14 results about "Cry1Ac" patented technology

The invention relates to a method for cultivating insect-resistant transgenic Xiaoblack poplar, in particular to a method for cultivating insect-resistant transgenic Xiaoblack poplar. The invention aims to solve the problem of low insect resistance of the populus similis. The cultivation method comprises the following steps: 1, synthesizing a Cry1Ac gene, wherein the nucleotide sequence of the gene is shown as Seq ID No: 1 in a sequence table; 2, a plant expression vector pBI121-Cry1Ac expressed by the Cry1Ac gene is constructed, and the plant expression vector pBI121-Cry1Ac is constructed; 3, preparing an infection solution; 4, cultivating the Cry1Ac-transgenic small black poplar by using an agrobacterium-mediated leaf disc transformation method, the Cry1Ac-transgenic poplar is created, the average concentration of leaf Cry1Ac protein is more than 200ng / g, the insect resistance of the transgenic poplar can be improved, all fall webworms die after being fed for 5 days, and the corrected death rate reaches 100%. The method is applied to the field of molecular breeding.

The invention discloses an insect-resistant transgenic cotton event BrsC35 as well as a detection method and application thereof. According to the invention, an epsps gene, a Cry1Ac gene and a Vip3A gene are transferred into a genome of Zhongmiansui 24 through an agrobacterium-mediated method, so that a transgenic cotton event BrsC35 of which the resistance to lepidoptera insects such as cotton bollworms, ostrinia nubilalis and spodoptera frugiperda is remarkably improved is obtained. The transgenic cotton event BrsC35 is preserved in China General Microbiological Culture Collection Center (CGMCC) on February 17, 2025, and the preservation number is CGMCC No.32181. The transgenic cotton event BrsC35 has been preserved in China General Microbiological Culture Collection Center (CGMCC). The invention also provides a method for identifying whether a plant sample is derived from the transgenic cotton event BrsC35 or an offspring of the transgenic cotton event BrsC35, the transgenic cotton event BrsC35 can be specifically detected, and the transgenic cotton event BrsC35 can be better supervised and managed. The method has an important application value.

The present invention belongs to the field of agricultural pest control and relates to a strain of Enterobacter asburiae Plutella xylostella (PxGF1) and its applications. The strain is classified and named Enterobacter asburiae PxGF1. The strain was deposited with the China Center for Type Culture Collection on January 10, 2023, with a deposit number of CCTCC NO: M2023056 at Wuhan University, Wuhan. Experiments have demonstrated that the strain PxGF1 of the present invention can improve the efficiency of Cry1Ac in killing Plutella xylostella. Furthermore, compared to feeding Plutella xylostella only with Cry1Ac protoxin, the pupation rate and pupal weight of the test insects were significantly reduced, and the intestinal tissue was severely damaged, making it impossible to observe intact intestinal epithelial cells. This suggests that PxGF1 promotes the killing ability of Cry1Ac against Plutella xylostella.

The invention relates to an in-situ binding method for evaluating binding of Bt toxin Cry1Ac and midgut epithelial cells of cotton bollworm larvae. The method comprises the following steps: incubating and culturing midgut tissues of larvae of lepidoptera pests and a Bt toxin-containing Sf-900 II cell culture solution, carrying out immunostaining, observing and photographing through a laser confocal microscope, and finally carrying out quantitative processing on a fluorescence image by utilizing ImageJ software so as to calculate an average unit area fluorescence signal value. And further evaluating the difference between the binding amounts of the midgut epithelial cells and the Bt toxins of the larvae of different strains. The method can intuitively and accurately observe and evaluate the binding condition of the Bt toxin and the midgut epithelial cells on the premise of retaining the original structure and physiological environment of the midgut tissue of the larvae, provides direct data support for Bt resistance research, assists in analyzing the pest Bt resistance mechanism, has important significance in formulating resistance early warning and treatment strategies, and has a wide application prospect. The method is suitable for the biotechnical fields of Bt toxin insecticidal mechanism research, lepidoptera pest drug resistance mechanism exploration and the like.

The invention provides three Cry1Ac modified toxin proteins with high insecticidal activity and application of the Cry1Ac modified toxin proteins, and relates to the field of genetic engineering and biological control. The amino acid sequences of the three modified proteins are sequentially shown as SEQ ID NO: 1, SEQ ID NO: 3 and SEQ ID NO: 5, and the amino acid sequences are sequentially shown as SEQ ID NO: 2, SEQ ID NO: 4 and SEQ ID NO: 6. The insect-resistant protein is a Cry1Ac toxin modified protein with improved insecticidal activity, which is screened from a Cry1Ac toxin key region saturation mutation library targeting Asiatic corn borer cadherin and ABC transporter, has higher insecticidal activity on lepidoptera pests, can be used for preventing and controlling agricultural pests and resisting the Bt resistance problem, and has important scientific and practical significance.

The present invention belongs to the field of biotechnology, and discloses a highly active Cry1Ac mutant and its application. The present invention uses site-directed mutagenesis technology to carry out site-directed mutagenesis on key amino acids, and constructs a series of Cry1Ac mutants. The phenylalanine at position 371 on the amino acid sequence of the Cry1Ac protein is mutated into glutamine, the asparagine at position 372 is mutated into glutamic acid, the isoleucine at position 373 is mutated into glutamine, or the isoleucine at position 375 is mutated into glutamine. The mutant protein obtained is 4.5, 2.7, 6.3 times and 3.4 times more toxic to the diamondback moth than the wild type. The present invention provides theoretical guidance for the transformation of Cry toxins and provides new ideas for insect resistance management.

The invention relates to the technical field of biological prevention and control of agricultural pests, and discloses application of dsRNA (dsRNA) of a calmodulin gene or a calmodulin inhibitor in cotton bollworm Bt protein resistance treatment. According to the invention, dsRNA is utilized to interfere with the calmodulin gene of the resistant cotton bollworm or a calmodulin inhibitor is used to feed Cry1Ac resistant line cotton bollworm, so that the activity of calmodulin in the midgut of the resistant cotton bollworm is obviously reduced, the sensitivity to Bt insecticidal protein is obviously improved, and an obvious resistance treatment effect is achieved. The method provided by the invention can be used for improving the insecticidal effect of Cry1Ac on resistant cotton bollworms, so that the sensitivity of Cry1Ac to Cry1Ac is recovered, and the effect of resistance treatment is achieved.

The present application relates to the technical field of pest control, in particular to a short peptide simulating a Cry1Ac protein anti-insect function epitope and application thereof. The present application takes a cotton bollworm middle intestine cadherin fragment (HaCad-TBR) as a target, and carries out screening on a phage display random 6-peptide library constructed by the laboratory, to obtain a 6-peptide LTETGK capable of combining with the cotton bollworm middle intestine cadherin, i.e. the 6-peptide has the effect of simulating a key anti-insect function epitope of the Cry1Ac protein, and a recombinant insecticidal protein C8CL2 having good insecticidal activity on the cotton bollworm can be obtained by replacing Loop2 of Domain II in the Cry1Ac. The present application expands the diversity of anti-insect materials of the Cry1Ac protein, and provides a potential alternative new material for preventing and controlling target pests, and can be used for the prevention and control of Lepidoptera pests, and provides technical support for the development of Lepidoptera pest control products.

The present invention relates to a chimeric insecticidal protein and a use thereof. The protein comprises domain I and domain II of Cry1Ac protein and domain III of Cry1Igg protein which are sequentially linked, and exhibits strong weight-inhibitory activity and lethal activity against various lepidopteran pests. The provided chimeric insecticidal protein broadens the insecticidal spectrum of the original protein, and also enhances toxicity against pests, effectively slowing the development of pest resistance in nature, and has good application prospects in agricultural production.

The present application relates to the technical field of pest control, in particular to a 7-peptide simulating a functional epitope of Cry1Ac protein and application thereof. The amino acid sequence of the 7-peptide is shown as SEQ ID NO. 1. The 7-peptide A42 capable of binding to HaCad-TBR is obtained by bio-panning of a Ph.D.-7 phage library (New England Biolabs) with HaCad-TBR as a target protein, and the 7-peptide A42 is used to replace part of the amino acids of Cry1Ac toxin to obtain a recombinant insecticidal protein A42D2L2 having good insecticidal activity against cotton bollworms. The present application enriches the Cry1Ac protein insect-resistant materials, provides a potential alternative new material for preventing and controlling target pests, and can be used for the prevention and control of pests in the order Lepidoptera, and provides technical support for the development of Lepidoptera pest control products.

The present invention provides modified Cry1Ac protein having enhanced toxicity towards resistant pink bollworm (PBW) species. Insertion of said specific, tightly binding gut peptides in Cry1Ac at one or more identified sites increases the retention of toxin in the PBW gut. resulting in solubilisation and pore formation leading to sepsis and larval death. The modified Bt toxin exhibits significant toxicity against resistant pink bollworm and thus can be used for generation of insect resistant transgenic plants.

The present invention discloses the application of the transcription factor SfGATAe in the green prevention and control of the Spodoptera frugiperda population. The present invention discovers that overexpressing SfGATAe of Spodoptera frugiperda in the Sf9 cell line insensitive to the Cry1Ac toxin can significantly improve the sensitivity of Sf9 cells to the Cry1Ac toxin; interfering with the expression of SfGATAe in the larvae of Spodoptera frugiperda can significantly increase the resistance of the larvae of Spodoptera frugiperda to the Cry1Ac toxin. Therefore, the transcription factor SfGATAe of Spodoptera frugiperda can be applied to the green prevention and control of the Spodoptera frugiperda population.

The invention belongs to the technical field of immunoassay, and particularly relates to a multicolor joint detection test strip for detecting transgenosis, which comprises a bottom plate, and a sample pad, a combination pad, an NC membrane and a water absorption pad which are fixed on the bottom plate and are connected in sequence, the conjugate pad is coated with a first microballoon-CP4mAb1 compound and a second microballoon-BTmAb1 compound; a detection line T1 and a detection line T2 are marked on the NC membrane by using CP4mAb2 and BTmAb2 respectively, and a quality control line is marked by using a goat anti-rabbit IgG antibody; wherein the first microballe-CP4mAb1 compound is a compound formed by coupling a first nano microsphere and a first antibody of a CP4EPSPS protein, and the second microballe-BTmAb1 compound is a compound formed by coupling a second nano microsphere and a first antibody of a BT-Cry1Ab / Ac protein; the CP4mAb2 is a second antibody of a CP4EPSPS protein, and the BTmAb2 is a second antibody of a BT-Cry1Ab / Ac protein. The test strip provided by the invention can be used for simultaneously detecting the BT-Cry1Ab protein, the BT-Cry1Ac protein and the CP4EPSPS protein, the detection result is accurate, the specificity and the stability are good, the sensitivity is relatively high, the screening efficiency can be remarkably improved, and the cross interference among the components is effectively reduced.

The present invention provides a monoclonal antibody for broad-spectrum detection of Bt Cry toxins and its application, belonging to the technical field of pesticide immunological detection. The cloned antibody JJF-GP-mAb provided in the present application can simultaneously and highly sensitively identify Bt Cry toxin subtypes such as Cry1Ab, Cry1Ac, Cry1Ah, Cry1B, Cry1C, Cry1F, Cry2Ab and Cry3Aa, and can be used for broad-spectrum and highly sensitive detection of Bt Cry toxins in food and environmental samples, and has broad application value.