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Peach aphid odorant binding protein and application thereof

A protein and green peach aphid technology, which can be used in instruments, measuring devices, scientific instruments, etc., can solve the problems of difficult to effectively use natural enemy insects to biologically control peach aphid and environmental pollution.

Active Publication Date: 2020-12-18
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The extensive use of chemical pesticides not only leads to aphid resistance, but also causes serious pollution to the environment. Therefore, it is necessary to develop a new effective and environmentally friendly sustainable development control method
[0003] In addition, when aphids are threatened by the outside world (such as predation by natural enemies), aphids can fall on the ground or escape through communication, which makes it difficult to effectively use natural enemies for biological control of peach aphids.

Method used

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  • Peach aphid odorant binding protein and application thereof
  • Peach aphid odorant binding protein and application thereof
  • Peach aphid odorant binding protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Construction of cDNA library

[0037] 1.1 Extraction of RNA from the apterous peach aphid, reverse transcription and synthesis of first-strand cDNA and ds cDNA

[0038] Get about 2000 green peach aphid adult tentacles, head, feet and body, utilize the Trizol method to carry out RNA extraction to these four parts respectively, electrophoresis detects RNA quality (see figure 1 ), and the concentrations of RNA measured by spectrophotometer were 172.5ng / μl, 244.9ng / μl, 145.6ng / μl, 2021.8ng / μl respectively. mRNA was isolated from total RNA using oligo(dT) magnetic beads and fragmented by incubating at 94°C for 5 minutes. Then use six random bases as random primers to synthesize the first-strand cDNA from the RNA obtained from the four parts of antennae, head, foot and body under the action of reverse transcriptase M-MuLV Reverse Transcriptase, and then use The first-strand cDNA of each part was used as a template, and dNTPs, DNA polymerase I and RNase H were added to quick...

Embodiment 2

[0053] Cloning, expression and purification of proteins

[0054] 2.1 Obtaining target genes and proteins from cDNA library

[0055] The constructed cDNA libraries of antennae, head, feet and body were sent to Beijing Thermo Lily Biotechnology Co., Ltd. for sequencing.

[0056] For the original sequencing sequences (Sequenced Reads or raw reads) obtained by sequencing, which contain adapters and low-quality reads, in order to ensure the quality of information analysis, the raw reads are filtered to obtain clean reads. Use Trinity software to assemble clean reads to generate unigenes. After assembly, all unigenes were compared with BLASTx and BLASTn in the NCBI database nr and nt, and those with E-value<1.0×10-5 were retained. In this application, a total of 110,059,204 raw reads were obtained. After cleaning adapter sequences and low-quality sequences, 107,199,360 cleanreads were obtained. After assembly, 50,352 unigenes were generated, ranging in length from 201bp to 27.17kb...

Embodiment 3

[0091] Fluorescence competition experiment

[0092] The principle of the fluorescence competition binding experiment is: after the fluorescent probe is combined with the odorant binding protein, it will emit fluorescence under the action of excitation light. At this time, a fluorescence value will be detected when the machine is used for detection. When the odorant compound is added, if the odorant compound Combined with protein, the odor compound will compete with the fluorescent probe, and the odor compound will bind to the protein. At this time, the detected fluorescence value will decrease. As the amount of the added odor compound increases, the fluorescence value will gradually decrease.

[0093] Carried out on F-380 fluorescence spectrophotometer (Tianjin Gangdong), the container used is a quartz cup with a width of 1 cm, a slit width of 10 nm, and a sensitivity of 2. In addition to the above-mentioned purified protein 3, protein 7 and protein 9, the reagents used also i...

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Abstract

The invention relates to an odorant-binding protein of Myzus persicae and an application of the odorant-binding protein. Amino acid sequence is at least one selected from amino acid sequences shown asSEQ ID No.2, SEQ ID No.4 and SEQ ID No.6.

Description

technical field [0001] The present application relates to an amino acid sequence and nucleotide sequence derived from peach aphid, in particular to the application of the amino acid sequence and nucleotide sequence in identifying odor compounds. Background technique [0002] The green peach aphid (Myzus persicae (Sulzer)), belonging to the Hemiptera Aphididae, is a destructive polyphagous pest. They can use more than 400 kinds of plants for asexual reproduction (Van Emden HF, Eastop VF, HughesRD, et al. The ecology of Myzus persicae. Annu. Rev. Entomol., 1969, 14:197-270), not only directly feed on plants sap, and Myzus persicae can transmit more than 100 viral diseases, such as potato leaf roll virus (RLRV) (Eskandari F, Sylvester ES and Richardson J. Evidence for lack of propagation of potato leaf roll virus in its aphid vector, Myzus persicae. Phytopathology, 1979,69:45-47), cucumber mosaic virus (CMV) (Bwye AM, Proudlove W, Berlandier FA, et al.Effects of applying insec...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11
CPCG01N21/6486
Inventor 谷少华王倩李显春
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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