1270 results about "Genetics transformation" patented technology

The present invention relates to the fields of genetic engineering and plant genetic breeding, and specifically to the application of wheat TaPND gene. In the present invention, by taking gene TaPND as a target, designing a sgRNA sequence based on CRISPR / Cas9, connecting a DNA fragment containing the encoding sgRNA sequence to a vector carrying CRISPR / Cas, a plant binary expression vector capable of knocking out the TaPND gene in wheat is constructed; by genetically transforming wheat, offspring with TaPND gene knockout are obtained, and wheat mutants with early flowering time are obtained. Experiments have confirmed that transgenic wheat with TaPND gene knockout achieves the advancement of wheat flowering time. The technical scheme of the present invention provides a practical method for realizing rapid wheat breeding using genetic engineering technology, which has important breeding application value and broad market application prospects.

The invention discloses application of a wheat WRKY transcription factor gene TaWRKY1, and belongs to the technical field of gene engineering. A nucleotide sequence of an open reading frame of the gene TaWRKY1 is shown as SEQ ID NO: 1, and the gene TaWRKY1 is infected by stripe rust to induce expression in interaction of wheat and stripe rust, and plays a negative regulation role in wheat stripe rust resistance. A TaWRKY1 gene editing mutant plant is created by using an agrobacterium tumefaciens-mediated genetic transformation method, the sporulation quantity on wheat leaves of the gene editing mutant is obviously reduced, and the allergic necrosis is increased, which indicates that the edited gene TaWRKY1 can improve the resistance of wheat to stripe rust. The invention provides a disease-resistant germplasm resource for the cultivation of the stripe rust-resistant wheat variety.

The invention belongs to the field of plant genetic engineering, and discloses application of a vitis amurensis cold-resistant gene VaWRKY65 in plant cold-resistant genetic improvement, the VaWRKY65 gene is a transcription factor separated and cloned from extremely cold-resistant vitis amurensis, and the sequence of the VaWRKY65 gene is shown as SEQ ID NO.1. The invention further discloses a preparation method of the vitis amurensis cold-resistant gene VaWRKY65. The gene is used for constructing an overexpression and interference vector, the overexpression and interference vector is respectively introduced into arabidopsis thaliana and vitis amurensis through agrobacterium tumefaciens-mediated genetic transformation, and biological function verification of the obtained transgenic plant shows that the cloned VaWRKY65 gene has the function of improving the cold resistance of the plant. The discovery of the gene provides a new gene resource for plant stress resistance molecular design breeding and provides a new genetic resource for implementation of green agriculture and water-saving agriculture, and development and utilization of the genetic resource are beneficial to reduction of agricultural production cost and realization of environmental friendliness.

The invention provides application of an ECT2 gene in regulating and controlling development of secondary xylem of a poplar. Transgenic poplar with overexpressed ECT2 genes is obtained through genetic transformation, it is found that overexpressed ECT2 genes can increase the width of secondary xylem of the poplar, and the nucleotide sequence of the ECT2 genes is shown as SEQ ID NO: 1. The ECT2 gene has a regulation effect on the development of the secondary xylem of the poplar.

The invention is applicable to the technical field of biotechnology and genetic engineering, and provides an agrobacterium rhizogenes mediated genetic transformation method for hairy roots of blueberries. According to the method, a blueberry perennial stem segment is used as a receptor material for infection, and rapid genetic transformation of the blueberry stem segment is realized through an agrobacterium tumefaciens-mediated method. The method ingeniously bypasses the lengthy process of differentiating the explants to form somatic embryos, and compared with a traditional leaf disc method genetic transformation system, the method has the advantage that the genetic transformation period is remarkably shortened. And the method can be used for mass infection. Even in a non-sterile environment, the method can realize high-efficiency transformation of the gene, and shows the characteristics of simplicity and high efficiency. Therefore, the invention not only provides a new way for genetic transformation of the blueberries, but also provides valuable reference for genetic transformation research and gene function research of the blueberries.

Disclosed is an innovative method for genetic transformation through injection of Agrobacterium containing plasmid DNA expressing PLETHORAs (PLT5) or / and WOUNDINDUCED DEDIFFERENTIATION 1 (WIND1) in multiple plant species including Snapdragon and tomato. In addition, this plasmid has also been tested that it can significantly improve the genetic transformation method is not genotype dependent. Genetic transformation can be achieved by promoting de-novo regeneration of shoots in adult plants, which will of significant impact on some perennial plant species such as citrus and blue berry etc. The transgenic de-novo shoot regenerated from adult plants can by-pass the juvenile phase of plants regenerated through the tissue culture method. Expression of PLT5, WIND1, or both can also greatly improve plant regeneration and transgenic transformation through in-vitro tissue culture in recalcitrant plant species.

The invention relates to the technical field of plant tissue culture, in particular to a Rhododendron pulchrum tissue culture method based on callus regeneration. According to the method, the tender cotyledons of aseptic seedlings are taken as explants, and an efficient Rhododendron pulchrum tissue culture and rapid propagation technical system is established through induction of calluses, subculture multiplication of the calluses, differentiation of adventitious buds, bud strengthening and rooting culture, so that complete tissue culture progeny plants are formed. The induction rate of the callus tissue of the system is 100.0%, the proliferation multiple of the callus tissue in a subculture cycle reaches 5.2 times, 28.5 adventitious buds can be differentiated from a callus tissue block with the size of about 0.5 mm * 0.5 mm * 0.5 mm, the rooting rate reaches 95.8%, the system is not limited by factors such as seasons, climate and regions, and efficient and rapid propagation can be achieved all the year round. Meanwhile, the establishment of the system also lays a technical foundation for genetic transformation improvement, mutant screening breeding and the like of the Rhododendron pulchrum based on the callus.

The invention relates to the field of microbial genetic transformation, and discloses a preparation method of Ilyonectria fungus protoplast and a construction method of a genetic transformation system.The preparation method comprises the following steps that S11, Ilyonectria fungus hyphae are prepared, Ilyonectria fungi are cultured on a PDA solid culture medium for 14 d, then the Ilyonectria fungi are inoculated to a CMC liquid culture medium to be subjected to shake-flask culture for 4 d and filtered, filtrate is centrifuged for 10 min at the speed of 10000 rpm, and spores are collected; diluting the concentration of spores to 107 pieces / mL, inoculating the spores into a YEPD liquid culture medium, carrying out shake-flask culture for 2 days, and filtering and collecting hyphae; and S12, preparing the Ilyonectria fungus protoplast, namely adding the hypha prepared in the step S11 into the enzymatic hydrolysate, performing enzymolysis in a shaker at the temperature of 30 DEG C at the rotating speed of 150-200rpm for 3-5 hours, filtering, centrifuging filtrate at the temperature of 4 DEG C, discarding supernate, adding 0.7 M NaCl for centrifuging, and discarding the supernate, thereby obtaining the Ilyonectria fungus protoplast. The genetic transformation constructed by the invention can be used for exploring the specific action and mechanism of a specific pathogenic gene in the pathogenic process of Ilyonectria fungi through gene knockout or overexpression experiments.

The invention discloses a rhodosporidium toruloides strain with high homologous recombination efficiency as well as a construction method and application thereof, and belongs to the field of oil-producing yeast engineering bacteria. According to the rhodosporidium toruloides strain, the rhodosporidium toruloides strain NP11-SaCas9-1 is used as an original strain, and an RAD52 gene is over-expressed. The modified yeast engineering bacterium provided by the invention can realize high homologous recombination through CRISPR system mediation, the homologous recombination efficiency reaches 90-100%, a multi-gene editing platform can be constructed on the basis, and the modified yeast engineering bacterium can be applied to the fields of synthetic biology, metabolic engineering, genetic engineering and the like, so that the modified yeast engineering bacterium is simpler, quicker, more convenient and more accurate in the genetic modification process. And the homologous recombination activity of the rhodosporidium toruloides can be improved by expressing the homologous recombination related protein Rad52, and the genetic engineering strategy can be used for modifying other unconventional yeasts.

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 discloses a watermelon transgenic screening expression cassette and application thereof, and belongs to the technical field of biology. According to the invention, an expression cassette of a TagRFP gene driven by a d35s promoter is constructed on a pCambia1300 vector, and a recombinant vector is obtained. A recombinant vector is introduced into watermelon cells by utilizing an agrobacterium-mediated genetic transformation technology, and a red fluorescent protein TagRFP gene is integrated into a watermelon genome, so that transgenic watermelons can be screened and identified by taking red fluorescence as an identifier in the whole growth period of watermelon genetic transformation. Compared with a traditional screening method, the screening method has the advantages that the screening process is greatly simplified, and the transgenic watermelon and the non-transgenic watermelon are clearly distinguished, so that the genetic transformation efficiency of the watermelon is remarkably improved.

The invention belongs to the technical field of biology, and particularly relates to application of a soybean GmSRO5 gene in promoting generation of leguminous plant root nodules. A pZP211 overexpression vector is utilized, a transgenic plant is rapidly obtained through agrobacterium tumefaciens-mediated soybean genetic transformation, GmSRO5 is successfully overexpressed, root nodule number statistics is carried out on a chimera transgenic plant, and a result shows that the GmSRO5 gene can positively regulate and control the number of soybean root nodules and has a remarkable effect. The invention proposes that SRO5 can positively regulate and control the soybean nodulation process for the first time, and proves that the soybean GmSRO5 gene overexpression can significantly increase the number of soybean root nodules, and the soybean GmSRO5 gene overexpression can be applied to soybean production and molecular breeding through transgenosis, molecular marking and other means in the future, and has very important application value.

The invention relates to the technical field of plant genetic engineering, in particular to a gene of leymus secalinus participating in adversity stress and application of the gene. The gene LsHSFA participating in adversity stress in leymus secalinus is found for the first time, the nucleotide sequence of the gene LsHSFA is shown as SEQ ID NO.1, and the amino acid sequence of the coded protein is shown as SEQ ID NO.2. Through cloning and identification of the adversity stress participating gene of the leymus secalinus, expression analysis of the gene, genetic transformation of the gene and other means, the function of the gene is analyzed and verified, and the result shows that the adversity stress participating gene of the leymus secalinus has important influence on promotion of upper development of the leymus secalinus and improvement of heat resistance, salt resistance, drought resistance and other stress resistance; the stress-resistant gene of the leymus secalinus is utilized to positively regulate and control the abiotic stress function, so that the leymus secalinus can cope with various stress adversities better, the leymus secalinus can adapt to more complex and harsh ecological environments, degenerated and desertified grassland is improved, and potential application prospects are provided for improved variety breeding and excellent population construction of the leymus secalinus in the future.

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 belongs to the technical field of genetic engineering, and particularly relates to an agrobacterium rhizogenes-mediated visualized genetic transformation method for cucurbitaceae plants, which comprises the step of transfecting the cucurbitaceae plants by using agrobacterium rhizogenes containing recombinant plasmids, and the recombinant plasmids are expression cassettes containing Ruby reporter genes. A visual agrobacterium rhizogenes-mediated transformation system of the cucurbitaceae plants is established by utilizing the RUBY gene, professional equipment or chemical treatment is not needed, red hairy roots can be visually screened to identify and induce the positive plants, and the method is simple, rapid and efficient and can be used for identifying and inducing the positive plants of the cucurbitaceae plants such as bottle gourds and the like. And the gene is transferred into an exogenous gene or is subjected to gene editing on the cucurbitaceae plant, so that the gene has important significance.

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 provides a tissue culture method for inducing cluster buds of paeonia delavayi embryo, and relates to the technical field of tissue culture of paeonia delavayi. The tissue culture method mainly comprises the steps of explant acquisition, embryo germination culture, embryo cluster bud induction culture, cluster bud multiplication culture, cluster bud rooting culture, acclimatization and transplantation and the like. According to the method, the defects in the prior art are overcome, the embryos in fresh and mature Paeonia delavayi seeds in the current year are used as explants, a regeneration system of an embryo cluster bud induction way is preliminarily established through embryo cluster bud induction, proliferation and rooting culture medium research, and a foundation is laid for establishment of a Paeonia delavayi genetic transformation system.

A novel maize variety designated X18V354 and seed, plants and plant parts thereof are produced by crossing inbred maize varieties. Methods for producing a maize plant by crossing hybrid maize variety X18V354 with another maize plant are disclosed. Methods for producing a maize plant containing in its genetic material one or more traits introgressed into X18V354 through backcrossing or genetic transformation, and to the maize seed, plant and plant part produced thereby are described. Maize variety X18V354, the seed, the plant produced from the seed, and variants, mutants, and minor modifications of maize variety X18V354 are provided. Methods for producing maize varieties derived from maize variety X18V354 and methods of using maize variety X18V354 are disclosed.

The invention relates to the field of plant genetic engineering, and discloses application of a ZmWOX5b gene in improvement of corn genetic transformation efficiency. By separating the ZmWOX5b gene and constructing a recombinant vector, an expression cassette or a recombinant bacterium containing the gene, genetic transformation is carried out on corn inbred lines B73, B104, Zheng 58, FSJ115 and Xiang 249 by utilizing an agrobacterium-mediated method, and a research result shows that the gene can remarkably improve the transformation efficiency. Compared with a traditional method, the method has the advantages that the efficiency of easily-transformed genotypes is effectively improved, meanwhile, the transformation effect of difficultly-transformed maize inbred lines is greatly improved, and the limitation of receptor genotypes in maize genetic transformation is effectively broken through. The achievement obviously widens the application range of corn genetic transformation, and provides a new tool for efficiently developing gene function research and application of high yield, high quality, disease and pest resistance, stress resistance and the like of corn.

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 discloses a gene RBOHB and application of the gene RBOHB in regulating and controlling waterlogging stress to induce cucumber adventitious root formation, a novel gene RBOHB for inducing cucumber adventitious root formation is identified, the adventitious root formation capability of an edited mutant of the gene RBOHB is weakened, the extension of the adventitious root is obviously inhibited, and the adventitious root formation capability of an overexpressed mutant is enhanced. The gene RBOHB disclosed by the invention is used for regulating and controlling the waterlogging tolerance of cucumbers, and has important application value in screening and improving waterlogging-tolerant germplasm resources of the cucumbers and breeding waterlogging-tolerant varieties of the cucumbers. The cucumber is subjected to transgenic genetic transformation through a gene editing technology and a gene overexpression technology, the cucumber waterlogging tolerance regulation gene RBOHB and application of the cucumber waterlogging tolerance regulation gene RBOHB in waterlogging tolerance cucumber varieties are provided, and the cucumber waterlogging tolerance regulation gene RBOHB has important application value in the field of breeding of resistant cucumbers.

The invention belongs to the technical field of genetic transformation, and particularly relates to application of tomato Halide linker dissociation enzyme, a method for improving crop bacterial wilt resistance and a method for inhibiting pathogenic bacteria through protein. The invention provides application of a tomato Halide linker dissociation enzyme. The amino acid sequence of the tomato Halide linker dissociation enzyme SlMOC1 is as shown in SEQ ID NO: 2. The nucleotide sequence of the coding gene SlMOC1 is as shown in SEQ ID NO: 1. The result of the embodiment shows that the bacterial wilt resistance of the plant can be improved by overexpression of the tomato Halide linker dissociation enzyme SlMOC1 gene.

A novel maize variety designated X08V033 and seed, plants and plant parts thereof are produced by crossing inbred maize varieties. Methods for producing a maize plant by crossing hybrid maize variety X08V033 with another maize plant are disclosed. Methods for producing a maize plant containing in its genetic material one or more traits introgressed into X08V033 through backcrossing or genetic transformation, and to the maize seed, plant and plant part produced thereby are described. Maize variety X08V033, the seed, the plant produced from the seed, and variants, mutants, and minor modifications of maize variety X08V033 are provided. Methods for producing maize varieties derived from maize variety X08V033 and methods of using maize variety X08V033 are disclosed.

The invention discloses application of a transcription factor BnaMYB52 in regulation and control of drought resistance of rape, and relates to the technical field of biology. The amino acid sequence of the transcription factor BnaMYB52 is as shown in any one of SEQ ID NO. 5 to 8. According to the invention, the negative regulation effect of the transcription factor BnaMYB52 in the drought resistance of the rape is disclosed for the first time, and a brand new molecular target and a genetic improvement strategy are provided for improving the drought resistance of the rape. Through in-depth study, it is found that knockout of the BnaMYB52 gene can significantly enhance the tolerance of rape plants to the drought environment. In addition, a new drought-resistant rape germplasm obtained after genetic modification of the gene shows excellent adaptive capacity in a drought stress environment. The technical breakthrough not only helps to solve practical problems faced by rape planting in arid regions, but also lays a solid theoretical foundation and technical support for cultivating high-yield and high-quality rape varieties adapting to extreme climate conditions.

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 belongs to the technical field of plant biology, and particularly relates to a genetic transformation method based on oat immature embryos, which comprises the following steps: preparing an infection solution containing a target gene; taking immature caryopses of the oats 21-23 days after pollination; infecting the immature embryos with infection liquid, and co-culturing the immature embryos after infection is completed; after co-culture is completed, immature embryos are sequentially transferred into an L3-T culture medium and an L3-B5 culture medium to induce formation of calluses, and after the calluses are formed, the calluses are sequentially induced to be differentiated, rooted and hardened. According to the method, efficient genetic transformation of exogenous genes is realized by selecting proper oat immature embryos, proper culture conditions and proper transformation conditions. By means of the genetic transformation method, the genetic transformation efficiency of the oats is improved to 25% from 2%-3%. And meanwhile, the immature embryo is used as a transformation acceptor material, and the complete plant can be efficiently and stably regenerated by the method disclosed by the invention.

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 relates to application of a GhRPS20B gene in improvement of insect resistance of cotton bollworm and insect-resistant breeding, and belongs to the technical field of agricultural biology. The invention provides an application of a GhRPS20B gene as shown in SEQ ID NO.1 in improving the disease resistance of cotton bollworm diseases of cotton. Experiments show that the defense mechanism of jasmonic acid (JA) in cotton can be activated by enhancing the transcriptional level of the GhRPS20B gene in a plant body. The GhRPS20B gene is over-expressed in cotton through an agrobacterium tumefaciens-mediated genetic transformation technology, so that the expression level of the GhRPS20B gene in a target plant is enhanced, the plant with enhanced resistance can be effectively cultivated, and the natural defense capability of the plant to cotton bollworms is improved.

The invention discloses lnc.3024.2-GbMYB2 for regulating and controlling synthesis of gingko flavonoids and application of the lnc.3024.2-GbMYB2, and belongs to the technical field of genetic engineering, aiming at analyzing the biological functions of lncRNA (lnc.3024.2) and MYB target gene (Gb06719) participating in gingko flavonoids anabolism, by utilizing the technical means of bioinformatics analysis, real-time fluorescence quantification, subcellular localization, prokaryotic expression, genetic transformation and the like, the GbMYB2 can be used for regulating and controlling synthesis of the gingko flavonoids. The regulation mechanism of the transcription factor lnc.3024.2 and the target gene GbMYB2 of the lnc.3024.2 participating in the synthesis of the flavonoids in the ginkgo leaves is discussed. And a gene regulation network model of gingko flavonoid anabolism is further enriched. On the basis, the molecular mechanism of regulating and controlling the synthesis of the gingko flavonoids by the lncRNA targeting MYB is further analyzed on the early-stage basis, the content of the gingko flavonoids is increased by utilizing a genetic engineering technical means, and a new way is provided for researching the synthesis way of the gingko flavonoids.