140 results about "Plant genomes" patented technology

The invention relates to the technical field of collaborative prediction, in particular to a plant polygene stress resistance collaborative prediction method and system based on a federal map neural network. The method comprises the following steps: modeling a multilayer heterogeneous graph according to acquired plant genome data; constructing a dynamic graph neural network model based on time sequence perception; training a dynamic graph neural network model by using the multi-layer heterogeneous graph, and performing distributed privacy calculation on the multi-layer heterogeneous graph by using a federated graph neural network; generalization training is carried out on the small sample scene data based on multi-task learning and a cross-species migration mechanism; and obtaining a gene function prediction result. Through a multi-layer heterogeneous graph fusion technology and a dynamic graph neural network design of time sequence perception, multi-dimensional biological information such as gene regulation, protein interaction, metabolic pathways and stress response can be captured at the same time, and time sequence change characteristics of gene expression in the plant stress response process can be captured at the same time.

The invention provides a method for realizing fixed-point activation of a plant genome by directionally inserting an enhancer, and particularly, by inserting a short enhancer (STE) in a specific region of a target gene promoter, a target gene can be efficiently activated, the expression level of the target gene can be enhanced by about 2000 times, and the enhancement multiple can be flexibly adjusted as required. The site-specific activation method for the plant genome has high efficiency, wide applicability and stable heredity, and has outstanding application value in plant research and breeding.

The invention discloses an RNA (Ribonucleic Acid) aptamer and application thereof in improving the salt tolerance of rice. The aptamer sequence eApt-2 with high affinity to eIF4A is obtained through enrichment by using a phylogenetic screening technology of the nucleic acid aptamer, and the aptamer sequence eApt-2 can inhibit the RNA helicase activity of eIF4A and selectively block a 5'cap-dependent translation initial process. After the eApt-2 coding sequence is introduced into a rice cell, the transgenic rice shows significantly enhanced salt tolerance under a salt stress condition: the growth condition is improved, and the biomass and the photosynthetic efficiency are both higher than those of a control plant. Compared with a traditional strategy of improving the salt tolerance through gene overexpression or knockout, the method does not need permanent modification of animal and plant genomes, accurate and reversible regulation and control of translation initiation factors are carried out through aptamers, optimization of stress response protein synthesis is achieved, and higher specificity and safety are achieved. The invention provides a novel and efficient technical approach for cultivating salt-tolerant crops, and has a wide application prospect.

The invention discloses a method for site-specific replacement or insertion of DNA fragments in a plant genome, and belongs to the technical field of biology. The method disclosed by the invention comprises the following steps: editing a genome of a plant protoplast by adopting a CRISPR / Cas9 system and taking a double-stranded DNA repair template as a donor template to obtain an edited protoplast; and carrying out culture regeneration on the edited protoplast by utilizing a plant protoplast transformation method to obtain a site-directed mutagenesis plant in the genome so as to realize site-directed mutagenesis of the target plant genome. The method disclosed by the invention can be used for accurately replacing DNA in a plant genome at a fixed point. According to the invention, a recognition sequence of recombinase can be accurately inserted into a genome at a fixed point through homologous recombination, and then accurate fixed-point replacement of a gene large fragment is realized by virtue of a specific recombinase system.

Compositions and methods are provided for high-efficiency large scale manipulation of genomic regions and chromosomal engineering of plant genomes. Enhancement of chromosomal modification includes the provision of one or more linker oligonucleotides complementary to the distinct chromosomal ends following double strand breaks, increasing the temperature effect and / or recurrent cutting by an endonuclease to facilitate higher chromosomal modification including large segment translocations and recombination. Site-specific directed DNA breaks under one or more of the experimental conditions (including CRISPR-Cas systems) disclosed enhance targeted recombination frequencies, crossover efficiency and movement of large chromosomal segments in crop plant cells.

Provided are a method for targeted modification of a specific sequence in a plant genome to a sequence of interest through a nuclease-reverse transcriptase fusion protein guided by a guide RNA, and genetically modified plants and progeny thereof produced by the method.

The invention relates to the field of biotechnology and biological breeding, and particularly relates to a method for constructing a single-chain guide RNA (sgRNA) vector for delivering CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) by utilizing a cucumber mosaic virus (CMV), carrying out site-specific editing on a target gene in a Cas9 transgenic plant, causing site-specific mutagenesis of plant genome DNA (Deoxyribose Nucleic Acid) by the vector to be inherited to a progeny, and not carrying the CMV in the progeny. The invention discloses a construction method of a vector pCB301R3-sgPDS and an application of the vector pCB301R3-sgPDS. The pCB301R3-sgPDS is a genetic gene editing vector based on mediation of a cucumber mosaic virus. According to the invention, sgRNA is delivered through CMV for the first time, so that fixed-point editing and seed transmission of plant target genes are efficiently realized in crops.

The invention belongs to the field of gene engineering, and relates to a plant genome directional editing tool based on transposon-coded nuclease. The invention aims to solve the technical problem that the application range of a plant genome editing tool based on CRISPR-Cas9 and Cas12a is limited to a great extent due to the fact that CRISPR-Cas9 and Cas12a are large in protein size and difficult to deliver. The invention provides transposon nuclease suitable for plant genome editing. The transposon nuclease is IsDge10, IsAam1 or enIscB subjected to codon preference optimization; and a gene editing system containing the three transposon nuclease is constructed. The gene editing system can be suitable for editing a coding region, a non-coding region and the like; the method can be widely applied to monocotyledonous plants, dicotyledonous plants, gymnosperm and the like.

The invention belongs to the technical field of gene engineering, and particularly relates to a negative-sense RNA virus plant genome editing system and application thereof. According to the genome editing system, a CRISPR-Cas9 gene editing technology is used, a plant cytoplasm rhabdovirus is used as a delivery carrier, the rhabdovirus is spread to a plant by means of entomophilous, a gene editing component is delivered to an axillary meristematic zone by utilizing RNA (Ribonucleic Acid) to move a motif, and a mutant lateral bud is obtained along with plant tillering. According to the method, plant gene editing can be completed without tissue culture, plant tissue culture dependence is overcome, the range of gene-editable strains is expanded, and pure mutants can be obtained only by one generation without hybridization and selfing processes.

The invention discloses application of novel reverse transcriptase and related fusion protein of the novel reverse transcriptase in plant-guided editing. Candidate RT protein is obtained through systematic mining from a public database, novel reverse transcriptase cl1 with activity is screened out in combination with a fluorescence report system, and a guided editing system suitable for plants is constructed. The system tests in wheat protoplast, and verifies the editing activity of cl1 to a plurality of endogenous targets. Further combining with protein structure prediction and rational design point mutation optimization, the cl1 mutant with higher editing efficiency is obtained, and the method can be applied to accurate and effective editing of plant genomes and plant breeding and improvement.

The invention discloses an insect-resistant and herbicide-resistant transgenic sorghum event BrsT01 as well as a detection method and application thereof. The insect-resistant and herbicide-resistant transgenic sorghum BrsT01 is preserved in the China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC No.46082. The invention also establishes a PCR detection method for detecting whether a plant sample to be detected is derived from the transgenic sorghum BrsT01, and the detection method can identify the junction region of the inserted exogenous DNA molecule and the plant genome DNA, and further can be used for accurately and rapidly identifying whether the plant sample contains the DNA molecule of the specific transgenic sorghum event BrsT01. Therefore, the breeding efficiency is improved, and a foundation is laid for research on cultivation of insect-resistant and herbicide-resistant sorghum varieties with excellent characters.

The disclosure relates to a plant that is tolerant or resistant to species of Ca. Liberibacter. Specifically exemplified are Citrus and solanaceous plants. Provided by the disclosure is a modified Citrus or solanaceous plant that is resistant or tolerant to Sec-dependent effectors secreted by bacteria. Also provided by the disclosure are methods of modifying a plant genome plant to provide tolerance or resistance to species of Ca. Liberibacter. Still further provided by the disclosure are methods conferring a population of plants with tolerance or resistance to species of Ca. Liberibacter and screening that population for the plants that are tolerant or resistant to species of Ca. Liberibacter.

The invention belongs to the field of gene engineering, and relates to nuclease generated based on AI and having a gene editing function and application of the nuclease. The technical problem to be solved by the invention is to develop more nuclease suitable for eukaryote gene editing. According to the technical scheme, nuclease AIGCprotein which is generated based on AI and has a gene editing function is obtained through the following steps that a plurality of protein amino acid sequences are generated through a known nuclease three-dimensional structure in a PDB database, the sequences are classified and compared, and a protein skeleton of the nuclease AIGCprotein is obtained through extraction; the known nuclease is Cas12. The invention provides nuclease which is generated on the basis of Protein MPNN AI and has a gene editing function, and the protein does not exist in the nature; the method can be used for constructing a plant genome editing system.

The present invention relates to a method for plant genome modification of at least one plant cell being in the developmental stage of a plant immature inflorescence meristem (IIM) cell, wherein the modification of the specific cell type is achieved by providing a genome modification or editing system, optionally together with at least one regeneration booster, preferably wherein the effector molecules are introduced by particle bombardment. To this end, new artificial and precisely controllable booster genes and proteins are provided. Further, the modified plant cells are regenerated in a direct or an indirect way. Finally, methods, tools, constructs and strategies are provided to effectively modify at least one genomic target site in a plant cell, to obtain the modified plant cell and to regenerate a plant tissue, organ, plant or seed from the modified plant cell.

The invention discloses a method for detecting an insertion site of agrobacterium T-DNA on a plant genome, which comprises the following steps: carrying out sequence merging on a T-DNA genome and a plant genome to obtain a merged genome G; performing sequencing read sequence alignment on the combined genome G to obtain an SAM file with an alignment result; performing classification analysis on the SAM file with the comparison result; sequentially carrying out blacklist filtering, merging and repeated area marking analysis on a classification analysis result; and finally outputting the number of the final integration sites and the number of the real integration events. According to the method disclosed by the invention, the sensitivity of T-DNA integration site detection is increased and the false positive rate of detection is reduced by taking measures of considering the special condition of double-end sequencing of an insertion site part, establishing a blacklist, carrying out secondary matching optimization on a specific repetitive sequence of a plant genome, carrying out multi-sequence similarity comparison on a result site and the like.

The invention discloses a cytochrome P450 gene CYP71AF43, a polypeptide, a carrier and application of the cytochrome P450 gene CYP71AF43, the polypeptide and the carrier, and the CYP71AF43 gene is derived from alopecurus macrophysalis and has the effect of resisting fenoxaprop-p-ethyl. The nucleotide sequence of the gene is SEQ ID NO: 1 or a sequence with not less than 80% of identity compared with the SEQ ID NO: 1. On the basis, the invention further discloses a protein polypeptide coded by the CYP71AF43 gene. The invention further deeply relates to construction of a carrier containing the CYP71AF43 gene and an all-around application strategy in the breeding process of herbicide-resistant transgenic plants. Through systematic optimization of vector design and transgenic process, it is ensured that the CYP71AF43 gene can be stably and efficiently integrated into a plant genome, the function of giving herbicide resistance to plants is fully exerted, and breeding, popularization and application of new lines of herbicide-resistant transgenic plants are promoted.

The invention provides a lysis solution for rapidly extracting animal and plant genomes and an application method thereof.The lysis solution is prepared from, by mass, 20-25 parts of Tris-Hcl, 10-15 parts of LICI, 5-10 parts of EDTA, 1-3 parts of 3-sulfopropyl hexadecyl dimethyl betaine, 1-3 parts of (NH4) 2SO4 and 30-40 parts of ddH2O. The 3-sulfopropyl hexadecyl dimethyl betaine and (NH4) 2SO4 in the formula of the lysis solution can effectively destroy cell membranes and nuclear membranes; the lysate prepared by the method is clear in component and simple to prepare, DNA precipitation or column purification can be directly carried out after lysis, meanwhile, the lysate is suitable for various sample types including animal tissues and plant tissues, a formula does not need to be adjusted according to different sample types, and compared with a traditional lysate, the lysate has the advantages that the cost is low, and the efficiency is high. The optimized formula has the advantages of high splitting efficiency, wide application range, high DNA purity, high extraction speed and good DNA integrity.

The present invention discloses a plant gene editing method for insufficiently editing a target gene, and belongs to the field of plant genetic engineering. The present invention provides a method for plant gene editing, which can achieve insufficient editing of the plant genome by using a promoter that functions after the callus enters the differentiation period to drive the expression of a gene editing functional element, thereby being able to be used to create a heterozygous mutant in the plant. The present invention also provides a promoter sequence for incomplete editing of the plant genome, a nucleic acid protein composition, and a method for creating a plant heterozygous mutant.

The present invention relates to a method for preparing a MAGEA8-Fc transgenic plant. Through a platform using a tobacco plant, a recombinant protein MAGEA8-Fc, in which a human IgG Fc fragment is fused to a MAGEA8 protein, can be stably expressed. The method of the present invention enables high-level expression efficiency and stable protein production by introducing a target gene into a plant genome using an Agrobacterium binary vector. By directly isolating and purifying the recombinant protein from the transgenic tobacco plant, large-scale production can be easily achieved. Compared to conventional microbial or animal cell-based expression systems, the method provides economic efficiency and scalability, and thus can be usefully applied as a platform for producing fusion proteins.

Compositions and methods are provided for introducing transgenic target sites for Site Specific Integration (SSI) and / or polynucleotides of interest into at least one double-strand break target site of a double-strand-break inducing agent in a genomic window of a plant genome. Also provided are methods and compositions for producing a complex trait locus in a genomic window of a plant comprising at least one transgenic target site for site specific integration integrated in at least double-strand-break target site. The double-strand-break target site can be, but is not limited to, a target site for a zinc finger endonuclease, an engineered endonuclease, a meganuclease, a TALENs and / or a Cas endonuclease. The genomic window of said plant can comprise at least one genomic locus of interest such as a trait cassette, a transgene, a mutated gene, a native gene, an edited gene or a site-specific integration (SSI) target site.

The disclosure relates to a plant that is tolerant or resistant to species of Ca. Liberibacter. Specifically exemplified are Citrus and solanaceous plants. Provided by the disclosure is a modified Citrus or solanaceous plant that is resistant or tolerant to Sec-dependent effectors secreted by bacteria. Also provided by the disclosure are methods of modifying a plant genome plant to provide tolerance or resistance to species of Ca. Liberibacter. Still further provided by the disclosure are methods conferring a population of plants with tolerance or resistance to species of Ca. Liberibacter and screening that population for the plants that are tolerant or resistant to species of Ca. Liberibacter.

Compositions containing chimeric RNA molecules which comprise meristem targeting sequences that are fused to RNA cargo sequences that include gene editing molecules are provided. Methods of using the compositions to efficiently edit plant genomes without intervening tissue culture steps are also provided. The solutions described here relate to engineered RNA molecules useful in producing plants with altered genomes. As such, it relates to substantially purified compositions, vectors, systems, as well as genomes of plants.

The application discloses a plant genome DNA extraction reagent, which comprises Buffer 1 lysate, Buffer 2 precipitate and Buffer 3 combination liquid; the Buffer 1 lysate comprises sodium chloride, PVP40, sodium metabisulfite and SDS; wherein the concentration of the sodium chloride is 250-1200 mM; the mass percentage of the PVP40 is 0.5%-3%; the concentration of the sodium metabisulfite is 50-150 mM; and the mass percentage of the SDS is 0.7%-2.5%; the Buffer 2 precipitate comprises acetate; wherein the concentration of the acetate is 2.2-4.3 M; and the Buffer 3 combination liquid comprises guanidine hydrochloride; wherein the concentration of the guanidine hydrochloride is 0.5-2.5 M. The extraction reagent does not contain organic solvents such as phenol and chloroform, and can improve the removal rate of polyphenols.

Methods arc provided for making a plant genome having a desired nucleic acid sequence. One example method includes identifying genomic segments of a plant that include at least one desired nucleic acid sequence, using a trained neural network, which is biology-informed through first pre-training connections between a genetic data layer and at least one intermediate layer and second pre-training connection between the at least one intermediate layer and a phenotypic layer. The method also includes recombining the at least one at least one desired nucleic acid sequence with a second nucleic acid sequence to form a target plant genome.

The invention belongs to the field of gene engineering. Specifically, the invention relates to a method for inserting an exogenous sequence in a genome at a fixed point. Specifically, on the basis of a guided editing system (PE), two adjacent pegRNAs with partially overlapped sequences on a reverse transcription template are used, and efficient and accurate exogenous sequence fixed-point insertion is realized in a genome, especially a plant genome. The system is further coupled with a recombinase system such as Cre / Lox or FLP / FRT and the like, and large-fragment exogenous sequence fixed-point insertion is realized in genomes, especially plant genomes.

The invention provides a kit for extracting plant genome DNA based on magnetic beads and application. The kit is composed of a lysis solution, a binding solution, a washing solution I, a washing solution II, an eluent and magnetic beads. The cracking combination solution comprises at least one of lauryl sodium sulfate, cetyltrimethylammonium bromide, guanidine hydrochloride, guanidine isothiocyanate, citric acid, trisodium citrate, Tween 20, polyvinylpyrrolidone, sodium chloride, ethylenediamine tetraacetic acid and Tris-HCl; the cracking combination solution comprises at least one of sodium dodecyl sulfate, cetyltrimethylammonium bromide, guanidine hydrochloride, guanidine isothiocyanate, citric acid, trisodium citrate, Tween 20, polyvinylpyrrolidone, sodium chloride, ethylenediamine tetraacetic acid and Tris-HCl; the binding solution comprises at least one of isopropanol, ethanol, guanidine hydrochloride and polyethylene glycol; the washing liquid I is at least one of guanidine hydrochloride, guanidine isothiocyanate, absolute ethyl alcohol, ethylenediaminetetraacetic acid, sodium chloride and Tris-HCl, the washing liquid II is an ethanol aqueous solution, the eluent is a Tris-HCl and ethylenediaminetetraacetic acid aqueous solution, and the magnetic beads are hydroxyl magnetic beads or carboxyl magnetic beads.

The invention belongs to the technical field of molecular biology, and particularly relates to a screening method of a plant broad-spectrum disease-resistant gene and application of the broad-spectrum disease-resistant gene HSP1 in prevention and treatment of plant diseases. The screening method comprises the following steps: comparing CDS sequences of N target pathogenic bacteria to obtain a homologous gene sequence; n is a natural number greater than or equal to 2; comparing the protein sequences of the N target pathogenic bacteria to obtain a homologous protein sequence; comparing the coding sequence of the homologous gene sequence with the coding sequence of the homologous protein sequence, and taking an intersection to obtain a conserved gene; and comparing the conserved gene with a plant genome DNA, and removing highly homologous genes to obtain the plant broad-spectrum disease-resistant gene. The screening method provided by the invention is a broad-spectrum anti-disease gene screening system suitable for different types of crops and coping with different types of crop pathogenic bacteria, and a brand new gene cloning thought is provided for plant anti-disease molecular breeding.

The invention discloses a cytosine deaminase mutant as well as a base editing system and application thereof, and relates to the technical field of molecular biology, the cytosine deaminase mutant is AncY16H, AncI50A, AncG149D, AncHA, AncHD or AncAD, the amino acid sequence is as shown in SEQ ID NO.4-9, and the amino acid sequence is as shown in SEQ ID NO.4-9. The cytosine base editing system comprises any one of the cytosine deaminase mutants, a Cas9 nick enzyme and a uracil-DNA (Deoxyribose Nucleic Acid) glycosylase inhibitor UGI (Uracil-Deoxyribose Nucleic Acid). The invention discloses a novel efficient cytosine deaminase variant without base preference, a novel accurate and efficient cytosine base editing system is established on the basis of the novel cytosine deaminase variant, and the cytosine deaminase variant has a good application prospect in plant genome adaptability transformation.

The invention provides a method for introducing a target DNA fragment into a plant and application of the method, and the method comprises the following steps: carrying out electric fusion on a plant protoplast and an erecta minium protoplast containing the target DNA fragment in a nuclear genome, so as to realize the purpose of introducing the target DNA fragment into the plant. According to the invention, through early condition exploration, a cell fusion system of the phytoneum palmatum and the plant is established, and cell fusion and stable culture of subsequent fusion cultures can be well realized. According to the method, the target DNA fragment is transferred by fusion of the strignum leptosphaereum and the arabidopsis thaliana, and the method is expected to play a key role in application fields such as plant genome synthesis and plant customized transformation research, and has a relatively important application value.

The application discloses a kind of plant whole genome RNA-chromatin interaction detection methods, the method includes the interaction state of fixed RNA and chromatin, preparation double link, obtain nuclear, chromatin DNA fragment, RNA / linker / DNA chimera, RNA-DNA double-stranded complex in turn;Tn5 transposase fragmentization cuts double-stranded complex;PCR amplification forms DNA library.The application applies Tn5 transposase to the original RADICL-seq method, successfully to RNA-DNA complex is cut and library, greatly simplifies the process, experimental procedure is reduced by at least half, experimental period is shortened from 4-5 days to 3 days, the fragment size of DNA library produced is between 250bp-400bp, increase the length of final effective RNA / DNA fragment pair, thereby greatly improve the efficiency when aligning to plant genome in subsequent bioinformatics analysis, provide effective means for plant whole genome RNA-chromatin interaction detection.