing the PIP-72Dc polypeptide encoded by SEQ ID NO: 14, and the PIP-72Dc polynucleotide encoded by SEQ ID NO: 13; *Pseudomonas moschata* strain containing the PIP-72Fa polypeptide encoded by SEQ ID NO: 18, and the PIP-72Fa polynucleotide encoded by SEQ ID NO: 17; *Pseudomonas aeruginosa* strain containing the PIP-72Ff polypeptide encoded by SEQ ID NO: 28, and the PIP-72Ff polynucleotide encoded by SEQ ID NO: 27; *Pseudomonas aeruginosa* strain containing the PIP-72Gb polypeptide encoded by SEQ ID NO: 32, and the PIP-72Gb polynucleotide encoded by SEQ ID NO: 32. 31 shows the PIP-72Gb polynucleotide; a *Pseudomonas aeruginosa* strain containing the PIP-72Ab polypeptide shown in SEQ ID NO: 927 and the PIP-72Ab polynucleotide shown in SEQ ID NO: 949; a *Pseudomonas brassicae* strain containing the PIP-72Ab polypeptide shown in SEQ ID NO: 928 and the PIP-72Bb polynucleotide shown in SEQ ID NO: 950;*Pseudomonas benzodiazepines* strains containing the polypeptide encoding PIP-72AFh (SEQ ID NO: 932) and the polynucleotide encoding PIP-72Fh (SEQ ID NO: 954); *Pseudomonas benzodiazepines* strains containing the polypeptide encoding PIP-72AFh (SEQ ID NO: 933) and the polynucleotide encoding PIP-72Fh (SEQ ID NO: 955); *Pseudomonas aeruginosa* strains containing the polypeptide encoding PIP-72Fj (SEQ ID NO: 934) and the polynucleotide encoding PIP-72Fj (SEQ ID NO: 956); *Pseudomonas aeruginosa* strains containing the polypeptide encoding PIP-72Fk (SEQ ID NO: 935) and the polynucleotide encoding PIP-72Fk (SEQ ID NO: 957); and *Burkholderia polyphaga* strains containing the polypeptide encoding PIP-72Fl (SEQ ID NO: 936) and the polynucleotide encoding PIP-72Fl (SEQ ID NO: 957). PIP-72Fl polynucleotide shown in SEQ ID NO: 958; *Pseudomonas aeruginosa* strain containing the PIP-72Gg polypeptide shown in SEQ ID NO: 939, and the PIP-72Gg polynucleotide shown in SEQ ID NO: 961; *Pseudomonas aeruginosa* strain containing the PIP-72Gh polypeptide shown in SEQ ID NO: 940, and the PIP-72Gh polynucleotide shown in SEQ ID NO: 962; *Pseudomonas moschata* strain containing the PIP-72Gi polypeptide shown in SEQ ID NO: 941, and the PIP-72Gi polynucleotide shown in SEQ ID NO: 963; *Pseudomonas biocontrolensis* strain containing the PIP-72Gk polypeptide shown in SEQ ID NO: 943, and the PIP-72Gk polynucleotide shown in SEQ ID NO: 965; *Pseudomonas proteobacterium* strain containing the PIP-72Gk polypeptide shown in SEQ ID NO: 943, and the PIP-72Gk polynucleotide shown in SEQ ID NO: 965; *Pseudomonas proteobacterium* strain containing the PIP-72Gk polypeptide shown in SEQ ID NO: 965. The PIP-72Gl polypeptide shown in SEQ ID NO: 944, and the PIP-72Gl polynucleotide shown in SEQ ID NO: 966;A strain of *Pseudomonas aeruginosa* containing the polypeptide encoding PIP-72Gn (SEQ ID NO: 946) and the polynucleotide encoding PIP-72Gn (SEQ ID NO: 968). These polynucleotide sequences are isolated from hosts such as *Haloxymonas*, *Luminobacterium*, *Pathobacterium*, *Burkholderia*, *Paludibacterium*, or *Pseudomonas*, and are therefore suitable for expression of the encoded PIP-72 polypeptide in other bacterial hosts. For example, SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:27 and SEQ ID NO:31, SEQ ID NO:949, SEQ ID NO:950, SEQ ID NO:955, SEQ ID NO:956, SEQ ID NO:957, SEQ ID NO:958, SEQ ID NO:961, SEQ ID NO:962, SEQ ID NO:963, SEQ ID NO:965, SEQ ID NO:966, SEQ ID NO:967, SEQ ID NO:968 can be used to express the PIP-72 polypeptide in bacterial hosts including but not limited to: Agrobacterium spp. (; Agrobacterium ), Bacillus spp., Escherichia spp. ( Escherichia Salmonella ( Salmonella ), Pseudomonas and Rhizobium ( Rhizobium Bacterial host cells. The polynucleotides can also be used as probes to isolate homologous or substantially homologous polynucleotides encoding the PIP-72 polypeptide or related proteins. Such probes can be used to identify homologous or substantially homologous polynucleotides derived from the following bacteria or other related bacteria: *Haliomonas*, *Luminobacter*, *Pathogenic Bacillus*, *Burkholderia*, etc. Paludibacterium Or Pseudomonas spp.

[0084] The polynucleotide encoding the PIP-72 polypeptide can also be synthesized de novo from the PIP-72 polypeptide sequence. The sequence of the polynucleotide gene can be deduced from the PIP-72 polypeptide sequence using the genetic code. Computer programs such as "BackTranslate" (GCG) can be used. ™The software package (Acclerys, Inc., San Diego, Calif.) converts peptide sequences into the corresponding nucleotide sequences encoding those peptides. Examples of PIP-72 polypeptide sequences that can be used to obtain the corresponding nucleotide coding sequence include, but are not limited to, PIP-72 polypeptides having the following sequences: SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 18, SEQ ID NO: 28 and SEQ ID NO: 32, SEQ ID NO: 927, SEQ ID NO: 928, SEQ ID NO: 932, SEQ ID NO: 933, SEQ ID NO: 934, SEQ ID NO: 935, SEQ ID NO: 936, SEQ ID NO: 939, SEQ ID NO: 940, SEQ ID NO: 941, SEQ ID NO: 943, SEQ ID NO: 944, SEQ ID NO: 945 or SEQ ID NO: 946. Furthermore, the synthetic PIP-72 polynucleotide sequence disclosed herein can be engineered for expression in plants. U.S. Patent 5,500,365 describes a method for synthesizing a plant gene to improve the expression level of a protein encoded by that synthetic gene. This method involves modifying the structural gene sequence of a foreign transgene to make it more efficiently transcribed, processed, translated, and expressed by the plant. A gene adequately expressed in a plant is characterized by the elimination of sequences that could cause undesirable intron splicing or polyadenylation in the coding region of the gene transcript, while substantially preserving the amino acid sequence of the toxic portion of the insecticidal protein. A similar method for obtaining enhanced expression of transgenes in monocotyledonous plants is disclosed in U.S. Patent 5,689,052.

[0085] In some embodiments, the nucleic acid molecule encoding the PIP-72 polypeptide is a polynucleotide having the sequences shown in SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:27, SEQ ID NO:31, SEQ ID NO:949, SEQ ID NO:950, SEQ ID NO:955, SEQ ID NO:956, SEQ ID NO:957, SEQ ID NO:958, SEQ ID NO:961, SEQ ID NO:962, SEQ ID NO:963, SEQ ID NO:965, SEQ ID NO:966, SEQ ID NO:967, and SEQ ID NO:968, as well as variants, fragments, and complementary sequences thereof. "Complementary sequence" is used herein to refer to a nucleic acid sequence that is sufficiently complementary to a given nucleic acid sequence such that it can hybridize to that given nucleic acid sequence to form a stable duplex. In this article, "polynucleotide sequence variant" refers to a nucleic acid sequence that encodes the same polypeptide except for genetic code degeneracy.

[0086] In some embodiments, the nucleic acid molecule encoding the PIP-72 polypeptide is a non-genomic nucleic acid sequence. As used herein, a “non-genomic nucleic acid sequence” or “non-genomic nucleic acid molecule” refers to a nucleic acid molecule that has one or more alterations in its nucleic acid sequence compared to a native or genomic nucleic acid sequence. In some embodiments, alterations in a native or genomic nucleic acid molecule include, but are not limited to: alterations in the nucleic acid sequence due to genetic code degeneracy; codon optimization of the nucleic acid sequence for expression in plants; alterations in the nucleic acid sequence compared to a native or genomic sequence to introduce at least one amino acid substitution, insertion, deletion, and / or addition; removal of one or more introns associated with a genomic nucleic acid sequence; insertion of one or more heterologous introns; deletion of one or more upstream or downstream regulatory regions associated with a genomic nucleic acid sequence; insertion of one or more heterologous upstream or downstream regulatory regions; deletion of 5' and / or 3' untranslated regions associated with a genomic nucleic acid sequence; insertion of heterologous 5' and / or 3' untranslated regions; and modification of polyadenylation sites. In some embodiments, the non-genomic nucleic acid molecule is cDNA. In some embodiments, the non-genomic nucleic acid molecule is a synthetic nucleic acid sequence. In some embodiments, the non-genomic nucleic acid molecule is not one of the following nucleic acid sequences: SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:27, SEQ ID NO:31, SEQ ID NO:949, SEQ ID NO:950, SEQ ID NO:955, SEQ ID NO:956, SEQ ID NO:957, SEQ ID NO:958, SEQ ID NO:961, SEQ ID NO:962, SEQ ID NO:963, SEQ ID NO:965, SEQ ID NO:966, SEQ ID NO:967, SEQ ID NO:968.

[0087] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the following amino acid sequences: SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 99%. 18, SEQ ID NO: 28, SEQ ID NO: 32, SEQ ID NO: 927, SEQ ID NO: 928, SEQ ID NO: 932, SEQ ID NO: 933, SEQ ID NO: 934, SEQ ID NO: 935, SEQ ID NO: 936, SEQ ID NO: 939, SEQ ID NO: 940, SEQ ID NO: 941, SEQ ID NO: 943, SEQ ID NO: 944, SEQ ID NO: 945 or SEQ ID NO: 946, wherein the polypeptide has insecticidal activity.

[0088] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO: 2, wherein the polypeptide has insecticidal activity.

[0089] In some embodiments, the amino acid sequence of the non-genomic nucleic acid molecule has at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO: 4, wherein the polypeptide has insecticidal activity.

[0090] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO: 6, wherein the polypeptide has insecticidal activity.

[0091] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO: 8, wherein the polypeptide has insecticidal activity.

[0092] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO: 10, wherein the polypeptide has insecticidal activity.

[0093] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO: 12, wherein the polypeptide has insecticidal activity.

[0094] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO: 14, wherein the polypeptide has insecticidal activity.

[0095] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO: 18, wherein the polypeptide has insecticidal activity.

[0096] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO: 28, wherein the polypeptide has insecticidal activity.

[0097] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO: 32, wherein the polypeptide has insecticidal activity.

[0098] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO: 927, wherein the polypeptide has insecticidal activity.

[0099] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO: 928, wherein the polypeptide has insecticidal activity.

[0100] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO: 932, wherein the polypeptide has insecticidal activity.

[0101] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO: 933, wherein the polypeptide has insecticidal activity.

[0102] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO: 934, wherein the polypeptide has insecticidal activity.

[0103] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO: 935, wherein the polypeptide has insecticidal activity.

[0104] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO: 936, wherein the polypeptide has insecticidal activity.

[0105] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO: 939, wherein the polypeptide has insecticidal activity.

[0106] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO: 940, wherein the polypeptide has insecticidal activity.

[0107] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO: 941, wherein the polypeptide has insecticidal activity.

[0108] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO: 943, wherein the polypeptide has insecticidal activity.

[0109] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO: 944, wherein the polypeptide has insecticidal activity.

[0110] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO: 945, wherein the polypeptide has insecticidal activity.

[0111] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence shown in SEQ ID NO: 946, wherein the polypeptide has insecticidal activity.

[0112] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that shares at least 50% identity with the following amino acid sequences: SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:18, SEQ ID NO:28, SEQ ID NO:32, SEQ ID NO:927, SEQ ID NO:928, SEQ ID NO:932, SEQ ID NO:933, SEQ ID NO:934, SEQ ID NO:935, SEQ ID NO:936, SEQ ID NO:939, SEQ ID NO:940, SEQ ID NO:941, SEQ ID NO:943, SEQ ID NO:944, SEQ ID NO:945, or SEQ ID NO:946, wherein the PIP-72 polypeptide has at least one amino acid change compared to the following sequences: SEQ ID NO:2, SEQ ID NO:32, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:18, SEQ ID NO:28, SEQ ID NO:32, SEQ ID NO:927, SEQ ID NO:928, SEQ ID NO:932, SEQ ID NO:933, SEQ ID NO:934, SEQ ID NO:935, SEQ ID NO:936, SEQ ID NO:939, SEQ ID NO:940, SEQ ID NO:941, SEQ ID NO:943, SEQ ID NO:944, SEQ ID NO:945, or SEQ ID NO:946, wherein the PIP-72 polypeptide has at least one amino acid change compared to the following sequences: SEQ ID NO:2, SEQ ID NO SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:18, SEQ ID NO:28, SEQ ID NO:32, SEQ ID NO:927, SEQ ID NO:928, SEQ ID NO:932, SEQ ID NO:933, SEQ ID NO:934, SEQ ID NO:935, SEQ ID NO:936, SEQ ID NO:939, SEQ ID NO:940, SEQ ID NO:941, SEQ ID NO:943, SEQ ID NO:944, SEQ ID NO:945 or SEQ ID NO:946, wherein the PIP-72 polypeptide has insecticidal activity.

[0113] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50% identity with the amino acid sequence shown in SEQ ID NO: 2, wherein the PIP-72 polypeptide has at least one amino acid change compared to SEQ ID NO: 2, and the PIP-72 polypeptide has insecticidal activity.

[0114] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50% identity with the amino acid sequence shown in SEQ ID NO: 4, wherein the PIP-72 polypeptide has at least one amino acid change compared to SEQ ID NO: 4, and the PIP-72 polypeptide has insecticidal activity.

[0115] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50% identity with the amino acid sequence shown in SEQ ID NO: 6, wherein the PIP-72 polypeptide has at least one amino acid change compared to SEQ ID NO: 6, and the PIP-72 polypeptide has insecticidal activity.

[0116] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50% identity with the amino acid sequence shown in SEQ ID NO: 8, wherein the PIP-72 polypeptide has at least one amino acid change compared to SEQ ID NO: 8, and the PIP-72 polypeptide has insecticidal activity.

[0117] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50% identity with the amino acid sequence shown in SEQ ID NO: 10, wherein the PIP-72 polypeptide has at least one amino acid change compared to SEQ ID NO: 10, and the PIP-72 polypeptide has insecticidal activity.

[0118] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50% identity with the amino acid sequence shown in SEQ ID NO: 12, wherein the PIP-72 polypeptide has at least one amino acid change compared to SEQ ID NO: 12, and the PIP-72 polypeptide has insecticidal activity.

[0119] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50% identity with the amino acid sequence shown in SEQ ID NO: 14, wherein the PIP-72 polypeptide has at least one amino acid change compared to SEQ ID NO: 14, and the PIP-72 polypeptide has insecticidal activity.

[0120] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 60% identity with the amino acid sequence shown in SEQ ID NO: 18, wherein the PIP-72 polypeptide has at least one amino acid change compared to SEQ ID NO: 18, and the PIP-72 polypeptide has insecticidal activity.

[0121] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 85% identity with the amino acid sequence shown in SEQ ID NO: 28, wherein the PIP-72 polypeptide has at least one amino acid change compared to SEQ ID NO: 28, and the PIP-72 polypeptide has insecticidal activity.

[0122] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 95% identity with the amino acid sequence shown in SEQ ID NO: 32, wherein the PIP-72 polypeptide has at least one amino acid change compared to SEQ ID NO: 32, and the PIP-72 polypeptide has insecticidal activity.

[0123] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 95% identity with the amino acid sequence shown in SEQ ID NO: 927, wherein the PIP-72 polypeptide has at least one amino acid change compared to SEQ ID NO: 927, and the PIP-72 polypeptide has insecticidal activity.

[0124] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 95% identity with the amino acid sequence shown in SEQ ID NO: 928, wherein the PIP-72 polypeptide has at least one amino acid change compared to SEQ ID NO: 928, and the PIP-72 polypeptide has insecticidal activity.

[0125] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 95% identity with the amino acid sequence shown in SEQ ID NO: 932, wherein the PIP-72 polypeptide has at least one amino acid change compared to SEQ ID NO: 932, and the PIP-72 polypeptide has insecticidal activity.

[0126] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that is at least 95% identical to the amino acid sequence shown in SEQ ID NO: 933, wherein the PIP-72 polypeptide has at least one amino acid change compared to SEQ ID NO: 933, and the PIP-72 polypeptide has insecticidal activity.

[0127] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 95% identity with the amino acid sequence shown in SEQ ID NO: 934, wherein the PIP-72 polypeptide has at least one amino acid change compared to SEQ ID NO: 934, and the PIP-72 polypeptide has insecticidal activity.

[0128] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that is at least 95% identical to the amino acid sequence shown in SEQ ID NO: 935, wherein the PIP-72 polypeptide has at least one amino acid change compared to SEQ ID NO: 935, and the PIP-72 polypeptide has insecticidal activity.

[0129] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 95% identity with the amino acid sequence shown in SEQ ID NO: 936, wherein the PIP-72 polypeptide has at least one amino acid change compared to SEQ ID NO: 936, and the PIP-72 polypeptide has insecticidal activity.

[0130] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 95% identity with the amino acid sequence shown in SEQ ID NO: 939, wherein the PIP-72 polypeptide has at least one amino acid change compared to SEQ ID NO: 939, and the PIP-72 polypeptide has insecticidal activity.

[0131] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 95% identity with the amino acid sequence shown in SEQ ID NO: 940, wherein the PIP-72 polypeptide has at least one amino acid change compared to SEQ ID NO: 940, and the PIP-72 polypeptide has insecticidal activity.

[0132] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 95% identity with the amino acid sequence shown in SEQ ID NO: 941, wherein the PIP-72 polypeptide has at least one amino acid change compared to SEQ ID NO: 941, and the PIP-72 polypeptide has insecticidal activity.

[0133] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 95% identity with the amino acid sequence shown in SEQ ID NO: 943, wherein the PIP-72 polypeptide has at least one amino acid change compared to SEQ ID NO: 943, and the PIP-72 polypeptide has insecticidal activity.

[0134] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 95% identity with the amino acid sequence shown in SEQ ID NO: 944, wherein the PIP-72 polypeptide has at least one amino acid change compared to SEQ ID NO: 944, and the PIP-72 polypeptide has insecticidal activity.

[0135] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 95% identity with the amino acid sequence shown in SEQ ID NO: 945, wherein the PIP-72 polypeptide has at least one amino acid change compared to SEQ ID NO: 945, and the PIP-72 polypeptide has insecticidal activity.

[0136] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 95% identity with the amino acid sequence shown in SEQ ID NO: 946, wherein the PIP-72 polypeptide has at least one amino acid change compared to SEQ ID NO: 946, and the PIP-72 polypeptide has insecticidal activity.

[0137] In some embodiments, the non-genomic nucleic acid molecule encodes the PIP-72 polypeptide, which comprises the amino acid sequences SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:18, SEQ ID NO:28, SEQ ID NO:32, SEQ ID NO:927, SEQ ID NO:928, SEQ ID NO:932, SEQ ID NO:933, SEQ ID NO:934, SEQ ID NO:935, SEQ ID NO:936, SEQ ID NO:939, SEQ ID NO:940, SEQ ID NO:941, SEQ ID NO:943, SEQ ID NO:944, SEQ ID NO:945, or SEQ ID NO:946, but not SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:946, SEQ ID NO:945, SEQ ID NO:946, but not SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:946, SEQ ID NO:947, SEQ ID NO:948, SEQ ID NO:949, SEQ ID NO:940, SEQ ID NO:941, SEQ ID NO:943, SEQ ID NO:944, SEQ ID NO:945, or SEQ ID NO:946. ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 18, SEQ ID NO: 28, SEQ ID NO: 32, SEQ ID NO: 927, SEQ ID NO: 928, SEQ ID NO: 932, SEQ ID NO: 933, SEQ ID NO: 934, SEQ ID NO: 935, SEQ ID NO: 936, SEQ ID NO: 939, SEQ ID NO: 940, SEQ ID NO: 941, SEQ ID NO: 943, SEQ ID NO: 944, SEQ ID NO: 945 or SEQ ID Compared to the natural amino acids at the corresponding positions in NO:946, it has 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions.

[0138] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing the amino acid sequence SEQ ID NO: 2, but having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions compared to the native amino acids at the corresponding positions in SEQ ID NO: 2.

[0139] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing the amino acid sequence SEQ ID NO: 4, but with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions compared to the native amino acids at the corresponding positions in SEQ ID NO: 4.

[0140] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing the amino acid sequence SEQ ID NO: 6, but with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions compared to the native amino acid at the corresponding position in SEQ ID NO: 6.

[0141] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing the amino acid sequence SEQ ID NO: 8, but with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions compared to the native amino acids at the corresponding positions in SEQ ID NO: 8.

[0142] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing the amino acid sequence SEQ ID NO: 10, but with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions compared to the native amino acids at the corresponding positions in SEQ ID NO: 10.

[0143] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing the amino acid sequence SEQ ID NO: 12, but having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions compared to the native amino acids at the corresponding positions in SEQ ID NO: 12.

[0144] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing the amino acid sequence SEQ ID NO: 14, but with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions compared to the native amino acids at the corresponding positions in SEQ ID NO: 14.

[0145] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing the amino acid sequence SEQ ID NO: 18, but with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 amino acid substitutions compared to the native amino acids at the corresponding positions in SEQ ID NO: 18.

[0146] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing the amino acid sequence SEQ ID NO: 28, but with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 amino acid substitutions compared to the native amino acid at the corresponding position in SEQ ID NO: 28.

[0147] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing the amino acid sequence SEQ ID NO: 32, but with 1, 2, 3, 4 or 5 amino acid substitutions compared to the native amino acid at the corresponding position in SEQ ID NO: 32.

[0148] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing the amino acid sequence SEQ ID NO: 927, but with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions compared to the native amino acids at the corresponding positions in SEQ ID NO: 927.

[0149] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing the amino acid sequence SEQ ID NO: 928, but with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions compared to the native amino acids at the corresponding positions in SEQ ID NO: 928.

[0150] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing the amino acid sequence SEQ ID NO: 932, but having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions compared to the native amino acids at the corresponding positions in SEQ ID NO: 932.

[0151] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing the amino acid sequence SEQ ID NO: 933, but with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions compared to the native amino acids at the corresponding positions in SEQ ID NO: 933.

[0152] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing the amino acid sequence SEQ ID NO: 934, but with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions compared to the native amino acids at the corresponding positions in SEQ ID NO: 934.

[0153] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing the amino acid sequence SEQ ID NO: 935, but having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions compared to the native amino acid at the corresponding position in SEQ ID NO: 935.

[0154] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing the amino acid sequence SEQ ID NO: 936, but with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions compared to the native amino acids at the corresponding positions in SEQ ID NO: 936.

[0155] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing the amino acid sequence SEQ ID NO: 939, but having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions compared to the native amino acids at the corresponding positions in SEQ ID NO: 939.

[0156] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing the amino acid sequence SEQ ID NO: 940, but with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions compared to the native amino acids at the corresponding positions in SEQ ID NO: 940.

[0157] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing the amino acid sequence SEQ ID NO: 941, but with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions compared to the native amino acids at the corresponding positions in SEQ ID NO: 941.

[0158] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing the amino acid sequence SEQ ID NO: 943, but with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions compared to the native amino acids at the corresponding positions in SEQ ID NO: 943.

[0159] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing the amino acid sequence SEQ ID NO: 944, but with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions compared to the native amino acids at the corresponding positions in SEQ ID NO: 944.

[0160] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing the amino acid sequence SEQ ID NO: 945, but having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions compared to the native amino acid at the corresponding position in SEQ ID NO: 945.

[0161] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide containing the amino acid sequence SEQ ID NO: 946, but with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions compared to the native amino acids at the corresponding positions in SEQ ID NO: 946.

[0162] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide comprising the amino acid sequence SEQ ID NO: 846, which, compared to the native amino acid at the corresponding position in SEQ ID NO: 2, has 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions at the positions indicated by Xaa.

[0163] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide comprising the amino acid sequence SEQ ID NO: 847, which, compared to the native amino acid at the corresponding position in SEQ ID NO: 2, has 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions at the positions indicated by Xaa.

[0164] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide comprising the amino acid sequence SEQ ID NO: 848, which, compared to the native amino acid at the corresponding position in SEQ ID NO: 2, has 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions at the positions indicated by Xaa.

[0165] In some embodiments, the non-genomic nucleic acid molecule encodes a PIP-72 polypeptide comprising the amino acid sequence SEQ ID NO: 849, which, compared to the native amino acid at the corresponding position in SEQ ID NO: 2, has 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions at the positions indicated by Xaa.

[0166] In some embodiments, the nucleic acid molecule encodes the PIP-72 polypeptide, which contains the amino acid sequence SEQ ID NO: 846, wherein Xaa at position 2 is Gly, Ala, Cys, Asp, Glu, Ile, Lys, Leu, Asn, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 3 is Ile or Trp; Xaa at position 4 is Thr, Ala, Asp, Glu, His, Ile, Lys, Leu, Arg, Ser, Val, Trp, or Tyr; Xaa at position 5 is Val, Ala, Cys, Gly, His, Ile, or Tyr; and Xaa at position 6 is Thr, Ala, Cys, Phe, Gly, His, Ile, Lys, Met, Pro, Gln, Arg, Ser, Trp, or Tyr; Xaa at position 7 is Asn, Ala, or Val; Xaa at position 8 is Asn, Ala, Cys, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Gln, Arg, Ser, Thr, or Val; Xaa at position 9 is Ser, Ala, Cys, Gly, or Thr; Xaa at position 10 is Ser, Ala, Glu, Phe, Gly, His, Ile, Lys, Leu, Asn, Pro, Gln, Arg, Thr, or Trp; Xaa at position 11 is Asn, Ala, C ys, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Gln, Ser, Thr, Val, or Tyr; Xaa at position 12 is Pro, Ala, Cys, Asp, Glu, Gly, His, Lys, Leu, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 13 is Ile, Asn, Gln, or Val; Xaa at position 14 is Glu, Ala, Cys, Phe, His, Lys, or Gln; Xaa at position 15 is Val, Ala, Cys, Ile, Met, or Arg; Xaa at position 17 is... The Xaa at position 1 is Ile, Glu, or Val; the Xaa at position 18 is Asn or Ser; the Xaa at position 19 is His, Ala, Glu, Lys, Leu, Pro, Arg, Ser, or Tyr; the Xaa at position 20 is Trp, Ala, or Thr; the Xaa at position 22 is Ser, Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Thr, Val, or Tyr; the Xaa at position 23 is Asp, Ala, Gly, His, Lys, Met, Asn, Gln, Ser, Thr, or Val.Xaa at position 24 is Gly, Asp, or Phe; Xaa at position 25 is Asp, Ala, Glu, Phe, Asn, or Gln; Xaa at position 26 is Thr, Glu, or Pro; Xaa at position 27 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Asn, Gln, Arg, or Thr; Xaa at position 28 is Phe, Pro, Trp, or Tyr; Xaa at position 29 is Phe, Ala, Cys, Ile, Leu, Gln, Arg, Trp, or Tyr; Xaa at position 30 is Ser, Ala, Cys, or Asp. The following are the possible values ​​for Xaa: Glu, Phe, Gly, His, Lys, Leu, Met, Asn, Pro, Gln, Arg, Thr, Val, Trp, or Tyr; Xaa at position 31 is Val, Ile, or Leu; Xaa at position 32 is Gly, Ala, Asp, Glu, Phe, His, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 33 is Asn, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Pro, Gln, Arg, Ser The Xaa at position 34 is Gly, Glu, Phe, His, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Tyr; the Xaa at position 35 is Lys, Ala, Cys, Asp, Gly, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Val; the Xaa at position 36 is Gln, Ala, Cys, Glu, Gly, His, Ile, Lys, Leu, Asn, Pro, Arg, Ser, Thr, or Val; the Xaa at position 37 is Glu, Ala, Cys, Asp, Phe, Gly, Ile, Lys, Leu, Met, Asn, Ser, Thr, or Val; Xaa at position 38 is Thr, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Val, Trp, or Tyr; Xaa at position 39 is Trp or Phe; Xaa at position 40 is Asp, Ala, Cys, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr;The Xaa at position 42 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Thr, Val, Trp, or Tyr; the Xaa at position 44 is Ser, Ala, Asp, Glu, Gly, Leu, Met, Asn, Pro, Gln, Thr, Val, or Tyr; the Xaa at position 45 is Arg, Lys, or Ser; the Xaa at position 46 is Gly, Ala, or Gln; the Xaa at position 47 is Phe, Cys, Val, or Tyr; the Xaa at position 48 is Val, Ile, or Leu; the Xaa at position 49 is... Xaa at position 9 is Leu, Cys, Phe, Met, Arg, or Tyr; Xaa at position 50 is Ser, Ala, Cys, Asp, Ile, Met, Pro, Gln, Thr, or Val; Xaa at position 51 is Leu, Ala, Cys, Met, or Val; Xaa at position 52 is Lys, Cys, Phe, His, Ile, Leu, Met, Asn, Arg, Ser, Thr, Trp, or Tyr; Xaa at position 53 is Lys, Ala, Cys, Asp, Glu, Phe, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Val. The Xaa at position 54 is either l or Tyr; the Xaa at position 54 is Asn, Cys, Asp, Glu, Phe, Gly, Lys, Met, Gln, Arg, Ser, or Trp; the Xaa at position 56 is Ala, Gly, Leu, Asn, Pro, Gln, Arg, Ser, or Thr; the Xaa at position 57 is Gln, Glu, Leu, Met, Ser, or Thr; the Xaa at position 58 is His, Ala, Asp, Phe, Leu, Met, Asn, Arg, Trp, or Tyr; the Xaa at position 60 is Tyr, Glu, or Phe; the Xaa at position 63 is Gln, Cys, Gly, Ile, Leu. Met, Asn, Thr, Val, or Tyr; Xaa at position 64 is Ala, Phe, Gly, His, Arg, Ser, or Tyr; Xaa at position 65 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Leu, Asn, Thr, or Val; Xaa at position 66 is Ser, Ala, or Gly; Xaa at position 67 is Lys, Ala, Cys, Asp, Phe, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 68 is Ile, Asp, Leu, or Val.Xaa at position 69 is Glu, Ala, Cys, Asp, Phe, His, Ile, Leu, Met, Gln, Arg, Ser, Thr, Val, or Tyr; Xaa at position 70 is Val, Cys, or Ile; Xaa at position 71 is Asp, Ala, Cys, Gly, His, Ile, Leu, Met, Asn, Ser, Thr, Val, or Tyr; Xaa at position 72 is Asn, Ala, Cys, Asp, Glu, Gly, Lys, Met, Pro, Gln, Arg, Ser, Thr, Val, or Trp; Xaa at position 73... The first Xaa is Asn, Ala, Cys, Asp, Phe, Gly, His, Ile, Leu, Ser, Thr, Val, or Tyr; the second Xaa at position 74 is Ala, Cys, Asp, Phe, Gly, His, Ile, Leu, Asn, Gln, Arg, Ser, Thr, Val, or Tyr; the third Xaa at position 75 is Val, Cys, Ile, or Leu; the fourth Xaa at position 76 is Lys, Ala, Cys, Phe, His, Ile, Leu, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; the fifth Xaa at position 77 is Asp or Tyr; the sixth Xaa at position 78 is Asn, Ala, Cys, Phe, His, Ile, Leu, Gln, Arg, Ser, Thr, Val, Trp, or Tyr. Xaa at position 78 is Gln, Ala, Cys, Asp, Phe, Gly, His, Ile, Leu, Met, Asn, Arg, Ser, Thr, Val, or Tyr; Xaa at position 79 is Gly, Arg, Ala, Cys, Asp, Glu, Phe, His, Lys, Leu, Asn, Gln, Arg, Ser, Thr, Trp, or Tyr; Xaa at position 80 is Arg, Ala, Cys, Asp, Phe, Gly, His, Ile, Leu, Asn, Ser, Thr, Val, or Tyr; Xaa at position 81 is Leu, Al a, Cys, Asp, Phe, Gly, His, Ile, Asn, Pro, Arg, Ser, Thr, or Val; Xaa at position 82 is Ile, Ala, Leu, Met, Arg, or Val; Xaa at position 83 is Glu, Ala, Cys, Asp, Phe, Gly, His, Ile, Lys, Leu, Asn, Pro, Arg, Ser, Thr, Val, or Tyr; Xaa at position 84 is Pro, Ala, Cys, Glu, Ile, Ser, Val, Trp, or Tyr; Xaa at position 85 is Leu, Cys, Gly, or Val.Furthermore, Xaa at position 86 is Ser, Ala, Ile, Thr, or Val, and optionally, 1 to 14 amino acids are deleted from the N-terminus and / or C-terminus of the PIP-72 polypeptide.

[0167] In some embodiments, the nucleic acid molecule encodes the PIP-72 polypeptide, which contains the amino acid sequence SEQ ID NO: 847, wherein Xaa at position 2 is Gly, Lys, or Ala; Xaa at position 3 is Ile or Leu; Xaa at position 4 is Thr or Ser; Xaa at position 5 is Val or Ile; Xaa at position 6 is Thr or Lys; Xaa at position 8 is Asn, Lys, Gly, or Ser; Xaa at position 9 is Ser or Ala; Xaa at position 11 is Asn, Lys, His, or Thr; Xaa at position 12 is Pro, Thr, Lys, or Ser; Xaa at position 13 is Ile or Val; Xaa at position 14 is Glu or Asp; Xaa at position 15 is... The Xaa at position 16 is Val, Ala, or Ile; the Xaa at position 17 is Ile or Val; the Xaa at position 18 is Asn or Ser; the Xaa at position 19 is His, Lys, Arg, Gln, or Ala; the Xaa at position 21 is Gly or Arg; the Xaa at position 22 is Ser, Lys, Asn, Asp, or Thr; the Xaa at position 25 is Asp or Asn; the Xaa at position 26 is Thr or Asp; the Xaa at position 27 is Ser, Thr, Asn, or Lys; the Xaa at position 28 is Phe, Tyr, or Pr. o; Xaa at position 29 is Phe or Tyr; Xaa at position 30 is Ser, Gly, or Lys; Xaa at position 31 is Val, Ile, or Met; Xaa at position 32 is Gly, Ala, or Asp; Xaa at position 33 is Asn, Ser, Gln, or Pro; Xaa at position 35 is Lys, Glu, or Ser; Xaa at position 36 is Gln, Asn, or Ser; Xaa at position 37 is Glu or Asp; Xaa at position 38 is Thr or Ser; Xaa at position 42 is Ser or Asn; Xaa at position 44 is Ser, Asp, or Ala. Or Leu; Xaa at position 47 is Phe or Tyr; Xaa at position 48 is Leu or Met; Xaa at position 49 is Leu or Met; Xaa at position 50 is Ser, Ala, or Tyr; Xaa at position 51 is Leu or Val; Xaa at position 52 is Lys or Gln; Xaa at position 53 is Lys, Arg, Met, or Leu; Xaa at position 54 is Asn, Lys, or Gly; Xaa at position 55 is Gly or Ser; Xaa at position 56 is Ala, Thr, Gln, or Ser; Xaa at position 57 is Gln, Val, or Ala;Xaa at position 58 is His, Ala, Lys, Tyr, or Thr; Xaa at position 59 is Pro or Thr; Xaa at position 62 is Val or Ile; Xaa at position 63 is Gln, Ser, or Leu; Xaa at position 64 is Ala, Gln, or Ser; Xaa at position 65 is Ser or Thr; Xaa at position 67 is Lys, Gln, Arg, or Asn; Xaa at position 69 is Glu, Lys, or Val; Xaa at position 70 is Val or Ile; Xaa at position 71 is Asp, Glu, or Tyr; Xaa at position 72 is Asn, His, Ser, or Asp; Xaa at position 73 is Asn, Ser, or Asp. Xaa at position 74 is Ala, Thr, Met, Ile, or Lys; Xaa at position 76 is Lys or Thr; Xaa at position 78 is Gln, His, or Ser; Xaa at position 80 is Arg, Glu, or Gln; Xaa at position 81 is Leu, Pro, Ala, or Thr; Xaa at position 82 is Ile or Leu; Xaa at position 83 is Glu, His, Asn, Gln, or Leu; Xaa at position 85 is Leu, Val, or Ala; and Xaa at position 86 is Ser, Ala, Tyr, or Asn, wherein optionally 1 to 14 amino acids are deleted from the N-terminus and / or C-terminus of the PIP-72 polypeptide, and / or an amino acid is inserted between residues at positions 24 and 25, compared to SEQ ID NO:847.

[0168] In some embodiments, the nucleic acid molecule encodes the PIP-72 polypeptide, which contains the amino acid sequence SEQ ID NO: 848, wherein Xaa at position 2 is Gly, Lys, Ala, or Arg; Xaa at position 3 is Ile, Leu, or Val; Xaa at position 4 is Thr or Ser; Xaa at position 5 is Val, Ile, or Leu; Xaa at position 6 is Thr, Lys, Ser, or Arg; Xaa at position 8 is Asn, Lys, Gly, Ser, Gln, Arg, Thr, or Ala; Xaa at position 9 is Ser, Ala, or Thr; Xaa at position 11 is Asn, Lys, Thr, Gln, Arg, His, or Ser; and Xaa at position 12 is Pro or Thr. The Xaa at position 13 is Ile, Val, or Leu; the Xaa at position 14 is Glu or Asp; the Xaa at position 15 is Val, Ala, Ile, or Leu; the Xaa at position 16 is Ala or Ser; the Xaa at position 17 is Ile, Val, or Leu; the Xaa at position 18 is Asn, Ser, Gln, or Thr; the Xaa at position 19 is His, Lys, Ala, Gln, Asn, or Arg; the Xaa at position 21 is Gly, Arg, or Lys; the Xaa at position 22 is Ser, Lys, Asn, Thr, Arg, A sp, Glu, or Gln; Xaa at position 25 is Asp, Asn, Glu, or Gln; Xaa at position 26 is Thr, Asp, Ser, or Glu; Xaa at position 27 is Ser, Thr, Lys, Asn, Gln, or Arg; Xaa at position 28 is Phe, Tyr, Pro, or Trp; Xaa at position 29 is Phe, Tyr, or Trp; Xaa at position 30 is Ser, Gly, Lys, Thr, or Arg; Xaa at position 31 is Val, Ile, Met, or Leu; Xaa at position 32 is Gly, Ala, Asp, or Glu; Xaa at position 33... The Xaa at position 35 is Asn, Ser, Gln, Pro, or Thr; the Xaa at position 36 is Gln, Ser, Asn, or Thr; the Xaa at position 37 is Glu or Asp; the Xaa at position 38 is Thr or Ser; the Xaa at position 42 is Ser, Asn, Thr, or Gln; the Xaa at position 44 is Ser, Asp, Ala, Leu, Thr, Glu, Ile, or Val; the Xaa at position 47 is Phe, Tyr, or Trp; the Xaa at position 48 is Leu, Met, Ile, or Val.Xaa at position 49 is Leu, Met, Ile, or Val; Xaa at position 50 is Ser, Ala, Tyr, or Thr; Xaa at position 51 is Leu, Val, or Ile; Xaa at position 52 is Lys, Gln, Arg, or Asn; Xaa at position 53 is Lys, Arg, Met, Leu, Ile, or Val; Xaa at position 54 is Asn, Lys, Gly, Gln, or Arg; Xaa at position 55 is Gly, Ser, or Thr; Xaa at position 56 is Ala, Thr, Gln, Ser, or Asn; Xaa at position 57 is Gln, Va The Xaa at position 58 is His, Ala, Asn, Leu, or Ile; the Xaa at position 59 is Pro, Thr, or Ser; the Xaa at position 62 is Val, Ile, or Leu; the Xaa at position 63 is Gln, Ser, Leu, Asn, Thr, Ile, or Val; the Xaa at position 64 is Ala, Gln, Ser, Asn, or Thr; the Xaa at position 65 is Ser or Thr; the Xaa at position 67 is Lys, Gln, Asn, or Arg; the Xaa at position 69 is Glu, Val, Asp, Lys, or Arg. The Xaa at position 70 is Val, Ile, or Leu; the Xaa at position 71 is Asp, Glu, Tyr, or Trp; the Xaa at position 72 is Asn, His, Ser, Asp, Gln, Thr, or Glu; the Xaa at position 73 is Asn, Ser, Asp, Gln, Thr, or Glu; the Xaa at position 74 is Ala, Thr, Met, Ile, Lys, Ser, Leu, Val, or Arg; the Xaa at position 76 is Lys, Thr, Arg, or Ser; the Xaa at position 78 is Gln, His, Ser, Asn, or Thr; the Xaa at position 79 is... Xaa at position 80 is Arg, Glu, Gln, Lys, Asp, or Asn; Xaa at position 81 is Leu, Pro, Thr, Ile, Val, Ala, or Ser; Xaa at position 82 is Ile, Leu, or Val; Xaa at position 83 is Glu, His, Asn, Leu, Gln, Ile, or Val; Xaa at position 85 is Leu, Val, or Ala; and Xaa at position 86 is Ser, Ala, Tyr, Asn, or Thr, wherein optionally 1 to 14 amino acids are deleted from the N-terminus and / or C-terminus of the PIP-72 polypeptide, and / or an amino acid is inserted between residues at positions 24 and 25, compared to SEQ ID NO: 848.

[0169] In some embodiments, the nucleic acid molecule encodes the PIP-72 polypeptide, which contains the amino acid sequence SEQ ID NO: 849, wherein Xaa at position 2 is Gly, Ala, Cys, Asp, Glu, Ile, Lys, Leu, Asn, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 3 is Ile, Leu, Val, or Trp; Xaa at position 4 is Thr, Ala, Asp, Glu, His, Ile, Lys, Leu, Arg, Ser, Val, Trp, or Tyr; Xaa at position 5 is Val, Ala, Cys, Gly, His, Ile, Leu, or Tyr; and Xaa at position 6 is Thr, Ala, Cys, Ph e, Gly, His, Ile, Lys, Met, Pro, Gln, Arg, Ser, Trp, or Tyr; the 7th position Xaa is Asn, Ala, or Val; the 8th position Xaa is Asn, Lys, Gly, Ser, Gln, Arg, Thr, Ala, Cys, Asp, Glu, His, Ile, Leu, Met, or Val; the 9th position Xaa is Ser, Ala, Cys, Gly, or Thr; the 11th position Xaa is Asn, Lys, Thr, Gln, Arg, Ser, Ala, Cys, Asp, Glu, Gly, or His The Xaa at position 12 is Pro, Thr, Lys, Ser, Arg, Ala, Cys, Asp, Glu, Gly, His, Leu, Asn, Gln, Arg, Val, Trp, or Tyr; the Xaa at position 13 is Ile, Asn, Gln, Leu, or Val; the Xaa at position 14 is Glu, Ala, Cys, Phe, His, Lys, Asp, or Gln; the Xaa at position 15 is Val, Ala, Ile, Leu, Cys, Met, or Arg; the Xaa at position 16 is Al a or Ser; Xaa at position 17 is Ile, Glu, Leu, or Val; Xaa at position 18 is Asn, Gln, Thr, or Ser; Xaa at position 19 is His, Lys, Ala, Arg, Glu, Leu, Pro, Ser, or Tyr; Xaa at position 20 is Trp, Ala, or Thr; Xaa at position 21 is Gly, Arg, or Lys; Xaa at position 22 is Ser, Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Thr, Val, or Tyr;The Xaa at position 23 is Asp, Ala, Gly, His, Lys, Met, Asn, Gln, Ser, Thr, or Val; the Xaa at position 24 is Gly, Asp, or Phe; the Xaa at position 25 is Asp, Ala, Glu, Phe, Asn, or Gln; the Xaa at position 26 is Thr, Glu, Asp, Ser, or Pro; the Xaa at position 27 is Ser, Thr, Lys, Arg, Ala, Cys, Asp, Glu, Phe, Gly, His, Asn, or Gln; the Xaa at position 28 is Phe, Tyr, Pro, or Trp; the 2 The Xaa at position 9 is Phe, Ala, Cys, Ile, Leu, Gln, Arg, Trp, or Tyr; the Xaa at position 30 is Ser, Gly, Lys, Thr, Arg, Ala, Cys, Asp, Glu, Phe, His, Leu, Met, Asn, Pro, Gln, Val, Trp, or Tyr; the Xaa at position 31 is Val, Ile, Met, or Leu; the Xaa at position 32 is Gly, Ala, Asp, Glu, Phe, His, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, or T rp or Tyr; Xaa at position 33 is Asn, Ser, Gln, Pro, Thr, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Arg, Val, or Tyr; Xaa at position 34 is Gly, Glu, Phe, His, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Tyr; Xaa at position 35 is Lys, Glu, Ala, Cys, Asp, Gly, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Val; at position 36... Xaa is Gln, Ala, Cys, Glu, Gly, His, Ile, Lys, Leu, Asn, Pro, Arg, Ser, Thr, or Val; Xaa at position 37 is Glu, Asp, Ala, Cys, Phe, Gly, Ile, Lys, Leu, Met, Asn, Ser, Thr, or Val; Xaa at position 38 is Thr, Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Gln, Arg, Val, Trp, or Tyr; Xaa at position 39 is Trp or Phe.The Xaa at position 40 is Asp, Ala, Cys, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; the Xaa at position 42 is Ser, Asn, Thr, Ala, Cys, Asp, Glu, Phe, Gly, Ile, Lys, Leu, Met, Arg, Val, Trp, Tyr, or Gln; the Xaa at position 44 is Ser, Asp, Ala, Leu, Thr, Glu, Ile, Ala, Gly, Leu, Met, Asn, Pro, Gln, Val, Tyr, or Va. The Xaa at position 45 is Arg, Lys, or Ser; the Xaa at position 46 is Gly, Ala, or Gln; the Xaa at position 47 is Phe, Tyr, Cys, Val, or Trp; the Xaa at position 48 is Leu, Met, Ile, Cys, Phe, Met, Arg, Tyr, or Val; the Xaa at position 49 is Leu, Met, Ile, or Val; the Xaa at position 50 is Ser, Ala, Tyr, Cys, Asp, Ile, Met, Pro, Gln, Val, or Thr; the Xaa at position 51 is Leu, Val, Ala, Cys, Met, or Ile; the Xaa at position 52... The first Xaa is Lys, Cys, Phe, His, Ile, Leu, Met, Asn, Arg, Ser, Thr, Gln, Trp, or Tyr; the second Xaa at position 53 is Lys, Arg, Met, Leu, Ile, Ala, Cys, Asp, Glu, Phe, His, Asn, Gln, Ser, Thr, Tyr, or Val; the third Xaa at position 54 is Asn, Cys, Asp, Glu, Phe, Gly, Lys, Met, Gln, Arg, Ser, or Trp; the fourth Xaa at position 55 is Gly, Ser, or Thr; the fifth Xaa at position 56 is Ala, Thr, Gln, Ser, or Gly. The Xaa at position 57 is Gln, Glu, Leu, Met, Ser, Val, Ala, Asn, Ile, or Thr; the Xaa at position 58 is His, Ala, Lys, Asp, Phe, Leu, Met, Asn, Arg, Trp, Tyr, or Thr; the Xaa at position 59 is Pro, Thr, or Ser; the Xaa at position 60 is Tyr, Glu, or Phe; the Xaa at position 62 is Val, Ile, or Leu; the Xaa at position 63 is Gln, Ser, Cys, Gly, Ile, Leu, Met, Asn, Thr, Val, or Tyr.Xaa at position 64 is Ala, Gln, Asn, Phe, Gly, His, Arg, Ser, or Tyr; Xaa at position 65 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Leu, Asn, Val, or Thr; Xaa at position 66 is Ser, Ala, or Gly; Xaa at position 67 is Lys, Gln, Asn, or Arg; Xaa at position 68 is Lys, Ala, Cys, Asp, Phe, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 68 is Ile... Asp, Leu, or Val; Xaa at position 69 is Glu, Ala, Cys, Asp, Phe, His, Ile, Leu, Met, Gln, Arg, Ser, Thr, Val, or Tyr; Xaa at position 70 is Val, Ile, Cys, or Leu; Xaa at position 71 is Asp, Glu, Tyr, Ala, Cys, Gly, His, Ile, Leu, Met, Asn, Ser, Thr, Val, or Trp; Xaa at position 72 is Asn, Ala, Cys, Asp, Glu, Gly, Lys, Met, Pro, Gln, Arg, Ser, Thr, Val, His, or Trp; Xaa at position 73 is Asn, Ser, Asp, Gln, Thr, Ala, Cys, Phe, Gly, His, Ile, Leu, Val, Tyr, or Glu; Xaa at position 74 is Ala, Thr, Met, Ile, Lys, Ser, Leu, Val, Cys, Asp, Phe, Gly, His, Asn, Gln, Tyr, or Arg; Xaa at position 75 is Val, Cys, Ile, or Leu; Xaa at position 76 is Lys, Ala, Cys, Phe, His, Ile, Leu, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; 77th position The Xaa at position 70 is Asp or Tyr; the Xaa at position 78 is Gln, His, Ser, Asn, Ala, Cys, Asp, Phe, Gly, Ile, Leu, Met, Asn, Arg, Val, Tyr, or Thr; the Xaa at position 79 is Gly, Arg, Ala, Cys, Asp, Glu, Phe, His, Lys, Leu, Asn, Gln, Arg, Ser, Thr, Trp, or Tyr; the Xaa at position 80 is Arg, Glu, Gln, Lys, Asp, Ala, Cys, Phe, Gly, His, Ile, Leu, Ser, Thr, Val, Tyr, or Asn.The Xaa at position 81 is Leu, Pro, Thr, Ile, Val, Ala, Cys, Asp, Phe, Gly, His, or Ser; the Xaa at position 82 is Ile, Ala, Leu, Met, Arg, and Val; the Xaa at position 83 is Glu, His, Asn, Leu, Gln, Ile, Ala, Cys, Asp, Phe, Gly, Lys, Pro, Arg, Ser, Thr, or Tyr. Or Val; Xaa at position 84 is Pro, Ala, Cys, Glu, Ile, Ser, Val, Trp, or Tyr; Xaa at position 85 is Leu, Val, Cys, Gly, or Ala; and Xaa at position 86 is Ser, Ala, Tyr, Asn, Ile, Val, or Thr, wherein optionally 1 to 14 amino acids are deleted from the N-terminus and / or C-terminus of the PIP-72 polypeptide, and / or an amino acid is inserted between residues at positions 24 and 25 compared to SEQ ID NO: 849.

[0170] In some embodiments, the nucleic acid molecule encodes a PIP-72 polypeptide, the amino acid motif of which is represented by positions 37 through 51 of the following sequences: SEQ ID NO:846, SEQ ID NO:847, SEQ ID NO:848, or SEQ ID NO:849.

[0171] In some embodiments, the nucleic acid molecule encodes a PIP-72 polypeptide containing an amino acid sequence that has at least 50% identity with the amino acid sequence shown in SEQ ID NO: 2.

[0172] In some embodiments, exemplary nucleic acid molecules encode the PIP-72 polypeptide shown in the following sequences: SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 18, SEQ ID NO: 28, SEQ ID NO: 32, any one of SEQ ID NO: 528 to SEQ ID NO: 768, any one of SEQ ID NO: 825 to SEQ ID NO: 844, SEQ ID NO: 771, SEQ ID NO: 772, SEQ ID NO: 852, any one of SEQ ID NO: 903 to SEQ ID NO: 914, SEQ ID NO: 927, SEQ ID NO: 928, SEQ ID NO: 932, SEQ ID NO: 933, SEQ ID NO: 934, SEQ ID NO: 935, SEQ ID NO: 936, SEQ ID NO: 768, SEQ ID NO: 825, SEQ ID NO: 936, SEQ ID NO: 937, SEQ ID NO: 938, SEQ ID NO: 932, SEQ ID NO: 933, SEQ ID NO: 934, SEQ ID NO: 93 ... SEQ ID NO: 939, SEQ ID NO: 940, SEQ ID NO: 941, SEQ ID NO: 943, SEQ ID NO: 944, SEQ ID NO: 945 or SEQ ID NO: 946, and amino acid substitutions, amino acid deletions, amino acid insertions, amino acid fragments, and combinations thereof in these polypeptides.

[0173] In some embodiments, the nucleic acid molecule encodes the PIP-72 polypeptide shown in Tables 14, 17, 20, 23, 24, 26, 28 and / or 29, and combinations thereof with amino acid substitutions, amino acid deletions and / or amino acid insertions.

[0174] This disclosure also provides nucleic acid molecules encoding transcription and / or translation products, which are subsequently spliced ​​to ultimately produce a functional PIP-72 polypeptide. Splicing can be performed in vitro or in vivo and can involve cis or trans splicing. The substrate for splicing can be a polynucleotide (e.g., RNA transcript) or a polypeptide. An example of polynucleotide cis splicing is the removal of introns from the inserted coding sequence, followed by splicing two flanking exon regions to obtain the PIP-72 polypeptide coding sequence. An example of trans splicing would be the encryption of polynucleotides by splitting the coding sequence into two or more fragments, which can be transcribed individually and then spliced ​​to form a full-length insecticidal coding sequence. The use of splicing enhancer sequences that can be introduced into the construct can facilitate cis or trans splicing of the polypeptide (US Patents 6,365,377 and 6,531,316). Therefore, in some embodiments, the polynucleotide does not directly encode the full-length PIP-72 polypeptide, but rather encodes one or more fragments of the PIP-72 polypeptide. Functional PIP-72 peptides can be expressed with these polynucleotides via mechanisms involving splicing, where splicing can occur at the polynucleotide (e.g., intron / exon) and / or peptide (e.g., inteptide / expeptide) levels. This can be used, for example, to control the expression of insecticidal activity, since the functional insecticidal peptide will only be expressed if all the necessary fragments are expressed in an environment that allows for splicing, thus producing a functional product. Furthermore, the introduction of one or more insert sequences into the polynucleotide can facilitate recombination with low-homology polynucleotides; the use of introns or intepids in the insert sequences facilitates the removal of the intercalated sequences, thereby restoring the function of the encoded variant.

[0175] Embodiments of this disclosure also cover nucleic acid molecules that are fragments of nucleic acid sequences encoding these PIP-72 polypeptides. As used herein, a “fragment” refers to a portion of a nucleic acid sequence encoding a PIP-72 polypeptide. Fragments of nucleic acid sequences may encode the biologically active portion of the PIP-72 polypeptide or may be used as hybridization probes or PCR primers when using the methods disclosed below. Nucleic acid molecules that are fragments of nucleic acid sequences encoding PIP-72 polypeptides contain at least about 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, or 260 consecutive nucleotides, the number of which is at most the total number of nucleotides present in the full-length nucleic acid sequence encoding the PIP-72 polypeptide disclosed herein, depending on the intended use. “Consecutive nucleotides” herein refers to nucleotide residues that are immediately adjacent to each other. Protein fragments encoded by fragments of nucleic acid sequences of embodiments of this disclosure will retain the biological activity of the PIP-72 polypeptide, and thus retain insecticidal activity. As used herein, "retaining PIP-72 activity" refers to an insecticidal activity of the peptide of at least about 10%, at least about 30%, at least about 50%, at least about 70%, 80%, 90%, 95%, or higher of the full-length PIP-72Aa peptide shown in SEQ ID NO: 2. In one embodiment, the insecticidal activity is lepidopteran activity. In one embodiment, the insecticidal activity is activity against Coleoptera species. In one embodiment, the insecticidal activity is activity against Root Leaf Beetles (… Diabrotica The insecticidal activity is the activity against one or more of the following insect pests in the corn rootworm species syndicate: Western corn rootworm (… Diabrotica virgifera virgifera Northern corn rootworm ( D. barberi ); Southern corn rootworm or cucumber leaf beetle root-eating subspecies ( Diabrotica undecimpunctata howardi ); and Mexican corn rootworm ( D. virgifera zeae In one implementation, the insecticidal activity is the killing of the western maize rootworm (…). Diabrotica virgifera virgifera ) activity.

[0176] In some embodiments, the fragment of the nucleic acid sequence encoding the PIP-72 polypeptide (the biologically active portion of the protein encoded by the PIP-72 polypeptide) encodes at least about 15, 20, 30, 40, 50, 60, 70, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84 or 85 consecutive amino acids, the number of consecutive nucleotides being at most the total number of amino acids present in the full-length PIP-72 polypeptide as shown in the embodiments of this disclosure. In some embodiments, the fragment is, for example, obtained by proteolytic digestion, insertion of a start codon, deletion of the codon encoding the missing amino acid accompanied by insertion of a stop codon, or insertion of a stop codon into the coding sequence, and is any of the sequences relative to SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 18, SEQ ID NO: 28, SEQ ID NO: 32, SEQ ID NO: 528 to SEQ ID NO: 768, any of the sequences SEQ ID NO: 825 to SEQ ID NO: 844, SEQ ID NO: 771, SEQ ID NO: 772, SEQ ID NO: 852, SEQ ID NO: 903 to SEQ ID NO: 914, SEQ ID NO: 927, SEQ ID NO: 928, SEQ ID NO: 932, SEQ ID NO: 914, SEQ ID NO: 927, SEQ ID NO: 928, SEQ ID NO: 932, SEQ ID NO: 928 ... The N-terminus and / or C-terminus of SEQ ID NO: 933, SEQ ID NO: 934, SEQ ID NO: 935, SEQ ID NO: 936, SEQ ID NO: 939, SEQ ID NO: 940, SEQ ID NO: 941, SEQ ID NO: 943, SEQ ID NO: 944, SEQ ID NO: 945 or SEQ ID NO: 946 or variants of these sequences are truncated by removing at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or more amino acids.In some implementations, the fragments covered herein are, for example, obtained by proteolytic digestion or by inserting a start codon into the coding sequence, from any of the sequences relative to SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 18, SEQ ID NO: 28, SEQ ID NO: 32, SEQ ID NO: 528 to SEQ ID NO: 768, any of the sequences SEQ ID NO: 825 to SEQ ID NO: 844, SEQ ID NO: 771, SEQ ID NO: 772, SEQ ID NO: 852, SEQ ID NO: 903 to SEQ ID NO: 914, SEQ ID NO: 927, SEQ ID NO: 928, SEQ ID NO: 932, SEQ ID NO: 933, SEQ ID NO: 934, SEQ ID NO: 935, SEQ ID NO: 936, SEQ ID NO: 939, SEQ ID NO: 940, SEQ ID NO: 941, SEQ ID NO: 943, SEQ ID NO: 944, SEQ ID NO: 945 or SEQ ID NO: 946 or variants of these sequences, obtained by removing 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or more amino acids from the N-terminus.In some implementations, the fragments covered herein are, for example, obtained by proteolytic digestion or by inserting a start codon into the coding sequence, from any of the sequences relative to SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 18, SEQ ID NO: 28 or SEQ ID NO: 32, SEQ ID NO: 528 to SEQ ID NO: 768, any of the sequences SEQ ID NO: 825 to SEQ ID NO: 844, any of the sequences SEQ ID NO: 771, SEQ ID NO: 772, SEQ ID NO: 852, SEQ ID NO: 903 to SEQ ID NO: 914, SEQ ID NO: 927, SEQ ID NO: 928, SEQ ID NO: 932, SEQ ID NO: 933, SEQ ID NO: 934, SEQ ID NO: SEQ ID NO: 935, SEQ ID NO: 936, SEQ ID NO: 939, SEQ ID NO: 940, SEQ ID NO: 941, SEQ ID NO: 943, SEQ ID NO: 944, SEQ ID NO: 945 or SEQ ID NO: 946 or variants of these sequences obtained by removing 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 amino acids from the N-terminus.

[0177] In some embodiments, the PIP-72 polypeptide is encoded by a nucleic acid sequence that is sufficiently homologous to the following nucleic acid sequences: SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:27, SEQ ID NO:31, SEQ ID NO:949, SEQ ID NO:950, SEQ ID NO:954, SEQ ID NO:955, SEQ ID NO:956, SEQ ID NO:957, SEQ ID NO:958, SEQ ID NO:961, SEQ ID NO:962, SEQ ID NO:963, SEQ ID NO:965, SEQ ID NO:966, SEQ ID NO:967, or SEQ ID NO:968. "Sufficient homology" herein refers to an amino acid or nucleic acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or higher sequence homology when compared with a reference sequence using one of the alignment procedures described herein and with standard parameters. Those skilled in the art will recognize that these values ​​can be appropriately adjusted to determine the corresponding homology of the proteins encoded by the two nucleic acid sequences by taking into account codon degeneracy, amino acid similarity, reading frame positioning, etc. In some embodiments, sequence homology is defined with respect to the full-length sequence of the polynucleotide encoding the PIP-72 polypeptide or to the full-length sequence of the PIP-72 polypeptide itself.In some embodiments, the PIP-72 peptide is associated with SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 18, SEQ ID NO: 28 or SEQ ID NO: 32, SEQ ID NO: 927, SEQ ID NO: 928, SEQ ID NO: 932, SEQ ID NO: 933, SEQ ID NO: 934, SEQ ID NO: 935, SEQ ID NO: 936, SEQ ID NO: 939, SEQ ID NO: 940, SEQ ID NO: 941, SEQ ID NO: 943, SEQ ID NO: 944, SEQ ID NO: 945 or SEQ ID NO: 946 has a sequence identity of at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or higher. In some embodiments, sequence identity is defined with respect to the full-length sequence of the polynucleotide encoding the PIP-72 polypeptide or with respect to the full-length sequence of the PIP-72 polypeptide. In some embodiments, sequence identity is defined using Vector NTI with all default parameters. ® ALIGNNX package (Invitrogen Corporation, Carlsbad, Calif.) ® The ClustalW algorithm in the module is used for calculation. In some implementations, sequence identity is calculated across the entire length of the peptide using the ClustalW algorithm in the ALIGNX module of the Vector NTI package (Invitrogen Corporation, Carlsbad, Calif.) with all default parameters.

[0178] To determine the percentage of identity between two amino acid sequences or two nucleic acid sequences, the sequences are aligned for optimal comparison purposes. The percentage of identity between the two sequences is a function of the number of common positions shared by the sequences (i.e., percentage of identity = number of common positions / total number of positions (e.g., overlapping positions) × 100). In one embodiment, the two sequences are of the same length. In another embodiment, the comparison spans the entire reference sequence (e.g., spans the entirety of one of SEQ ID NO:1 and SEQ ID NO:2). The percentage of identity between the two sequences can be determined using techniques similar to those described below, with or without gaps. In calculating the percentage of identity, exact matches are typically counted.

[0179] Mathematical algorithms can be used to determine the percentage of identity between two sequences. A non-restrictive example of a mathematical algorithm for comparing two sequences is Karlin and Altschul (1990). Proc. Natl. Acad. Sci. USA The algorithm proposed in 87:2264, Karlin and Altschul, (1993). Proc. Natl.Acad. Sci . USA 90:5873-5877 improved the algorithm. This algorithm was incorporated into Altschul et al. (1990). J. Mol. Biol. The BLASTN and BLASTX procedures proposed in 215:403 are used. BLAST nucleotide searches can be performed using the BLASTN procedure with a score of 100 and a word length of 12 to obtain nucleic acid sequences homologous to the insecticidal nucleic acid molecule of the described embodiment. BLAST protein searches can be performed using the BLASTX procedure with a score of 50 and a word length of 3 to obtain amino acid sequences homologous to the insecticidal protein molecule of the described embodiment. For comparison of vacancy alignment results, Altschul et al. (1997) can be consulted. Nucleic Acids Res As described in 25:3389, vacancy BLAST (BLAST 2.0) is used. Alternatively, PSI-Blast can be used for iterative searching, which can detect distant relationships between molecules. See Altschul et al. (1997) (ibid.). When using BLAST, Gapped BLAST, and PSI-Blast programs, the default parameters of each program (e.g., BLASTX and BLASTN) can be used. Sequence alignment can also be performed manually.

[0180] Another non-restricted example of a mathematical algorithm for sequence comparison is the ClustalW algorithm (Higgins et al., (1994)). Nucleic Acids Res (22:4673-4680). ClustalW compares sequences and aligns amino acids or DNA sequences as a whole, thus providing data on the sequence conservation of entire amino acid sequences. The ClustalW algorithm is used in several commercial DNA / amino acid analysis software packages, such as Vector NTI. ® ALIGNX of the package (Invitrogen Corporation, Carlsbad, Calif.) ® The module allows for the assessment of amino acid identity percentages after alignment using ClustalW. A non-limiting example of a software program that can be used to analyze ClustalW alignments is GENEDOC. ™ GENEDOC ™ (Karl Nicholas) was able to assess the amino acid (or DNA) similarity and identity of a variety of proteins. Another non-limiting example of a mathematical algorithm for sequence comparison is Myers and Miller, (1988). CABIOS The algorithm presented in 4:11-17. This type of algorithm is incorporated into the ALIGN program (version 2.0), which is part of the GCG Wisconsin Genetics software package version 10 (Accelrys, Inc., 9685 Scranton Rd., San Diego, Calif., USA). When comparing amino acid sequences using the ALIGN program, the PAM120 weighted residue table, vacancy length penalty 12, and vacancy penalty 4 can be used.

[0181] Another non-restricted example of a mathematical algorithm for sequence comparison is Needleman and Wunsch (1970). J. Mol. Biol The algorithm proposed in .48(3):443-453 uses GAP version 10 software and determines sequence identity or similarity using the following default parameters: identity % and similarity % of nucleic acid sequences are determined using GAP weight 50 and length weight 3 and the nwsgapdna.cmpii scoring matrix; identity % or similarity % of amino acid sequences are determined using GAP weight 8 and length weight 2 and the BLOSUM62 scoring program. An equivalence procedure can also be used. “Equivalence procedure” is used herein to refer to any such sequence comparison procedure that, for any two sequences under consideration, produces alignments with the same nucleotide residue matches and the same percentage of sequence identity as the corresponding alignments produced by GAP version 10.

[0182] This disclosure also covers nucleic acid molecules encoding variants of the PIP-72 polypeptide. "Variants" of nucleic acid sequences encoding the PIP-72 polypeptide include those sequences encoding the PIP-72 polypeptide disclosed herein but exhibiting conserved differences due to genetic code degeneracy, as well as those sequences having sufficiently high identity with the sequences described above. Naturally occurring allelic variants can be identified using well-known molecular biology techniques, such as polymerase chain reaction (PCR) and hybridization techniques outlined below. Nucleic acid sequence variants also include synthetically obtained nucleic acid sequences, for example, generated by site-directed mutagenesis, but still encoding the disclosed PIP-72 polypeptide, as described below.

[0183] This disclosure provides isolated or recombinant polynucleotides encoding any of the PIP-72 polypeptides disclosed herein. Those skilled in the art will readily understand that, due to the degeneracy of the genetic code, there are numerous nucleotide sequences encoding the PIP-72 polypeptides of this disclosure. Table 1 is a codon table providing synonymous codons for each amino acid. For example, codons AGA, AGG, CGA, CGC, CGG, and CGU all encode the amino acid arginine. Therefore, at each position in the nucleic acid of this disclosure designated as arginine by a certain codon, that codon can be changed to any of the corresponding codons mentioned above without altering the encoded polypeptide. It should be understood that U in the RNA sequence corresponds to T in the DNA sequence.

[0184] Table 1 .

[0185] Those skilled in the art will also understand that alterations can be introduced by mutating nucleic acid sequences, thereby causing changes in the amino acid sequence of the encoded PIP-72 polypeptide without altering the protein's biological activity. Therefore, variant nucleic acid molecules can be formed by introducing one or more nucleotide substitutions, additions, and / or deletions into the corresponding nucleic acid sequences disclosed herein, such that one or more amino acid substitutions, additions, or deletions are introduced into the encoded protein. Mutations can be introduced using standard techniques such as site-directed mutagenesis and PCR-mediated mutagenesis. Such variant nucleic acid sequences are also covered by this disclosure.

[0186] Alternatively, mutant nucleic acid sequences can be obtained by randomly introducing mutations along all or part of the coding sequence, such as through saturation mutagenesis, and the resulting mutants can be screened for their insecticidal activity to identify those that retain the activity. After mutagenesis, the encoded protein can be expressed in a recombinant manner, and the activity of the protein can be determined using standard assay techniques.

[0187] In addition to the standard cloning methods described by Ausubel, Berger, and Sambrook, the polynucleotides and fragments thereof disclosed herein are optionally used as substrates for a variety of recombination and recurrent recombination reactions, i.e., to generate additional insecticidal polypeptide homologs and fragments thereof with desired properties. A variety of such reactions are known, including those developed by the inventors and colleagues. Methods for generating variants of any of the nucleic acids listed herein include recurrent recombination of such polynucleotides with a second (or more) polynucleotide to form a library of variant polynucleotides, as are embodiments of this disclosure, the resulting library, the cell containing the library, and any recombinant polynucleotides generated by such methods. Additionally, such methods optionally include selecting variant polynucleotides from such libraries based on insecticidal activity when such recurrent recombination is performed in vitro or in vivo.

[0188] A variety of diversity generation schemes, including nucleic acid recursive recombination schemes, are available and have been well described in the art. These procedures can be used alone and / or in combination to generate one or more variants of nucleic acids or nucleic acid sets, as well as variants of the proteins they encode. These procedures, individually and collectively, provide robust and widely applicable methods for generating diverse nucleic acids and nucleic acid sets, including, for example, nucleic acid libraries, which can be used for, for example, the engineering or rapid evolution of nucleic acids, proteins, pathways, cells, and / or organisms to acquire new and / or improved properties.

[0189] Although distinctions and classifications have been made in the subsequent discussion for clarity, it should be understood that these techniques are generally not mutually exclusive. In fact, the various methods described can be used alone or in combination, in parallel or sequentially, to obtain a wide variety of sequence variants.

[0190] The result of any diversity generation procedure described herein may be the production of one or more nucleic acids, which may be selected or screened to obtain nucleic acids having or conferring desired properties or nucleic acids encoding proteins having or conferring desired properties. Diversification can be performed using one or more methods described herein, or other methods available to those skilled in the art, and then any of the produced nucleic acids can be selected for desired activity or property (e.g., insecticidal activity or such insecticidal activity at a desired pH, etc.). This may include identifying any activity that can be detected by any assay in the art in, for example, an automated or automatable mode, see, for example, the discussion of screening for insecticidal activity below. Multiple related (or even unrelated) properties may be evaluated by the operator at their discretion, either continuously or in parallel.

[0191] A description of the diverse generation procedures for producing modified nucleic acid sequences (e.g., those nucleic acid sequences encoding polypeptides or fragments thereof with insecticidal activity) can be found in the following publications and references cited therein: Soong et al., (2000). Nat Genet 25(4):436-439; Stemmer et al., (1999) Tumor Targeting 4:1-4; Ness et al., (1999) Nat Biotechnol 17:893-896; Chang et al., (1999) Nat Biotechnol 17:793-797; Minshull and Stemmer, (1999) Curr Opin Chem Biol 3:284-290; Christians et al., (1999) Nat Biotechnol 17:259-264; Crameri et al., (1998) Nature 391:288-291; Crameri et al., (1997) Nat Biotechnol 15:436-438; Zhang et al., (1997) PNAS USA 94:4504-4509; Patten et al., (1997) Curr Opin Biotechnol 8:724-733; Crameri et al., (1996) Nat Med 2:100-103; Crameri et al., (1996) Nat Biotechnol 14:315-319; Gates et al., (1996) J Mol Biol 255:373-386; Stemmer, (1996) "Sexual PCR and Assembly PCR", The Encyclopedia of Molecular Biology VCH Publishers, New York. pp. 447-457; Crameri and Stemmer, (1995) BioTechniques 18:194-195; Stemmer et al., (1995) Gene , 164:49-53; Stemmer, (1995) Science 270: 1510; Stemmer, (1995) Bio / Technology 13:549-553; Stemmer, (1994) Nature 370:389-391; Stemmer, (1994) PNAS USA 91:10747-10751.

[0192] Mutation methods that generate diversity include (for example): site-directed mutagenesis (Ling et al., (1997)). Anal Biochem 254(2):157-178; Dale et al., (1996) Methods Mol Biol 57:369-374; Smith, (1985) Ann Rev Genet 19:423-462; Botstein and Shortle, (1985) Science 229:1193-1201; Carter, (1986) Biochem J 237:1-7; and Kunkel, (1987) "The efficiency of oligonucleotide directed mutagenesis", Nucleic Acids&Molecular Biology (Eckstein and Lilley, eds., Springer Verlag, Berlin); mutagenesis using uracil-containing templates (Kunkel, (1985)). PNAS USA 82:488-492; Kunkel et al., (1987) Methods Enzymol 154:367-382; and Bass et al., (1988) Science 242:240-245); site-directed mutagenesis of oligonucleotides (Zoller and Smith, (1983) Methods Enzymol 100:468-500; Zoller and Smith, (1987) Methods Enzymol 154:329-350 (1987); Zoller and Smith, (1982) Nucleic Acids Res 10:6487-6500); DNA mutagenesis induced by phosphate thioester modification (Taylor et al., (1985)). Nucl Acids Res 13:8749-8764; Taylor et al., (1985) Nucl Acids Res 13:8765-8787 (1985); Nakamaye and Eckstein, (1986) Nucl Acids Res 14:9679-9698; Sayers et al., (1988) Nucl Acids Res 16:791-802; and Sayers et al., (1988) Nucl Acids Res 16:803-814); mutagenesis using double-stranded DNA with vacancies (Kramer et al., (1984)). Nucl Acids Res 12:9441-9456; Kramer and Fritz, (1987) Methods Enzymol 154:350-367; Kramer et al., (1988) Nucl Acids Res 16:7207; and Fritz et al., (1988) Nucl Acids Res 16:6987-6999).

[0193] Other suitable methods include point mismatch repair (Kramer et al., (1984)). Cell 38:879-887), mutagenesis using repair-deficient host strains (Carter et al., (1985)). Nucl Acids Res 13:4431-4443; and Carter, (1987) Methods in Enzymol 154:382-403), deletion-induced mutagenesis (Eghtedarzadeh and Henikoff, (1986)). Nucl Acids Res 14:5115), restricted selection and restricted purification (Wells et al., (1986) Phil TransR Soc Lond A 317:415-423), mutagenesis via whole-genome synthesis (Nambiar et al., (1984)). Science 223:1299-1301; Sakamar and Khorana, (1988) Nucl Acids Res 14:6361-6372; Wells et al., (1985) Gene 34:315-323; and Grundström et al., (1985) Nucl Acids Res 13:3305-3316), double-strand fracture repair (Mandecki, (1986) PNAS USA , 83:7177-7181; and Arnold, (1993) Curr Opin Biotech 4:450-455). For more details on many of the methods described above, please refer to [reference needed]. Methods Enzymol 154, which also describes a control group among various mutagenesis methods that can effectively eliminate uncertainties.

[0194] Further details regarding various methods of generating diversity can be found in the following U.S. patents, PCT publications and applications, and EPO publications: U.S. Patent 5,723,323, U.S. Patent 5,763,192, U.S. Patent 5,814,476, U.S. Patent 5,817,483, U.S. Patent 5,824,514, U.S. Patent 5,976,862, U.S. Patent 5,605,793, U.S. Patent 5,811,238, U.S. Patent 5,830,721, U.S. Patent 5,834,252, U.S. Patent 5,837,458, WO 1995 / 22625, WO 1996 / 33207, WO 1997 / 20078, WO 1997 / 35966, WO 1999 / 41402, WO 1999 / 41383, WO 1999 / 41369, WO 1999 / 41368、EP 752008、EP 0932670、WO 1999 / 23107、WO 1999 / 21979、WO 1998 / 31837、WO 1998 / 27230、WO 1998 / 27230、WO 2000 / 00632、WO 2000 / 09679、WO 1998 / 42832、WO1999 / 29902、WO 1998 / 41653、WO 1998 / 41622、WO 1998 / 42727、WO 2000 / 18906、WO 2000 / 04190、WO 2000 / 42561、WO 2000 / 42559、WO 2000 / 42560, WO 2001 / 23401 and PCT / US01 / 06775.

[0195] The nucleotide sequences of the embodiments disclosed herein can also be used to isolate corresponding sequences from other organisms, particularly other bacteria, especially species of the genus *Pseudomonas*, and even more particularly *Pseudomonas putida*. Pseudomonas putida ), Pseudomonas aeruginosa ( Pseudomonas fulva (e.g., *Pseudomonas aeruginosa* strains). In this manner, such sequences can be identified based on their sequence identity with the sequences shown herein, using methods such as PCR or hybridization. This disclosure covers sequences selected based on their sequence identity with the complete sequences or fragments thereof shown herein. Such sequences include sequences that are orthologs of the disclosed sequences. The term "ortholog" refers to a gene derived from a common ancestor and present in different species due to speciation. Genes present in different species are considered orthologs when their nucleotide sequences and / or the protein sequences they encode have substantial identity as defined elsewhere herein. The function of orthologs is often highly conserved across species.

[0196] In PCR methods, oligonucleotide primers can be designed for PCR reactions to amplify corresponding DNA sequences from cDNA or genomic DNA extracted from any organism of interest. The methods for designing PCR primers and PCR cloning are well-known in the art and are disclosed in the following literature: Sambrook et al., (1989). Molecular Cloning: A Laboratory Manual (2nd edition, Cold Spring Harbor Laboratory Press, Plainview, New York), hereinafter referred to as "Sambrook". See also Innis et al., eds. (1990), PCR Protocols: A Guide to Methods and Applications (Academic Press, New York); edited by Innis and Gelfand (1995) PCR Strategies (Academic Press, New York); edited by Innis and Gelfand (1999) PCR MethodsManual (AcademicPress, New York). Known PCR methods include, but are not limited to, methods using paired primers, nested primers, single-specific primers, degenerate primers, gene-specific primers, vector-specific primers, and partially mismatched primers.

[0197] To identify potential PIP-72 peptides from bacterial deposits, bacterial cell lysates can be screened using Western blotting and / or ELISA methods with antibodies generated from PIP-72 peptides as antigens. This type of assay can be performed in high-throughput mode. Various techniques, such as antibody-based protein purification and identification techniques, can be used to further analyze positive samples. Methods for generating antibodies are well known in the art and are discussed below.

[0198] Alternatively, mass spectrometry-based protein identification methods can be used to identify homologs of the PIP-72 peptide using the protocol described in the literature (Scott Patterson, (1998), 10.22, 1-24, Current Protocol in Molecular Biology, John Wiley & Son Inc). Specifically, using LC-MS / MS-based protein identification methods, MS data of a given cell lysate or a sample rich in the desired molecular weight (cleaved from the PIP-72-related molecular weight band in an SDS-PAGE gel) are correlated with the sequence information of PIP-72 (e.g., SEQ ID NO: 2) and its homologs. Any match in the peptide sequence indicates the possible presence of a homolog in the sample. Further techniques (protein purification and molecular biology) can be used to separate the protein and identify the sequences of homologs.

[0199] When using hybridization methods, a portion or the entire insecticidal nucleic acid sequence can be used to screen cDNA or genomic libraries. Methods for constructing such cDNA and genomic libraries are well-known in the art and disclosed in Sambrook and Russell's 2001 publication (ibid.). Hybridization probes can be genomic DNA fragments, cDNA fragments, RNA fragments, or other oligonucleotides, and can be labeled with detectable groups such as 32P or any other detectable markers, such as other radioisotopes, fluorescent compounds, enzymes, or enzyme cofactors. Hybridization probes can be prepared by labeling synthetic oligonucleotides based on the known PIP-72 polypeptide-encoded nucleic acid sequence disclosed herein. Alternatively, degenerate primers designed based on conserved nucleotides or amino acid residues in the nucleic acid sequence or the encoded amino acid sequence can be used. The probe typically contains a nucleic acid sequence region that, under stringent conditions, hybridizes to at least about 12, at least about 25, at least about 50, 75, 100, 125, 150, 175, or 200 consecutive nucleotides of a nucleic acid sequence or fragment or variant thereof encoding the disclosed PIP-72 polypeptide. Methods for preparing the hybridization probe are well known in the art and are disclosed in Sambrook and Russell's 2001 publication (ibid.), which is incorporated herein by reference.

[0200] For example, the entire nucleic acid sequence encoding the PIP-72 polypeptide disclosed herein, or one or more portions thereof, can be used as a probe capable of specifically hybridizing to a corresponding nucleic acid sequence and messenger RNA encoding a PIP-72 polypeptide-like sequence. To achieve specific hybridization under various conditions, such probes comprise unique sequences and are preferably at least about 10 nucleotides or at least about 20 nucleotides in length. Such probes can be used to amplify the corresponding insecticidal sequence from selected organisms by PCR. This technique can be used to isolate additional coding sequences from desired organisms or as a diagnostic assay to determine the presence of coding sequences in organisms. Hybridization techniques include hybridization screening of DNA libraries inoculated on plates (plaques or colonies; see, for example, Sambrook et al., (1989) Molecular Cloning: A Laboratory Manual (2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY).

[0201] Hybridization of these sequences can be performed under stringent conditions. "Stringent conditions" or "stringent hybridization conditions" are used herein to refer to conditions where the probe hybridizes to a detectably higher degree with its target sequence than it hybridizes to other sequences (e.g., at least 2-fold higher than background). Stringent conditions are sequence-dependent and will vary in different situations. By controlling the stringency of hybridization and / or washing conditions, target sequences that are 100% complementary to the probe can be identified (homologous detection). Alternatively, stringent conditions can be adjusted to allow for some mismatches in the sequences, thereby detecting a lower degree of similarity (heterologous detection). Probe length is typically less than about 1000 nucleotides, preferably less than 500 nucleotides.

[0202] Typically, stringent conditions are as follows: a salt concentration below about 1.5 M sodium ions (typically about 0.01 to 1.0 M sodium ion concentration (or other salts)), a pH of 7.0 to 8.3, a usage temperature of at least about 30 °C for short probes (e.g., 10 to 50 nucleotides), and a usage temperature of at least about 60 °C for long probes (e.g., more than 50 nucleotides). Stringent conditions can also be achieved by adding a destabilizing agent such as formamide. An exemplary low-stringent condition involves hybridization at 37 °C with a buffer of 30% to 35% formamide, 1 M NaCl, and 1% SDS (sodium dodecyl sulfate), followed by washing with 1 to 2 times the amount of SSC (20 times SSC = 3.0 M NaCl / 0.3 M trisodium citrate) at 50 to 55 °C. An exemplary medium-stringent condition involves hybridization at 37 °C with 40% to 45% formamide, 1.0 M NaCl, and 1% SDS, followed by washing with 0.5 to 1 times the amount of SSC at 55 to 60 °C. Exemplary high-toughness conditions include hybridization at 37°C in 50% formamide, 1M NaCl, and 1% SDS, followed by washing with 0.1-fold SSC at 60°C to 65°C. The wash buffer may optionally contain about 0.1% to about 1% SDS. The hybridization duration is typically less than about 24 hours, generally about 4 to about 12 hours.

[0203] Specificity typically varies with washing conditions after hybridization, with key factors being the ionic strength and temperature of the final washing solution. For DNA-DNA hybrids, this can be referenced according to Meinkoth and Wahl (1984). Anal. Biochem Tm is calculated using the equation Tm = 81.5℃ + 16.6 (log M) + 0.41 (% GC) - 0.61 (% form) - 500 / L, as proposed in .138:267-284; where M is the molar concentration of the monovalent cation, %GC is the percentage of guanine and cytosine nucleotides in the DNA, %form is the percentage of formamide in the hybridization solution, and L is the length of the hybrid (in base pairs). Tm is the temperature at which 50% of the complementary target sequence hybridizes with a perfectly matched probe (at a defined ionic strength and pH). For every 1% increase in mismatch rate, Tm decreases by approximately 1℃; therefore, hybridization with sequences possessing the desired identity can be achieved by adjusting Tm, hybridization conditions, and / or washing conditions. For example, if seeking sequences with ≥90% identity, Tm can be reduced by 10℃. Typically, stringent conditions are chosen to be approximately 5℃ lower than the pyrolysis temperature (Tm) of the specific sequence and its complement at a defined ionic strength and pH. However, extremely stringent conditions can be achieved using hybridization and / or washing at temperatures 1, 2, 3, or 4 °C lower than the thermal desorption temperature (Tm); moderately stringent conditions can be achieved using hybridization and / or washing at temperatures 6, 7, 8, 9, or 10 °C lower than the thermal desorption temperature (Tm); and low stringent conditions can be achieved using hybridization and / or washing at temperatures 11, 12, 13, 14, 15, or 20 °C lower than the thermal desorption temperature (Tm). Using the formulas described above, the hybridization and washing compositions, and the desired Tm, those skilled in the art will recognize that variations in the stringency of the hybridization and / or washing solutions are inherently described. If the desired degree of mismatch results in a Tm below 45 °C (aqueous solution) or 32 °C (formamide solution), it is preferable to increase the SSC concentration to allow for the use of higher temperatures. Detailed instructions on nucleic acid hybridization can be found in Tijssen, (1993) Laboratory Techniques in Biochemistry and Molecular Biology - Hybridization with Nucleic Acid Probes, Part I, Chapter 2 (Elsevier, NY); and Ausubel et al., eds., Current Protocols in Molecular Biology, Chapter 2 (Greene Publishing and Wiley-Interscience, New York). See also Sambrook et al., (1989) Molecular Cloning: A Laboratory Manual (2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY).

[0204] In some embodiments, a nucleic acid molecule encoding a polypeptide is provided, wherein the polypeptide comprises an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or higher sequence identity with the following amino acid sequences: SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 30, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 929, SEQ ID NO: 930, SEQ ID NO: 931, SEQ ID NO: 937, SEQ ID NO: 938, SEQ ID NO: 942, SEQ ID NO: 947 or SEQ ID NO: 948, wherein the polypeptide has insecticidal activity.

[0205] Proteins and their variants and fragments This disclosure also covers the PIP-72 polypeptide. The terms “Pseudomonas insecticidal protein-72,” “PIP-72 polypeptide,” or “PIP-72 protein” are used interchangeably herein and refer to a polypeptide with insecticidal activity that is sufficiently homologous to the protein shown in SEQ ID NO: 2, wherein the insecticidal activity includes, but is not limited to, insecticidal activity against one or more insect pests in the order Coleoptera. This disclosure envisions various PIP-72 polypeptides. The sources of polynucleotides encoding the PIP-72 polypeptide or related proteins include, but are not limited to: *Pseudomonas aeruginosa* strains containing the PIP-72Aa polypeptide encoded by SEQ ID NO: 2, or the PIP-72Aa polynucleotide shown in SEQ ID NO: 1; *Pseudomonas repens* strains containing the PIP-72Ba polypeptide encoded by SEQ ID NO: 4, or the PIP-72Ba polynucleotide shown in SEQ ID NO: 3; *Pseudomonas aeruginosa* strains containing the PIP-72Ca polypeptide encoded by SEQ ID NO: 6, or the PIP-72Ca polynucleotide shown in SEQ ID NO: 5; *Pseudomonas meningitidis* strains containing the PIP-72Cb polypeptide encoded by SEQ ID NO: 8, or the PIP-72Cb polynucleotide shown in SEQ ID NO: 7; and *Pseudomonas icariina* strains containing the PIP-72Da polypeptide encoded by SEQ ID NO: 10, or the PIP-72Cb polynucleotide shown in SEQ ID NO: 10. PIP-72Da polynucleotide shown in SEQ ID NO:9; *Pseudomonas meningitidis* strain containing the PIP-72Db polypeptide shown in SEQ ID NO:12, and the PIP-72Db polynucleotide shown in SEQ ID NO:11; *Pseudomonas serrata* strain containing the PIP-72Dc polypeptide shown in SEQ ID NO:14, and the PIP-72Dc polynucleotide shown in SEQ ID NO:13; *Pseudomonas moschata* strain containing the PIP-72Fa polypeptide shown in SEQ ID NO:18, and the PIP-72Fa polynucleotide shown in SEQ ID NO:17; *Pseudomonas aeruginosa* strain containing the PIP-72Ff polypeptide shown in SEQ ID NO:28, and the PIP-72Ff polynucleotide shown in SEQ ID NO:27; *Pseudomonas aeruginosa* strain containing the PIP-72Gb polypeptide shown in SEQ ID NO:32, and the PIP-72Db polynucleotide shown in SEQ ID NO:32. 31 shows the PIP-72Gb polynucleotide; a Pseudomonas aeruginosa strain containing the PIP-72Ab polypeptide shown in SEQ ID NO: 927, and the PIP-72Ab polynucleotide shown in SEQ ID NO: 949;A *Pseudomonas rapeus* strain containing the polypeptide encoding PIP-72Ab (SEQ ID NO: 928) and the polynucleotide encoding PIP-72Bb (SEQ ID NO: 950); a *Pseudomonas entomopathogenicus* strain containing the polypeptide encoding PIP-72AFh (SEQ ID NO: 932) and the polynucleotide encoding PIP-72Fh (SEQ ID NO: 954); a *Pseudomonas entomopathogenicus* strain containing the polypeptide encoding PIP-72AFh (SEQ ID NO: 933) and the polynucleotide encoding PIP-72Fh (SEQ ID NO: 955); a *Pseudomonas aeruginosa* strain containing the polypeptide encoding PIP-72Fj (SEQ ID NO: 934) and the polynucleotide encoding PIP-72Fj (SEQ ID NO: 956); and a *Pseudomonas aeruginosa* strain containing the polypeptide encoding PIP-72Fk (SEQ ID NO: 935). PIP-72Fk polynucleotide shown in SEQ ID NO: 957; Burkholderia polyphaga strain containing PIP-72Fl polypeptide shown in SEQ ID NO: 936, PIP-72Fl polynucleotide shown in SEQ ID NO: 958; Pseudomonas aeruginosa strain containing PIP-72Gg polypeptide shown in SEQ ID NO: 939, PIP-72Gg polynucleotide shown in SEQ ID NO: 961; Pseudomonas aeruginosa strain containing PIP-72Gh polypeptide shown in SEQ ID NO: 940, PIP-72Gh polynucleotide shown in SEQ ID NO: 962; Pseudomonas moschata strain containing PIP-72Gi polypeptide shown in SEQ ID NO: 941, PIP-72Gi polynucleotide shown in SEQ ID NO: 963; Biocontrol Pseudomonas strain containing PIP-72Gi polypeptide shown in SEQ ID NO: 941, PIP-72Gi polynucleotide shown in SEQ ID NO: 963; The following are listed: PIP-72Gk polypeptide shown in SEQ ID NO: 943, PIP-72Gk polynucleotide shown in SEQ ID NO: 965; *Pseudomonas proteus* strain containing PIP-72Gl polypeptide shown in SEQ ID NO: 944, PIP-72Gl polynucleotide shown in SEQ ID NO: 966; *Pseudomonas aeruginosa* strain containing PIP-72Gn polypeptide shown in SEQ ID NO: 946, PIP-72Gn polynucleotide shown in SEQ ID NO: 968. In some embodiments, the insecticidal activity is the killing of *Hemiberlesia lataniae* (Western corn rootworm). Diabrotica virgifera virgifera ) activity.

[0206] In some embodiments, the PIP-72 polypeptide is fully homologous to the following amino acid sequences: SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 18, SEQ ID NO: 28, SEQ ID NO: 32, SEQ ID NO: 927, SEQ ID NO: 928, SEQ ID NO: 932, SEQ ID NO: 933, SEQ ID NO: 934, SEQ ID NO: 935, SEQ ID NO: 936, SEQ ID NO: 939, SEQ ID NO: 940, SEQ ID NO: 941, SEQ ID NO: 943, SEQ ID NO: 944, SEQ ID NO: 945, or SEQ ID NO: 946. As used herein, "sufficient homology" means that, when compared with a reference sequence using one of the alignment procedures described herein with standard parameters, a given amino acid sequence has at least approximately 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher sequence homology with a reference sequence. Those skilled in the art will recognize that these values ​​can be appropriately adjusted to take into account amino acid similarity, etc., to determine the corresponding homology of a protein. In some embodiments, sequence homology is homology relative to the full-length sequence of the PIP-72 polypeptide.In some embodiments, this PIP-72 peptide is compared to SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 18, SEQ ID NO: 28, SEQ ID NO: 32, SEQ ID NO: 927, SEQ ID NO: 928, SEQ ID NO: 932, SEQ ID NO: 933, SEQ ID NO: 934, SEQ ID NO: 935, SEQ ID NO: 936, SEQ ID NO: 939, SEQ ID NO: 940, SEQ ID NO: 941, SEQ ID NO: 943, SEQ ID NO: 944, SEQ ID NO: 945 or SEQ ID NO: 946, having at least about 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher sequence identity. In some embodiments, sequence identity is identity relative to the full-length sequence of the PIP-72 polypeptide. In some embodiments, sequence identity is determined using Vector NTI with all default parameters. ® ALIGNNX package (Invitrogen Corporation, Carlsbad, Calif.) ® The ClustalW algorithm in the module is used for computation. In some implementations, sequence identity is calculated using Vector NTI with all default parameters. ® ALIGNNX package (Invitrogen Corporation, Carlsbad, Calif.) ® The ClustalW algorithm in the module performs calculations across the entire length of the polypeptide.

[0207] As used herein, the terms “protein,” “peptide molecule,” or “polypeptide” include any molecule comprising five or more amino acids. It is well known in the art that protein, peptide, or polypeptide molecules can be modified, including post-translational modifications, such as, but not limited to, the formation of disulfide bonds, glycosylation, phosphorylation, or oligomerization. Therefore, as used herein, the terms “protein,” “peptide molecule,” or “polypeptide” include any protein modified by any biological or non-biological process. The term “amino acid” refers to all naturally occurring L-amino acids.

[0208] “Recombinant protein” is used herein to refer to a protein that is no longer in its natural environment, such as in vitro or in the cells of a recombinant bacterial or plant host. PIP-72 polypeptides that are substantially free of cellular material include protein preparations containing less than about 30%, 20%, 10%, or 5% (on dry weight) of non-insecticidal proteins (also referred to herein as “polluting proteins”).

[0209] The “fragment” or “bioactive portion” includes polypeptide fragments that contain an amino acid sequence with sufficient identity to the PIP-72 polypeptide and exhibit insecticidal activity. The “fragments” or “bioactive portions” of the PIP-72 polypeptide include the following fragments: the amino acid sequences of these fragments are identical to any one of the sequences specified in SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 18, SEQ ID NO: 28, SEQ ID NO: 32, SEQ ID NO: 528 to SEQ ID NO: 768, any one of the sequences specified in SEQ ID NO: 825 to SEQ ID NO: 844, SEQ ID NO: 771, SEQ ID NO: 772, SEQ ID NO: 852, SEQ ID NO: 903 to SEQ ID NO: 914, SEQ ID NO: 927, SEQ ID NO: 928, SEQ ID NO: 932, SEQ ID NO: 933, SEQ ID NO: 934, SEQ ID NO: 935, SEQ ID NO: 936, SEQ ID NO: 937, SEQ ID NO: 93 ...9, SEQ ID NO: 938, SEQ ID NO: 939, SEQ ID NO: 939, SEQ ID NO: 939, SEQ ID NO: 939, SEQ ID NO: 939, SEQ ID NO: 939, SEQ ID NO: 939, SEQ ID NO: 939, SEQ ID NO: 939, SEQ ID NO: 939, SEQ ID NO: 939, SEQ ID NO: 939, The amino acid sequences shown in SEQ ID NO: 939, SEQ ID NO: 940, SEQ ID NO: 941, SEQ ID NO: 943, SEQ ID NO: 944, SEQ ID NO: 945, or SEQ ID NO: 946 have sufficiently high identity. The bioactive portion of the PIP-72 peptide can be a peptide of length (e.g.) 10, 25, 50, 55, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, or 85 amino acids. Such bioactive portions can be prepared using recombinant techniques and their insecticidal activity can be evaluated. As used herein, the fragment contains at least eight consecutive amino acids of the PIP-72 peptide.In some embodiments, the PIP-72 polypeptide fragment comprises at least eight consecutive amino acids from the following sequences: SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 18, SEQ ID NO: 28, SEQ ID NO: 32, any one of SEQ ID NO: 528 to SEQ ID NO: 768, any one of SEQ ID NO: 825 to SEQ ID NO: 844, SEQ ID NO: 771, SEQ ID NO: 772, SEQ ID NO: 852, any one of SEQ ID NO: 903 to SEQ ID NO: 914, SEQ ID NO: 927, SEQ ID NO: 928, SEQ ID NO: 932, SEQ ID NO: 933, SEQ ID NO: 934, SEQ ID NO: 935, SEQ ID NO: 936, SEQ ID NO: 937, SEQ ID NO: 928, SEQ ID NO: 932, SEQ ID NO: 933, SEQ ID NO: 934, SEQ ID NO: 935, SEQ ID NO: 936, SEQ ID NO: 937, SEQ ID NO: 938, SEQ ID NO: 939 ... 936, SEQ ID NO:939, SEQ ID NO:940, SEQ ID NO:941, SEQ ID NO:943, SEQ ID NO:944, SEQ ID NO:945 or SEQ ID NO:946.In some embodiments, the PIP-72 polypeptide fragment is, for example, obtained by proteolytic digestion, insertion of a start codon, deletion of the codon encoding the missing amino acid accompanied by the insertion of a start codon, and / or insertion of a stop codon, from any of the sequences relative to SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 18, SEQ ID NO: 28, SEQ ID NO: 32, SEQ ID NO: 528 to SEQ ID NO: 768, any of the sequences relative to SEQ ID NO: 825 to SEQ ID NO: 844, any of the sequences relative to SEQ ID NO: 771, SEQ ID NO: 772, SEQ ID NO: 852, SEQ ID NO: 903 to SEQ ID NO: 914, SEQ ID NO: 927, SEQ ID NO: 928, SEQ ID NO: 932, SEQ ID NO: 914, SEQ ID NO: 927, SEQ ID NO: 928, SEQ ID NO: 932, SEQ ID NO: 928 ... The N-terminus and / or C-terminus of SEQ ID NO: 933, SEQ ID NO: 934, SEQ ID NO: 935, SEQ ID NO: 936, SEQ ID NO: 939, SEQ ID NO: 940, SEQ ID NO: 941, SEQ ID NO: 943, SEQ ID NO: 944, SEQ ID NO: 945 or SEQ ID NO: 946 are obtained by truncating at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or more amino acids.

[0210] In some embodiments, the PIP-72 polypeptide fragments covered herein are, for example, obtained by proteolytic digestion, insertion of a start codon, deletion of the codon encoding the missing amino acid, and the insertion of a start codon, and are any of the sequences relative to SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 18, SEQ ID NO: 28, SEQ ID NO: 32, SEQ ID NO: 528 to SEQ ID NO: 768, any of the sequences relative to SEQ ID NO: 825 to SEQ ID NO: 844, any of the sequences relative to SEQ ID NO: 771, SEQ ID NO: 772, SEQ ID NO: 852, SEQ ID NO: 903 to SEQ ID NO: 914, SEQ ID NO: 927, SEQ ID NO: 928, SEQ ID NO: 932, SEQ ID NO: 933, SEQ ID NO: 944, SEQ ID NO: 952, SEQ ID NO: 903 to SEQ ID NO: 914, SEQ ID NO: 927, SEQ ID NO: 928, SEQ ID NO: 932, SEQ ID NO: 933, SEQ ID NO: 944, SEQ ID NO: 771, SEQ ID NO: 772, SEQ ID NO: 852, SEQ ID NO: 903 to SEQ ID NO: 914, SEQ ID NO: 927, SEQ ID NO: 928, SEQ ID NO: 932, SEQ ID NO: 933, SEQ ID NO: 944, SEQ ID NO: 952, SEQ ID NO: 952, SEQ ID NO: 903 The N-terminus of SEQ ID NO: 934, SEQ ID NO: 935, SEQ ID NO: 936, SEQ ID NO: 939, SEQ ID NO: 940, SEQ ID NO: 941, SEQ ID NO: 943, SEQ ID NO: 944, SEQ ID NO: 945 or SEQ ID NO: 946 is obtained by removing 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acids.

[0211] In some embodiments, the PIP-72 polypeptide fragments covered herein are obtained by removing 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids from the N-terminus of the following fragments: SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, or variations thereof, including but not limited to any of SEQ ID NO: 528 to SEQ ID NO: 768, any of SEQ ID NO: 825 to SEQ ID NO: 844, SEQ ID NO: 771, SEQ ID NO: 772, SEQ ID NO: 852, any of SEQ ID NO: 903 to SEQ ID NO: 914, SEQ ID NO: 927, SEQ ID NO: 928, SEQ ID NO: 932, SEQ ID NO: 933, SEQ ID NO: 934, SEQ ID NO: 935, SEQ ID NO: 936, SEQ ID NO: 939 ... NO: 940, SEQ ID NO: 941, SEQ ID NO: 943, SEQ ID NO: 944, SEQ ID NO: 945 or SEQ ID NO: 946.In some embodiments, the first four amino acids of the following sequences are truncated: SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 18, SEQ ID NO: 28, SEQ ID NO: 32, or variations thereof, including but not limited to any of SEQ ID NO: 528 to SEQ ID NO: 768, any of SEQ ID NO: 825 to SEQ ID NO: 844, SEQ ID NO: 771, SEQ ID NO: 772, SEQ ID NO: 852, any of SEQ ID NO: 903 to SEQ ID NO: 914, SEQ ID NO: 927, SEQ ID NO: 928, SEQ ID NO: 932, SEQ ID NO: 933, SEQ ID NO: 934, SEQ ID NO: 935, SEQ ID NO: 936, SEQ ID NO: 937, SEQ ID NO: 928, SEQ ID NO: 932, SEQ ID NO: 933, SEQ ID NO: 934, SEQ ID NO: 935, SEQ ID NO: 936, SEQ ID NO: 937, SEQ ID NO: 93 ...9, SEQ ID NO: 932, SEQ ID NO: 933, SEQ ID NO: 934, SEQ ID NO: 935, SEQ ID NO: 936, SEQ ID NO: 937, 936, SEQ ID NO: 939, SEQ ID NO: 940, SEQ ID NO: 941, SEQ ID NO: 943, SEQ ID NO: 944, SEQ ID NO: 945 or SEQ ID NO: 946.

[0212] As used herein, “variants” refers to proteins or polypeptides that have an amino acid sequence identity with their parental amino acid sequence of at least about 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%. The term “about” as used herein with respect to the percentage of sequence identity (%) means a deviation limit of ±0.5%, in increments of 0.1%. For example, “approximately 90%” sequence identity includes sequence identity of 89.5%, 89.6%, 89.7%, 89.8%, 89.9%, 90%, 90.1%, 90.2%, 90.3%, 90.4%, and 90.5%.

[0213] In some embodiments, the PIP-72 polypeptide shares at least about 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the full length of the following amino acid sequences: SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 18, SEQ ID NO: 28, SEQ ID NO: 99%. NO: 32, SEQ ID NO: 927, SEQ ID NO: 928, SEQ ID NO: 932, SEQ ID NO: 933, SEQ ID NO: 934, SEQ ID NO: 935, SEQ ID NO: 936, SEQ ID NO: 939, SEQ ID NO: 940, SEQ ID NO: 941, SEQ ID NO: 943, SEQ ID NO: 944, SEQ ID NO: 945 or SEQ ID NO: 946.

[0214] In some embodiments, the PIP-72 polypeptide has at least about 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity with the full length of the amino acid sequence shown in SEQ ID NO: 2.

[0215] In some embodiments, the PIP-72 polypeptide has at least about 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity with the full length of the amino acid sequence shown in SEQ ID NO: 4.

[0216] In some embodiments, the PIP-72 polypeptide has at least about 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity with the full length of the amino acid sequence shown in SEQ ID NO: 6.

[0217] In some embodiments, the PIP-72 polypeptide has at least about 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity with the full length of the amino acid sequence shown in SEQ ID NO: 8.

[0218] In some embodiments, the PIP-72 polypeptide has at least about 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity with the full length of the amino acid sequence shown in SEQ ID NO: 10.

[0219] In some embodiments, the PIP-72 polypeptide has at least about 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity with the full length of the amino acid sequence shown in SEQ ID NO: 12.

[0220] In some embodiments, the PIP-72 polypeptide has at least about 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity with the full length of the amino acid sequence shown in SEQ ID NO: 14.

[0221] In some embodiments, the PIP-72 polypeptide has at least about 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity with the full length of the amino acid sequence shown in SEQ ID NO: 18.

[0222] In some embodiments, the PIP-72 polypeptide has at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity with the full length of the amino acid sequence shown in SEQ ID NO: 28.

[0223] In some embodiments, the PIP-72 polypeptide has at least about 95%, 96%, 97%, 98%, 99% or higher identity with the full length of the amino acid sequence shown in SEQ ID NO: 32.

[0224] In some embodiments, the PIP-72 polypeptide has at least about 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity with the full length of the amino acid sequence shown in SEQ ID NO: 927.

[0225] In some embodiments, the PIP-72 polypeptide has at least about 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity with the full length of the amino acid sequence shown in SEQ ID NO: 928.

[0226] In some embodiments, the PIP-72 polypeptide has at least about 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity with the full length of the amino acid sequence shown in SEQ ID NO: 932.

[0227] In some embodiments, the PIP-72 polypeptide has at least about 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity with the full length of the amino acid sequence shown in SEQ ID NO: 933.

[0228] In some embodiments, the PIP-72 polypeptide has at least about 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity with the full length of the amino acid sequence shown in SEQ ID NO: 934.

[0229] In some embodiments, the PIP-72 polypeptide has at least about 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity with the full length of the amino acid sequence shown in SEQ ID NO: 935.

[0230] In some embodiments, the PIP-72 polypeptide has at least about 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity with the full length of the amino acid sequence shown in SEQ ID NO: 936.

[0231] In some embodiments, the PIP-72 polypeptide has at least about 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity with the full length of the amino acid sequence shown in SEQ ID NO: 939.

[0232] In some embodiments, the PIP-72 polypeptide has at least about 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity with the full length of the amino acid sequence shown in SEQ ID NO: 940.

[0233] In some embodiments, the PIP-72 polypeptide has at least about 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity with the full length of the amino acid sequence shown in SEQ ID NO: 941.

[0234] In some embodiments, the PIP-72 polypeptide has at least about 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity with the full length of the amino acid sequence shown in SEQ ID NO: 943.

[0235] In some embodiments, the PIP-72 polypeptide has at least about 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity with the full length of the amino acid sequence shown in SEQ ID NO: 944.

[0236] In some embodiments, the PIP-72 polypeptide has at least about 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity with the full length of the amino acid sequence shown in SEQ ID NO: 945.

[0237] In some embodiments, the PIP-72 polypeptide has at least about 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 78%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity with the full length of the amino acid sequence shown in SEQ ID NO: 946.

[0238] In some embodiments, the PIP-72 polypeptide contains an amino acid sequence that is at least 50% identical to the following amino acid sequences: SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12 or SEQ ID NO: 14, wherein the polypeptide has insecticidal activity.

[0239] In some embodiments, the PIP-72 polypeptide contains an amino acid sequence that is at least 70% identical to the following amino acid sequences: SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12 or SEQ ID NO: 14, wherein the polypeptide has insecticidal activity.

[0240] In some embodiments, the PIP-72 peptide contains an amino acid sequence that is at least 50% identical to the amino acid sequence shown in SEQ ID NO: 2, wherein the peptide has insecticidal activity.

[0241] In some embodiments, the PIP-72 peptide contains an amino acid sequence that is at least 50% identical to the amino acid sequence shown in SEQ ID NO: 4, wherein the peptide has insecticidal activity.

[0242] In some embodiments, the PIP-72 peptide contains an amino acid sequence that is at least 50% identical to the amino acid sequence shown in SEQ ID NO: 6, wherein the peptide has insecticidal activity.

[0243] In some embodiments, the PIP-72 peptide contains an amino acid sequence that is at least 50% identical to the amino acid sequence shown in SEQ ID NO: 8, wherein the peptide has insecticidal activity.

[0244] In some embodiments, the PIP-72 peptide contains an amino acid sequence that is at least 50% identical to the amino acid sequence shown in SEQ ID NO: 10, wherein the peptide has insecticidal activity.

[0245] In some embodiments, the PIP-72 peptide contains an amino acid sequence that is at least 50% identical to the amino acid sequence shown in SEQ ID NO: 12, wherein the peptide has insecticidal activity.

[0246] In some embodiments, the PIP-72 peptide contains an amino acid sequence that is at least 50% identical to the amino acid sequence shown in SEQ ID NO: 14, wherein the peptide has insecticidal activity.

[0247] In some implementations, sequence identity is achieved using Vector NTI with all default parameters. ® ALIGNNX package (Invitrogen Corporation, Carlsbad, Calif.) ® The ClustalW algorithm in the module performs calculations across the entire length of the polypeptide.

[0248] In some embodiments, the amino acid motif of the PIP-72 polypeptide is represented by amino acid residues from position 37 to position 51 of the following sequences: SEQ ID NO: 846, SEQ ID NO: 847, SEQ ID NO: 848, or SEQ ID NO: 849.

[0249] In some embodiments, the PIP-72 polypeptide comprises the amino acid sequence SEQ ID NO: 2, but selected from the 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd, 23rd, 24th, 25th, 26th, 27th, 28th, 29th, 30th, 31st, 32nd, 33rd, 34th, 35th, 36th, 37th, 38th, 39th, 40th, 42nd, 44th, 45th, 46th, 47th, 48th, 49th, 50th, 51st, 5th... The PIP-72 polypeptide has an amino acid substitution at one or more residues at positions 2, 53, 54, 56, 57, 58, 59, 60, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, or 86, and optionally, compared to SEQ ID NO: 2, a deletion of 1 to 5 amino acids, an insertion of 1 to 5 amino acids, an addition of one or more amino acids at the N-terminus, and / or an addition of one or more amino acids at the C-terminus (these changes can be combined in any way).

[0250] In some embodiments, the PIP-72 polypeptide comprises the amino acid sequence SEQ ID NO: 2, but selected from the 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 17th, 18th, 19th, 20th, 22nd, 23rd, 24th, 25th, 26th, 27th, 28th, 29th, 30th, 31st, 32nd, 33rd, 34th, 35th, 36th, 37th, 38th, 39th, 40th, 42nd, 44th, 45th, 46th, 47th, 48th, 49th, 50th, and 51st amino acids in SEQ ID NO: 2. The PIP-72 polypeptide has an amino acid substitution at one or more residues at positions 52, 53, 54, 56, 58, 60, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, or 86, and optionally, compared to SEQ ID NO: 2, a deletion of 1 to 5 amino acids, an insertion of 1 to 5 amino acids, an addition of one or more amino acids at the N-terminus, or an addition of one or more amino acids at the C-terminus (these changes can be combined in any way).

[0251] In some embodiments, the PIP-72 polypeptide comprises the amino acid sequence SEQ ID NO: 2, but selected from the 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 16th, 17th, 18th, 19th, 20th, 21st, 22nd, 23rd, 24th, 25th, 26th, 27th, 28th, 29th, 30th, 31st, 32nd, 33rd, 34th, 35th, 36th, 37th, 38th, 39th, 40th, 42nd, 44th, 45th, 46th, 47th, 48th, 49th, 50th, 51st, 5th... The PIP-72 polypeptide has amino acid substitutions at residues 2, 53, 54, 56, 57, 58, 59, 60, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, and 86, and optionally, compared to SEQ ID NO: 2, 1 to 5 amino acids are deleted, 1 to 5 amino acids are inserted, one or more amino acids are added at the N-terminus, and / or one or more amino acids are added at the C-terminus (these changes can be combined in any way).

[0252] In some embodiments, the PIP-72 polypeptide comprises the amino acid sequence SEQ ID NO: 2, which, compared to the natural amino acid shown in SEQ ID NO: 2, is selected from amino acids 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 17, 18, 19, 20, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 42, 44, 45, 46, 47, 48, 49, 50, 51, ... The PIP-72 polypeptide has amino acid substitutions at residues 52, 53, 54, 56, 58, 60, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, and 86, in any combination of 1 to 45 residues, and optionally, compared to SEQ ID NO: 2, the PIP-72 polypeptide also has 1 to 5 amino acids missing, 1 to 5 amino acids inserted, one or more amino acids added at the N-terminus, and / or one or more amino acids added at the C-terminus (these changes are in any combination).

[0253] In specific implementations, the substitution is made by replacing the natural amino acid with alanine at the listed positions. Also covered are nucleic acid sequences encoding variant proteins or polypeptides.

[0254] In some embodiments, the PIP-72 polypeptide comprises the amino acid sequence SEQ ID NO: 846, wherein Xaa at position 2 is Gly, Ala, Cys, Asp, Glu, Ile, Lys, Leu, Asn, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 3 is Ile or Trp; Xaa at position 4 is Thr, Ala, Asp, Glu, His, Ile, Lys, Leu, Arg, Ser, Val, Trp, or Tyr; Xaa at position 5 is Val, Ala, Cys, Gly, His, Ile, or Tyr; and Xaa at position 6 is Thr, Ala, Cys, Phe, Gly, His, Ile, Lys, Met, Pro, Gln, Arg, Ser, Trp, or Tyr; Xaa at position 7 is Asn, Ala, or Val; Xaa at position 8 is Asn, Ala, Cys, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Gln, Arg, Ser, Thr, or Val; Xaa at position 9 is Ser, Ala, Cys, Gly, or Thr; Xaa at position 10 is Ser, Ala, Glu, Phe, Gly, His, Ile, Lys, Leu, Asn, Pro, Gln, Arg, Thr, or Trp; Xaa at position 11 is Asn, Ala, C ys, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Gln, Ser, Thr, Val, or Tyr; Xaa at position 12 is Pro, Ala, Cys, Asp, Glu, Gly, His, Lys, Leu, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 13 is Ile, Asn, Gln, or Val; Xaa at position 14 is Glu, Ala, Cys, Phe, His, Lys, or Gln; Xaa at position 15 is Val, Ala, Cys, Ile, Met, or Arg; Xaa at position 17 is... The Xaa at position 1 is Ile, Glu, or Val; the Xaa at position 18 is Asn or Ser; the Xaa at position 19 is His, Ala, Glu, Lys, Leu, Pro, Arg, Ser, or Tyr; the Xaa at position 20 is Trp, Ala, or Thr; the Xaa at position 22 is Ser, Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Thr, Val, or Tyr; the Xaa at position 23 is Asp, Ala, Gly, His, Lys, Met, Asn, Gln, Ser, Thr, or Val.Xaa at position 24 is Gly, Asp, or Phe; Xaa at position 25 is Asp, Ala, Glu, Phe, Asn, or Gln; Xaa at position 26 is Thr, Glu, or Pro; Xaa at position 27 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Asn, Gln, Arg, or Thr; Xaa at position 28 is Phe, Pro, Trp, or Tyr; Xaa at position 29 is Phe, Ala, Cys, Ile, Leu, Gln, Arg, Trp, or Tyr; Xaa at position 30 is Ser, Ala, Cys, or Asp. The following are the possible values ​​for Xaa: Glu, Phe, Gly, His, Lys, Leu, Met, Asn, Pro, Gln, Arg, Thr, Val, Trp, or Tyr; Xaa at position 31 is Val, Ile, or Leu; Xaa at position 32 is Gly, Ala, Asp, Glu, Phe, His, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 33 is Asn, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Pro, Gln, Arg, Ser The Xaa at position 34 is Gly, Glu, Phe, His, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Tyr; the Xaa at position 35 is Lys, Ala, Cys, Asp, Gly, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Val; the Xaa at position 36 is Gln, Ala, Cys, Glu, Gly, His, Ile, Lys, Leu, Asn, Pro, Arg, Ser, Thr, or Val; the Xaa at position 37 is Glu, Ala, Cys, Asp, Phe, Gly, Ile, Lys, Leu, Met, Asn, Ser, Thr, or Val; Xaa at position 38 is Thr, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Val, Trp, or Tyr; Xaa at position 39 is Trp or Phe; Xaa at position 40 is Asp, Ala, Cys, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr;The Xaa at position 42 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Thr, Val, Trp, or Tyr; the Xaa at position 44 is Ser, Ala, Asp, Glu, Gly, Leu, Met, Asn, Pro, Gln, Thr, Val, or Tyr; the Xaa at position 45 is Arg, Lys, or Ser; the Xaa at position 46 is Gly, Ala, or Gln; the Xaa at position 47 is Phe, Cys, Val, or Tyr; the Xaa at position 48 is Val, Ile, or Leu; the Xaa at position 49 is... Xaa at position 9 is Leu, Cys, Phe, Met, Arg, or Tyr; Xaa at position 50 is Ser, Ala, Cys, Asp, Ile, Met, Pro, Gln, Thr, or Val; Xaa at position 51 is Leu, Ala, Cys, Met, or Val; Xaa at position 52 is Lys, Cys, Phe, His, Ile, Leu, Met, Asn, Arg, Ser, Thr, Trp, or Tyr; Xaa at position 53 is Lys, Ala, Cys, Asp, Glu, Phe, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Val. The Xaa at position 54 is either l or Tyr; the Xaa at position 54 is Asn, Cys, Asp, Glu, Phe, Gly, Lys, Met, Gln, Arg, Ser, or Trp; the Xaa at position 56 is Ala, Gly, Leu, Asn, Pro, Gln, Arg, Ser, or Thr; the Xaa at position 57 is Gln, Glu, Leu, Met, Ser, or Thr; the Xaa at position 58 is His, Ala, Asp, Phe, Leu, Met, Asn, Arg, Trp, or Tyr; the Xaa at position 60 is Tyr, Glu, or Phe; the Xaa at position 63 is Gln, Cys, Gly, Ile, Leu. Met, Asn, Thr, Val, or Tyr; Xaa at position 64 is Ala, Phe, Gly, His, Arg, Ser, or Tyr; Xaa at position 65 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Leu, Asn, Thr, or Val; Xaa at position 66 is Ser, Ala, or Gly; Xaa at position 67 is Lys, Ala, Cys, Asp, Phe, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 68 is Ile, Asp, Leu, or Val.Xaa at position 69 is Glu, Ala, Cys, Asp, Phe, His, Ile, Leu, Met, Gln, Arg, Ser, Thr, Val, or Tyr; Xaa at position 70 is Val, Cys, or Ile; Xaa at position 71 is Asp, Ala, Cys, Gly, His, Ile, Leu, Met, Asn, Ser, Thr, Val, or Tyr; Xaa at position 72 is Asn, Ala, Cys, Asp, Glu, Gly, Lys, Met, Pro, Gln, Arg, Ser, Thr, Val, or Trp; Xaa at position 73... The first Xaa is Asn, Ala, Cys, Asp, Phe, Gly, His, Ile, Leu, Ser, Thr, Val, or Tyr; the second Xaa at position 74 is Ala, Cys, Asp, Phe, Gly, His, Ile, Leu, Asn, Gln, Arg, Ser, Thr, Val, or Tyr; the third Xaa at position 75 is Val, Cys, Ile, or Leu; the fourth Xaa at position 76 is Lys, Ala, Cys, Phe, His, Ile, Leu, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; the fifth Xaa at position 77 is Asp or Tyr; the sixth Xaa at position 78 is Asn, Ala, Cys, Phe, His, Ile, Leu, Gln, Arg, Ser, Thr, Val, Trp, or Tyr. Xaa at position 78 is Gln, Ala, Cys, Asp, Phe, Gly, His, Ile, Leu, Met, Asn, Arg, Ser, Thr, Val, or Tyr; Xaa at position 79 is Gly, Arg, Ala, Cys, Asp, Glu, Phe, His, Lys, Leu, Asn, Gln, Arg, Ser, Thr, Trp, or Tyr; Xaa at position 80 is Arg, Ala, Cys, Asp, Phe, Gly, His, Ile, Leu, Asn, Ser, Thr, Val, or Tyr; Xaa at position 81 is Leu, Al a, Cys, Asp, Phe, Gly, His, Ile, Asn, Pro, Arg, Ser, Thr, or Val; Xaa at position 82 is Ile, Ala, Leu, Met, Arg, or Val; Xaa at position 83 is Glu, Ala, Cys, Asp, Phe, Gly, His, Ile, Lys, Leu, Asn, Pro, Arg, Ser, Thr, Val, or Tyr; Xaa at position 84 is Pro, Ala, Cys, Glu, Ile, Ser, Val, Trp, or Tyr; Xaa at position 85 is Leu, Cys, Gly, or Val.Furthermore, Xaa at position 86 is Ser, Ala, Ile, Thr, or Val, and optionally, 1 to 14 amino acids are deleted from the N-terminus and / or C-terminus of the PIP-72 polypeptide.

[0255] In some embodiments, the PIP-72 polypeptide comprises the amino acid sequence SEQ ID NO: 846, which, compared to the native amino acid at the corresponding position in SEQ ID NO: 2, has any combination of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions at the residues marked by Xaa in SEQ ID NO: 846.

[0256] In some embodiments, the PIP-72 polypeptide comprises the amino acid sequence SEQ ID NO: 846, which, compared to the native amino acid at the corresponding position in SEQ ID NO: 2, has any combination of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, or 29 amino acid substitutions at the residues marked Xaa in SEQ ID NO: 846.

[0257] In some embodiments, the PIP-72 peptide comprises the amino acid sequence SEQ ID NO: 847, wherein Xaa at position 2 is Gly, Lys, or Ala; Xaa at position 3 is Ile or Leu; Xaa at position 4 is Thr or Ser; Xaa at position 5 is Val or Ile; Xaa at position 6 is Thr or Lys; Xaa at position 8 is Asn, Lys, Gly, or Ser; Xaa at position 9 is Ser or Ala; Xaa at position 11 is Asn, Lys, His, or Thr; Xaa at position 12 is Pro, Thr, Lys, or Ser; Xaa at position 13 is Ile or Val; Xaa at position 14 is Glu or Asp; and Xaa at position 15 is Val. The Xaa at position 16 is Ala or Ile; the Xaa at position 17 is Ile or Val; the Xaa at position 18 is Asn or Ser; the Xaa at position 19 is His, Lys, Arg, Gln, or Ala; the Xaa at position 21 is Gly or Arg; the Xaa at position 22 is Ser, Lys, Asn, Asp, or Thr; the Xaa at position 25 is Asp or Asn; the Xaa at position 26 is Thr or Asp; the Xaa at position 27 is Ser, Thr, Asn, or Lys; the Xaa at position 28 is Phe, Tyr, or Pro; the Xaa at position 29 is Phe Or Tyr; Xaa at position 30 is Ser, Gly, or Lys; Xaa at position 31 is Val, Ile, or Met; Xaa at position 32 is Gly, Ala, or Asp; Xaa at position 33 is Asn, Ser, Gln, or Pro; Xaa at position 35 is Lys, Glu, or Ser; Xaa at position 36 is Gln, Asn, or Ser; Xaa at position 37 is Glu or Asp; Xaa at position 38 is Thr or Ser; Xaa at position 42 is Ser or Asn; Xaa at position 44 is Ser, Asp, Ala, or Leu; Xaa at position 47 is Phe or Tyr. The Xaa at position 48 is Leu or Met; the Xaa at position 49 is Leu or Met; the Xaa at position 50 is Ser, Ala, or Tyr; the Xaa at position 51 is Leu or Val; the Xaa at position 52 is Lys or Gln; the Xaa at position 53 is Lys, Arg, Met, or Leu; the Xaa at position 54 is Asn, Lys, or Gly; the Xaa at position 55 is Gly or Ser; the Xaa at position 56 is Ala, Thr, Gln, or Ser; the Xaa at position 57 is Gln, Val, or Ala; the Xaa at position 58 is His, Ala, Lys, Tyr, or Thr.Xaa at position 59 is Pro or Thr; Xaa at position 62 is Val or Ile; Xaa at position 63 is Gln, Ser, or Leu; Xaa at position 64 is Ala, Gln, or Ser; Xaa at position 65 is Ser or Thr; Xaa at position 67 is Lys, Gln, Arg, or Asn; Xaa at position 69 is Glu, Lys, or Val; Xaa at position 70 is Val or Ile; Xaa at position 71 is Asp, Glu, or Tyr; Xaa at position 72 is Asn, His, Ser, or Asp; Xaa at position 73 is Asn, Ser, or Asp; Xaa at position 74 is Ala, T The Xaa at position 76 is Lys or Thr; the Xaa at position 78 is Gln, His, or Ser; the Xaa at position 80 is Arg, Glu, or Gln; the Xaa at position 81 is Leu, Pro, Ala, or Thr; the Xaa at position 82 is Ile or Leu; the Xaa at position 83 is Glu, His, Asn, Gln, or Leu; the Xaa at position 85 is Leu, Val, or Ala; and the Xaa at position 86 is Ser, Ala, Tyr, or Asn, wherein optionally 1 to 14 amino acids are deleted from the N-terminus and / or C-terminus of the PIP-72 polypeptide, and / or an amino acid is inserted between residues at positions 24 and 25, compared to SEQ ID NO: 847.

[0258] In some embodiments, the PIP-72 polypeptide comprises the amino acid sequence SEQ ID NO: 847, which, compared to the native amino acid at the corresponding position in SEQ ID NO: 2, has any combination of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions at the residues marked by Xaa in SEQ ID NO: 847.

[0259] In some embodiments, the PIP-72 polypeptide comprises the amino acid sequence SEQ ID NO: 847, which, compared to the native amino acid at the corresponding position in SEQ ID NO: 2, has any combination of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, or 29 amino acid substitutions at the residues marked Xaa in SEQ ID NO: 847.

[0260] In some embodiments, the PIP-72 peptide comprises the amino acid sequence SEQ ID NO: 848, wherein Xaa at position 2 is Gly, Lys, Ala, or Arg; Xaa at position 3 is Ile, Leu, or Val; Xaa at position 4 is Thr or Ser; Xaa at position 5 is Val, Ile, or Leu; Xaa at position 6 is Thr, Lys, Ser, or Arg; Xaa at position 8 is Asn, Lys, Gly, Ser, Gln, Arg, Thr, or Ala; Xaa at position 9 is Ser, Ala, or Thr; Xaa at position 11 is Asn, Lys, Thr, Gln, Arg, His, or Ser; and Xaa at position 12 is Pro, Thr, Lys, S The Xaa at position 13 is Ile, Val, or Leu; the Xaa at position 14 is Glu or Asp; the Xaa at position 15 is Val, Ala, Ile, or Leu; the Xaa at position 16 is Ala or Ser; the Xaa at position 17 is Ile, Val, or Leu; the Xaa at position 18 is Asn, Ser, Gln, or Thr; the Xaa at position 19 is His, Lys, Ala, Gln, Asn, or Arg; the Xaa at position 21 is Gly, Arg, or Lys; the Xaa at position 22 is Ser, Lys, Asn, Thr, Arg, Asp, Glu, or Gln; the Xaa at position 2... Xaa at position 5 is Asp, Asn, Glu, or Gln; Xaa at position 26 is Thr, Asp, Ser, or Glu; Xaa at position 27 is Ser, Thr, Lys, Asn, Gln, or Arg; Xaa at position 28 is Phe, Tyr, Pro, or Trp; Xaa at position 29 is Phe, Tyr, or Trp; Xaa at position 30 is Ser, Gly, Lys, Thr, or Arg; Xaa at position 31 is Val, Ile, Met, or Leu; Xaa at position 32 is Gly, Ala, Asp, or Glu; Xaa at position 33 is Asn, Ser, Gln, or Pro. Or Thr; Xaa at position 35 is Lys, Glu, Ser, Arg, or Thr; Xaa at position 36 is Gln, Ser, Asn, or Thr; Xaa at position 37 is Glu or Asp; Xaa at position 38 is Thr or Ser; Xaa at position 42 is Ser, Asn, Thr, or Gln; Xaa at position 44 is Ser, Asp, Ala, Leu, Thr, Glu, Ile, or Val; Xaa at position 47 is Phe, Tyr, or Trp; Xaa at position 48 is Leu, Met, Ile, or Val; Xaa at position 49 is Leu, Met, Ile, or Val;Xaa at position 50 is Ser, Ala, Tyr, or Thr; Xaa at position 51 is Leu, Val, or Ile; Xaa at position 52 is Lys, Gln, Arg, or Asn; Xaa at position 53 is Lys, Arg, Met, Leu, Ile, or Val; Xaa at position 54 is Asn, Lys, Gly, Gln, or Arg; Xaa at position 55 is Gly, Ser, or Thr; Xaa at position 56 is Ala, Thr, Gln, Ser, or Asn; Xaa at position 57 is Gln, Val, Ala, Asn, Leu, or Ile; Xaa at position 5... The Xaa at position 8 is His, Ala, Lys, Tyr, or Thr; the Xaa at position 59 is Pro, Thr, or Ser; the Xaa at position 62 is Val, Ile, or Leu; the Xaa at position 63 is Gln, Ser, Leu, Asn, Thr, Ile, or Val; the Xaa at position 64 is Ala, Gln, Ser, Asn, or Thr; the Xaa at position 65 is Ser or Thr; the Xaa at position 67 is Lys, Gln, Asn, or Arg; the Xaa at position 69 is Glu, Val, Asp, Lys, Arg, Ile, or Leu; the Xaa at position 70... The Xaa at position 71 is Val, Ile, or Leu; the Xaa at position 72 is Asn, His, Ser, Asp, Gln, Thr, or Glu; the Xaa at position 73 is Asn, Ser, Asp, Gln, Thr, or Glu; the Xaa at position 74 is Ala, Thr, Met, Ile, Lys, Ser, Leu, Val, or Arg; the Xaa at position 76 is Lys, Thr, Arg, or Ser; the Xaa at position 78 is Gln, His, Ser, Asn, or Thr; the Xaa at position 80... a is Arg, Glu, Gln, Lys, Asp, or Asn; Xaa at position 81 is Leu, Pro, Thr, Ile, Val, Ala, or Ser; Xaa at position 82 is Ile, Leu, or Val; Xaa at position 83 is Glu, His, Asn, Leu, Gln, Ile, or Val; Xaa at position 85 is Leu, Val, or Ala; and Xaa at position 86 is Ser, Ala, Tyr, Asn, or Thr, wherein optionally 1 to 14 amino acids are deleted from the N-terminus and / or C-terminus of the PIP-72 polypeptide, and / or an amino acid is inserted between residues at positions 24 and 25, compared to SEQ ID NO:848.

[0261] In some embodiments, the PIP-72 polypeptide comprises the amino acid sequence SEQ ID NO: 848, which, compared to the native amino acid at the corresponding position in SEQ ID NO: 2, has any combination of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions at the residues marked by Xaa in SEQ ID NO: 848.

[0262] In some embodiments, the PIP-72 polypeptide comprises the amino acid sequence SEQ ID NO: 848, which, compared to the native amino acid at the corresponding position in SEQ ID NO: 2, has any combination of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, or 29 amino acid substitutions at the residues marked Xaa in SEQ ID NO: 848.

[0263] In some embodiments, the PIP-72 polypeptide comprises the amino acid sequence SEQ ID NO: 849, wherein Xaa at position 2 is Gly, Ala, Cys, Asp, Glu, Ile, Lys, Leu, Asn, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 3 is Ile, Leu, Val, or Trp; Xaa at position 4 is Thr, Ala, Asp, Glu, His, Ile, Lys, Leu, Arg, Ser, Val, Trp, or Tyr; Xaa at position 5 is Val, Ala, Cys, Gly, His, Ile, Leu, or Tyr; and Xaa at position 6 is Thr, Ala, Cys, Phe, Gly, His, Ile, Lys, Met, Pro, Gln, Arg, Ser, Trp, or Tyr; the 7th position Xaa is Asn, Ala, or Val; the 8th position Xaa is Asn, Lys, Gly, Ser, Gln, Arg, Thr, Ala, Cys, Asp, Glu, His, Ile, Leu, Met, or Val; the 9th position Xaa is Ser, Ala, Cys, Gly, or Thr; the 11th position Xaa is Asn, Lys, Thr, Gln, Arg, Ser, Ala, Cys, Asp, Glu, Gly, His, Ile, Leu, Met, Val, or Tyr; the 12th position Xaa is... Xaa is Pro, Thr, Lys, Ser, Arg, Ala, Cys, Asp, Glu, Gly, His, Leu, Asn, Gln, Arg, Val, Trp, or Tyr; Xaa at position 13 is Ile, Asn, Gln, Leu, or Val; Xaa at position 14 is Glu, Ala, Cys, Phe, His, Lys, Asp, or Gln; Xaa at position 15 is Val, Ala, Ile, Leu, Cys, Met, or Arg; Xaa at position 16 is Ala or Ser; Xaa at position 17 is Ile, Glu, Leu, or Val; Xaa at position 18... The Xaa at position 19 is Asn, Gln, Thr, or Ser; the Xaa at position 20 is Trp, Ala, or Thr; the Xaa at position 21 is Gly, Arg, or Lys; the Xaa at position 22 is Ser, Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Thr, Val, or Tyr; the Xaa at position 23 is Asp, Ala, Gly, His, Lys, Met, Asn, Gln, Ser, Thr, or Val.Xaa at position 24 is Gly, Asp, or Phe; Xaa at position 25 is Asp, Ala, Glu, Phe, Asn, or Gln; Xaa at position 26 is Thr, Glu, Asp, Ser, or Pro; Xaa at position 27 is Ser, Thr, Lys, Arg, Ala, Cys, Asp, Glu, Phe, Gly, His, Asn, or Gln; Xaa at position 28 is Phe, Tyr, Pro, or Trp; Xaa at position 29 is Phe, Ala, Cys, Ile, Leu, Gln, Arg, Trp, or Tyr; Xaa at position 30 is Ser, Gly, ... Lys, Thr, Arg, Ala, Cys, Asp, Glu, Phe, His, Leu, Met, Asn, Pro, Gln, Val, Trp, or Tyr; Xaa at position 31 is Val, Ile, Met, or Leu; Xaa at position 32 is Gly, Ala, Asp, Glu, Phe, His, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 33 is Asn, Ser, Gln, Pro, Thr, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, L The Xaa at position 34 is Gly, Glu, Phe, His, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Tyr; the Xaa at position 35 is Lys, Glu, Ala, Cys, Asp, Gly, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Val; the Xaa at position 36 is Gln, Ala, Cys, Glu, Gly, His, Ile, Lys, Leu, Asn, Pro, Arg, Ser, Thr, or Val; the Xaa at position 37 is Glu. The Xaa at position 38 is Thr, Ser, Ala, Cys, Phe, Gly, Ile, Lys, Leu, Met, Asn, Ser, Thr, or Val; the Xaa at position 39 is Trp or Phe; the Xaa at position 40 is Asp, Ala, Cys, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Gln, Arg, Val, Trp, or Tyr.The Xaa at position 42 is Ser, Asn, Thr, Ala, Cys, Asp, Glu, Phe, Gly, Ile, Lys, Leu, Met, Arg, Val, Trp, Tyr, or Gln; the Xaa at position 44 is Ser, Asp, Ala, Leu, Thr, Glu, Ile, Ala, Gly, Leu, Met, Asn, Pro, Gln, Val, Tyr, or Val; the Xaa at position 45 is Arg, Lys, or Ser; the Xaa at position 46 is Gly, Ala, or Gln; the Xaa at position 47 is Phe, Tyr, Cys, Val, or Trp; the Xaa at position 48 is Leu, Me The Xaa at position 49 is Leu, Met, Ile, Phe, Met, Arg, Tyr, or Val; the Xaa at position 50 is Ser, Ala, Tyr, Cys, Asp, Ile, Met, Pro, Gln, Val, or Thr; the Xaa at position 51 is Leu, Val, Ala, Cys, Met, or Ile; the Xaa at position 52 is Lys, Cys, Phe, His, Ile, Leu, Met, Asn, Arg, Ser, Thr, Gln, Trp, or Tyr; the Xaa at position 53 is Lys, Arg, Met, Leu, Ile, Ala, Cys, As p, Glu, Phe, His, Asn, Gln, Ser, Thr, Tyr, or Val; Xaa at position 54 is Asn, Cys, Asp, Glu, Phe, Gly, Lys, Met, Gln, Arg, Ser, or Trp; Xaa at position 55 is Gly, Ser, or Thr; Xaa at position 56 is Ala, Thr, Gln, Ser, Gly, Leu, Pro, Arg, or Asn; Xaa at position 57 is Gln, Glu, Leu, Met, Ser, Val, Ala, Asn, Ile, or Thr; Xaa at position 58 is His, Ala, Lys, Asp, Phe, Leu, Me t, Asn, Arg, Trp, Tyr, or Thr; Xaa at position 59 is Pro, Thr, or Ser; Xaa at position 60 is Tyr, Glu, or Phe; Xaa at position 62 is Val, Ile, or Leu; Xaa at position 63 is Gln, Ser, Cys, Gly, Ile, Leu, Met, Asn, Thr, Val, or Tyr; Xaa at position 64 is Ala, Gln, Asn, Phe, Gly, His, Arg, Ser, or Tyr; Xaa at position 65 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Leu, Asn, Val, or Thr;The Xaa at position 66 is Ser, Ala, or Gly; the Xaa at position 67 is Lys, Gln, Asn, or Arg; the Xaa at position 67 is Lys, Ala, Cys, Asp, Phe, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; the Xaa at position 68 is Ile, Asp, Leu, or Val; the Xaa at position 69 is Glu, Ala, Cys, Asp, Phe, His, Ile, Leu, Met, Gln, Arg, Ser, Thr, Val, or Tyr; the Xaa at position 70 is Val, Ile, Cys, or Leu; Xaa at position 71 is Asp, Glu, Tyr, Ala, Cys, Gly, His, Ile, Leu, Met, Asn, Ser, Thr, Val, or Trp; Xaa at position 72 is Asn, Ala, Cys, Asp, Glu, Gly, Lys, Met, Pro, Gln, Arg, Ser, Thr, Val, His, or Trp; Xaa at position 73 is Asn, Ser, Asp, Gln, Thr, Ala, Cys, Phe, Gly, His, Ile, Leu, Val, Tyr, or Glu; Xaa at position 74 is Ala, Thr, Met, Ile, Lys, Ser, Leu, Val, Cys, Asp, Phe, Gly, His, Asn, Gln, Tyr, or Arg; Xaa at position 75 is Val, Cys, Ile, or Leu; Xaa at position 76 is Lys, Ala, Cys, Phe, His, Ile, Leu, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 77 is Asp or Tyr; Xaa at position 78 is Gln, His, Ser, Asn, Ala, Cys, Asp, Phe, Gly, Ile, Leu, Met, Asn, Arg, Val, Tyr, or Thr; Xaa at position 79... The Xaa at position 1 is Gly, Arg, Ala, Cys, Asp, Glu, Phe, His, Lys, Leu, Asn, Gln, Arg, Ser, Thr, Trp, or Tyr; the Xaa at position 80 is Arg, Glu, Gln, Lys, Asp, Ala, Cys, Phe, Gly, His, Ile, Leu, Ser, Thr, Val, Tyr, or Asn; the Xaa at position 81 is Leu, Pro, Thr, Ile, Val, Ala, Cys, Asp, Phe, Gly, His, or Ser; the Xaa at position 82 is Ile, Ala, Leu, Met, Arg, or Val.Xaa at position 83 is Glu, His, Asn, Leu, Gln, Ile, Ala, Cys, Asp, Phe, Gly, Lys, Pro, Arg, Ser, Thr, Tyr, or Val; Xaa at position 84 is Pro, Ala, Cys, Glu, Ile, Ser, Val, Trp, or Tyr; Xaa at position 85 is Leu, Val, Cys, Gly, or Ala; and Xaa at position 86 is Ser, Ala, Tyr, Asn, Ile, Val, or Thr, wherein, compared to SEQ ID NO:849, 1 to 14 amino acids are optionally deleted from the N-terminus and / or C-terminus of the PIP-72 polypeptide, and / or an amino acid is inserted between residues 24 and 25.

[0264] In some embodiments, the PIP-72 polypeptide comprises the amino acid sequence SEQ ID NO:849, which, compared to the native amino acid at the corresponding position in SEQ ID NO:2, has any combination of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 amino acid substitutions at the residues marked by Xaa in SEQ ID NO:849.

[0265] In some embodiments, the PIP-72 polypeptide comprises the amino acid sequence SEQ ID NO:849, which, compared to the native amino acid at the corresponding position in SEQ ID NO:2, has any combination of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, or 29 amino acid substitutions at the residues marked by Xaa in SEQ ID NO:849.

[0266] In some embodiments, the exemplary PIP-72 peptide is composed of any one of SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 17, SEQ ID NO: 27, SEQ ID NO: 31, SEQ ID NO: 287 to SEQ ID NO: 527, any one of SEQ ID NO: 796 to SEQ ID NO: 815, SEQ ID NO: 769, SEQ ID NO: 770, SEQ ID NO: 850, SEQ ID NO: 852, SEQ ID NO: 853 to SEQ ID NO: 864, any one of SEQ ID NO: 915 to SEQ ID NO: 926, SEQ ID NO: 949, SEQ ID NO: 950, SEQ ID NO: 954, SEQ ID NO: 955, SEQ ID NO: 956, SEQ ID NO: 927, SEQ ID NO: 957, SEQ ID NO: 958, SEQ ID NO: 959 ... The polynucleotide sequences shown in SEQ ID NO: 956, SEQ ID NO: 957, SEQ ID NO: 958, SEQ ID NO: 961, SEQ ID NO: 962, SEQ ID NO: 963, SEQ ID NO: 965, SEQ ID NO: 966, SEQ ID NO: 967 or SEQ ID NO: 968 encode the polynucleotide sequences.

[0267] In some embodiments, the PIP-72 polypeptide consists of SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 17, SEQ ID NO: 27, SEQ ID NO: 31, SEQ ID NO: 769, SEQ ID NO: 770, SEQ ID NO: 850, SEQ ID NO: 852, SEQ ID NO: 949, SEQ ID NO: 950, SEQ ID NO: 954, SEQ ID NO: 955, SEQ ID NO: 956, SEQ ID NO: 957, SEQ ID NO: 958, SEQ ID NO: 961, SEQ ID NO: 962, SEQ ID NO:963, SEQ ID NO: 965, SEQ ID NO: 966, SEQ ID NO: 967 or SEQ ID NO: The polynucleotide code shown in 968.

[0268] In some embodiments, exemplary PIP-72 polypeptides are set forth in SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 18, SEQ ID NO: 28, SEQ ID NO: 32, SEQ ID NO: 528, SEQ ID NO: 529, SEQ ID NO: 530, SEQ ID NO: 531, SEQ ID NO: 532, SEQ ID NO: 533, SEQ ID NO: 534, SEQ ID NO: 535, SEQ ID NO: 536, SEQ ID NO: 537, SEQ ID NO: 538, SEQ ID NO: 539, SEQ ID NO: 540, SEQ ID NO: 541, SEQ ID NO: 542, SEQ ID NO: 543, SEQ ID NO: 544, SEQ ID NO: 545, SEQ ID NO: 546, SEQ ID NO: 547, SEQ ID NO: 548, SEQ ID NO: 549, SEQ ID NO: 550, SEQ ID NO: 551, SEQ ID NO: 552, SEQ ID NO: 553, SEQ ID NO: 554, SEQ ID NO: 555, SEQ ID NO: 556, SEQ ID NO: 557, SEQ ID NO: 558, SEQ ID NO: 559, SEQ ID NO: 560, SEQ ID NO: 561, SEQ ID NO: 562, SEQ ID NO: 563, SEQ ID NO: 564, SEQ ID NO: 565, SEQ ID NO: 566, SEQ ID NO: 567, SEQ ID NO: 568, SEQ ID NO: 569, SEQ ID NO: 570, SEQ ID NO: 571, SEQ ID NO: 572, SEQ ID NO: 573, SEQ ID NO: 574, SEQ ID NO: 575, SEQ ID NO: 576, SEQ ID NO: 577, SEQ ID NO: 578, SEQ ID NO: 579, SEQ ID NO: 580, SEQ ID NO: 581, SEQ ID NO: 582, SEQ ID NO: 583, SEQ ID NO:584、SEQ ID NO: 585、SEQ IDNO: 586、SEQ ID NO: 587、SEQ ID NO: 588、SEQ ID NO: 589、SEQ ID NO: 590、SEQ IDNO: 591、SEQ ID NO: 592、SEQ ID NO: 593、SEQ ID NO: 594、SEQ ID NO: 595、SEQ IDNO: 596、SEQ ID NO: 597、SEQ ID NO: 598、SEQ ID NO: 599、SEQ ID NO: 600、SEQ IDNO: 601、SEQ ID NO: 602、SEQ ID NO: 603、SEQ ID NO: 604、SEQ ID NO: 605、SEQ IDNO: 606、SEQ ID NO: 607、SEQ ID NO: 608、SEQ ID NO: 609、SEQ ID NO: 610、SEQ IDNO: 611、SEQ ID NO: 612、SEQ ID NO: 613、SEQ ID NO: 614、SEQ ID NO: 615、SEQ IDNO: 616、SEQ ID NO: 617、SEQ ID NO: 618、SEQ ID NO: 619、SEQ ID NO: 620、SEQ IDNO: 621、SEQ ID NO: 622、SEQ ID NO: 623、SEQ ID NO: 624、SEQ ID NO: 625、SEQ IDNO: 626、SEQ ID NO: 627、SEQ ID NO: 628、SEQ ID NO: 629、SEQ ID NO: 630、SEQ IDNO: 631、SEQ ID NO: 632、SEQ ID NO: 633、SEQ ID NO: 634、SEQ ID NO: 635、SEQ IDNO: 636、SEQ ID NO: 637、SEQ ID NO: 638、SEQ ID NO: 639、SEQ ID NO: 640、SEQ IDNO: 641、SEQ ID NO: 642、SEQ ID NO: 643、SEQ ID NO: 644、SEQ ID NO: 645、SEQ IDNO: 646、SEQ ID NO: 647、SEQ ID NO: 648、SEQ ID NO: 649、SEQ ID NO: 650、SEQ IDNO: 651、SEQID NO: 652、SEQ ID NO: 653、SEQ ID NO: 654、SEQ ID NO: 655、SEQ IDNO: 656、SEQ ID NO: 657、SEQ ID NO: 658、SEQ ID NO: 659、SEQ ID NO: 660、SEQ IDNO: 661、SEQ ID NO: 662、SEQ ID NO: 663、SEQ ID NO: 664、SEQ ID NO: 665、SEQ IDNO: 666、SEQ ID NO: 667、SEQ ID NO: 668、SEQ ID NO: 669、SEQ ID NO: 670、SEQ IDNO: 671、SEQ ID NO: 672、SEQ ID NO: 673、SEQ ID NO: 674、SEQ ID NO: 675、SEQ IDNO: 676、SEQ ID NO: 677、SEQ ID NO: 678、SEQ ID NO: 679、SEQ ID NO: 680、SEQ IDNO: 681、SEQ ID NO: 682、SEQ ID NO: 683、SEQ ID NO: 684、SEQ ID NO: 685、SEQ IDNO: 686、SEQ ID NO: 687、SEQ ID NO: 688、SEQ ID NO: 689、SEQ ID NO: 690、SEQ IDNO: 691、SEQ ID NO: 692、SEQ ID NO: 693、SEQ ID NO: 694、SEQ ID NO: 695、SEQ IDNO: 696、SEQ ID NO: 697、SEQ ID NO: 698、SEQ ID NO: 699、SEQ ID NO: 700、SEQ IDNO: 701、SEQ ID NO: 702、SEQ ID NO: 703、SEQ ID NO: 704、SEQ ID NO: 705、SEQ IDNO: 706、SEQ ID NO: 707、SEQ ID NO: 708、SEQ ID NO: 709、SEQ ID NO: 710、SEQ IDNO: 711、SEQ ID NO: 712、SEQ ID NO: 713、SEQ ID NO: 714、SEQ ID NO: 715、SEQ IDNO: 716、SEQ ID NO: 717、SEQ ID NO: 718、SEQ ID NO:719、SEQ ID NO: 720、SEQ IDNO: 721、SEQ ID NO: 722、SEQ ID NO: 723、SEQ ID NO: 724、SEQ ID NO: 725、SEQ IDNO: 726、SEQ ID NO: 727、SEQ ID NO: 728、SEQ ID NO: 729、SEQ ID NO: 730、SEQ IDNO: 731、SEQ ID NO: 732、SEQ ID NO: 733、SEQ ID NO: 734、SEQ ID NO: 735、SEQ IDNO: 736、SEQ ID NO: 737、SEQ ID NO: 738、SEQ ID NO: 739、SEQ ID NO: 740、SEQ IDNO: 741、SEQ ID NO: 742、SEQ ID NO: 743、SEQ ID NO: 744、SEQ ID NO: 745、SEQ IDNO: 746、SEQ ID NO: 747、SEQ ID NO: 748、SEQ ID NO: 749、SEQ ID NO: 750、SEQ IDNO: 751、SEQ ID NO: 752、SEQ ID NO: 753、SEQ ID NO: 754、SEQ ID NO: 755、SEQ IDNO: 756、SEQ ID NO: 757、SEQ ID NO: 758、SEQ ID NO: 759、SEQ ID NO: 760、SEQ IDNO: 761、SEQ ID NO: 762、SEQ ID NO: 763、SEQ ID NO: 764、SEQ ID NO: 765、SEQ IDNO: 766、SEQ ID NO: 767、SEQ ID NO: 768、SEQ ID NO: 771、SEQ ID NO: 772、SEQ IDNO: 825、SEQ ID NO: 826、SEQ ID NO: 827、SEQ ID NO: 828、SEQ ID NO: 829、SEQ IDNO: 830、SEQ ID NO: 831、SEQ ID NO: 832、SEQ ID NO: 833、SEQ ID NO: 834、SEQ IDNO: 835、SEQ ID NO: 836、SEQ ID NO: 837、SEQ ID NO: 838、SEQ ID NO: 839、SEQ IDNO: 840、SEQID NO: 841, SEQ ID NO: 842, SEQ ID NO: 843, SEQ ID NO: 844, SEQ ID NO: 852, SEQ ID NO: 853, SEQ ID NO: 854, SEQ ID NO: 855, SEQ ID NO: 856, SEQ ID NO: 857, SEQ ID NO: 858, SEQ ID NO: 859, SEQ ID NO: 860, SEQ ID NO: 861, SEQ ID NO: 862, SEQ ID NO: 863, SEQ ID NO: 864, SEQ ID NO: 903, SEQ ID NO: 904, SEQ ID NO: 905, SEQ ID NO: 906, SEQ ID NO: 907, SEQ ID NO: 908, SEQ ID NO: 909, SEQ ID NO: 910, SEQ ID NO: 911, SEQ ID NO: 912, SEQ ID NO: 913, SEQ ID NO: 914, SEQ ID NO: 927, SEQ ID NO: 928, SEQ ID NO: 932, SEQ ID NO: 933, SEQ ID NO: 934, SEQ ID NO: 935, SEQ ID NO: 936, SEQ ID NO: 939, SEQ ID NO: 940, SEQ ID NO: 941, SEQ ID NO: 943, SEQ ID NO: 944, SEQ ID NO: 945 and SEQ ID NO: 946 are shown.

[0269] In some embodiments, the PIP-72 polypeptide comprises the amino acid sequence shown in SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 18, SEQ ID NO: 28, SEQ ID NO: 32, SEQ ID NO: 927, SEQ ID NO: 928, SEQ ID NO: 932, SEQ ID NO: 933, SEQ ID NO: 934, SEQ ID NO: 935, SEQ ID NO: 936, SEQ ID NO: 939, SEQ ID NO: 940, SEQ ID NO: 941, SEQ ID NO: 943, SEQ ID NO: 944, SEQ ID NO: 945, or SEQ ID NO: 946.

[0270] In some implementations, the exemplary PIP-72 polypeptide is any combination of the polypeptides shown in Tables 14, 17, 20, 23, 24, 26, 28 and / or 29, as well as their amino acid substitutions, deletions and / or insertions and fragments.

[0271] In some embodiments, the calculated molecular weight of the PIP-72 peptide is between about 6 kDa and about 13 kDa, between about 7 kDa and about 12 kDa, between about 8 kDa and about 11 kDa, between about 9 kDa and about 10 kDa, about 8.75 kDa, about 9 kDa, about 9.25 kDa, about 9.5 kDa, about 9.75 kDa, about 10 kDa, about 10.25 kDa, and about 10.5 kDa.

[0272] As used herein, the term "about" is used where the molecular weight of the PIP-72 peptide is an average ± 0.25 kDaltons. In some embodiments, the PIP-72 peptide has modified physical properties. As used herein, the term "physical property" refers to any parameter suitable for describing the physicochemical characteristics of a protein. As used herein, "physical property of interest" and "property of interest" are used interchangeably and refer to the physical properties of the protein being studied and / or modified. Examples of physical properties include, but are not limited to, net surface charge and charge distribution on the protein surface, net hydrophobicity and distribution of hydrophobic residues on the protein surface, surface charge density, surface hydrophobicity density, total number of ionizable groups on the surface, surface tension, protein size and its distribution in solution, melting temperature, heat capacity, and second virial coefficient. Examples of physical properties also include, but are not limited to, solubility, foldability, stability, and digestibility. In some embodiments, the PIP-72 peptide enhances the digestibility of the hydrolyzed fragments in the insect gut. Models of digestion by simulating gastric juice are known to those skilled in the art (Fuchs, RL, and JDAstwood). Food Technology 50: 83-88, 1996; Astwood, JD et al. Nature Biotechnology 14: 1269-1273, 1996; Fu TJ et al., J. Agric Food Chem . 50: 7154-7160, 2002).

[0273] In some embodiments, the variants include polypeptides whose amino acid sequences differ due to mutagenesis. The variant proteins covered by this disclosure possess biological activity, meaning they retain the desired biological activity (i.e., insecticidal activity) of the natural protein. In one embodiment, the variant will have at least about 10%, at least about 30%, at least about 50%, at least about 70%, at least about 80% or more of the insecticidal activity of the natural protein. In some embodiments, the variant may have improved activity compared to the natural protein.

[0274] Bacterial genes often have multiple methionine start codons near the start of the open reading frame. Typically, translation initiation at one or more of these start codons leads to the production of a functional protein. These start codons may include the ATG codon. However, bacteria such as Bacillus species also recognize the GTG codon as a start codon, and proteins translated at the GTG codon contain a methionine as their first amino acid. In rare cases, translation in bacterial systems may begin at the TTG codon, although in this case TTG encodes methionine. Furthermore, it is generally not possible to a priori determine which of these codons are naturally used in bacteria. Therefore, it should be understood that using one of the alternative methionine codons can also lead to the production of insecticidal proteins. These insecticidal proteins are covered in this disclosure and can be used in the methods of this disclosure. It should be understood that when expressed in plants, it is necessary to change the alternative start codon to ATG for correct translation.

[0275] In another aspect, the PIP-72 peptide can be expressed as a precursor protein with intercalation sequences that catalyze multi-step post-translational protein splicing. Protein splicing involves the excision of the intercalation sequence from the peptide, accompanied by the joining of flanking sequences, thereby generating a new peptide (Chong et al., (1996)). J. Biol. Chem (Evans et al., 271:22159-22168). This intercalated sequence or protein splicing element is called an intipeptide, which catalyzes its own cleavage at the N-terminal and C-terminal splice junctions through three coordinated reactions: acyl rearrangement of the N-terminal cysteine ​​or serine; transesterification between the two ends to form a branched ester or thioester intermediate; and peptide bond cleavage accompanied by cyclization of the C-terminal asparagine of the intipeptide, thereby releasing the intipeptide (Evans et al., (2000)). J. Biol. Chem ., 275:9091-9094). The elucidation of this protein splicing mechanism has spurred a variety of intepitide-based applications (Comb et al., U.S. Patent 5,496,714; Comb et al., U.S. Patent 5,834,247; Camarero and Muir, (1999)). J. Amer. Chem Soc. 121:5597-5598; Chong et al., (1997) Gene 192:271-281; ​​Chong et al., (1998) Nucleic Acids Res 26:5109-5115; Chong et al., (1998) J. Biol. Chem 273:10567-10577; Cotton et al., (1999) J. Am. Chem. Soc .121:1100-1101; Evans et al., (1999) J. Biol. Chem 274:18359-18363; Evans et al., (1999) J. Biol. Chem 274:3923-3926; Evans et al., (1998) Protein Sci 7:2256-2264; Evans et al., (2000) J. Biol. Chem. 275:9091-9094; Iwai and Pluckthun, (1999) FEBS Lett 459:166-172; Mathys et al., (1999) Gene 231:1-13; Mills et al., (1998) Proc. Natl. Acad. Sci. USA 95:3543-3548; Muir et al., (1998) Proc. Natl. Acad. Sci. USA 95:6705-6710; Otomo et al., (1999) Biochemistry 38:16040-16044; Otomo et al., (1999) J. Biolmol. NMR 14:105-114; Scott et al., (1999) Proc. Natl. Acad. Sci. USA 96:13638-13643; Severinov and Muir, (1998) J. Biol. Chem 273:16205-16209; Shingledecker et al., (1998) Gene 207:187-195; Southworth et al., (1998) EMBO J 17:918-926; Southworth et al., (1999) Biotechniques 27:110-120; Wood et al., (1999) Nat. Biotechnol 17:889-892; Wu et al., (1998a) Proc. Natl. Acad. Sci. USA 95:9226-9231; Wu et al., (1998b) Biochim Biophys Acta 1387:422-432; Xu et al., (1999) Proc. Natl. Acad. Sci. USA 96:388-393; Yamazaki et al., (1998) J. Am. Chem. Soc ., 120:5591-5592). For information on the application of integrins in plant transgenics, see Yang et al., (Transgene Res 15:583-593 (2006)) and Evans et al., ( Annu. Rev. Plant Biol . 56:375-392 (2005)).

[0276] In another aspect, the PIP-72 polypeptide can be encoded by two separate genes, in which the inteins of the precursor protein (called the fragmented inteins) originate from these two genes, and the two parts of the precursor are linked together by peptide bond formation. This peptide bond formation is achieved through intein-mediated trans-splicing. For this purpose, the first and second expression cassettes containing these two separate genes also encode inteins capable of mediating protein trans-splicing. Through trans-splicing, the protein and polypeptide encoded by the first and second fragments are linked together by peptide bond formation. The trans-splicing inteins can be selected from the nucleolar and organelle genomes of various organisms, including eukaryotes, archaea, and bacteria. Usable inteins are listed at neb.com / neb / inteins.html (accessible via the World Wide Web using the "www" prefix). The nucleotide sequence encoding the inteins can be split into 5' and 3' portions encoding the 5' and 3' portions of the inteins, respectively. Sequence portions not required for intein splicing (e.g., homing endonuclease domains) can be deleted. The inpeptide-coding sequence is broken to allow for trans-splicing of the 5' and 3' portions. To select a suitable break site for the inpeptide-coding sequence, considerations can be followed as published by Southworth et al., (1998) EMBO J. 17:918-926. The construction of the first and second expression cassettes is as follows: the 5' inpeptide-coding sequence is ligated to the 3' end of a first fragment encoding the N-terminal portion of the PIP-72 polypeptide, and the 3' inpeptide-coding sequence is ligated to the 5' end of a second fragment encoding the C-terminal portion of the PIP-72 polypeptide.

[0277] Generally, any fragmented inteptide (including any naturally occurring or artificially fragmented fragmented inteptides) can be used to design trans-splicing chaperones. Several naturally occurring fragmented inteptides are known, for example, species of the genus *Syntrophus* (…). Synechocystis sp .) The fragmented inteins of the DnaE gene in PCC6803 (see Wu et al., (1998)). Proc Natl Acad Sci USA . 95(16):9226-31 and Evans et al., (2000) J Biol Chem . 275(13):9091-4), and from Nostoc punctata ( Nostoc punctiforme The fragmented inteins of the DnaE gene (see Iwai et al., (2006) FEBS Lett. 580(7):1853-8). New fragmented inteins have been artificially fragmented in the laboratory to form new fragmented inteins, for example: the artificially fragmented Ssp DnaB intein (see Wu et al., (1998)). Biochim Biophys Acta 1387:422-32), fragmented Sce VMA inteptides (see Brenzel et al., (2006)). Biochemistry . 45(6):1571-8), and artificially fractured fungal microintegral peptides (see, Elleuche et al., (2007)). Biochem Biophys Res Commun 355(3):830-4). There are also databases of inteins that contain known inteins available for use (see, for example, the online database: bioinformatics.weizmann.ac.il / ˜pietro / inteins / Inteinstable.html, accessible on the World Wide Web with the prefix "www").

[0278] Naturally occurring non-fragmented intepids may possess endonuclease or other enzymatic activities, which can typically be removed when designing artificially fractured intepids. Such miniature intepids or minimized fractured intepids are well known in the art and are typically less than 200 amino acid residues in length (see, Wu et al., (1998)). Biochim Biophys Acta (1387:422-32). Suitable fragmented integrins may have other polypeptide elements added to their structure to achieve purification, provided that such elements do not inhibit the splicing of the fragmented integrin or are added in a manner that allows them to be removed before splicing. Protein splicing using proteins containing bacterial integrin-like (BIL) domains has been reported (see, Amitai et al., (2003)). Mol Microbiol. 47:61-73), Hedgehog protein (Hog) self-processing domain (the latter combined with inteins is called the Hog / inteins superfamily or HINT family (see, Dassa et al., (2004)). J Biol Chem (279:32001-7), and such domains can also be used to prepare artificially cleaved intepids. Specifically, non-splicing members of this family can be modified using molecular biology methods to introduce or restore splicing activity in such related substances. Recent studies have shown that splicing can be observed when N-terminal cleaved intepid components are reacted with C-terminal cleaved intepid components that are not naturally occurring as their "partners." For example, splicing has been observed when using partners with as little as 30% to 50% homology to "natural" splicing partners (see, Dassa et al., (2007)). Biochemistry. 46(1):322-30). It has been shown that other such mixtures of dissimilar fragmented integument chaperones do not react with each other (see, Brenzel et al., (2006)). Biochemistry 45(6):1571-8). However, it is within the capabilities of those skilled in the art that it can be determined, using conventional methods and without the need for original techniques, whether a specific pair of polypeptides can be linked together to provide a functional integrin.

[0279] In another respect, the PIP-72 peptide is a variant with a cyclic arrangement. In some embodiments, the PIP-72 peptide is a variant of the cyclic arrangement of the following peptides: SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 18, SEQ ID NO: 28, SEQ ID NO: 32, any one of SEQ ID NO: 528 to SEQ ID NO: 768, any one of SEQ ID NO: 825 to SEQ ID NO: 844, SEQ ID NO: 771, SEQ ID NO: 772, SEQ ID NO: 852, any one of SEQ ID NO: 903 to SEQ ID NO: 914, SEQ ID NO: 927, SEQ ID NO: 928, SEQ ID NO: 932, SEQ ID NO: 933, SEQ ID NO: 934, SEQ ID NO: 935, SEQ ID NO: 936, SEQ ID NO: 939, SEQ ID NO: 940, SEQ ID NO: 941, SEQ ID NO: 943, SEQ ID NO: 944, SEQ ID NO: 945 or SEQ ID NO: 946.

[0280] The development of recombinant DNA methods has enabled the study of the effects of sequence transposition on protein folding, structure, and function. The methods used to form new sequences are similar to those related to the linear reconstruction of naturally occurring protein pairs with their amino acid sequences (Cunningham et al., (1979)). Proc. Natl. Acad. Sci USA 76:3218-3222; Teather and Erfle, (1990) J. Bacteriol 172:3837-3841; Schimming et al., (1992) Eur. J. Biochem .204:13-19; Yamiuchi and Minamikawa, (1991) FEBS Lett 260:127-130; MacGregor et al., (1996) FEBS Lett . 378:263-266). This type of rearrangement was first applied in vitro to proteins such as Goldenenberg and Creighton ( ). J. Mol. BiolAs described in 165:407-413, 1983. In variants forming a circular arrangement, a new N-terminus is selected at an internal site (breakpoint) of the initial sequence, from which the new sequence has the same amino acid sequence as the initial sequence until it reaches an amino acid at or near the initial C-terminus. At this point, the new sequence is directly or via another part of the sequence (linker) connected to the amino acid at or near the initial N-terminus, and the new sequence continues to have the same sequence as the initial sequence until it reaches a point at or near the N-terminus of the initial sequence breakpoint, where the residue forms the new C-terminus of the chain. The amino acid sequence length of the linker can be selected empirically, or under the guidance of structural information, or by using a combination of both methods. When no structural information is available, a small series of linkers can be prepared for testing using a design where the length of the design varies across the range of 0 to 50 Å, and the sequence of the design is selected to conform to surface exposure (hydrophilicity, Hopp and Woods, (1983)). Mol. Immunol 20:483-489; Kyte and Doolittle, (1982) J. Mol. Biol 157:105-132; Solvent-exposed surface areas, Lee and Richards, (1971) J. Mol. Biol. 55:379-400) and the ability to adopt the desired conformation without disturbing the configuration of the insecticidal peptide (conformal flexibility; Karplus and Schulz, (1985)). Naturwissenschaften 72:212-213). Assuming an average translation of 2.0 to 3.8 Å per residue, this means the length to be tested will be between 0 and 30 residues, with 0 to 15 residues being the preferred range. An example of such empirical series would be using cassette sequences such as Gly-Gly-Gly-Ser repeated n times (where n is 1, 2, 3, or 4) to construct linkers. Those skilled in the art will recognize that many such sequences exist that vary in length or composition and can be used as linkers, with the primary consideration being that they are neither too long nor too short (see Sandhu, (1992)). Critical Rev. Biotech(12:437-462); If they are too long, entropy effects may disrupt the stability of three-dimensional folding and may also make folding kinetically impractical, and if they are too short, they may disrupt molecular stability due to torsion or spatial strain. Technicians in protein structure information analysis will recognize that the distance between chain ends (defined as the distance between c-α carbons) can be used to define the length of the sequence to be used, or at least to limit the number of possibilities that must be tested in empirical selection of linkers. They will also recognize that sometimes the position of the polypeptide chain ends is not clear in the structural model derived from X-ray diffraction or NMR spectroscopy data, and when this is the case, this situation must therefore be taken into account in order to correctly estimate the required linker length. From those residues whose positions are clear, two residues adjacent to the chain ends in the sequence are selected, and the approximate length of the linker between them is calculated using the distance between their c-α carbons. The calculated length is then used as a guide to select linkers with a range of residue numbers (calculated using 2 to 3.8 Å per residue). These linkers can be composed of an initial sequence, which can be shortened or lengthened as needed, and when lengthened, additional residues can be selected as flexible and hydrophilic as described above; alternatively, a series of linkers can be used to replace the initial sequence, an example being the Gly-Gly-Gly-Ser box method described above; or alternatively, a combination of the initial sequence and a new sequence with an appropriate total length can be used. Sequences of insecticidal peptides capable of folding into a biologically active state can be prepared by appropriately selecting start (amino terminus) and end (carboxyl terminus) positions from within the initial polypeptide chain, while using the linker sequences described above. The amino and carboxyl terms are selected from a common segment of the sequence (called the breakpoint region) using the guidelines below. Thus, by selecting the amino and carboxyl terms from the same breakpoint region, novel amino acid sequences are generated. In many cases, the selection of the new terminus will result in the initial position of the carboxyl terminus immediately preceding the initial position of the amino terminus. However, those skilled in the art will recognize that selecting the terminus at any position within this region can be effective, and these will effectively cause the deletion or addition of the amino or carboxyl portion of the new sequence. A core principle of molecular biology is that the primary amino acid sequence of a protein determines its folding into the three-dimensional structure required to express its biological function. Methods for obtaining and interpreting three-dimensional structural information using X-ray diffraction of a single protein crystal or nuclear magnetic resonance spectroscopy of a protein solution are known to those skilled in the art.Examples of structural information associated with breakpoint identification include the location and type of protein secondary structures (α-helices and 3-10 helices, parallel and antiparallel β-sheets, chain inversions and turns, and loops; Kabsch and Sander, (1983) Biopolymers 22:2577-2637); the degree of solvent exposure of amino acid residues, the degree and type of interaction between residues (Chothia, (1984) Ann. Rev. Biochem. 53:537-572); and the static and dynamic conformational distribution along the polypeptide chain (Alber and Mathews, (1987)). Methods Enzymol . 154:511-533). In some cases, additional information regarding residue solvent exposure is known; an example is the need for post-translational linking sites of carbohydrates on the protein surface. When experimental structural information is unavailable or impossible to obtain, various methods can be used to analyze the primary amino acid sequence to predict protein tertiary and secondary structures, solvent accessibility, and the presence of turns and loops. When direct structural methods are not feasible, biochemical methods can sometimes be applied empirically to determine surface exposure. For example, the identification of chain break sites after restriction protease digestion can be used to infer surface exposure (Gentile and Salvatore, (1993) Eur. J. Biochem. 218:603-621). Therefore, experimentally derived structural information or predictive methods (e.g., Srinivisan and Rose, (1995)) can be used. Proteins: Struct., Funct . &Genetics (22:81-99) Examine the parental amino acid sequences to classify regions based on their indispensability for maintaining secondary and tertiary structures. Sequences appearing in regions known to involve periodic secondary structures (α-helices and 3-10 helices, parallel and antiparallel β-sheets) are regions to be avoided. Similarly, regions observed or predicted to have amino acid sequences with low solvent exposure are more likely to be the so-called hydrophobic core of the protein and should also be avoided when selecting amino and carboxyl terms. In contrast, those regions known or predicted to be in surface turns or loops, especially those not known to be required for biological activity, are preferred sites for polypeptide chain terminal localization. The continuous segments of amino acid sequences preferred based on the above criteria are called breakpoint regions. Polynucleotides encoding a cyclically arranged PIP-72 polypeptide, having a novel N-terminus / C-terminus containing a linker region separating the initial C-terminus and N-terminus, can be prepared essentially according to the methods described in the following literature: Mullins et al., (1994). J. Am. Chem. Soc.116:5529-5533. The DNA sequence encoding the primary amino acid sequence of a protein is rearranged using multiple steps of polymerase chain reaction (PCR) amplification. A polynucleotide encoding a circularly arranged PIP-72 polypeptide having a novel N-terminus / C-terminus containing a linker region separating the initial C-terminus and N-terminus can be prepared based on the tandem repeat method described in the following literature: Horlick et al., (1992). Protein Eng 5:427-431. Polymerase chain reaction (PCR) amplification of novel N-terminal / C-terminal genes using template DNA with tandem repeats.

[0281] In another aspect, a fusion protein is provided, the fusion protein comprising, within its amino acid sequence, an amino acid sequence constituting a PIP-72 polypeptide, said PIP-72 polypeptide including, but not limited to, any one of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 18, SEQ ID NO: 28, SEQ ID NO: 32, any one of SEQ ID NO: 528 to SEQ ID NO: 768, any one...

Claims

1. A DNA construct comprising a heterologous nucleic acid molecule, said heterologous nucleic acid molecule encoding a target for the western maize rootworm (Maize Rootworm). Diabrotica virgifera virgifera PIP-72 polypeptide with insecticidal activity.

2. The DNA construct according to claim 1, wherein the encoded PIP-72 polypeptide comprises an amino acid sequence having at least 50% identity with SEQ ID NO:

2.

3. The DNA construct according to claim 1 or 2, wherein the encoded PIP-72 polypeptide, compared with the corresponding amino acid shown in SEQ ID NO: 2, is present at amino acid positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 42, 44, 45. Positions 46, 47, 48, 49, 50, 51, 52, 53, 54, 56, 57, 58, 59, 60, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, or 86 contain 1 to 45 amino acid substitutions.

4. The DNA construct according to claim 1 or 2, wherein the encoded PIP-72 polypeptide is associated with SEQ ID NO: Compared to the corresponding amino acids shown in Figure 2, the amino acids include 1 to 45 amino acid substitutions at positions 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 17, 18, 19, 20, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 42, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 56, 58, 60, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, or 86.

5. The DNA construct according to claim 1, 2, or 4, wherein the encoded PIP-72 polypeptide comprises the amino acid sequence shown in the following formula: in The second Xaa is Gly, Ala, Cys, Asp, Glu, Ile, Lys, Leu, Asn, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 3 is either Ile or Trp; The fourth Xaa is Thr, Ala, Asp, Glu, His, Ile, Lys, Leu, Arg, Ser, Val, Trp, or Tyr; The Xaa in the 5th position is Val, Ala, Cys, Gly, His, Ile, or Tyr; The Xaa at position 6 is Thr, Ala, Cys, Phe, Gly, His, Ile, Lys, Met, Pro, Gln, Arg, Ser, Trp, or Tyr; Xaa at position 7 is either Asn, Ala, or Val; The Xaa at position 8 is Asn, Ala, Cys, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Gln, Arg, Ser, Thr, or Val; The Xaa at position 9 is either Ser, Ala, Cys, Gly, or Thr; The Xaa at the 10th position is Ser, Ala, Glu, Phe, Gly, His, Ile, Lys, Leu, Asn, Pro, Gln, Arg, Thr, or Trp; The Xaa at position 11 is Asn, Ala, Cys, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Gln, Ser, Thr, Val, or Tyr; The Xaa at position 12 is Pro, Ala, Cys, Asp, Glu, Gly, His, Lys, Leu, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 13 is Ile, Asn, Gln, or Val; Xaa at position 14 is Glu, Ala, Cys, Phe, His, Lys, or Gln; Xaa at position 15 is Val, Ala, Cys, Ile, Met, or Arg; Xaa at position 17 is Ile, Glu, or Val; Xaa at position 18 is either Asn or Ser; The Xaa at position 19 is His, Ala, Glu, Lys, Leu, Pro, Arg, Ser, or Tyr; Xaa at position 20 is Trp, Ala, or Thr; Xaa at position 22 is Ser, Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Thr, Val, or Tyr; Xaa at position 23 is Asp, Ala, Gly, His, Lys, Met, Asn, Gln, Ser, Thr, or Val; Xaa at position 24 is Gly, Asp, or Phe; Xaa at position 25 is Asp, Ala, Glu, Phe, Asn, or Gln; Xaa at position 26 is Thr, Glu, or Pro; Xaa at position 27 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Asn, Gln, Arg, or Thr; Xaa at position 28 is Phe, Pro, Trp, or Tyr; Xaa at position 29 is Phe, Ala, Cys, Ile, Leu, Gln, Arg, Trp, or Tyr; The Xaa at position 30 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Lys, Leu, Met, Asn, Pro, Gln, Arg, Thr, Val, Trp, or Tyr; Xaa at position 31 is Val, Ile, or Leu; Xaa at position 32 is Gly, Ala, Asp, Glu, Phe, His, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 33 is Asn, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Pro, Gln, Arg, Ser, Thr, Val, or Tyr; Xaa at position 34 is Gly, Glu, Phe, His, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Tyr; Xaa at position 35 is Lys, Ala, Cys, Asp, Gly, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Val; Xaa at position 36 is Gln, Ala, Cys, Glu, Gly, His, Ile, Lys, Leu, Asn, Pro, Arg, Ser, Thr, or Val; Xaa at position 37 is Glu, Ala, Cys, Asp, Phe, Gly, Ile, Lys, Leu, Met, Asn, Ser, Thr, or Val; Xaa at position 38 is Thr, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Val, Trp, or Tyr; Xaa at position 39 is either Trp or Phe; The Xaa at position 40 is Asp, Ala, Cys, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 42 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, Ile, Lys, Leu, Met, Asn, Gln, Arg, Thr, Val, Trp, or Tyr; Xaa at position 44 is Ser, Ala, Asp, Glu, Gly, Leu, Met, Asn, Pro, Gln, Thr, Val, or Tyr; Xaa at position 45 is Arg, Lys, or Ser; Xaa at position 46 is Gly, Ala, or Gln; Xaa at position 47 is Phe, Cys, Val, or Tyr; Xaa at position 48 is Val, Ile, or Leu; Xaa at position 49 is Leu, Cys, Phe, Met, Arg, or Tyr; Xaa at position 50 is Ser, Ala, Cys, Asp, Ile, Met, Pro, Gln, Thr, or Val; Xaa at position 51 is Leu, Ala, Cys, Met, or Val; Xaa at position 52 is Lys, Cys, Phe, His, Ile, Leu, Met, Asn, Arg, Ser, Thr, Trp, or Tyr; Xaa at position 53 is Lys, Ala, Cys, Asp, Glu, Phe, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, or Tyr; Xaa at position 54 is Asn, Cys, Asp, Glu, Phe, Gly, Lys, Met, Gln, Arg, Ser, or Trp; Xaa at position 56 is Ala, Gly, Leu, Asn, Pro, Gln, Arg, Ser, or Thr; Xaa at position 57 is Gln, Glu, Leu, Met, Ser, or Thr; Xaa at position 58 is His, Ala, Asp, Phe, Leu, Met, Asn, Arg, Trp, or Tyr; Xaa at position 60 is Tyr, Glu, or Phe; Xaa at position 63 is Gln, Cys, Gly, Ile, Leu, Met, Asn, Thr, Val, or Tyr; Xaa at position 64 is Ala, Phe, Gly, His, Arg, Ser, or Tyr; Xaa at position 65 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Leu, Asn, Thr, or Val; Xaa at position 66 is Ser, Ala, or Gly; Xaa at position 67 is Lys, Ala, Cys, Asp, Phe, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 68 is Ile, Asp, Leu, or Val; Xaa at position 69 is Glu, Ala, Cys, Asp, Phe, His, Ile, Leu, Met, Gln, Arg, Ser, Thr, Val, or Tyr; Xaa at position 70 is Val, Cys, or Ile; Xaa at position 71 is Asp, Ala, Cys, Gly, His, Ile, Leu, Met, Asn, Ser, Thr, Val, or Tyr; Xaa at position 72 is Asn, Ala, Cys, Asp, Glu, Gly, Lys, Met, Pro, Gln, Arg, Ser, Thr, Val, or Trp; Xaa at position 73 is Asn, Ala, Cys, Asp, Phe, Gly, His, Ile, Leu, Ser, Thr, Val, or Tyr; Xaa at position 74 is Ala, Cys, Asp, Phe, Gly, His, Ile, Leu, Asn, Gln, Arg, Ser, Thr, Val, or Tyr; Xaa at position 75 is Val, Cys, Ile, or Leu; Xaa at position 76 is Lys, Ala, Cys, Phe, His, Ile, Leu, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 77 is Asp or Tyr; Xaa at position 78 is Gln, Ala, Cys, Asp, Phe, Gly, His, Ile, Leu, Met, Asn, Arg, Ser, Thr, Val, or Tyr; Xaa at position 79 is Gly, Arg, Ala, Cys, Asp, Glu, Phe, His, Lys, Leu, Asn, Gln, Arg, Ser, Thr, Trp, or Tyr; Xaa at position 80 is Arg, Ala, Cys, Asp, Phe, Gly, His, Ile, Leu, Asn, Ser, Thr, Val, or Tyr; Xaa at position 81 is Leu, Ala, Cys, Asp, Phe, Gly, His, Ile, Asn, Pro, Arg, Ser, Thr, or Val; Xaa at position 82 is Ile, Ala, Leu, Met, Arg, or Val; Xaa at position 83 is Glu, Ala, Cys, Asp, Phe, Gly, His, Ile, Lys, Leu, Asn, Pro, Arg, Ser, Thr, Val, or Tyr; Xaa at position 84 is Pro, Ala, Cys, Glu, Ile, Ser, Val, Trp, or Tyr; Xaa at position 85 is Leu, Cys, Gly, or Val; and Xaa at position 86 is either Ser, Ala, Ile, Thr, or Val.

6. The DNA construct according to claim 1, 2 or 3, wherein the encoded PIP-72 polypeptide comprises the amino acid sequence shown in the following formula: in The second Xaa is Gly, Ala, Cys, Asp, Glu, Ile, Lys, Leu, Asn, Arg, Ser, Thr, Val, Trp, or Tyr; The Xaa in the third position is Ile, Leu, Val, or Trp; The fourth Xaa is Thr, Ala, Asp, Glu, His, Ile, Lys, Leu, Arg, Ser, Val, Trp, or Tyr; The Xaa in the 5th position is Val, Ala, Cys, Gly, His, Ile, Leu, or Tyr; The Xaa at position 6 is Thr, Ala, Cys, Phe, Gly, His, Ile, Lys, Met, Pro, Gln, Arg, Ser, Trp, or Tyr; Xaa at position 7 is either Asn, Ala, or Val; The Xaa at position 8 is Asn, Lys, Gly, Ser, Gln, Arg, Thr, Ala, Cys, Asp, Glu, His, Ile, Leu, Met, or Val; The Xaa at position 9 is either Ser, Ala, Cys, Gly, or Thr; The Xaa at position 11 is Asn, Lys, Thr, Gln, Arg, Ser, Ala, Cys, Asp, Glu, Gly, His, Ile, Leu, Met, Val, or Tyr; The Xaa at position 12 is Pro, Thr, Lys, Ser, Arg, Ala, Cys, Asp, Glu, Gly, His, Leu, Asn, Gln, Arg, Val, Trp, or Tyr; Xaa at position 13 is Ile, Asn, Gln, Leu, or Val; Xaa at position 14 is Glu, Ala, Cys, Phe, His, Lys, Asp, or Gln; The Xaa at position 15 is Val, Ala, Ile, Leu, Cys, Met, or Arg; Xaa at position 16 is either Ala or Ser; Xaa at position 17 is Ile, Glu, Leu, or Val; Xaa at position 18 is Asn, Gln, Thr, or Ser; The Xaa at position 19 is His, Lys, Ala, Arg, Glu, Leu, Pro, Ser, or Tyr; Xaa at position 20 is Trp, Ala, or Thr; Xaa at position 21 is Gly, Arg, or Lys; Xaa at position 22 is Ser, Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Thr, Val, or Tyr; Xaa at position 23 is Asp, Ala, Gly, His, Lys, Met, Asn, Gln, Ser, Thr, or Val; Xaa at position 24 is Gly, Asp, or Phe; Xaa at position 25 is Asp, Ala, Glu, Phe, Asn, or Gln; Xaa at position 26 is Thr, Glu, Asp, Ser, or Pro; Xaa at position 27 is Ser, Thr, Lys, Arg, Ala, Cys, Asp, Glu, Phe, Gly, His, Asn, or Gln; Xaa at position 28 is Phe, Tyr, Pro, or Trp; Xaa at position 29 is Phe, Ala, Cys, Ile, Leu, Gln, Arg, Trp, or Tyr; The Xaa at position 30 is Ser, Gly, Lys, Thr, Arg, Ala, Cys, Asp, Glu, Phe, His, Leu, Met, Asn, Pro, Gln, Val, Trp, or Tyr; Xaa at position 31 is Val, Ile, Met, or Leu; Xaa at position 32 is Gly, Ala, Asp, Glu, Phe, His, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 33 is Asn, Ser, Gln, Pro, Thr, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Arg, Val, or Tyr; Xaa at position 34 is Gly, Glu, Phe, His, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Tyr; Xaa at position 35 is Lys, Glu, Ala, Cys, Asp, Gly, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Val; Xaa at position 36 is Gln, Ala, Cys, Glu, Gly, His, Ile, Lys, Leu, Asn, Pro, Arg, Ser, Thr, or Val; Xaa at position 37 is Glu, Asp, Ala, Cys, Phe, Gly, Ile, Lys, Leu, Met, Asn, Ser, Thr, or Val; Xaa at position 38 is Thr, Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Gln, Arg, Val, Trp, or Tyr; Xaa at position 39 is either Trp or Phe; The Xaa at position 40 is Asp, Ala, Cys, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 42 is Ser, Asn, Thr, Ala, Cys, Asp, Glu, Phe, Gly, Ile, Lys, Leu, Met, Arg, Val, Trp, Tyr, or Gln; Xaa at position 44 is Ser, Asp, Ala, Leu, Thr, Glu, Ile, Ala, Gly, Leu, Met, Asn, Pro, Gln, Val, Tyr, or Val; Xaa at position 45 is Arg, Lys, or Ser; Xaa at position 46 is Gly, Ala, or Gln; Xaa at position 47 is Phe, Tyr, Cys, Val, or Trp; Xaa at position 48 is Leu, Met, Ile, Cys, Phe, Met, Arg, Tyr, or Val; Xaa at position 49 is Leu, Met, Ile, or Val; Xaa at position 50 is Ser, Ala, Tyr, Cys, Asp, Ile, Met, Pro, Gln, Val, or Thr; Xaa at position 51 is Leu, Val, Ala, Cys, Met, or Ile; Xaa at position 52 is Lys, Cys, Phe, His, Ile, Leu, Met, Asn, Arg, Ser, Thr, Gln, Trp, or Tyr; Xaa at position 53 is Lys, Arg, Met, Leu, Ile, Ala, Cys, Asp, Glu, Phe, His, Asn, Gln, Ser, Thr, Tyr, or Val; Xaa at position 54 is Asn, Cys, Asp, Glu, Phe, Gly, Lys, Met, Gln, Arg, Ser, or Trp; Xaa at position 55 is Gly, Ser, or Thr; Xaa at position 56 is Ala, Thr, Gln, Ser, Gly, Leu, Pro, Arg, or Asn; Xaa at position 57 is Gln, Glu, Leu, Met, Ser, Val, Ala, Asn, Ile, or Thr; Xaa at position 58 is His, Ala, Lys, Asp, Phe, Leu, Met, Asn, Arg, Trp, Tyr, or Thr; Xaa at position 59 is Pro, Thr, or Ser; Xaa at position 60 is Tyr, Glu, or Phe; Xaa at position 62 is Val, Ile, or Leu; Xaa at position 63 is Gln, Ser, Cys, Gly, Ile, Leu, Met, Asn, Thr, Val, or Tyr; Xaa at position 64 is Ala, Gln, Asn, Phe, Gly, His, Arg, Ser, or Tyr; Xaa at position 65 is Ser, Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Leu, Asn, Val, or Thr; Xaa at position 66 is Ser, Ala, or Gly; Xaa at position 67 is Lys, Gln, Asn, or Arg; Xaa at position 67 is Lys, Ala, Cys, Asp, Phe, His, Ile, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 68 is Ile, Asp, Leu, or Val; Xaa at position 69 is Glu, Ala, Cys, Asp, Phe, His, Ile, Leu, Met, Gln, Arg, Ser, Thr, Val, or Tyr; Xaa at position 70 is Val, Ile, Cys, or Leu; Xaa at position 71 is Asp, Glu, Tyr, Ala, Cys, Gly, His, Ile, Leu, Met, Asn, Ser, Thr, Val, or Trp; Xaa at position 72 is Asn, Ala, Cys, Asp, Glu, Gly, Lys, Met, Pro, Gln, Arg, Ser, Thr, Val, His, or Trp; Xaa at position 73 is Asn, Ser, Asp, Gln, Thr, Ala, Cys, Phe, Gly, His, Ile, Leu, Val, Tyr, or Glu; Xaa at position 74 is Ala, Thr, Met, Ile, Lys, Ser, Leu, Val, Cys, Asp, Phe, Gly, His, Asn, Gln, Tyr, or Arg; Xaa at position 75 is Val, Cys, Ile, or Leu; Xaa at position 76 is Lys, Ala, Cys, Phe, His, Ile, Leu, Gln, Arg, Ser, Thr, Val, Trp, or Tyr; Xaa at position 77 is Asp or Tyr; Xaa at position 78 is Gln, His, Ser, Asn, Ala, Cys, Asp, Phe, Gly, Ile, Leu, Met, Asn, Arg, Val, Tyr, or Thr; Xaa at position 79 is Gly, Arg, Ala, Cys, Asp, Glu, Phe, His, Lys, Leu, Asn, Gln, Arg, Ser, Thr, Trp, or Tyr; Xaa at position 80 is Arg, Glu, Gln, Lys, Asp, Ala, Cys, Phe, Gly, His, Ile, Leu, Ser, Thr, Val, Tyr, or Asn; Xaa at position 81 is Leu, Pro, Thr, Ile, Val, Ala, Cys, Asp, Phe, Gly, His, or Ser; Xaa at position 82 is Ile, Ala, Leu, Met, Arg, and Val; Xaa at position 83 is Glu, His, Asn, Leu, Gln, Ile, Ala, Cys, Asp, Phe, Gly, Lys, Pro, Arg, Ser, Thr, Tyr, or Val; Xaa at position 84 is Pro, Ala, Cys, Glu, Ile, Ser, Val, Trp, or Tyr; Xaa at position 85 is Leu, Val, Cys, Gly, or Ala; and Xaa at position 86 is either Ser, Ala, Tyr, Asn, Ile, Val, or Thr.

7. The DNA construct according to any one of claims 1, 2, 3 and 4, wherein the encoded PIP-72 polypeptide comprises an amino acid motif represented by positions 37 to 51 of the following sequences: SEQ ID NO: 846, SEQ ID NO: 847, SEQ ID NO: 848 or SEQ ID NO:

849.

8. A DNA construct comprising: A polynucleotide encoding a polypeptide having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher sequence identity with the following amino acid sequences: SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 30, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 929, SEQ ID NO: 930, SEQ ID NO: 931, SEQ ID NO: 937, SEQ ID NO: 938, SEQ ID NO: 942, SEQ ID NO: 947 or SEQ ID NO: 948; and A heterologous regulatory sequence, wherein the polynucleotide is operatively linked to the heterologous regulatory sequence.

9. A DNA construct comprising a chimeric nucleic acid molecule encoding a chimeric PIP-72 polypeptide, the chimeric PIP-72 polypeptide comprising at least a first portion and a second portion, the first portion comprising a first PIP-72 polypeptide moiety, the second portion comprising a complementary second PIP-72 polypeptide moiety, wherein the first PIP-72 polypeptide and the second PIP-72 polypeptide have different amino acid sequences in the respective portions.

10. An isolated polynucleotide comprising a nucleic acid molecule, said nucleic acid molecule encoding a receptor for the western maize rootworm (…). Diabrotica virgifera virgifera PIP-72 polypeptide with insecticidal activity.

Images