64 results about "Genetically modified crops" patented technology

The invention discloses a herbicide-resistant protein GAT hybridoma cell strain, an antibody generated by the herbicide-resistant protein GAT hybridoma cell strain and an application of the herbicide-resistant protein GAT hybridoma cell strain. The herbicide-resistant protein GAT hybridoma cell strain is preserved in the China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC No.46333. The preparation method comprises the following steps: a) purifying prokaryotic expression to obtain GAT recombinant protein; b) immunizing animals: immunizing BALB / c mice by taking the GAT recombinant protein as an antigen; c) cell fusion: splenocytes of the immune BALB / c mice are collected and fused with SP2 / 0 cells; and d) cell strain establishment: carrying out subcloning through a limited dilution method, carrying out ELISA detection after 5-7 days of subcloning until a hybridoma cell strain capable of stably secreting a positive antibody is screened out, and carrying out amplification, re-culture and storage. The monoclonal antibody secreted by the hybridoma cell strain lays a foundation for detection of herbicide-resistant protein GAT in transgenic crops, so that development, seed production and seed conservation of transgenic gat crops are supported, and stable industrial promotion of the transgenic gat crops and sustainable application of related transformants are guaranteed.

Using chemical genetic screening, we discovered a small molecule Cellulosin (aka endosidin20 or ES20) that causes cell swollen and inhibits plant growth, but does not disrupt global vesicle trafficking. By doing mutant screening, we obtained multiple alleles of Arabidopsis thaliana that are resistant to Cellulosin inhibition in growth. Those mutated amino acid residues are conserved across plant species. Cellulosin targets a group of cellulose synthases (CesAs) of Arabidopsis thaliana by binding to a conserved domain essential for the catalytic activity of CesA. Cellulosin may target and inhibit all subtypes of CesAs in plants. The present invention relates to Cellulosin, a cellulose synthase inhibitor, its analogs or derivatives as a broad-spectrum herbicide. The mutated genes, their protein products and a cell or a plant having those mutated genes or expressing those protein products are within the scope of this disclosure.

The invention belongs to the technical field of biological detection, and discloses a transgenic crop intelligent identification method based on high-throughput sequencing, which comprises the following steps: acquiring multi-platform high-throughput sequencing original data and sample metadata, evaluating quality by platforms, dividing according to a sample partitioning rule, and generating a quality-controlled data block set; distributing data blocks to a plurality of parallel computing nodes, synchronously executing technical detection to obtain technical fact fragments, aggregating and supplementing batch statistical information, and generating a technical fact report; matching the judgment rule set, carrying out parallel judgment, triggering expert rechecking on boundary cases, and integrating to generate a supervision judgment conclusion set; the method comprises the following steps: extracting original sequencing reads of all samples, generating a unique hash for each read, constructing a processing track chain, integrating metadata and visual evidence, generating a credible authentication report, dividing different scene versions, and performing targeted distribution; and extracting identification cases for mining and clustering, generating optimization suggestions, and feeding back the optimization suggestions to the quality control rule base and the judgment rule base.

The invention discloses a multi-index fusion-based biotechnology product risk dynamic identification method and system, and relates to the technical field of biological safety management, and the method comprises the steps: S1, obtaining experimental data and environmental data from each stage of a transgenic crop life cycle, constructing a dynamic data set containing a molecular structure sequence, a function performance index and an ecological impact factor, and carrying out the construction of a dynamic data set; extracting the mutation frequency of the molecular structure sequence along with the time change, the abnormal fluctuation of the function expression index and the dynamic change trend of the ecological influence factor; s2, according to molecular structure sequence mutation frequency and functional performance index abnormal fluctuation, extracting a nucleotide replacement mode and a functional abnormality data set of the gene editing product in a research and development stage and an application stage, and determining a nucleotide replacement mode and a functional abnormality association mode; according to the biotechnology product risk dynamic identification method and system based on multi-index fusion, accurate prediction and dynamic management of ecological risks of transgenic crops are realized, and the environmental release safety is remarkably improved.

This invention provides an siRNA nucleic acid pesticide targeting the DICER2 gene to control rice blast fungus, belonging to the field of biotechnology. The siRNA molecule can delay and inhibit the formation of rice blast fungus appressoria, thereby reducing the pathogenicity of the fungus and providing a new method for controlling rice blast. When used in combination with tricyclazole or clotrimazole, it has a synergistic effect. The siRNA molecule targeting the DICER2 gene is prepared into a nucleic acid pesticide and sprayed on rice leaves, effectively reducing rice blast lesions and their area, thus enabling its use in rice blast control. Furthermore, it avoids the cumbersome process of cultivating genetically modified crops and addresses consumer concerns.

The application belongs to the technical field of molecular biology, and particularly relates to a nucleic acid rapid extraction kit for transgenic detection of high-oil crops and application, and is particularly suitable for fluorescent quantitative PCR detection of transgenic components of high-oil crops. The kit comprises a lysis solution, a precipitation solution, a neutralization solution and an elution solution. The application can be completed within 15 minutes, effectively removes PCR inhibitors such as oil and protein, is specifically used for high-oil crops, and is a low-cost, safe and non-toxic nucleic acid rapid extraction method, and meets the requirement of on-site rapid detection of transgenic crops.

The present invention discloses the application of a plant APT2 protein and the encoding nucleic acid in the breeding of tobacco crops. The present invention provides the application of any one of the following substances 1-3 in regulating the resistance of tobacco plants to pathogens and regulating the height of tobacco plants: 1. the protein APT2; 2. the nucleic acid molecule encoding the protein APT2; 3. a recombinant vector, expression cassette, transgenic crop, transgenic cell line or recombinant bacterium containing the nucleic acid molecule encoding the protein APT2. The experiments of the present invention prove that APT2 can be used to regulate the height of tobacco plants and to control tobacco mosaic virus or other pathogens by affecting the efficiency of viral cell-to-cell movement.

The invention discloses a herbicide-resistant protein GAT hybridoma cell strain, an antibody generated by the herbicide-resistant protein GAT hybridoma cell strain and application of the herbicide-resistant protein GAT hybridoma cell strain and the antibody. The herbicide-resistant protein GAT hybridoma cell strain is preserved in the China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC No.46332. The preparation method comprises the following steps: a) purifying prokaryotic expression to obtain GAT recombinant protein; b) immunizing animals: immunizing BALB / c mice by taking the GAT recombinant protein as an antigen; c) cell fusion: splenocytes of the immune BALB / c mice are collected and fused with SP2 / 0 cells; and d) cell strain establishment: carrying out subcloning through a limited dilution method, carrying out ELISA detection after 5-7 days of subcloning until a hybridoma cell strain capable of stably secreting a positive antibody is screened out, and carrying out amplification, re-culture and storage. The monoclonal antibody secreted by the hybridoma cell strain lays a foundation for detection of herbicide-resistant protein GAT in transgenic crops, so that development, seed production and seed conservation of transgenic gat crops are supported, and stable industrial promotion of the transgenic gat crops and sustainable application of related transformants are guaranteed.

The application belongs to the field of agricultural biotechnology, and discloses a database construction and analysis method for soybean transgenic crops, which comprises the following steps: obtaining soybean high-throughput sequencing original data and metadata, performing soybean specificity quality control, storing in a database sub-database according to a scene label, and generating standardized sub-database data; identifying the priority of each sub-database scene, allocating hardware according to a priority-resource matching rule, generating resource configuration records, synchronously executing multi-dimensional feature extraction, and generating a multi-dimensional feature dataset; dynamically adjusting the feature importance degree according to the scene, performing feature collaborative analysis, combining an enhanced decision knowledge base to retrieve and match results and solving feature conflicts, and generating a dynamic confidence identification report; extracting key feature data and identification process information to generate a hash value, writing the hash value into a consortium chain, and associating a soybean sample ID to form a digital traceability certificate; dynamically optimizing the feature importance degree, the enhanced decision knowledge base and the database sub-database rule, and forming a full-link iteration.

The invention discloses an active polypeptide of protoporphyrinogen oxidase and application, the polypeptide has at least 85%, 90%, 95%, 99% and 100% sequence identity of an amino acid sequence as shown in one of SEQ ID NO: 1-42, and has protoporphyrinogen oxidase activity. The active polypeptide disclosed by the invention shows relatively high tolerance to PPO inhibitor herbicides, and has a wide application prospect in plants. According to the weed control method provided by the invention, the occurrence of resistant weeds can be effectively reduced, and transgenic crops obtained by the method can resist various herbicides such as PPO inhibitors, ACC enzyme inhibitors, ALS inhibitors, EPSPS inhibitors and synthetic auxin at the same time, and have broad-spectrum herbicide resistance; in the weed control process, the PPO inhibitor herbicide is used in combination with one or more other herbicides, so that the cost can be effectively reduced, the control efficiency can be improved, and the weed resistance can be greatly avoided.

The invention provides a crop allergen dynamic baseline construction and trusted sharing system based on a multi-dimensional heterogeneous data cube, and the system comprises a collection layer which is used for collecting physical information metadata of a to-be-detected sample based on an agricultural environment monitoring terminal, the method comprises the following steps: performing physical detection on a to-be-detected sample through a high-throughput sequencing terminal to generate original mass spectrum data and transcriptome data, and calling meteorological metadata of a region where the to-be-detected sample is located through a third-party meteorological server; the core processing layer is used for generating a dynamic baseline by adopting a central processing server cluster based on the physical information metadata, the original mass spectrum data, the transcriptome data and the meteorological metadata; and the application layer is used for checking the dynamic baseline based on the user query terminal and checking the GLP log based on the checking and auditing terminal. By constructing the multi-dimensional heterogeneous data cube and the dynamic baseline model, standardized data calculation service is provided for safety evaluation of transgenic crops and biotechnology products.

This invention relates to the field of plant transgenic detection technology, and discloses a method for rapid batch detection of transgenic crops, comprising: Step 1, placing centrifuge tubes containing crop samples to be tested in an array into a multi-well plate to complete batch plate arrangement of samples; Step 2, performing overall mechanized processing based on the multi-well plate, so that the crop samples in the centrifuge tubes are simultaneously ground into powder under mechanical force. This integrated operation process, which places independent centrifuge tubes in an array within a multi-well plate and uses a tissue homogenizer, batch liquid addition equipment, and a centrifuge to simultaneously process all samples, achieves a significant improvement in detection throughput and efficiency. Compared to existing technologies that rely on manual, one-by-one operation, this method overcomes the shortcomings of low detection efficiency and the inability to simultaneously process multiple samples, thus failing to meet the needs of large-scale crop sample screening.

The present disclosure relates to compositions comprising nitroreductase enzymes; plants, plant calluses, plant seeds, and vegetables incorporating genes expressing nitroreductase enzymes; methods for stably and operably incorporating genes expressing nitroreductase enzymes into plants and plant tissues; wherein the nitroreductase enzymes are capable of reducing nitro groups on phytotoxic and / or otherwise harmful compounds such as, for example, thaxtomins, wherein the thaxtomins are secreted by plant pathogenic bacteria and / or exogenously applied as an agricultural composition such as an herbicide, and wherein reducing the nitro groups renders the thaxtomins non-damaging or reduces the level of damage from the thaxtomins to the plants expressing the nitroreductase enzymes.

The invention provides an insect-resistant and herbicide-resistant gene of corn and application of the insect-resistant and herbicide-resistant gene to cultivation of insect-resistant and herbicide-resistant corn, and belongs to the field of plant disease resistance. Experiments prove that the expression vector for simultaneously expressing the Cry1Ab, Cry2Ab and cp4-epsps genes is constructed for the first time, transgenic crops obtained by transforming crops with the vector can simultaneously resist Lepidoptera pests such as ostrinia nubilalis and cotton bollworm and have good glyphosate resistance, the insect-resistant and herbicide-resistant application of the transgenic crops is widened, and the transgenic crops have broad application prospects. The method has a wide application prospect in corn genetic breeding improvement.

The present invention discloses a tobacco calcium ion sensor-related gene and its application. The gene is named tobacco NtCBL4 gene, and its nucleotide sequence is: the nucleotide sequence shown in SEQ ID NO.1; a nucleotide sequence having the same function as the nucleotide sequence shown in SEQ ID NO.1 by substitution, deletion, and / or addition of one or more nucleotides; and a nucleotide sequence that hybridizes with the sequence shown in SEQ ID NO.1 under stringent conditions, wherein the stringent conditions are hybridization in 0.1×SSPE containing 0.1% SDS or 0.1×SSC containing 0.1% SDS at 65°C, and the membrane is washed with the solution. The present invention can control the salt tolerance of tobacco by regulating the expression level of the NtCBL4 gene, and obtain transgenic crops with different NtCBL4 gene expression levels and different salt tolerances through genetic engineering. Overexpression of NtCBL4 can reduce tobacco's tolerance to high salt.

Provided in the present invention is a hypermorphic allele ETD1 against rice blast, wherein the ETD1 gene has a nucleotide sequence as set forth in SEQ ID NO: 1, and a cDNA sequence thereof is as set forth in SEQ ID NO: 2. Further provided is an encoded protein of the gene, wherein the protein has an amino acid sequence as set forth in SEQ ID NO: 3. The gene is a hypermorphic allele of OsCNGC13, which exhibits enhanced calcium-ion transport capacity and has the function of significantly accelerating the influx of calcium ions into cells. Therefore, the normal expression or overexpression of the protein in a gene knockout plant can enhance resistance of rice, and thus the protein has a positive regulatory effect on rice blast resistance. The present invention can be used for the rapid identification of transgenic crops harboring ETD1 mutant genes, thereby improving the efficiency of molecular biological breeding.

This invention belongs to the field of genetic engineering pest control technology, specifically relating to an insect-resistant protein Cry166 that enhances plant resistance to pests and its applications. The amino acid sequence of the insect-resistant protein Cry166 is shown in SEQ ID NO:1, and the application is in plant pest control. After expression in plants, the insect-resistant protein Cry166 provided by this invention significantly enhances the resistance of transgenic crops containing it to the target pest, the fall armyworm. It also expands the insect resistance spectrum of this protein, solving the problems of limited insect resistance mechanisms, narrow insect resistance spectrum, and easy development of resistance in insects in existing technologies. Furthermore, it improves the breeding efficiency of insect-resistant plants to a certain extent and reduces breeding costs.

The present invention belongs to the field of plant genetic engineering technology and specifically provides the use of the GhGLK1 protein and its encoding gene in regulating cotton callus growth and differentiation. The protein meets the following conditions: B1) a protein with an amino acid sequence of SEQ ID NO. 1; B2) a fusion protein with the same function obtained by attaching a tag to the N-terminus and / or C-terminus of B1). GhGLK1 After the gene was cloned, overexpression vectors and CRISPR vectors were constructed, they were transformed into cotton to obtain overexpression plants and knockout plants. GhGLK1 Overexpression plants can significantly improve callus growth and differentiation, greatly shortening the transformation cycle of transgenic crops. This is of great significance for breaking the dependence on the genotype of the recipient material and accelerating the cycle of plant transformation and regeneration.

Various examples relate to a transgenic crop plant where a sequence of a cis-regulatory element in the promoter of certain genes is genomically changed in order to alter the flowering time or flowering rate of the plant. In one example, the cis-regulatory element in the promoter of such certain genes may be modified from TGAATG(A / T) (A / T / C) to TGAGGG(A / T) (A / T / C). The certain genes may be genes that encode SbEhd1, OsFTL10 / FT8, SbFT10, ZmELF3.2, or combinations thereof. And, the crop plant can be for example, sorghum, maize or rice. In another example, the present transgenic crop plant is a progeny plant. In yet another example, flowering time of the transgenic crop plant is regulated by a method that includes changing the sequence of the cis-regulatory element in the promoter of certain genes.

The invention belongs to the technical field of gene engineering pest control, and particularly relates to an insect-resistant protein Cry166 capable of improving insect pest resistance of plants and application of the insect-resistant protein Cry166, the amino acid sequence of the insect-resistant protein Cry166 is shown as SEQ ID NO: 1, and the application refers to application in the aspect of plant pest control. After the insect-resistant protein Cry166 provided by the invention is expressed in a plant body, the resistance of a target pest spodoptera frugiperda of a transgenic crop containing the insect-resistant protein Cry166 is greatly improved, meanwhile, the insect-resistant spectrum of the insect-resistant protein is expanded, and the problems that in the prior art, the insect-resistant mechanism is few, the insect-resistant spectrum is narrow, insects easily generate resistance and the like are solved; the breeding efficiency of insect-resistant plants is improved at a certain level, and the breeding cost is reduced.

The invention discloses a transgenic crop detection method in a one-step method signal enhancement mode, aminated CdSe / ZnS quantum dots (QDs) are coupled with an antigen to be detected, and an electrochemical luminescence detection signal can be enhanced by adopting a one-step incubation method, so that the detection time is remarkably shortened, and an antibody is saved.

The present invention relates to a method for the relative quantification of GM crops using digital PCR without using certified reference materials (CRMs). More specifically, the present invention relates to a primer and probe set composition for digital PCR for the relative quantification of transgenes in GM crops without using CRMs, which comprises a first primer and probe set capable of specifically detecting genes present in a single copy or in 2-3 copies within the genome of GM (genetically modified) crops; and a second primer and probe set capable of specifically detecting transgenes in GM crops, and uses thereof.

The invention relates to a single-chain antibody for recognizing Vip3Aa toxin of bacillus thuringiensis and application of the single-chain antibody. The single-chain antibody is constructed by taking a mouse hybridoma cell strain secreting a monoclonal antibody for resisting the Vip3Aa toxin as a template; the nucleotide sequence and the amino acid sequence are respectively shown as SEQ ID NO. 1 and SEQ ID NO. 2; the single-chain antibody can be used as a detection antibody, is applied to detection of Vip3Aa toxin by a double-sandwich ELISA (enzyme-linked immuno sorbent assay) method, is high in sensitivity and specificity, and can be used for rapid large-scale screening of Vip3Aa toxin in transgenic crops.

The present invention discloses a method for quantitatively evaluating the safety of transgenic crops against non-target organisms based on transcriptome steady-state. The method comprises the following steps: selecting a transgenic insect-resistant crop and a control crop; exposing non-target organisms and target organisms to the transgenic insect-resistant crop and control crop for feeding; collecting the treated non-target organisms and target organisms, extracting total RNA to construct libraries, and sequencing the libraries to obtain corresponding transcriptome data; performing bioinformatics analysis on the transcriptome data to calculate transcriptome diversity parameters, which include information entropy, normalized information entropy, and KL divergence; and performing a one-way analysis of variance on the transcriptome diversity parameters of insects feeding on the transgenic insect-resistant crop and the control crop to assess the impact of the transgenic insect-resistant crop on non-target organisms. The present invention achieves highly sensitive, quantitative, and efficient detection of the impact of transgenic crops on non-target organisms, providing a new approach for environmental safety assessment of transgenic crops.

The invention relates to a method for evaluating the influence of insect-resistant transgenic crops on the diversity of symbiotic bacteria in target and non-target pests, and aims to solve the problems of inaccuracy, incompleteness and the like of the existing evaluation method. The method is suitable for insect-resistant transgenic crops with various construction modes, overcomes the defects of a traditional culture technology by means of a high-throughput sequencing technology, reduces the workload and shortens the evaluation period. The method comprises the specific steps of transgenic crop planting, experimental pest selection, pest sample treatment and collection, sample genome DNA extraction and sequencing, diversity analysis and key flora quantitative verification. The method can comprehensively detect the diversity of the symbiotic bacteria in the pests, provides a scientific basis for evaluating the safety of insect-resistant transgenic crops, and also provides a reference for evaluating the ecological environment influence of other transgenic crops.

The invention discloses a method for evaluating the influence of insect-resistant transgenic crops on the diversity of enteric microorganisms of natural enemy insects based on a 16S rRNA high-throughput sequencing technology. The method comprises the following steps: (1) space-time gradient sampling: collecting natural enemy insect samples according to a crop growth cycle in a transgenic crop planting area or a laboratory simulation environment; (2) sequencing intestinal microorganisms: extracting DNA (deoxyribonucleic acid) of intestinal contents, amplifying a V3-V4 region of the 16S rRNA gene through a universal primer, and sequencing; (3) bioinformatics analysis: an alpha diversity (Shannon, Chao1 index) and a beta diversity (PCoA) are calculated, and the microflora structure difference is analyzed; and (4) screening of core bacteria: identifying the core bacteria significantly related to the health indexes (survival rate and egg laying amount) of the natural enemy insects based on Spearman correlation analysis (rgt, 0.7, plt, 0.05), and establishing a prediction model to evaluate ecological risks. According to the method, the non-target effect of the transgenic crop is quantified through the dynamic change of the microbiome, and a new strategy is provided for ecological safety evaluation.

The invention belongs to the technical field of plant genetic engineering, and particularly provides application of GhGLK1 protein and a coding gene thereof in regulation and control of cotton callus growth and differentiation. The protein meets the following conditions: B1) a protein with an amino acid sequence of SEQ ID NO.1; and B2) a fusion protein with the same function obtained by connecting a tag to the N end and / or C end of B1). According to the invention, the GhGLK1 gene is cloned to construct an overexpression vector and a CRISPR (clustered regularly interspaced short palindromic repeats) vector, and then is transformed into cotton to obtain an overexpression plant and a knockout plant. GhGLK1 overexpression plants can significantly improve the growth and differentiation of calluses and greatly shorten the transformation cycle of transgenic crops, which has important significance in breaking the dependence on the genotype of receptor materials and accelerating the circulation of plant transformation and regeneration.

The present invention discloses a method for producing a modified crop plant with enhanced tolerance to abiotic stress-induced oxidative stress. The method comprises introducing at least one genomic modification in the endogenous ATR7 gene using targeted genome editing techniques. A modified crop plant exhibiting improved resilience to oxidative stress conditions induced by abiotic factors, due to the disruption or knockout of the ATR7 gene is also disclosed.

The invention discloses a gene related to synthesis of amygdalin from crop seed endosperm and application of the gene, and belongs to the technical field of gene engineering. According to the invention, gene sequences of CYP79D16, CYP71AN24, UGT94AF3 and UGT94AF2 are optimized, CYP79D16 and CYP71AN24 are connected by using a linker peptide I to obtain a fusion gene I, and the fusion gene I is fused with a breeding crop endosperm strong expression promoter to construct a gene expression cassette I; connecting UGT94AF3 and UGT94AF2 by using a connecting peptide II to obtain a fusion gene II, and fusing the fusion gene II with a breeding crop endosperm strong expression promoter to construct a gene expression cassette II; transferring the gene expression cassette into an expression vector to construct a recombinant vector, and introducing the recombinant vector into a plant to obtain a transgenic crop. According to the invention, synthesis of amygdalin in crop seed endosperm is realized, the storability of crop seeds is improved, and the product can be used as a raw material for producing amygdalin, and has guiding significance and production application value for crop breeding.

The present invention discloses a method and system for dynamically identifying risks of biotechnology products based on multi-indicator fusion, which relates to the field of biosafety management technology, including: S1, obtaining experimental data and environmental data from various stages of the life cycle of genetically modified crops, constructing a dynamic data set including molecular structure sequences, functional performance indicators and ecological impact factors, and extracting the mutation frequency of molecular structure sequences over time, abnormal fluctuations in functional performance indicators and dynamic change trends of ecological impact factors; S2, based on the mutation frequency of molecular structure sequences and abnormal fluctuations in functional performance indicators, extracting nucleotide substitution patterns and functional abnormality data sets of gene editing products in the research and development stage and the application stage, and determining the correlation pattern between nucleotide substitution patterns and functional abnormalities; this method and system for dynamically identifying risks of biotechnology products based on multi-indicator fusion realizes accurate prediction and dynamic management of ecological risks of genetically modified crops, and significantly improves the safety of environmental release.