20results about How to "High induction rate" patented technology

The invention belongs to the technical field of plant tissue culture, and particularly relates to a tissue culture and rapid propagation culture medium and a tissue culture and rapid propagation method for culturing Shaniodendron subaequalis. The invention provides a tissue culture and rapid propagation culture medium and a tissue culture and rapid propagation method for culturing Shaniodendron subaequalis. Kinetin KT (Kinetin) in the tissue culture and rapid propagation culture medium provided by the invention can promote the generation of buds and the growth of adventitious buds. 6-BA is 6-benayl aminopurine and is used for promoting the growth of buds; tDZ is thiabendazole, and growth and development of buds are promoted. Under the combined action of KT, 6-BA and TDZ, the bud induction rate and the bud multiplication coefficient of the stem with the bud are improved. The invention further provides a tissue culture and rapid propagation method of the Shaniodendron subaequalis, stem segments subjected to induction culture are low in pollution rate, high in survival rate and low in browning rate; the lateral bud induction rate of the stem section is as high as 91%, and the multiplication coefficient, the rooting rate and the transplanting survival rate are high.

The invention relates to the field of plant tissue culture, and discloses a method for culturing tissue culture seedlings of small angel viridae, which comprises the following steps: S1, explant selection and primary culture; s2, carrying out subculture multiplication culture; s3, rooting culture; s4, domesticating and transplanting; in the primary culture stage, MS is used as a basic culture medium, a proper amount of 6-BA, NAA, sucrose and agar are added, and induction of cluster buds is promoted; in the subculture multiplication culture stage, an optimal multiplication culture medium is screened through an orthogonal test, so that the multiplication coefficient is improved; in the rooting stage, key factors such as inorganic salt concentration, NAA and banana puree in MS are optimized, and the number and quality of root systems are improved; in the domestication transplanting stage, the optimal proportion of river sand, coco coir and peat soil is researched, and the transplanting survival rate and the tissue culture seedling quality are effectively improved. By means of the method, efficient propagation of the small angel vitis vinifera and large-scale production of high-quality seedlings are achieved, and important technical support is provided for industrial popularization of the plant.

The invention relates to an anti-browning method in a coconut microspore somatic embryo culture process, and belongs to the technical field of coconut tissue culture. The anti-browning method in the coconut microspore somatic embryo culture process comprises the following steps: S1, anther collection and pretreatment; s2, anther observation, disinfection and separation; s3, anther culture and embryo culture. According to the anti-browning method in the coconut microspore somatic embryo culture process, PVP is added into the culture medium to resist the browning phenomenon in the coconut microspore embryo induction process; when the explant is cut, a citric acid solution is dipped to reduce browning caused by physical damage; ascorbic acid (vitamin C) is also added into the culture medium to adjust the pH value and resist the browning phenomenon; in addition, low-concentration TDZ (thidiazuron) is used as an induction hormone, so that the embryo induction efficiency is improved.

The invention provides a culture medium for reducing the content of broussonetia papyrifera esterase and a tissue culture method, and belongs to the technical field of broussonetia papyrifera tissue culture. The culture medium for reducing the content of the broussonetia papyrifera esterase provided by the invention comprises an induction culture medium, a proliferation culture medium, a differentiation proliferation culture medium and a rooting culture medium, wherein the induction culture medium is composed of an MS culture medium, 6-BA and IAA; the proliferation culture medium is composed of a 1 / 2 MS culture medium, NAA, KT and IBA; the differentiation and proliferation culture medium consists of a B5 culture medium, folic acid, TDZ, 2, 4-D, sucrose and agar; the rooting culture medium consists of an N6 culture medium, gibberellin 3, ethylene, IAA, cane sugar and carrageenan. The culture medium and the tissue culture method provided by the invention can promote more than 98% of adventitious buds to root, ensure that the survival rate is more than 98% after acclimatization and transplantation, not only can effectively reduce the content of lignin in paper mulberry tissue culture seedlings, but also cannot influence the normal growth of the paper mulberry tissue culture seedlings.

The invention provides a culture method for obtaining high-quality hemsley rockvine root calluses, belongs to the technical field of tissue culture, and aims to solve the problems of slow growth, high adventitious root occurrence rate, premature aging of the calluses, serious browning, high death rate after subculture and the like in the hemsley rockvine root callus growth process. The culture method comprises the following steps: step 1, pretreatment of explants; step 2, callus induction culture: putting the treated explant into a callus induction culture medium, and putting the callus induction culture medium in an environment with a day-night temperature difference for callus induction culture; and step 3, subculture and enlarged culture of the callus: stripping the callus generated on the stem segment in the step 2, transferring the callus to a subculture multiplication medium, adjusting the light quality ratio of red light to blue light, and carrying out subculture in an environment with gradually changed day temperature to obtain the high-quality hemsley rockvine root callus. In the subculture process, the content of zeatin is increased, the light quality ratio is adjusted, and the high-quality hemsley rockvine root callus can be obtained in a circulation management mode that the daily temperature is gradually changed.

The present application relates to the technical field of polyploid breeding of shellfish, and specifically discloses a high-efficiency breeding method for triploid Ruditapes philippinarum based on asynchronous acquisition of sperm and egg and early induction. The method comprises the following steps: identifying and separating male and female parent shellfish before inducing parturition; allowing female parent shellfish to naturally release eggs; dissecting male parent shellfish to obtain sperm; artificially mixing and fertilizing; adding cytochalasin B to inhibit the release of the second polar body when the first polar body release rate of the fertilized egg reaches 30%-40%; and washing and hatching to obtain triploid larvae. The present application realizes controllable starting time of fertilization by asynchronous acquisition of sperm and egg, and has a triploid induction rate as high as 92.90% and a standard deviation of only ±1.60% in combination with an early induction window, and has the advantages of stable operation, good repeatability and suitability for large-scale production, thereby providing a reliable technical solution for the genetic improvement and sustainable development of the Ruditapes philippinarum industry.

The present application relates to the technical field of breeding and seedling raising of forest trees, and discloses a method for realizing in vitro regeneration of Rosa roxburghii by hypocotyl culture, which comprises the following steps: taking seedling germinated from cold storage of Rosa roxburghii seeds as material, removing cotyledon and root, cutting hypocotyl segments with a length of 0.6-1.5 cm as explants, and sterilizing the explants; inoculating the sterilized explants into primary induction medium, and carrying out dark culture until callus is generated and primary adventitious buds are induced; inoculating the cultured callus into differentiation medium, and carrying out dark culture until secondary adventitious buds are regenerated; inoculating the obtained secondary adventitious buds with a height of 1.5-2.5 cm into rooting medium until rooted plants are obtained; and hardening the obtained rooted plants, and then transplanting them into mixed substrate for culture.

The invention provides a preparation method and application of centella asiatica stem cells for high-yield madecassoside, and belongs to the technical field of stem cells. The preparation method comprises the steps of disinfection, induction, proliferation, culture of centella asiatica stem cells with high madecassoside yield and the like, and has the unexpected technical effects that the pollution rate is low, the induction rate is high, the proliferation multiple is high, and the madecassoside content in the obtained centella asiatica stem cells with high madecassoside yield is high. The product is beneficial to industrial production and can be used as a skin care component in a product with anti-inflammatory, soothing and other effects.

The application belongs to the technical field of blueberry cultivation, and discloses a blueberry callus induction method, which comprises the following steps: (1) selecting blueberry flower buds 1-3 days before field flowering, removing petals and stamens, and placing the ovary with the stigma and pedicel retained in a 75% ethanol solution for sterilization for 10-15 min; (2) under aseptic conditions, the ovary with the stigma and pedicel retained is cut to remove the stigma, 1 / 4 of the ovary and the receptacle, and the ovule surrounded on the placenta is carefully taken out with sterilized tweezers and inoculated into a callus induction medium; after 30 days of dark culture, callus is obtained; the application provides a blueberry callus induction method, and has the technical effect of improving the callus induction rate of blueberries.

The invention relates to the field of plant genetic engineering, and discloses a recalcitrant rice variety genetic transformation method based on agrobacterium tumefaciens mediation, which comprises the following steps: 1) carrying out illumination culture on rice seeds in an induction culture medium to obtain calluses; 2) culturing the activated agrobacterium in a suspension culture medium, and adding the callus to obtain an infected callus; (3) carrying out dark culture on the infected callus in a screening culture medium; 4) selecting resistant infection calluses without browning death, and carrying out light-dark alternate culture in a differential medium; and 5) after the resistant infection callus forms a regeneration seedling, transferring the regeneration seedling into a rooting culture medium for alternate light and dark culture to obtain a plant. The efficient, stable and repeatable agrobacterium tumefaciens-mediated genetic transformation method suitable for recalcitrant rice varieties is established by systematically optimizing culture conditions, culture media and the like at the stages of induction, screening, differentiation, rooting and the like, and the transformation efficiency can be remarkably improved.

The invention relates to a culture medium combination and method for constructing a zantedeschia aethiopica regeneration system by tubers, the culture medium combination comprises a callus induction culture medium, a proliferation culture medium and a differentiation culture medium, the induction culture medium takes MS as a basic culture medium, and also contains 6.5-7.0 g / L of agar, 30 g / L of sucrose, 1.0-2.0 mg / L of 6-BA and 0.5-1.0 mg / L of NAA; or the culture medium also contains the following substances in concentration: 6.5-7.0 g / L of agar, 30 g / L of cane sugar, 0.5 mg / L of 6-BA and 0.8 mg / L of NAA. According to the method, the sterile bud-free tubers of the colored callalily growing to the specific period are used as the explants, the specific induction culture medium, the proliferation culture medium and the differentiation culture medium are matched to obtain the cluster buds, the callus induction rate is high, and technical support is provided for establishment of a colored callalily tissue culture rapid propagation and genetic transformation system.

The present invention discloses a method for producing tetraploid largemouth black bass by inducing a double hydrostatic shock method. The method first obtains high-quality fertilized eggs through natural fertilization: the steps of injecting hormones and squeezing the abdomen to obtain sperm and eggs in the traditional method are avoided, thereby significantly reducing the damage to the parent fish. Combined with the innovative double hydrostatic shock method, by optimizing the pressure intensity and processing time, the damage to the fertilized eggs is further reduced, the second cleavage is successfully inhibited, and the tetraploid induction rate can be as high as 24.44%. The method can be applied to the tetraploid production of largemouth black bass: the operating method of the present invention is simple and easy, has strong repeatability, and has a high induction rate. These characteristics make the method of the present invention have a high application value in largemouth black bass breeding, and provide strong support for the breeding of tetraploid largemouth black bass.

This invention relates to the field of plant tissue culture technology, specifically to a culture medium and method for inducing and regenerating tiger nut callus. This invention creatively combines growth regulators (2,4-D, 6-BA, NAA, and ZT) in the culture medium for three different stages of tiger nut callus induction, regeneration, and differentiation, and optimizes the selection of the best explants. This allows each component to fully synergistically enhance its effect at each stage, resulting in high callus induction rate, good growth status, low browning degree, good proliferation effect, and high differentiation rate and differentiation coefficient. Furthermore, no growth regulators are required during rooting induction, resulting in lower costs. This invention lays the foundation for tiger nut gene function research, variety genetic improvement, and germplasm resource optimization.

The invention discloses a method for producing rice haploids and application of the method in breeding, by editing one or more of OsFEB1, OsFEB2 and OsFEB3 genes in rice, the functions of the OsFEB1, OsFEB2 and / or OsFEB3 genes are lost, a rice haploid induction line is obtained, generation of haploids is induced through sexual reproduction, the gene sequence of the OsFEB1 is as shown in SEQ ID NO.1, the gene sequence of the OsFEB2 is as shown in SEQ ID NO.2, the gene sequence of the OsFEB2 is as shown in SEQ ID NO.1, the gene sequence of the OsFEB2 is as shown in SEQ ID NO.2, and the gene sequence of the OsFEB3 is as shown in SEQ ID NO.2. And the gene sequence of the OsFEB3 is as shown in SEQ ID NO. 3. The haploid induction line obtained by the method provided by the invention has the capability of inducing generation of haploid plants for breeding. An RHG1 haploid induction line is combined with an MIME system, and an apomixis material is artificially created to fix the heterosis of rice.

The invention discloses a rapid propagation method for tissue culture of phalaenopsis amabilis, which comprises the following steps: selection and treatment of explants: selecting healthy and disease-free explants, and properly treating the explants to improve the success rate of subsequent culture; primary induction culture: putting the treated explant into an induction culture medium to promote primary growth and differentiation; cluster bud induction and subculture multiplication culture: inducing the explant to form a plurality of cluster buds by optimizing culture conditions, and carrying out subculture multiplication culture so as to improve the propagation coefficient; rooting culture: carrying out rooting culture on the formed buds to promote the development of root systems and improve the transplanting survival rate; transplanting and seedling hardening: transplanting the plants with good rooting into a proper matrix, and carrying out seedling hardening to enhance the adaptive capacity of the plants; through the steps, rapid propagation of phalaenopsis amabilis can be achieved, the output period is remarkably shortened, and the production cost is reduced.

The invention relates to a genome editing system and method for melilotus albiflora based on a hairy root genetic transformation system, and belongs to the technical field of plant genetic engineering. According to the invention, an editing carrier system of the MaPDS gene is constructed according to two target sites on the MaPDS gene of the sweet clover, and the editing system is CRISPR2.0-MtLAP1-KO-MaPDS; according to the editing system, sweet clover hypocotyl is used as an explant material, hairy roots are obtained through an agrobacterium rhizogenes-mediated hypocotyl explant infection method, and meanwhile, the hairy roots are obtained on the basis of the principle that anthocyanin is over-expressed and accumulated based on an MtLAP1 transcription factor for color development. Positive hairy roots are obtained through visual, rapid and efficient screening, and efficient editing of the MaPDS gene of the red positive hairy roots is achieved. The method provided by the invention can effectively reduce the occurrence of solid roots in the inducing process of the sweet clover hairy roots, prevent agrobacterium pollution, reduce the detection workload and realize efficient gene editing of target genes.

The invention relates to the technical field of tissue culture of dysosma versipellis, in particular to a tissue culture medium combination and a tissue culture rapid propagation method of dysosma versipellis. The invention provides a dysosma versipellis tissue culture medium combination and a tissue culture rapid propagation method.The dysosma versipellis tissue culture medium combination comprises a germination culture medium, a callus induction culture medium, a cluster bud proliferation culture medium and a rooting culture medium, the callus induction rate can be increased, and the tissue culture rapid propagation rate can be increased. The inductivity and the multiplication coefficient of the multiple shoots of the dysosma versipellis are improved, and the rooting rate of tissue culture seedlings of the dysosma versipellis is improved. According to the tissue culture rapid propagation method for the dysosma versipellis, the dysosma versipellis is high in propagation speed, short in production period, simple in equipment and convenient for industrialized production, and guarantee conditions are provided for production of authentic medicinal materials and postpartum processing and selling.

The invention discloses a method for inducing establishment of a rapid asexual propagation system of helleborus thibetanus, and relates to the technical field of plant tissue culture, and the method comprises the following steps: obtaining axillary buds: taking current-year axillary buds as explants, and pretreating the explants; performing induction culture: inoculating the obtained pretreated explants to an induction culture medium for induction culture, and culturing for a period of time to obtain plantlets; multiplication culture: preparing a multiplication culture medium in advance, inoculating the obtained plantlets into the multiplication culture medium for multiplication culture, and culturing for a period of time to obtain rootless plantlets; and rooting culture: preparing a rooting culture medium in advance, inoculating the obtained rootless plantlets into the rooting culture medium for rooting culture, and culturing for a period of time to obtain a complete plant. According to the method for establishing the system for inducing the rapid vegetative propagation of the helleborus thibetanus, MS is used as a basic culture medium, and the bud induction rate of the helleborus thibetanus is increased to 30% or above by adjusting the concentrations of TDZ and 6-BA in the induction culture medium.

The application provides a kind of festuca ovina mature seed callus induction medium, induction method and the method for establishing regeneration system, belong to plant tissue culture technical field.The application provides a kind of callus induction medium, with MS medium as base medium, further include the following components with the following content: 2, 4-dichlorophenoxyacetic acid 4~6 mg / L, hydrolyzed casein 40~60 mg / L, abscisic acid 1~3 mg / L and kinetin 0.04~0.07 mg / L.The callus induction medium described in the application can efficiently induce mature seeds to form callus, and the induction rate of callus is more than 81%.The application selects festuca ovina mature seeds as explants for callus induction culture, which is convenient and low in cost, and lays a foundation for the research of festuca ovina gene editing.

This invention discloses a method for improving the induction rate of prothallus and sporophyte in *Cibotium barometz*, comprising: S1, sterilizing the substrate with a fungicide, wherein the fungicide includes thiophanate-methyl and tebuconazole, and after sterilization, reducing the substrate moisture content to 40-50%, obtaining a sterilized substrate; S2, contacting the bottom of the substrate with a hormone aqueous solution, obtaining a substrate absorbent with the hormone aqueous solution; S3, sowing *Cibotium barometz* spores on the substrate surface for cultivation. This invention shortens the spore germination time of *Cibotium barometz*, improves the induction rate of prothallus and sporophyte, and is simple and easy to implement. Using this method to cultivate *Cibotium barometz* seedlings, prothallus are induced in an average of 22 days, and sporophyte are induced in 66 days. This method can be applied on a large scale to the sowing and seedling cultivation of *Cibotium barometz*, saving seedling time and labor costs.