38 results about "Somatic Cell Hybridization" patented technology

The provided specific site can integrates exogenous gene to stable express and affect little to the acceptor. Besides, it also provides a composite gene structure, which can be integrated by rice gene DNA sequence near specific site and DNA sequence of exogenous gene, and be used to culture new rice series by sexual hybridization and somatic hybridization.

The invention belongs to the technical field of plant gene engineering, in particular to a modified glyphosate-resistant gene and a glyphosate-resistant rice cultivation method and comprises modification of glyphosate genes, building of plant expression vectors of the glyphosate-resistant gene and a glyphosate-resistant transgenosis rice cultivation method. Artificially-synthesized modified glyphosate-resistant gene serves as a target gene, the sequence of the gene is as shown in SEQ ID NO: 1, and the target gene is led into rice receptors by an agrobacterium tumefaciens mediated transformation method to obtain high glyphosate-resistant transgenosis rice GT28. the glyphosate-resistant gene of GT28 is recombined by the aid of sexual hybridization and somatic hybridization technique to obtain new rice germplasm, and new glyphosate-resistant varieties (strains) of rice are cultivated.

The invention discloses a rapid propagation method for suspension cell culture of Millettia speciosa Champ. According to the rapid propagation method, young and tender cotyledons taken from Millettia speciosa Champ pods 50-60 days after flowering are utilized as culture materials, subcultured callus tissue is obtained through inducing culture and subculture, and a stable Millettia speciosa Champ suspension cell culture fluid is obtained through the suspension cell culture. With the adoption of the method, stable Millettia speciosa Champ suspension cells with good quality and high propagation yield can be obtained, and a novel way and method are provided for development and utilization of effective components of the Millettia speciosa Champ and creation of new germplasm resources from the Millettia speciosa Champ with biotechnology methods such as somatic hybridization, genetic transformation, mutant induction and the like.

The invention relates to a preparation method of catalpa fargesii leaf blade protoplasts. The catalpa fargesii leaf blade protoplasts are obtained by controlling the ratios and the concentrations of cellulase Onozuka RS and Macerozyme R-10, the concentration of mannitol in enzymatic hydrolysate, enzymolysis temperature, enzymolysis time and other factors. Free protoplasts obtained by the method disclosed by the invention are high in quantity and vitality, and can lay a foundation for researching catalpa fargesii protoplast culture, plant regeneration, genetic transformation, somatic hybridization, protoplast fusion breeding and the like.

The invention provides a cultivating method of a new species of an anti-late blight and anti-beetle potato, relating to a cultivating method of a new species of the potato with a cell culture technology. The method comprises the following steps of: electrically fusing a protoplast (A) which has anti-late blight gene, anti-beetle gene and anti-oxidation gene, and is separated from the wild species of the potato, with a protoplast (B) of the potato culturing species with wide adaptability to form into a fusion cell with the protoplast A and the protoplast B; generating a callus by means of cell culture, and generating a hybrid test tube plantlet of a regenerated somatic cell by means of induction; and further culturing the tissue culture test tube plantlet, transplanting the cultured tissue culture test tube plantlet into a glasshouse or a land for growing field crops, and generating a test tube first generation somatic cell hybridized potato to obtain the anti-diseases and insect cell hybrid potato, wherein the somatic cell hybridized potato has the good agronomic trait of the potato cultivating species and the anti-late blight and anti-beetle trait. The cultivating method provides a new available gene resource which can be used for improving the quality of the potato and improving the antioxidant activity of the potato cultivating species with the wild species.

The invention discloses a rubber tree protolast culture plant regeneration method which comprises the following steps of: establishing an embryonic cell suspension system, separating and purifying protoplast, nursing and culturing the protoplast, carrying out mulitiplication in protoplast regeneration cell clone, inducing somatic embryos and regenerating plants. The rubber tree protolast culture plant regeneration method disclosed by the invention is simple and easy to carry out, the components of a nursing and culture medium employed to nursing and culturing the protoplast are greatly simplified, and the suspension cell systems of the varieties of the rubber tree self are used as the nursing cells, so that the production cost is greatly lowered, the efficiency that the rubber tree protolast is split to form cell clone and the efficiency that the somatic embryos are generated subsequently are effectively improved. A good receptor system is provided for rubber tree variety improvement and quality innovation by utilizing biotechnological breeding modes with significant application prospects, such as protoplast fusion (namely somatic hybridization) and genetic transformation.

The invention belongs to the technical field of rice genetic engineering, and particularly relates to a cultivating method for transgenic lepidoptera pest resistant rice. The method comprises the following steps: using an artificially synthesized novel pesticidal gene cry1C as a target gene, wherein the nucleotide sequence is shown as SEQ ID NO: 1; using a bar gene as a selective marker gene and adopting an agrobacterium mediated genetic transformation method to introduce the target gene into rice excellent restorer minghui 63 to acquire a novel transgenic rice material T1C-19 highly resisting rice lepidoptera pests. Besides, a pest-resistant gene of T19-1 can be recombined into a new rice germplasm through sexual hybridization and somatic hybridization technologies to cultivate a new rice pest resistant product. The invention discloses the verification tests of processes of transformation vecter construction, rice genetic transformation, molecular identification of transformed plants, insertion site sequence analysis, indoor and field pest resistant experiments and the like.

The process of transferring Tibetan medicinal effective component into receptor cell by means of somatic cell cross breeding technology includes the following main steps: 1) selecting parent material gene types; 2) dissociating the receptor protoplaster; 3) UV irradiating donor protoplaster; 4) fusing the donor protoplaster and the receptor protoplaster and culturing; 5) identifying hybrid callus and regenerated plant; and 6) detecting target characteristic in somatic cell hybrid. The present invention obtains cell line with high medicinal component content and fast growth by means of somatic cell cross breeding technology and may be used in obtaining famous and rare Tibetan medicinal effective component.

The invention discloses a seaweed breeding method based on an induction polyploid gametocyte, which is characterized by comprising the following steps: firstly processing haploid seaweed gametophyte cells by utilizing mitosis hinder agent to lead the chromosome in the cells to be doubled, so as to induce the haploid seaweed gametophyte cells into polyploid seaweed gametophyte cells, and then breeding polyploid seaweed thalluses by utilizing a seaweed gametophyte cell hybridization technique. The method of the invention has the advantages that the seaweed gene content can be improved through polyploid breeding; and the seaweed physiological activity can be strengthened by the increased quantitative character gene, so as to improve economic character thereof. The chromosome doubling method of the invention can be applied for breeding homology polyploids and allopolyploids.

The invention discloses a method for establishing a hevea brasiliensis suspension cell line. The method comprises material selection and disinfection, suspension culture induction process for the callus tissue and a subculture screening culture process. The method for rapidly establishing the hevea brasiliensis suspension cell line, which is provided by the invention, is simple in technological process and relatively short in time consumption, is capable of establishing a stable, uniform, good-dispersity and rapid-growth suspension cell line within only 60-90 days which is reduced by at least a half in comparison with that spent in establishing the hevea brasiliensis suspension cell line by use of a conventional method, greatly lowering the production cost, shortening the experiment period and rapidly providing favorable material sources for technical systems of synchronously inducing regeneration plants from a batch of somatic embryos, or performing protolast culture and somatic hybridization and performing direct conversion on foreign genes by utilizing protoplast as a receptor, and the like.

The invention relates to a plant in-vivo somatic cell hybridization method which comprises the following steps: selecting a female organ of a good variety of a plant type expected to be obtained and a somatic cell cluster of a reproduction unit as a hybridization female parent receptor, performing treatment before insemination to remove reproduction isolation, so that the hybridization female parent receptor is easy to fertilize; individuals, organs, tissues, cells, chromosomes or chromosome segments, DNA or DNA segments and genes of animals or plants, microorganisms and other organisms with the characters of the expected variety are selected as male parent donors, a somatic cell hybridization agent is added and stirred evenly, a sperm is obtained, the sperm is used for insemination of a receptor under mediation of the somatic cell hybridization agent, and the variety is obtained. And embryo rescue and seedling culture are carried out after the fertilization embryo is taken back. The bottleneck of distant hybridization is broken through, high-yield and high-quality varieties can be cultivated, and the method has wide application prospects.

The present invention discloses a sterile line rapid transformation breeding method, and sterile line rapid transformation breeding is performed through combining of somatic cell hybridization and molecular marker selection. The sterile line rapid transformation breeding method belongs to the technical field of crop breeding. First, donor (sterile) cytoplasm and receptor (fertile) protoplast are separated; then iodoacetamide (IOA) is used for processing the receptor protoplast for inactivation of the cytoplasm, then the donor cytoplasm and receptor protoplast are mixed in the ratio of 2: 1 (v / v ), an electric fusion method or a chemical fusion method is used for donor and receptor fusion; and finally, under appropriate culture conditions, the fusion body is regenerated, a sterile regenerated plant with genotype and phenotype consistent with that of the receptor is selected, and the plant which can seed normally after pollinated by receptor pollen is a receptor sterile line.

The invention disclose a method for increasing the production efficiency of hybrid maize seeds. The method includes the steps of S1, culturing maize seeds to obtain maize calluses; S2, separating themaize calluses into a maize protoplast solution; S3, removing the supernate of the maize protoplast solution, and performing precipitation and resuspension; S4, adding maize protoplast into PEG1000, slowly adding 9.5ml of Hanks solution into the test tube 90 seconds later, standing the test tube in a water bath pot, centrifuging and abandoning supernate, adding 1ml of cell culture liquid into thecell precipitates, gently evenly mixing, and culturing in water bath of 37 DEG C for 30 minutes to obtain activated maize protoplast fused cells; S5, using a plant in-vitro tissue culture method to culture the maize protoplast fused cells into regenerated hybrid maize plants. The method has the advantages that maize seeds are bred through a somatic hybridization method, breeding cycle is shortenedgreatly, breeding quality is increased, breeding time is shortened, and the targeting performance of breeding objects is increased.

The invention belongs to the technical field of plant somatic cell hybridization, in particular to a method for electrofusion of sugarcane protoplasts, comprising the following steps: (1) preparationof parent protoplasts; (2) protoplast inactivation; (3) the addition of protoplast fusion inducer: agglutinating and binding plants to the surface of one parent protoplast and binding biotin to the surface of another parent protoplast; (4) protoplast fusion. The invention utilizes the principle that plant lectin can combine with biotin, plant agglutination and biotin binding to the surface of thetwo parent protoplasts. The two parental protoplasts could be fused by lectin and biotin to improve one-to-one fusion rate. The hybrid progeny possessed the genes of ROC22 sugarcane with high yield, high sugar content and high drought tolerance, and also possessed the genes of GT28 sugarcane with high sugar content and cold tolerance.

The invention belongs to the technical field of plant somatic cell hybridization, in particular to a method for sugarcane somatic cell hybridization, comprising the following steps: (1) preparation ofparent protoplast; (2) protoplast inactivation; (3) the addition of protoplast fusion inducer: one parent protoplast was cultured in silicate solution and mixed with another parent protoplast to obtain mixed protoplast; (4) fusion of protoplasts: electrofusion of the mixed protoplasts obtained in step (3). The invention utilizes the principle that silicon has high affinity with glycolipid on thesurface of plant cell membrane, The two parent protoplasts fused under the induction of silicon to improve the one-to-one fusion rate. The hybrid progenies had the genes of ROC22 sugarcane with high yield, high sugar content and high drought tolerance, and had the genes of GT28 sugarcane with high sugar content and cold tolerance.

The invention discloses an application that low celiac disease epitope coding DNA wheat can be created using couch grass with long ear. The application screens out distant grass of couch grass with long ear for low celiac disease epitope coding DNA wheat using a homologous sequencing; gene group or DNA of distant grass part is transferred into wheat using somatic cell hybridization technology before low celiac disease epitope coding DNA wheat somatic hybrid is screened out using the homologous sequencing. The somatic hybrid can be used for breeding work of low celiac disease epitope wheat and lay the foundation for food problem during therapy period for celiac disease patients.

The invention discloses a method for preparing high-quality rehmannia protoplast, which comprises the following steps: (1) rehmannia leaf pretreatment: taking rehmannia seedling leaves, cutting the rehmannia seedling leaves into thin strips, and soaking the thin strips in a buffer solution in a dark place; (2) enzymolysis: transferring the pretreated rehmannia leaves into an enzymatic hydrolysate, and oscillating in a dark place for enzymolysis; (3) protoplast separation; (4) purifying the protoplast: resuspending the separated protoplast by using a buffer solution; adding the resuspended liquid to the upper layer of the sucrose salt solution, and centrifuging; after centrifugation, the middle layer is a protoplast band. The method is simple, the prepared protoplast is regular in shape, complete in structure, high in living protoplast content and good in quality, and a foundation is laid for research in the fields of heredity and molecular biology, such as genetic transformation, rapid propagation, somatic cell hybridization and breeding of good varieties by using the rehmannia protoplast as an experimental material.

The invention provides a method for preparing amaranthus protoplast. The method is characterized in that tender true leaves of amaranthus sterile seedlings are used as materials, filter residues are extruded during filtration after enzymolysis, a large number of amaranthus protoplast with normal form, few impurities and no damage is prepared, the experimental requirements of subsequent conversion, subcellular localization and the like can be met, and important technical support can be provided for fundamental researches and application researches such as dual-fluorescence complementation, somatic cell hybridization and genetic transformation, so that the development of genetic breeding and fundamental researches of the edible amaranthus is promoted.