457 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 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 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 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 belongs to the technical field of plant genetic engineering, and particularly discloses application of an important member of a potato mitogen-activated protein kinase (MAPK) family, namely an StMKK9 gene in regulation and control of maturity and cold resistance of potatoes. CDNA of a cultivar Desiree leaf is used as a template, the potato StMKK9 gene is separated and cloned, an over-expression vector is constructed, and the over-expression vector is used for expression of the potato. Genetic transformation of the potatoes is further carried out, it is found that the overexpressed StMKK9 gene can promote prematurity of the potatoes, meanwhile, the cold resistance of potato plants can be improved, and a new genetic material and a theoretical basis are provided for analysis of a potato tuberization molecular mechanism and improved breeding of the potatoes.

The invention provides application of a potato StMYB4 gene in regulation and control of solanine accumulation in potato tubers, the StMYB4 gene is cloned through a sequence published in a potato genome database by taking cDNA of leaves of cultivated species Hubei potato No.3 and AC142 as a template, an overexpression and knockout vector is constructed, genetic transformation is performed on the Hubei potato No.3 and AC142 respectively for transgenic function verification, and the solanine accumulation in the potato tubers is regulated and controlled. It is found that excessive StMYB4 genes can significantly promote synthesis of solanine in potato tubers, and knockout of the StMYB4 genes can significantly inhibit synthesis of solanine in potato tubers. The application of the potato StMYB4 gene to regulation and control of tuber solanine can be used for molecular marker development and low solanine variety breeding.

The invention relates to the technical field of plant genetic engineering, in particular to application of an algae NySAT2 gene in regulation and control of alfalfa crude protein and detergent fiber. According to the invention, a sequence after codon optimization of an NySAT2 gene coding region is obtained by using gene synthesis and codon optimization technologies, an algae NySAT2 gene overexpression vector is constructed, a target gene is introduced into an alfalfa genome by using an agrobacterium rhizogenes mediated genetic transformation method, and a transgenic plant is obtained by using a genetic transformation system. Molecular identification and phenotypic analysis show that on the premise of keeping normal growth vigor (plant height and biomass have no significant difference from those of a wild type), the crude protein content of a medicago sativa plant with the overexpressed NySAT2 gene is remarkably increased compared with that of the wild type medicago sativa plant, and meanwhile, the contents of acid detergent fibers and neutral detergent fibers are remarkably reduced, so that the content of the crude protein in the medicago sativa plant with the overexpressed NySAT2 gene is remarkably increased. The forage quality and the feeding value of the medicago sativa are effectively improved.

This invention discloses the application of the BnaTOE1 gene in regulating the flowering time of Brassica napus and its molecular marker primers for identification. The DNA nucleotide sequence of the BnaTOE1 gene is shown in SEQ ID NO.1, with a sequence length of 1254 bp, and the encoded amino acid sequence is shown in SEQ ID NO.2. The molecular marker primers used to identify the flowering time of Brassica napus are shown in SEQ ID NO.15 and SEQ ID NO.16. Functional analysis of genetic transformation in Brassica napus shows that BnaTOE1 participates in regulating the flowering time of Brassica napus. The development of molecular markers is of great significance for promoting the genetic improvement and breeding of Brassica napus.

The invention relates to the technical field of plant genetic engineering, in particular to a ZmbHLH166 gene related to drought resistance in the flowering stage of corn, an encoding protein and application of the ZmbHLH166 gene. The key effect of the ZmbHLH166 gene in the drought resistance of the corn in the flowering period is identified for the first time on the basis of whole genome association analysis (GWAS) and transgenic function verification, and through experiments, through genetic transformation and continuous selfing screening of a Ubi1: ZmbHLH166 overexpression vector, the obtained T2-generation corn strain of the overexpression ZmbHLH166 gene has the advantages that compared with a wild type corn strain, the yield is increased, and the yield is increased. The ZmbHLH166 gene has a shorter pollen scattering-spinning interval, which indicates that under drought stress, the overexpression of the ZmbHLH166 gene can effectively promote the synchronous development of male and female flowers of corn, thereby enhancing the drought resistance of crops. Therefore, the application of the ZmbHLH166 gene and the protein coded by the ZmbHLH166 gene in regulating and controlling the drought resistance of the corn in the flowering period is feasible, and a new and reliable gene resource and an improvement direction are provided for drought-resistant breeding of the corn.

The invention discloses application of a tea tree CsGT-3b gene in improving the disease resistance of plants and a method of the tea tree CsGT-3b gene. The CsGT-3b gene is a Trihelx transcription factor family gene, the CsGT-3b gene is constructed to a plant expression vector and is transferred into tobacco, a transgenic tobacco plant is obtained through stable genetic transformation, and the CsGT-3b gene shows relatively strong fungal inhibitory activity on botrytis cinerea inoculation; according to the present invention, the CsGT-3b gene on the tea tree leaf is subjected to instantaneous silencing by using the antisense oligonucleotides (AsODNs) technology, such that the disease resistance of the plant on the lasiodiplodia theobroma is significantly weakened, and the CsGT-3b gene on the tea tree leaf is subjected to the enzyme-linked immunosorbent assay so as to significantly reduce the disease resistance of the plant on the lasiodiplodia theobroma; therefore, the CsGT-3b gene has the effect of improving the disease resistance of the plants, can be used as a disease-resistant gene, can improve the disease resistance of the plants by being introduced into the plants such as tobacco, tea trees or vegetables, and has a wide market application prospect.

The invention relates to the technical field of plant genetic engineering, and discloses a limonium bicolor gene LbYAB1 and application thereof. The new function of the LbYAB1 is found, firstly, the expression of the LbYAB1 in salt glands and leaf primordia is proved through RNA in-situ hybridization and a promoter driven GUS tissue staining technology, and the expression of the LbYAB1 is induced by treatment of 6-benzylaminopurine. Secondly, a genetic transformation technology is updated, a stable overexpressed LbYAB1 strain is obtained, and by exploring the LbYAB1 function of an overexpressed and silent strain, it is found that compared with a wild type, development of salt glands of the overexpressed strain is inhibited, the salt secreting capacity is weakened, and the salt tolerance is reduced; a silent strain shows an opposite phenotype, and a result proves that the LbYAB1 is a negative regulation factor for salt gland development. Overexpression of the gene in limonium bicolor and arabidopsis thaliana both leads to enhancement of salt sensitivity. The invention not only lays a foundation for further exploring and revealing the development mechanism of salt glands, but also opens up a new path for cultivating salt-tolerant crops.

The application discloses application of SbWOX gene and SbBBM gene in corn breeding and a corn genetic transformation method, and relates to a bioengineering technology.The application provides a nucleic acid molecule with co-expression of SbWOX protein gene and SbBBM protein gene, and corresponding gene co-expression vectors and microbial transformants.The application also provides application of the co-expression of the SbWOX gene and the SbBBM gene in the corn genetic transformation method.The application obtains corresponding genes and mutant proteins based on the SbWOX gene and the SbBBM gene of sorghum, the two genes are co-expressed in the corn genetic transformation process, even without adding plant growth hormones in the antibiotic screening medium in the tissue culture process, the plant genetic transformation process can be normally completed, and the transformation efficiency and the regeneration efficiency of the corn can be remarkably improved, the transformation rate reaches 10.42%, and the differentiation rate reaches 32.89%.

A novel maize variety designated X05T063 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 X05T063 with another maize plant are disclosed. Methods for producing a maize plant containing in its genetic material one or more traits introgressed into X05T063 through backcrossing or genetic transformation, and to the maize seed, plant and plant part produced thereby are described. Maize variety X05T063, the seed, the plant produced from the seed, and variants, mutants, and minor modifications of maize variety X05T063 are provided. Methods for producing maize varieties derived from maize variety X05T063 and methods of using maize variety X05T063 are disclosed.

This invention discloses the application of the GhATG18a gene in accelerating callus induction in cotton. The invention constructs a pCAMBIA2300-GhATG18a overexpression vector from the full-length CDS fragment of this gene, then introduces it into Agrobacterium, and transforms cotton using Agrobacterium-mediated transformation to obtain transgenic lines. qRT-PCR results show that, compared with the CCRI24 control material, the expression level of the GhATG18a gene in the overexpression lines OE-GhATG18a-1 and OE-GhATG18a-2 is significantly upregulated. Phenotypic observation and histological sections both show that the OE-GhATG18a line overexpressing this gene accelerates the process of somatic cell dedifferentiation and callus induction in cotton, thereby improving the genetic transformation efficiency of cotton. Therefore, overexpressing this gene and applying it to commercial cotton varieties that are difficult to genetically transform can help accelerate the callus induction process, thereby improving the genetic transformation efficiency and providing technical support for breeding high-yield and stress-resistant high-quality cotton germplasm.

The invention belongs to the technical field of microbial genetic transformation, and particularly relates to a method for high-yield production of pleurotus ostreatus transformants, a genetic transformation system and application of the genetic transformation system. Through optimization of protoplast preparation conditions, the number of the obtained protoplast is 4.95 * 10 < 8 > CFU.mL <-1 >, and the regeneration rate of the protoplast in an OM-YEPS regeneration culture medium is 12.7%. A method for producing high-yield pleurotus ostreatus transformants is researched by using a PEG-mediated protoplast transformation method, a set of efficient and stable genetic transformation and screening method is established, about 200-300 positive transformants with stably expressed exogenous genes are obtained, and the transformation efficiency is 65.38%. The genetic transformation system of the high-yield Pleurotus ostreatus transformant, constructed by the invention, lays a solid technical foundation for subsequent gene function verification, metabolic engineering transformation and molecular breeding with excellent characters, and further accelerates the biological research and industrial application process of Pleurotus ostreatus and even other macro fungi.

This invention discloses a genetic transformation and gene editing method applicable to multiple sesquiterpene species, belonging to the field of plant genetic transformation and gene editing technology. The method uses the cotyledonary nodes of sterile seedlings of the sesquiterpene genus as explants, and introduces an optimized pScEF1α-Cas9 gene editing transformation vector using Agrobacterium infection. Regenerated plants are obtained through recovery culture, selection culture, and rooting culture. The vector contains four gene expression units: pScEF1α:Cas9, pGmU6:sgRNA, pGmUBI:DsRed2, and pCaMV35S:HygR. In vivo screening is further performed using the DsRed2 fluorescent reporter gene, and the gene-edited plants are identified by molecular detection. This method is applicable to common sesquiterpene, stem-nodled sesquiterpene, and spiny sesquiterpene, and has advantages such as strong species versatility, high transformation efficiency, short cycle, visualized screening, and high positive rate, providing efficient technical support for gene function research and genetic improvement of sesquiterpene species.

This invention discloses a method for establishing a high-efficiency leaf regeneration system for aseptic seedlings of Populus tomentosa 'Bofeng No. 3', belonging to the field of agricultural biotechnology. This invention, through 5-AU combined with dark culture treatment, not only induces early leaf differentiation in aseptic seedlings of Populus tomentosa 'Bofeng No. 3', but also significantly increases the number of adventitious buds and the rooting rate. Through chemical treatment and optimized culture conditions, this invention successfully established a high-efficiency plant regeneration system for aseptic leaves of Populus tomentosa 'Bofeng No. 3', laying the foundation for the genetic transformation and gene editing of Populus tomentosa 'Bofeng No. 3', and also providing a reference for the establishment of tissue culture systems for other tree species.

The present disclosure provides an application method in rice callus differentiation based on an Oryza sativa Leafy Cotyledon 1 (OsLEC1) gene, including the steps of: selecting guide ribonucleic acid (gRNA) target sites; cloning tandem fragments including gRNA; ligating each gRNA fragment; performing polymerase chain reaction (PCR) amplification on a ligation product; performing enzyme digestion on the purified product and a target vector; transforming the ligated vector; performing Agrobacterium-mediated genetic transformation of rice; and screening and identifying transgenetic plants. In the present disclosure, the differentiation of callus directly affects the emergence efficiency of transgenic plants. The knockout of OsLEC1 can promote the differentiation of rice callus, suggesting that OsLEC1 can serve as an important target gene for improving the transformation efficiency of rice and even gramineous crops. OsLEC1 can be used as a starting point to construct various molecular tools to enhance transformation efficiency.

A novel maize variety designated X08T074 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 X08T074 with another maize plant are disclosed. Methods for producing a maize plant containing in its genetic material one or more traits introgressed into X08T074 through backcrossing or genetic transformation, and to the maize seed, plant and plant part produced thereby are described. Maize variety X08T074, the seed, the plant produced from the seed, and variants, mutants, and minor modifications of maize variety X08T074 are provided. Methods for producing maize varieties derived from maize variety X08T074 and methods of using maize variety X08T074 are disclosed.

The invention belongs to the technical field of genetic transformation, and particularly relates to a method for improving apple leaf disc regeneration and genetic transformation efficiency. And culturing the apple leaf disc in the apple leaf disc regeneration culture medium to obtain a leaf disc regeneration adventitious bud. The amino acid sequence of the MdREF1 small peptide is as shown in SEQ ID NO. 1. The apple homologous peptide MdREF1 is identified in an NCBI (National Center of Biotechnology Information) database on the basis of AtPROPEP1-8 and AtPep1-8 sequences of arabidopsis thaliana. The synthetic peptide of the apple MdREF1 is added into a basic regeneration culture medium, so that the leaf disc callus growth amount and the adventitious bud differentiation number can be remarkably increased. Wherein by adding 50nmol / L of MdREF1 peptide, the regeneration frequency can be improved by 1.74 times, the average regeneration bud number can be increased by 2.16 times, and the conversion efficiency can be improved by 3.14 times.

A novel maize variety designated X87T010 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 X87T010 with another maize plant are disclosed. Methods for producing a maize plant containing in its genetic material one or more traits introgressed into X87T010 through backcrossing or genetic transformation, and to the maize seed, plant and plant part produced thereby are described. Maize variety X87T010, the seed, the plant produced from the seed, and variants, mutants, and minor modifications of maize variety X87T010 are provided. Methods for producing maize varieties derived from maize variety X87T010 and methods of using maize variety X87T010 are disclosed.

The application discloses a method for improving genetic transformation rate of high-altitude meadow grass, comprising the following steps: (1) taking mature seeds of high-altitude meadow grass as explants to induce callus, and obtaining the callus; (2) adding a surfactant Silwet L-77 with a mass fraction of 3‰ of the bacterial liquid to the agrobacterium bacterial liquid carrying a hygromycin resistance gene, and obtaining a culture bacterial liquid; (3) placing the regenerated plant in a screening culture medium containing 40 mg / L of hygromycin resistance for culture, screening the survived plants, and obtaining the genetically transformed plants. The method can improve the transformation efficiency of the high-altitude meadow grass by about 4.54 times through adding the surfactant in the genetic transformation process and cooperating with the screening of the obtained differentiation culture medium.

The invention belongs to the technical field of plant genetic engineering, and relates to a GhAL5 gene and application thereof in cultivation of heat-resistant cotton and rice germplasm. Researches find that expression of the AL5 gene is up-regulated by using an agrobacterium tumefaciens-mediated genetic transformation technology, so that heat resistance of cotton and rice can be enhanced, plant anther fertility can be enhanced, and leaf damage degree can be reduced. The invention provides a new approach and method for improving the heat resistance of cotton and rice, and has wide agricultural application prospect and market value.

This invention discloses a method for inducing and culturing burdock stem cells and stem cells, belonging to the field of plant biotechnology. Using burdock roots as explants, the cambium is determined by staining, and the cambium is obtained using enzymatic digestion. The cambium is then inoculated into a specific ratio of induction solid medium and cultured in the dark to efficiently induce primary stem cells. These primary stem cells are then transferred to a proliferation solid medium for subculture and selection to obtain and maintain a vigorous, soft stem cell line. The solid stem cell line is further transferred to a liquid medium for shaking culture to establish a suspension stem cell line. This invention provides high induction efficiency, stable stem cell characteristics, and good reproducibility, offering a high-quality starting material and technical platform for in vitro preservation of burdock germplasm, large-scale production of secondary metabolites, and genetic transformation.

The invention relates to a non-tissue-culture-dependent hevea brasiliensis high-altitude layering genetic transformation method, and relates to the technical field of plant genetic engineering. The method comprises the following steps: S1, providing an agrobacterium rhizogenes K599 bacterial solution containing a target gene, and loading the target gene in a pCAMBIA2301 vector; s2, preparing an infection material (infection liquid or lawn) by using the agrobacterium rhizogenes K599; s3, carrying out girdling treatment on stems of the rubber tree seedlings, and carrying out pretreatment on girdling wounds; s4, the pretreated girdling wound is infected with the infection material; s5, covering and mounting the girdling wound with a wet culture medium so as to keep the wound wet and incubate the strain; wherein the target gene is a GUS gene, an RUBY gene or a JcFT gene. The method can avoid a series of complicated operations such as tissue culture, and is simple, convenient, rapid, low in cost, not limited by seasons, easy to operate and independent of genotypes; particularly, by means of the method, the high positive rate can be obtained.

The invention discloses a method for realizing efficient in-vitro regeneration of tea tree cotyledons through a somatic embryo indirect generation way, and belongs to the technical field of plant tissue culture. The tea tree somatic embryo indirect generation system is successfully constructed by screening culture conditions, a culture medium formula and other methods, and a foundation is laid for construction of a tea tree genetic transformation system and industrialized seedling production; according to the method, immature cotyledons of tea trees are used as explants, the cotyledons are induced to generate calluses, then the calluses are proliferated and differentiated to generate somatic embryos, and the somatic embryos are further differentiated to form complete plants, so that efficient and synchronous propagation of tea tree seedlings is realized, and technical support is provided for high-quality variety propagation and new variety breeding of tea trees; the problems of low propagation coefficient, long growth cycle, serious descendant degeneration and the like in traditional seed propagation of tea trees are solved, and the technical bottlenecks of serious explant browning, high bacterial contamination rate, poor plant growth synchronism and the like in other tissue culture technologies are solved.

This invention relates to a rapid induction culture medium and method for inducing bud formation from kiwifruit leaves to complete plants, belonging to the field of biotechnology. The callus induction culture medium for rapidly inducing bud formation from kiwifruit leaves includes MS medium, sucrose, 6-benzylaminopurine, naphthaleneacetic acid, thidiazuron, and agar, with the following weight ratios: MS medium 0.002%-0.006%, sucrose 0.01%-0.05%, 6-benzylaminopurine 0.000001%-0.000003%, naphthaleneacetic acid 0.0000002%-0.0000008%, thidiazuron 0.000001%-0.000003%, agar 0.006%-0.01%, and the balance being water. The formula for the callus induction bud culture medium is as follows: MS medium 0.002%-0.006%, sucrose 0.01%-0.05%, 6-benzylaminopurine 0.0000005%-0.000003%, naphthaleneacetic acid 0.00000005%-0.0000002%, agar 0.006%-0.01%, with the balance being water. This invention provides a rapid induction culture medium and method for kiwifruit leaves to complete plants, effectively inhibiting problems such as callus waterlogging, browning, and vitrification. It not only increases the number of buds but also improves the quality of buds, which is beneficial to the growth and differentiation of callus tissue, making it easier for it to differentiate into buds, thus laying a solid foundation for subsequent genetic transformation experiments.