67results 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 discloses a method for in vitro regeneration of a Feizixiao litchi variety. The method comprises the following steps: step 1, taking the anther of the Feizixiao litchi variety as an explant for callus induction and culture, and screening to obtain embryogenic callus; the culture medium is added with 2, 4-D with final concentration of 3 mg / l, BA with final concentration of 0.5 mg / l, NAA with final concentration of 0.5 mg / l and sugar with final concentration of 30 g / L on the basis of a culture medium MS; step 2, carrying out somatic embryo induction and culture to the embryogenic callus to obtain a somatic embryo; the culture medium is added with NAA with final concentration of 0.1 mg / l, LKT with final concentration of 5 mg / l, inositol with final concentration of 100 mg / l and sucrose with final concentration of 50 g / L on the basis of the culture medium MS; step 3, carrying out maturation to the somatic embryo to obtain a mature embryo, culture mediums are MS, 50 ml / L coconut milk and 60 g / L sucrose; and step 4, carrying out regeneration culture to the mature embryo, the culture medium is added with sucrose with final concentration of 30 g / L and agar with final concentration of 7 g / L on the culture medium MS. The invention establishes a method for in vitro plant regeneration of a Feizixiao litchi variety, achieves the high induction rate, improves the production efficiency, reduces the cost and lays a good foundation for litchi variety improvement and biotechnology breeding.

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 discloses a method for improving the perennial ryegrass callus regeneration rate, and relates to the technical field of plant tissue culture. The method comprises the following steps of: (1) screening seeds; (2) treating the seeds; (3) disinfecting the seeds; (4) preparing a culture medium, comprising inducing of a culture medium, subculturing of the culture medium, differentiating of the culture medium, and maintaining of the culture medium for plants; (5) inducing callus; (6) subculturing; (7) differentiating the callus tissue; (8) maintaining and culturing plants of regenerated seedlings; and (9) inducing the callus tissue with a good regeneration performance. The method for improving the perennial ryegrass callus regeneration rate not only optimizes the genetic transformation program, greatly shortens the transformation cycle, guarantees higher transformation rate, but also provides a favorable acceptor material for the subsequent genetic transformation.

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 relates to the technical field of cell engineering tissue culture, and aims to provide a method for remarkably improving the callus inductivity of iris pseudacorus L. The method comprises the following steps: taking materials, configuring a callus induction culture medium, disinfecting an explant, inoculating the disinfected explant, and inducing a callus and an embryonic callus. Adopting a tender embryo of iris pseudacorus L. to serve as the explant, the method has the characteristics of low pollution rate and high material availability, and has the outstanding advantages of being high in callus induction efficiency, low in explant tender viruses, simple and convenient to operate, and high in obtained callus embryo quality.

The invention discloses an induction method for embryogenesis and plant regeneration of in vitro haploid of macleaya cordata anther, which belongs to the field of plant tissue culture techniques. The method comprises the following steps: (1) preparation of a medium; (2) somatic embryogenesis; (3) germination and seedling formation of a somatic embryo; and (4) domestication, transplantation and the like of a tissue culture seedling. According to the invention, the induction method is a tissue culture method established with the macleaya cordata anther as an explant and can realize rapid, high-efficiency and large-scale production of high-quality macleaya cordata haploid plants; after field screening, the macleaya cordata haploid plants can be directly used as breeding materials for research on breeding and genomics.

The invention discloses a wolfberry anther induction medium and an anther breeding method and belongs to the field of tissue culture of wolfberry. The wolfberry anther induction medium contains major elements, minor elements, iron salt and complex agent, organic ingredients, inorganic additive, plant growth regulator, physiological activator, carbon source, coagulator and other additives. The wolfberry anther breeding method includes steps of bud taking and low-temperature treatment, preparation of the medium, anther inoculation, temperature-variable induction culture, differentiation and rooting, domestication and transplanting. By adopting the wolfberry anther induction medium and the anther breeding method for induction culture of wolfberry anther, anther callus induction rate can be increased effectively, obtained wolfberry anther callus is high in quality, and a lot of high-quality test tube seedlings can be formed in a short time, so that wolfberry breeding process is accelerated greatly. The wolfberry anther induction medium and the anther breeding method can be used for wolfberry anther breeding and can also be used for the field of wolfberry in-vitro rapid propagation and germplasm preservation.

The invention relates to a kit and a method for detecting an anthurium andraeanum SERK gene expression status, and belongs to the field of plant molecular biology. According to the invention, a just-expended young leaf of an anthurium andraeanum potted variety Alabama mature plant is adopted as an explant, and anthurium andraeanum embryogenic callus induction is carried out; non-embryogenic callus and embryogenic callus obtained in two induction stages are subjected to cytological observation; a full-length sequence of cDNA gene of anthurium andraeanum somatic embryo SERK gene is obtained by cloning; primers with relatively high specificity are designed; and researches and analysis upon changes of expression amount of SERK gene transcriptional level of anthurium andraeanum somatic embryo induction and development stages are carried out by using a real-time fluorescent quantitative PCR technology, such that a low-cost and high-specificity kit used for detecting anthurium andraeanum SERK gene expression status is obtained, and a method thereof is provided.

The invention discloses a culture method capable of promoting cyclocarya paliurus callus growth and secondary metabolite accumulation. The culture method includes: taking materials, and disinfecting;selecting semi-lignified stems with buds and top-end leaflets of cyclocarya paliurus, and disinfecting; performing callus induced culture: culturing stems and leaves in an induced culture medium to generate callus, wherein culture conditions are continuous dark treatment, and culture temperature is set at 20 DEG C; performing callus multiplication culture: transferring the callus into a transgenerational multiplication culture medium for transgenerational multiplication culture, and using a sealing film to sea, and putting into a culture chamber for culture, wherein culture conditions are continuous dark treatment, and culture temperature is set at 20 DEG C. Induction and multiplication culture of the callus are performed at temperature of 20 DEG C, so that generation of phenolic substances is inhibited while normal growth of cyclocarya paliurus is ensured, and callus growth and secondary metabolite accumulation are promoted at low temperature.

The invention relates to a method for inducing rapid callusing of phoebe zhennan. The method comprises the following steps: by taking tender phoebe zhennan leaves as explants, collecting the explants,cleaning, sterilizing, cutting, inoculating, and carrying out dark culture and light culture, thereby completing rapid induced callusing of the phoebe zhennan. The callusing induction rate is up to 98.1% at most, a remarkable greening degrees and a callusing growth amount are achieved, and bases and related technical support can be provided for future phoebe zhennan callus induction, tissue culture for nursery stock reproduction, stock breeding acceleration and market requirement meeting.

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 belongs to the technical field of plant biotechnology. Tender stem and young leaves of aralia chinensis bud are dedifferentiated, callus differentiates a great deal of embryoid, the embryoid efficiently differentiates to form seedlings, and rootage and seedling strengthening are carried out. The process includes the steps that a dedifferentiation culture medium is inoculated with an aralia chinensis bud explants, the callus induction rate is 100%, the aralia chinensis bud explants are transferred to a culture medium for inducing the callus to differentiate the embryoid, three times of subculturing is carried out, and the propagation coefficient of propagating third-time embryoid through the embryoid reaches 11.337; the embryoid is transferred to a culture medium for differentiating the embryoid into seedlings and is cultured for 60 days, and the seedling differentiation multiple is 3.889; the seedlings are transferred to a culture medium for rootage and seedling strengthening and are cultured for 45 days, the rooting rate is 98.6%, the average root length is 11.5 cm, and the average number of main roots is 3.2. The culture period of the method is short, few materials are used, wild resources can be effectively protected, the culture medium cost and labor cost are saved, and the method is suitable for planting aralia chinensis bud on a large scale.

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 pedicles 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 the pedicles are used as the explants, the pollution rate can be lowered to 0, the callus inductivity of up to 100% can be obtained, the callus conglomeration reproducibility is up to 100%, the embryogenic property is high, and every 1cm of pedicle explant can obtain 60-80 small plants.

The invention discloses an induction method of ixora flower anther callus. The method comprises the following steps: performing low-temperature pretreatment on ixora flower buds in a mononuclear period at 4 DEG C, soaking the ixora flower buds in ethanol, soaking the ixora flower buds in a sodium hypochlorite solution, washing the ixora flower buds with sterile water, peeling off the anther, inoculating the anther into an induction culture medium, sealing with a sealing film, and putting into an incubator at 27 + / -1 DEG C for dark culture; the induction culture medium is composed of a basic culture medium and a plant growth regulator, the basic culture medium is formed by adding cane sugar, activated carbon and agar into an MS culture medium, and plant growth agents are 2, 4-D and KT. According to the method, the ixora flower anther is pretreated at the low temperature of 4 DEG C and then is subjected to induction culture in the MS culture medium + 4 mg L<-1> of 2,4-D + 2 mg L<-1> of KT, the induction rate of the ixora flower anther callus is increased, and reference is provided for ixora flower anther culture and breeding.

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 culture method of an ellcalyplus grandis Hill ex Maid tissue culture seedling, which comprises the following operation steps of taking an ellcalyplus grandis Hill ex Maid tissue culture seedling leaf as an explant, inducing a callus tissue of the ellcalyplus grandis Hill ex Maid leaf, and performing adventitious bud differentiation culture and rootage culture on the callus tissue of the leaf, and is characterized in that the callus tissue of the ellcalyplus grandis Hill ex Maid leaf is induced under the irradiation of red light and blue light, the adventitious bud differentiation culture is performed on the callus tissue of the leaf under the irradiation of the red light or the blue light, adventitious bud growth culture is performed under the irradiation of green light, and then the callus tissue is transferred to be under LED (light-emitting diode) yellow light for the rootage culture. According to the method, light sources are adjusted timely according to difference in requirements of the ellcalyplus grandis Hill ex Maid leaf on the light in different tissue culture stages, and the induction rate, the adventitious bud differentiation rate and the rootage rate of the callus tissue are increased effectively. In addition, the method is low in energy consumption, and can greatly increase the yield of the ellcalyplus grandis Hill ex Maid tissue culture seedling and effectively shorten the seedling emergence time in comparison to the traditional tissue culture method.

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 provides a method for increasing the content of benzyl isoquinoline alkaloids (BAIs) in Macleaya cordata (Willd.) R.Br. through over-expressed berberine bridging enzyme (BBE) and application of the method. The method comprises the steps: a complete encoding sequence of a key gene, namely the BBE participating in alkaloid metabolism in the Macleaya cordata (Willd.) R.Br. is amplifiedthrough a PCR technology, by combining with an In-Fusion homologous recombination method efficient vector construction technology, the BBE gene in the Macleaya cordata (Willd.) R.Br. is successfully integrated into a plant expression vector pCAMBIA2301D, and then the expression vector is guided into an explant of the Macleaya cordata (Willd.) R.Br. through an agrobacterium tumefaciens mediated method to increase the content of the BAIs in the Macleaya cordata (Willd.) R.Br.. The content of the certain BAIs, such as (S)-reticuline, (S)-scoulerine, (S)-cheilanthifolin and (S)-tetrahydrocolumbamine, in over-expressed Macleaya cordata (Willd.) R.Br. positive plants obtained through the method is remarkably increased by about three times, and the early foundation can be provided for deeply researching the BBE gene function and increasing the content of the BAIs in the Macleaya cordata (Willd.) R.Br..

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 discloses a method for in vitro regeneration and genetic transformation of immature bamboo embryos of Moso bamboo, the method comprising: induction of callus of immature moso bamboo embryos, proliferation of callus of moso bamboo, construction of transformation vector pER8-WUS-RdCodA, and Agrobacterium Infection, selection of resistant callus of Moso bamboo and differentiation of adventitious buds, regeneration of transgenic plants, transplanting of hardened seedlings and detection of transgenic regenerated plants of Moso bamboo. Wherein, the construction of the transformation vector pER8-WUS-RdCodA includes: carrying out Xho I / Spe I double enzyme digestion of the estradiol-inducible vector pER8; adding vector homologous sequences to both ends of the corn WUS gene, and inserting it into pER8 after digestion In the vector, the complete expression frame of the CodA gene induced by the Rd29A promoter was inserted into the EcoR V site to construct the transformation vector pER8‑WUS‑RdCodA. The method of the invention can shorten the transformation period and improve the transformation efficiency.

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.

The invention relates to the technical field of cell engineering tissue culture, and aims to provide a method for effectively preserving embryogenic callus of Acorus calamus. The method for effectively preserving the embryogenic callus of Acorus iris includes the steps of material collection, callus induction culture, screening and preservation of the embryogenic callus. The invention uses the young embryos of calamus calamus as explants, which has extremely low pollution rate, high callus induction rate, good embryogenicity of callus, and high reproduction coefficient, and at the same time solves the problem of easy browning and differentiation in the preservation of calamus calamus callus Serious problems, strong practicability, easy operation and good promotion.