853 results about "Genetics transformation" patented technology

The invention discloses a method for improving the inductivity and genetic transformation efficiency of hairy roots of toona ciliata. According to the method, agrobacterium rhizogenes carrying RUBY-TcGRF8-GIF2 chimeric protein are injected into stems and petioles of aseptic seedlings of the toona ciliata, and the TcGRF8-GIF2 chimeric gene is over-expressed, so that the induction efficiency and the genetic transformation efficiency of the toona ciliata hairy roots are remarkably improved, and the transformation period of the toona ciliata hairy roots is remarkably shortened. Specifically, after overexpression of the TcGRF8-GIF2 gene, the induction time of the hairy roots is shortened from 70 days of a RUBY-XbaI no-load plasmid control group to 25 days, the induction rate and conversion rate of the hairy roots mediated by agrobacterium tumefaciens are improved, the operation efficiency of genetic transformation of the toona ciliata is effectively improved, and the method has important significance for promoting genetic improvement of the toona ciliata.

The invention relates to the technical field of plant drought tolerance, in particular to a plant drought tolerance related protein FYVE4 and application of a coding gene thereof. By inhibiting expression or protein activity of the FYVE4 gene in the plant, the survival rate of the plant under drought stress is increased; according to the method, in genetic improvement of arabidopsis thaliana, after genetic transformation is conducted through an agrobacterium-mediated method or a gene gun method, the stomata closing response speed and chlorophyll accumulation amount of crops can be effectively increased, and therefore plants with stable genetic drought resistance characters are established; the technical scheme is particularly suitable for constructing a water-saving agricultural system, has very important significance for improving crops and cultivating drought-resistant crops, and is suitable for popularization and application.

The application discloses an NLR plug-in gene for improving plant disease resistance and application thereof, and belongs to the field of plant molecular biology and biotechnology.The NLR plug-in gene StEM1 has a whole gene sequence as shown in SEQ ID NO.1 and a CDS sequence as shown in SEQ ID NO.2, and the amino acid sequence of the encoded protein is as shown in SEQ ID NO.3.The gene plays a key role in improving plant disease resistance, especially resistance to field late blight variant bacteria.In the NLR background, overexpression of the gene significantly promotes the disease resistance of tobacco and potato to pathogenic pythium, and the gene is a gene for enhancing plant disease resistance by improving ETI immunity.Overexpression of the gene in the NLR background is obtained by a genetic transformation method, and a strain line is obtained to resist field variant strains, so as to cope with field diseases of crops.The application provides an excellent gene resource for green prevention and control of late blight and disease-resistant genetic engineering breeding.

The invention discloses an application of a BnaMYB7 gene or an encoded protein thereof in regulating and controlling the oil content of rape, and belongs to the fields of genetic engineering and crop genetic breeding. The nucleotide sequence of the BnaMYB7 gene is shown as SEQ ID NO: 1 or SEQ ID NO: 2, an agrobacterium-mediated genetic transformation method is used, an overexpression vector of the BnaMYB7 gene and a CRISPR / Cas9 gene editing vector are respectively transformed into a genome of the rape, the rape germplasm with overexpression and function deletion of the BnaMYB7 gene is obtained, and the rape germplasm with overexpression and function deletion of the BnaMYB7 gene is determined by measuring the oil content of seeds of overexpression and mutant materials. It is found that the BnaMYB7 gene positively regulates the oil content of the rape, so that rape varieties with different oil contents can be obtained by using the gene to perform molecular breeding on the rape. The feed additive is of great significance in relieving grain and oil tension and guaranteeing feed safety.

The invention discloses application of a TaAGD gene in promoting wheat regeneration and genetic transformation, and belongs to the technical field of plant genetic engineering. According to the present invention, the first research discovers that the wheat regeneration efficiency can be improved by overexpressing the TaAGD gene in the wheat immature embryo explant, such that the wheat genetic transformation process is promoted, the new regeneration gene resource is provided for the wheat biological breeding, and the important significance is provided for the promotion of the wheat genetic improvement.

The invention belongs to the technical field of genetic engineering of edible mushrooms, and particularly relates to a construction method of an efficient and stable genetic transformation system of flammulina velutipes. The construction method comprises the following steps: culturing in an MM culture medium by taking a flammulina velutipes strain 0747 as a receptor; the pFgnpt vector is converted into agrobacterium AGL1 (AGL1); inoculating hyphae into an IM liquid culture medium containing acetosyringone and MES, mixing with agrobacterium tumefaciens, culturing, and transferring the hyphae to an IM solid flat plate paved with filter paper; hyphae are transferred to a PDA resistance plate containing G418 and cephalosporin for resistance screening, and an efficient and stable flammulina velutipes genetic transformation system is obtained. The new selection marker G418 is developed and applied, limitation of a traditional selection marker is broken, more choices are provided for flammulina velutipes genetic transformation, a new thought is provided for genetic transformation research of other edible mushrooms, and the application field of an agrobacterium tumefaciens-mediated transformation technology is expanded.

The invention provides an AbAIL5 gene and application of the AbAIL5 gene in improvement of genetic transformation efficiency of amorphophallus bulbifer, and belongs to the technical field of biology. The invention provides an AbAIL5 gene. The nucleotide sequence of the AbAIL5 gene is shown as SEQ ID NO: 1. Genetic transformation is performed on the amorphophallus bulbifer somatic embryo by constructing an overexpression vector of the AbAIL5 gene, so that the genetic transformation efficiency of the amorphophallus bulbifer callus can be improved, and candidate genes are provided for application of a large-scale breeding technology and a genetic transformation technology of the amorphophallus bulbifer callus in good varieties.

The invention provides a CsERECTA gene-based tea tree leaf shape regulation and control method and a tea tree light utilization rate improving method, which are characterized in that a sequence of a tea tree gene is cloned to obtain a complete sequence; then, the CsERECTA is subjected to bioinformatics analysis, expression of a GUS reporter gene is started by constructing a plant expression vector and connecting a target fragment, and a recombinant plant vector is used for genetic transformation of tobacco, so that the function of the CsERECTA is verified; the character of the CsERECTA transgenic line is analyzed, the leaf shape is determined, the method capable of improving the leaf shape of the tea tree is obtained, and a foundation is laid for cultivating a new variety and improving the light utilization rate of the tea tree by using the CsERECTA gene.

The invention discloses a TaIMPA1 gene and application of the TaIMPA1 gene in improvement of plant genetic transformation efficiency. The nucleotide sequence of the gene is shown as SEQ ID No: 1 in a sequence table. According to the invention, the key gene TaIMPA1 is cloned from a plant body to delivery for the first time, and the genetic transformation performance is enhanced by remarkably improving the delivery efficiency of exogenous DNA (Deoxyribose Nucleic Acid). The discovery fills up the research blank of plant endogenous delivery regulatory factors, and an original solution is provided for overcoming the core problem of genotype dependence.

The invention belongs to the technical field of ryegrass plant tillering regulation and control methods, and discloses a ryegrass LpZAT12 gene for regulating and controlling plant tillering and application, and the nucleotide sequence of the LpZAT12 gene is as shown in SEQ ID NO: 1; the amino acid sequence is shown as SEQ ID NO: 2; and the nucleotide sequence of the corresponding promoter is as shown in SEQ ID NO: 3. Transgenic ryegrass obtained through a perennial ryegrass genetic transformation system is subjected to phenotypic analysis, and the result shows that the gene inhibits tillering of perennial ryegrass. The discovery of the LpZAT12 gene provides a new gene resource for cultivating a new perennial ryegrass variety with strong tillering ability. The overexpression of the gene can reduce the management cost of the lawn and prolong the management period; knockout of the gene can promote plant tillering, increase plant biomass and improve industrial benefits, and application of the gene has important production practice significance.

The invention provides an application of a sunflower HaSnRK2 gene in regulating and controlling salt tolerance and drought tolerance of plants, and belongs to the technical field of plant genetic engineering. The invention provides an overexpression vector of a sunflower HaSnRK2 gene. The overexpression vector is applied to regulation and control of salt tolerance and drought tolerance of plants after genetic transformation is carried out on the plants by using a flower dipping method. Through overexpression of the sunflower HaSnRK2 gene, the salt stress tolerance and the drought stress tolerance of plants are remarkably enhanced, and damage caused by reactive oxygen free radicals induced by salt stress and drought stress can be effectively relieved.

The present application relates to the technical field of elicitor peptide MsPep and its application, and discloses the use of elicitor peptide MsPep in improving the genetic transformation efficiency of alfalfa, wherein the application takes the leaf of alfalfa as an explant, introduces an exogenous gene through Agrobacterium tumefaciens, and adds the elicitor peptide MsPep in each stage of genetic transformation, so as to significantly improve the dedifferentiation rate, the out-growth rate, the differentiation rate, the out-bud rate, the rooting rate and the transformation efficiency, and shorten the transformation period, and the present application has application prospect in improving the genetic transformation of alfalfa and other plants.

This invention discloses the TaJDR1 gene and its application in improving plant resistance to Fusarium head blight and drought. The nucleotide sequence of the TaJDR1 gene is shown in SEQ ID NO.1, and its encoded amino acid sequence is shown in SEQ ID NO.2. This invention is the first to clone the TaJDR1 gene from the local wheat variety Wangshuibai and achieve overexpression of this gene in wheat using gene gun-mediated genetic transformation technology. The results show that the transgenic wheat exhibits significantly improved resistance to Fusarium head blight and drought resistance. Since the TaJDR1 gene originates from wheat itself, overexpression does not affect food safety; therefore, this invention can be widely applied in the field of disease and drought resistance breeding for various plants.

The invention discloses application and a method of wheat demethylase TaALKBH23 in regulation and control of drought stress resistance of plants, a coding sequence of the demethylase TaALKBH23 is shown as SEQ ID NO: 1, and an amino acid sequence coded by the demethylase TaALKBH23 is shown as SEQ ID NO: 2. The invention provides demethylase TaALKBH23 and a function and application of a coding gene of the demethylase TaALKBH23 in regulation and control of plant drought resistance, and it is found through construction of a TaALKBH23 wheat genetic transformation material that the gene negatively regulates and controls wheat drought resistance. It is found that TaALKBH23 plays an important role in the process of regulating and controlling drought stress resistance of plants, and a new gene resource is provided for improving drought resistance of crops.

The invention belongs to the technical field of plant biology, and discloses a wild soybean genetic transformation method which comprises the following steps: S1, disinfecting wild soybean seeds; s2, the backs of the disinfected wild soybean seeds are cut once, the cut wild soybean seeds are placed on a culture medium, and imbibition seeds are obtained; s3, seed coats of the wild soybean seeds are removed, growing points of embryo tips are exposed through dissection, and explants are obtained; s4, putting the explant into an agrobacterium infection solution containing target plasmids, and carrying out vacuum infiltration in a vacuum environment; and S5, after releasing vacuum, standing and infecting in an agrobacterium infection solution for 3-16 hours. The glycine soja genetic transformation method can effectively improve the transformation efficiency of glycine soja.

The invention provides application of wheat cysteine receptor kinase TaCRK25-1B and a coding gene thereof in regulation and control of stripe rust resistance of plants, and belongs to the technical field of plant genetic engineering. The TaCRK25-1B provided by the invention is subjected to induced expression of stripe rust infection in wheat, a VIGS technology instantaneous silencing experiment shows that the resistance of the wheat to stripe rust CYR31 is enhanced by silencing the TaCRK25-1B, the TaCRK25-1B is over-expressed in the wheat by a genetic transformation technology, the resistance of the wheat to stripe rust is weakened, and an RNAi interference plant infection pathogen experiment of the TaCRK25-1B created by an RNAi technology shows that the TaCRK25-1B has the advantages that the TaCRK25-1B can be used for expressing the stripe rust CYR31 in the wheat; the TACRK25-1B plays a negative regulation role in wheat stripe rust resistance. Therefore, the invention provides a new means for wheat stripe rust resistance and resistant wheat breeding.

The invention discloses an application of a tobacco cytochrome protein P450 gene, and particularly relates to an application of the tobacco cytochrome protein P450 gene in regulation and control of tobacco dwarfing, and the tobacco cytochrome protein P450 gene is an NtCYP71AU50 gene and / or an NtCYP78A7 gene; the nucleotide sequence of the NtCYP71AU50 gene is as shown in SEQ ID NO. 1, and the nucleotide sequence of the NtCYP78A7 gene is as shown in SEQ ID NO. 3. According to the invention, an editing vector for knocking out NtCYP71AU50 and NtCYP78A7 genes is constructed by virtue of a CRISPR / Cas9 mediated gene editing technology, NtCYP71AU50 and NtCYP78A7 gene editing plants are obtained after genetic transformation, and the plant height, stem girth, waist leaf length, waist leaf width and leaf area of the two gene editing plants are lower than those in a control group. By regulating and controlling the growth and development process of the tobacco plants, the tobacco plants are promoted to be dwarfed, and the problem of shortage of tobacco dwarfing resources is solved, so that application of the tobacco plants in dwarfing close planting cultivation is promoted, the lodging resistance of the tobacco is enhanced, and possibility is provided for increasing the yield per unit area.

The invention relates to an application of a genetic transformation method of populus sibirica and a CRISPR / Cas9 gene editing method in plant breeding, and belongs to the technical field of plant molecular breeding. The method comprises the following steps: (1) soaking an explant in an infection bacterium solution; (2) spreading on a CM1 culture medium; (3) transferring the explant to a CM2 culture medium for 4-5 weeks; (4) transferring the calluses to a CM3 culture medium until adventitious buds are obtained; (5) transferring the callus with the adventitious buds to a CM4 culture medium until cluster buds are obtained; and (6) cutting off clustered seedlings, and inserting the clustered seedlings on a CM5 culture medium. According to the application of the genetic transformation and CRISPR / Cas9 gene editing method of the poplar in plant breeding, a target gene is transferred into a poplar body to obtain a positive plant, meanwhile, a poplar phytoene dehydrogenase (PDS) gene is taken as an example, a poplar mutant of which the PDS gene is knocked out is successfully obtained by applying a CRISPR / Cas9 gene editing technology, and the method is suitable for plant breeding. And a technical support is provided for application of a CRISPR / Cas9 gene editing technology in molecular breeding of the populus sibirica.

The invention provides an agrobacterium rhizogenes-mediated forsythia suspensa hairy root genetic transformation method and application thereof, and belongs to the technical field of plant genetic engineering. The method comprises the following steps: introducing an expression vector into agrobacterium rhizogenes, and mixing with an infection solution to prepare an agrobacterium rhizogenes infection solution; the forsythia suspensa stem is put into the agrobacterium rhizogenes infection liquid to be infected for 0.5-1 h through a vacuum infiltration method, and the infected forsythia suspensa stem is cut into a matrix to be subjected to dark culture and then continues to grow till hairy roots are generated; the OD600 value of the agrobacterium rhizogenes infection liquid is 0.5 to 1.0. The method is suitable for an agrobacterium rhizogenes mediated genetic transformation system of the fructus forsythiae, and the system provides technical support for functional gene research, medicinal component analysis and molecular breeding of the fructus forsythiae, and has important significance for promoting genetic improvement and resource utilization of the fructus forsythiae.

The invention specifically discloses a salix cheilophila in-situ genetic transformation method independent of a tissue culture system, and belongs to the field of plant genetic engineering. According to the method, semi-lignified branches of salix cheilophila are taken as receptors, vacuumizing treatment is performed by utilizing agrobacterium rhizogenes infection liquid, hairy roots are directly induced under a water culture condition after short-term dark culture, and positive identification is performed through reporter genes. According to the method, tedious sterile tissue culture operation is completely avoided, the transformation period is shortened to about 22 days, and the technical threshold and the time cost are remarkably reduced. By adopting the system, the positive rate can reach 54.33% when the pBI121 vector is used for transformation, and the efficiency is remarkably improved compared with that of an existing method. The method provides an efficient and reliable technical platform for root system gene function research, expression analysis and genetic improvement of salix cheilophila and related species thereof, and has high application potential.

The invention discloses an application of an alfalfa gene MsCBF4 in improving the cold resistance of plants, and belongs to the technical field of gene engineering. In order to excavate a gene with a clear anti-freezing function from medicago sativa, the medicago sativa gene MsCBF4 is successfully cloned, and the expression of the MsCBF4 gene is found to be induced by cold stress and freezing stress through the expression specificity and space-time expression analysis of the gene in the medicago sativa. Besides, a plant overexpression vector containing the alfalfa MsCBF4 gene and recombinant agrobacterium are constructed, transgenic arabidopsis thaliana overexpressing the MsCBF4 gene is obtained through a genetic transformation experiment, freezing tolerance analysis on the transgenic arabidopsis thaliana overexpressing the MsCBF4 gene shows that the overexpression of the MsCBF4 gene enhances the tolerance of the arabidopsis thaliana to freezing stress, and the transgenic arabidopsis thaliana overexpressing the MsCBF4 gene can be used for preventing freezing stress. Therefore, the MsCBF4 gene can be used for improving the cold resistance of the plant.

The invention provides a pogostemon cablin cold stress response transcription factor and application thereof in resisting cold and increasing pogostemon cablin alcohol content, and belongs to the technical field of gene engineering. The terpene metabolism cold response transcription factor PatbZIP6 and / or PatMYB5 is obtained through screening, the transcription factor obtained through screening can remarkably improve synthesis accumulation of patchouli alcohol through a genetic transformation method, the cold resistance of patchouli is improved, and the transcription factor can be used for breeding new varieties of patchouli.

The invention relates to a method for inhibiting rumen gas production by using a genetically modified laver filament as a ruminant feed additive and a method for inhibiting methane production by using the genetically modified laver filament as the ruminant feed additive. A porphyra yezoensis bromine peroxidase gene is cloned, an overexpression vector of the gene is constructed, porphyra yezoensis thallus is used as a receptor for genetic transformation, and a porphyra algal strain of which the key enzyme expression quantity is remarkably improved is obtained through screening. The method comprises the following steps: inducing thalli to generate protonema, crushing the protonema by using a crusher, carrying out ventilated large-scale culture in a seawater culture medium added with PES by using a column bioreactor to obtain a large number of protonema materials, collecting the protonema materials, crushing the protonema materials by using the crusher, and drying at low temperature. The obtained laver protonema powder is put into a vacuum bag for low-temperature preservation in dark light, and can be applied to preparation of ruminant feed as a dairy cow feed additive. The filamentous can accumulate halogenated compounds such as bromidogen and the like, can inhibit the activity of a key enzyme generated by methane in rumen, and further inhibits the generation of methane.

The invention belongs to the technical field of plant genetic engineering and soybean breeding, and discloses application of a GmCDPKb gene in regulation and control of the number of soybean symbiotic nodules, the nucleotide sequence of the GmCDPKb gene is shown as SEQ ID NO.1, and the amino acid sequence of encoded protein is shown as SEQ ID NO.2. According to the invention, a GmCDPKb gene mutation and overexpression soybean stable genetic transformation plant is constructed, and character investigation shows that the nodulation number of a mutant plant is obviously lower than that of a wild type, and the nodulation number of an overexpression plant is obviously higher than that of the wild type. The result shows that the GmCDPKb gene plays a role in regulating and controlling the number of soybean nodules. The invention reveals that the GmCDPKb gene is a key regulatory factor for regulating the number of soybean root nodules symbiotic nodules, the number of soybean nodules can be stably and obviously regulated by regulating the expression quantity of the gene, and important gene resources and germplasm materials are provided for analyzing a soybean symbiotic nitrogen fixation molecular mechanism and cultivating a new efficient nitrogen fixation soybean variety.

The invention relates to the technical field of genetic engineering, in particular to an agrobacterium tumefaciens-mediated radix pseudostellariae hairy root genetic transformation method and application thereof. The method comprises the following steps: preparing a radix pseudostellariae explant; activating and culturing agrobacterium rhizogenes; constructing an engineering bacterium containing a target gene recombinant vector; infecting explants and inducing hairy roots; carrying out resistance screening and sterilization on transgenic hairy roots; carrying out amplification culture on hairy roots; and carrying out RT-PCR detection. The hairy root genetic transformation method provided by the invention is simple and rapid to operate and high in gene expression level, and can provide technical support for gene function research. Meanwhile, by using the genetic transformation method provided by the invention, through overexpression of the radix pseudostellariae cyclic peptide coding gene, a transgenic material with significantly improved content of the pharmacodynamic substance cyclic peptide of the radix pseudostellariae can be obtained, and a research material is provided for researching a biosynthetic pathway of the cyclic peptide.

The present application belongs to the technical field of genetic engineering, and particularly relates to a methyltransferase gene CmCMT2 and CmDRM2 application in regulating chrysanthemum flowering. The present application constructs an RNAi silencing expression vector by isolating CmCMT2 and CmDRM2 genes in a chrysanthemum variety "Jinma", and obtains CmCMT2 -RNAi and CmDRM2 -RNAi transgenic "Jinma" plants by using a genetic transformation method, then performs phenotype observation and statistics on the transgenic plants, and explores CmCMT2 and CmDRM2 the mechanism of the genes affecting early flowering of chrysanthemum by using an RNA-seq method. The CmCMT2 and CmDRM2 genes are cloned from the chrysanthemum variety "Jinma", and spatiotemporal expression analysis shows that CmCMT2 and CmDRM2 the genes are highly expressed during the process of "Jinma" from bud development to color change; and CmCMT2 and CmDRM2 the expression level in leaves gradually decreases during the growth and development of "Jinma". The research results show that CmCMT2 and CmDRM2 the content of GA1 is changed by negatively regulating the expression of CmG20ox2a and CmG20ox2b , so that the flowering period of chrysanthemum is advanced by 15 and 8 days, respectively.

The invention discloses a stropharia rugoso-annulata SRU6 gene promoter, a silent vector containing the promoter and application of silent stropharia rugoso-annulata endogenous genes, and relates to the field of molecular biology and gene engineering. According to the principle of shRNA mediated gene silencing, a silencing vector suitable for gene silencing of stropharia rugoso-annulata is constructed, and meanwhile, a stropharia rugoso-annulata efficient genetic transformation system is established; by utilizing the silent vector and the transformation system established by the invention, the expression quantity of the target gene can be reduced by about 80%, the purpose of gene function research is achieved, and the silent vector and the transformation system have very strong theoretical research and application values.

The invention relates to the technical field of plant genetic engineering, in particular to an application of a poplar PtbHLH47 gene and an encoding protein thereof in improving the salt tolerance of plants. The invention discloses application of a PtbHLH47 gene to improvement of plant salt tolerance and improvement of chlorophyll content of plant leaves. The nucleotide sequence of the PtbHLH47 gene is as shown in SEQ ID NO. 1, and the amino acid sequence of the expression protein of the PtbHLH47 gene is as shown in SEQ ID NO. 2. PtbHLH47 genes are cloned from populus tomentosa leaves, overexpressed PtbHLH47 transgenic populus tomentosa is obtained through an agrobacterium tumefaciens-mediated populus genetic transformation system, salt tolerance evaluation shows that the salt tolerance of the overexpressed PtbHLH47 transgenic populus tomentosa is remarkably improved, particularly, the chlorophyll content in the populus tomentosa leaves under salt stress can be remarkably improved, and the chlorophyll content in the populus tomentosa leaves under salt stress can be remarkably reduced. And gene resources are provided for cultivating salt-tolerant plants.

The invention discloses a rubus cochinchinensis efficient genetic transformation method based on RUBY reporter gene and agrobacterium rhizogenes mediation, which realizes that the transformation period is shortened to be within 15 days and the transformation efficiency is improved to be more than 60% through direct explant treatment under a non-tissue culture condition. The method comprises the following specific steps: 1) preparing an explant material; (2) constructing an RUBY carrier and transforming agrobacterium rhizogenes; (3) infecting the explant by utilizing agrobacterium rhizogenes carrying a target gene; (4) co-culturing after infection and carrying out RUBY visual screening; 5) positive root molecular biology verification; (6) inducing positive roots into buds; according to the method, induction of the positive callus, the positive root and the positive bud of the rubus coreanus stem segment under the non-tissue culture condition is successfully achieved, a stable and efficient genetic transformation system is constructed, the method has the remarkable advantages of being easy and convenient to operate, low in cost, wide in applicability, high in reproducibility and the like, and a key technical support is provided for molecular breeding of the rubus coreanus.

The invention discloses a BnaPSK26 gene and application thereof in promoting callus formation, and belongs to the field of plant genetic engineering. The gene can regulate and control the callus of the rape, and a new way is provided for improving the genetic transformation efficiency of the rape. According to the invention, the coding sequence of the brassica napus BnaPSK26 is synthesized by a company, and overexpression of the BnaPSK26 in the brassica napus is beneficial to formation of plant calluses, so that the function and the application way of the gene are proved.