66results about How to "High rate of callus induction" patented technology

The invention belongs to the technical field of plant biology, in particular relates to a method of tissue culture of Medicago sativa L. By way of developing a novel tissue culture technical method, the invention focuses on improvement and innovation of two technical nodes (seed disinfection and callus formation and differentiation) in culture steps. With adoption of a mercury bichloride and hydrogen peroxide compound disinfection method, the invention realizes the purposes of thorough disinfection effect and low pollution rate and minimum damage to seeds. Finally, the invention solves the problem of severe pollution to current explants. Meanwhile, a method of TDZ and 6-BA is adopted, thereby, the callus induction rate and the formation rate are remarkably increased, the callus quality is improved, the differentiation capacity of the callus is reserved to a maximum extent, the formation of embryoid sprouts is promoted so that the novel technical method of the tissue culture of the Medicago sativa L. which is efficient, saves time, and has low toxicity and low browning, is obtained.

The invention relates to the technical field of quick reproduction of plants, and aims to provide a method for establishing a lily embryogenic callus regeneration system by using stamens as explants. The method comprises the following steps: material acquisition, callus-induced culture medium preparation, explant inoculation and callus induction, small plant regeneration, successive transfer culture of small plants, and transplanting of small plants. The method has the characteristics of low pollution rate, high callus inductivity, large callus conglomeration, favorable callus embryogenic property, high regeneration capacity and high reproducibility; especially, as antheral filaments in the young bud are used as the explants, the pollution rate can be lowered to 0, the embryogenic callus inductivity of up to 15% can be obtained, the callus conglomeration reproducibility is 100%, the embryogenic property is high, and 6 stamens contained in each bud can obtain 60-120 small plants on the whole.

The invention relates to a method for establishing an eremochloa ophiuroides highly efficient regeneration system with a young spike as an explant. The method includes the steps of explant selection and sterilization, callus induction and proliferation, callus differentiation, and rooting induction. The method provided by the invention is simple in explant sterilization, high in callus induction rate, easy to implement, and short in cycle of callus induction, obtained callus adventitious buds have high differentiation rate, test-tube plantlets are easy to root, and the method is suitable for eremochloa ophiuroides germplasm innovation, genetic inheritance transformation and other modern biological technology operations.

The invention discloses a method for somatic embryogenesis and plant regeneration of macleaya cordata, which belongs to the field of plant tissue culture techniques. The method comprises the following steps: (1) preparation of a medium; (2) somatic embryogenesis of macleaya cordata; (3) germination and seedling formation of a somatic embryo; and (4) transplantation, culture and the like of a regenerated seedling. According to the invention, the method is a tissue culture method established with filament of macleaya cordata as an explant for induction of embryoid and can realize rapid, high-efficiency and large-scale production of high-quality macleaya cordata seeds and seedlings.

The invention relates to a callus induction method of anther of small fruit tomatoes in the field of agricultural breeding technology. Steps of this method are demonstrated as follows. First, pick up the buds from a number of small fruit tomatoes at one time, classify them in terms of length, and preserve them in standard fixed fluid. Then take out pollens at the mid-late mononuclear stage from small fruit tomatoes and sterilize them. After that pick up anther from buds in an sterilized environment, and inoculate them with culture medium. Finally, cultivate the inoculated anther in dark so as to obtain calluses of small fruit tomatoes. In this invention, the key to successfully obtaining calluses lies in the identification of the developmental stages of pollens. Since the callus induction rates vary in terms of different lengths of buds, and by the comparison of the development stages of the pollen, the best cultivation stage of the small fruit tomatoes can be identified.

A method for genetic transformation of mature embryo callus of indica rice Chuanxiang 29B, which includes processes such as callus induction, subculture and differentiation, and genetic transformation. The induced callus has a high induction rate and good quality, and the subculture is not easy to brown and differentiate. High efficiency, and good applicability in other four widely used indica rice varieties (9311, Minghui 63, Zhong 9B, Ⅱ-32B), the regeneration rate is between 60-80%, and its genetic transformation efficiency can be Up to 26% or more.

The invention discloses methods for callus suspension culture and protoplast separation of camellia oleifera and belongs to the field of plant tissue culture. According to the method for callus suspension culture of the camellia oleifera, a massive number of stem segments are taken as explants, compact callus obtained through induction is subjected to subculture on a culture medium containing NAA,2,4-D and 6-BA, and loose granular callus easily establishing a suspension cell system is obtained. By means of the method, the problems that induced callus of the stem segments of the camellia oleifera in the prior art is relatively compact and is not suitable for suspension cell culture are solved. According to the method for protoplast separation of the camellia oleifera, callus protoplast ofthe camellia oleifera is suspended in a sucrose solution with higher density, and impurities such as chips with larger density sink and the protoplast of the camellia oleifera floats through centrifugation, so that separation and purification of protoplast are realized. By means of the method, the problem that the callus protoplast of the camellia oleifera cannot be collected and purified throughcentrifugation is solved, the purification efficiency of the method is high, and the obtained protoplast is clear, visible and high in purity.

The invention belongs to the technical field of plant tissue culture and particularly relates to a tissue culture rapid propagation method of pelargonium graveolens. The method comprises selecting pelargonium graveolens tender stems in the good growth condition as explants, disinfecting the explants, cutting the explants into small sections, carrying out callus induction, transferring the inducedcallus into a differentiation medium, inducing the generation of buds, inoculating a novel differentiation medium with the buds, inducing to produce bud leaves, inoculating a rooting medium with the bud with leaves, carrying out rooting culture, and carrying out seeding hardening and transplantation. The tissue culture rapid propagation method greatly improves the propagation speed, reduces the cost, can better maintain the pelargonium graveolens advantages and can provide an effective way for the rapid propagation of pelargonium graveolens. Through compounding and selection of various kinds of culture mediums, the browning and material pollution in the tissue culture are significantly reduced, the success rate of callus induction is high and fast propagation is realized. Through the cooperation of various conditions, the rooting rate and survival rate are improved.

The invention discloses a callus induction and subculture medium for cyclocarya paliurus, and also discloses a culture method based on the callus induction and subculture medium for the cyclocarya paliurus. The culture method includes explant selection, explant sterilization, establishment of a sterile anti-browning system, and proliferation culture. The callus induction and subculture medium andthe culture method have the advantages that the anti-browning abilities of calluses of the cyclocarya paliurus can be effectively improved, a large quantity of non-browning calluses with high growth speeds can be obtained in a short time, the problem of resource shortage of the cyclocarya paliurus can be solved through biotechnological measures, and accordingly, the foundation is laid for industrial extraction of secondary metabolites of the cyclocarya paliurus, such as triterpenes, flavonoids and polysaccharides, in later period.

The invention belongs to the technical field of plant genetic engineering, and provides an agrobacterium-mediated genetic transformation method using the mature embryo of sorghum as an explant to induce callus for the disadvantageous that in traditional sorghum genetic transformation systems, material drawing by using young embryos and young ears as explants is seasonally restricted and also for the tedious operation that embryo stripping is needed for other crops in which the mature crops are used as the explants. Sorghum seeds which are germinated and then appear white are utilized as objects for agrobacterium infection and the explants for callusogenesis induction, resistant callus are obtained through screening and cultivation, and then a regenerated and transformed plant is obtained through subculture, differentiation, rooting, and transplantation. The disadvantages that in an existing sorghum genetic transformation system, the material drawing by using the young embryos and youngears as the explants is seasonally restricted is avoided, the tedious operation that the embryo stripping is needed for other crops in which the mature crops are used as the explants is overcome, andthe sorghum genetic transformation efficiency is greatly improved. According to the agrobacterium-mediated genetic transformation method using the mature embryo of sorghum as the explant to induce the callus, references and technical approaches are provided for performing genetic improvement of sorghum by using biotechnology.

The invention relates to a method for transforming rape. The method comprises the following steps: obtaining hypocotyls or segments thereof after rape seeds germinate; infecting the hypocotyls or segments thereof by using agrobacterium strains; performing callus induction culturing on the infected hypocotyls or segments thereof; screening and culturing rape calluses by using a selecting agent, and performing bud differentiation culturing on screened rape resistance calluses on a ZT-containing differentiation culturing medium; culturing young rape seedlings growing from the rape buds on a first rooting culture medium which contains cytokinin and auxin and a second rooting culture medium which does not contain any hormone. The method for transforming the rape is high in callus induction rate, high in bud differentiation rate and high in plant survival rate, and facilitates rooting; the transforming efficiency is up to 8 percent.

The invention discloses a tissue culture method of coconut mature zygotic embryos. The coconut mature zygotic embryos are used as explants; a culture medium formula is strictly controlled at each phase of tissue culture; and after the tissue culture is carried out, the callus induction rate can reach 98.0 percent, the highest embryo induction rate can reach 98.4 percent, the highest embryo regeneration rate can reach 98.7 percent and the highest transplanting survival rate can reach 98.1 percent. The culture medium formula and the method, disclosed by the invention has a board spectrum, is applicable to various varieties including Hainan local high-height seeds, Wenye No. 2, Wenye No. 3 and the like, and has good repeatability and a short period; and industrialized and commercial popularization and application are facilitated.

The invention discloses a coconut immature inflorescence tissue culture method. The method adopts immature inflorescence of coconuts as explants, and the formula of a culture medium is strictly controlled in each stage of tissue culture. The highest rate of callus induction can reach 97.2%, the highest rate of somatic embryo induction can reach 98.0%, the highest rate of regeneration of somatic embryos can reach 99.1%, and the highest transplantation survival rate can reach 97.5% after the tissue culture. The formula and the method of the culture medium provided by the invention are more broad-spectrum, and are suitable for various varieties such as Hainan local high varieties, Wenye No.2 and Wenye No.3. The culture method has good repeatability and short cycle, and is beneficial to factory and commercial application.

The invention relates to a tissue culture method of lycoris incarnata, and belongs to the technical field of plant tissue culture and rapid propagation. The tissue culture method of the lycoris incarnata comprises the following steps of 1, inoculating sterilized lycoris incarnata bulbs into a cluster-bud induction culture medium for induction culture to obtain first bulblets; 2, inoculating the first bulblets into a callus induction culture medium for inducing callus culture to obtain callus lumps; 3, inoculating the callus lumps to a callus differential culture medium for differentiation culture to obtain second bulblets; 4, inoculating the second bulblets to a bud strengthening and rooting culture medium for bud strengthening and rooting culture. The culture method is high in safety andstrong in differentiation capacity, and efficient breeding of the lycoris incarnata can be achieved.

The invention provides an inducing method of a Buchloe dactyloides callus. The inducing method adopts a Buchloe dactyloides seed as an explant, the Buchloe dactyloides seed is disinfected to be arranged on a sterile filter paper for germination, selecting germinated seeds with radicle, and placing into a callus induction medium for culturing for 20-25 days at temperature of 25 +/-1 DEG C, wherein the callus induction medium is a solid medium with pH value of 5.8-6.0 and is prepared from the following components: MS medium, 3.0-3.5mg/L of 2,4-D (2,4-Dicholrophenoxyacetic acid), 30g/L of saccharose and 7g/L of agar. The method can eliminate the phenomenon that the seeds are not disinfected thoroughly during disinfection process and cause contamination during callus induction, also eliminates interference of dead seeds on the callus induction, reduces the operation difficulty, removes a step of vertically cutting a mature embryo, improves the callus induction rate up to more than 90% under a condition with simplicity and convenience in operation.