16results about How to "High differentiation rate" patented technology

The invention provides a method for sterile sowing and efficient propagation of cymbidium faberi. The method comprises the following steps: pretreatment of cymbidium faberi seeds, sterile germination of the cymbidium faberi seeds, protocorm proliferation, differentiation, seedling strengthening and acclimatization and transplantation. The pretreatment comprises the steps of refrigerating hybrid seeds of cymbidium orchid at 4 DEG C, performing secondary disinfection with sodium hypochlorite, performing dark culture for 10-30 days after sowing, and then performing light culture, and the germination culture medium is prepared by taking 1 / 4 MS as a basic culture medium and adding 100-200ml / L of coconut water, 0.5-1.0 mg / L of 6-benayl aminopurine, 0.1-0.5 mg / L of naphthylacetic acid, 0.5-2.0 g / L of peptone and 0.5-1.0 g / L of activated carbon. According to the method, the seed germination rate can be effectively increased, the 60d seed germination rate reaches up to 85%, the pollution rate is reduced to 5% or below, 50.0 g / L inositol, 2.0 mg / L glycine and 2.0 mg / L nicotinic acid are added into a differentiation culture formula, the differentiation rate can be effectively increased, the leaf bud differentiation rate reaches up to 90%, meanwhile, rooting can be induced, the growth time of cymbidium faberi from seeds to seedlings is greatly shortened, a large number of high-quality seedlings can be obtained within a short time, and the method is suitable for large-scale popularization and application. Technical data are accumulated for cymbidium resource protection and new variety cultivation.

The invention relates to motherwort cultivation. The leonurus japonicus rooting induction method is characterized by comprising rooting induction: when adventitious buds grow to 1-3 cm, the adventitious buds are cut off from explants and transferred into a rooting culture medium to be cultured, the rooting culture medium takes an MS culture medium as a basic culture medium, and IBA with the concentration of 0.1 mg.L <-1 > is added into the rooting culture medium. The components of the culture medium are selected, and the rooting rate of the selected rooting culture medium is high.

The invention discloses a zanthoxylum bungeanum tissue culture method, which belongs to the technical field of tissue culture, and comprises the following steps: (1) callus induction: taking zanthoxylum bungeanum tender stems, disinfecting, and carrying out callus induction culture; (2) multiplication culture: inoculating the callus into a multiplication liquid culture medium for oscillation illumination culture, and spraying a dispersing agent in the culture process; (3) adventitious bud induction: carrying out adventitious bud induction culture on the embryogenic callus obtained by multiplication culture; (4) adventitious bud elongation: cutting the induced clumpy buds into small clumpy buds for adventitious bud elongation culture; (5) performing rooting culture: when the adventitious buds extend to 3-4cm, performing single-plant separation on the adventitious bud clusters, and performing rooting culture; and (6) seedling hardening and transplanting: opening the bottle opening, hardening the seedlings for 3-5 days, taking out the rooted seedlings, and transplanting, so that the problem that high differentiation rate, high density, high quality and high survival rate are difficult to achieve at the same time in the pepper tissue culture process is solved.

The invention provides a callus induction, differentiation and plant regeneration method with rhododendron petals as explants and application of the callus induction, differentiation and plant regeneration method, and belongs to the technical field of tissue culture. S2, performing callus induction; s3, carrying out callus subculture; s4, adventitious bud differentiation; s5, adventitious root differentiation. In the method, the limiting conditions of each step and the components of the culture medium are specifically limited, and by adopting the culture medium and the related condition control method provided by the invention, the pollution rate of rhododendron petal tissue culture can be effectively reduced, the callus rate and differentiation rate of calluses can be improved, and the survival rate of regenerated plants can be further improved.

The invention belongs to the field of plant tissue culture, and particularly relates to a culture medium composition and method for inducing plant regeneration through peony petioles, and the culture medium composition is characterized in that a callus induction tissue culture medium and an adventitious bud differentiation culture medium are MS culture mediums containing 0.5-1.5 mg / L TDZ and 0.05-0.25 mg / L NAA; the rooting induction culture medium is a 1 / 2 MS culture medium containing 2.0 to 4.0 mg / L of IBA, 0.3 to 0.7 mg / L of NAA and 0.4 to 0.6 g / L of TA. Petioles of peony sterile test-tube plantlets are used as explants; inoculating the explant into the callus induction culture medium for dedifferentiation culture to obtain a callus; inoculating the callus into the adventitious bud differentiation culture medium to induce bud differentiation to obtain adventitious buds; dividing the adventitious buds, and inoculating the adventitious buds into the rooting induction culture medium to induce rooting, so as to obtain regenerated seedlings. The culture medium is combined with a tissue culture method, petioles of peony seedlings are adopted for tissue culture, plant seedlings are obtained, the tissue culture seedling obtaining efficiency is high, the adventitious bud and adventitious root induction effect is good, and the transplanting survival rate is high.

The present application relates to the technical field of plant tissue culture, and particularly relates to a method for promoting scale differentiation of Lilium auratum and reducing scale browning. ‑1 When NAA is 0.05 mg / L ‑1 , TDZ is 0.25 mg / L ‑1 , and AC is 6 g / L, the scale differentiation rate of Lilium auratum can reach 71.66%±6.00ab, the browning rate is reduced to 26.66%±6.00c, and the bulb growth is significantly promoted, and the bulb size can reach (0.52 cm±0.01ab)×(0.4 cm±0.02a), which can provide support for efficient propagation of Lilium auratum bulbs.

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 present invention relates to the technical field of cell bioreactors, and in particular to a double-layer hard plastic fluidized bed cell bioreactor with a four-rocker shaker. The reactor includes a polygonal insulation tank body, a double-layer cell reaction tank and a four-rocker shaker; the four-rocker shaker of the present invention adopts a cross-axis structure with the upper and lower ends of the rocker, and its vertical load is directly guided to the ground through the rocker, which can eliminate the overturning moment acting on the shaker and the ground, reduce the vibration and noise generated during the entire culture process, and greatly reduce the manufacturing cost and weight of the shaker, and this shaker can withstand a large load; the double-layer reaction tank of the present invention and the filter screen thereon can be used to culture cells by perfusion process when high-density microcarriers are used to culture adherent cells, and the cells grow in a true microgravity environment, so that the cells grow in three dimensions and maintain a high differentiation rate and amplification. The reaction tank is particularly suitable for culturing mesenchymal stem cells, which is conducive to promotion and application.

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 present invention relates to the field of biotechnology and cell therapy, and discloses a method and application of inducing gallbladder-derived stem cells to differentiate into functional hepatocytes; the method comprises the steps of: culturing gallbladder-derived stem cells in a culture system in the presence of a first culture medium, a second culture medium, and a third culture medium, thereby obtaining functional hepatocytes. The functional hepatocytes obtained by the present invention have a high differentiation efficiency and have the effect of treating liver-related diseases.

This invention provides a method for rapid propagation of *Sedum aizoon* via tissue culture, belonging to the field of plant tissue culture technology. The method includes the following steps: 1) Using *Sedum aizoon* stem segments as explants, the stem segments are disinfected, cut, and inoculated into a callus induction medium for callus induction culture to obtain callus tissue; 2) The obtained callus tissue is inoculated into a shoot induction medium for shoot induction culture to obtain adventitious shoots; 3) The obtained adventitious shoots are inoculated into a shoot proliferation medium for shoot proliferation culture to obtain regenerated seedlings; 4) The regenerated seedlings are inoculated into a rooting medium for root induction culture to obtain transplanted seedlings; 5) The transplanted seedlings are transplanted into soil for cultivation. The method provided by this invention can significantly improve the differentiation rate of callus tissue and the survival rate of regenerated seedlings.

The application belongs to the technical field of flower cultivation, and discloses a constant-temperature culture device for tulipa gesneriana and a culture method thereof. The constant-temperature culture device for tulipa gesneriana comprises a box body, double-layer box doors are mounted on the side of the box body, a plurality of supporting rods are mounted in the inner cavity of the box body, and a lattice plate is fixed to the side wall of each supporting rod. The cooperation of the structures such as the air diffuser plate, the ventilation pipe, the limiting rod and the air inlet joint improves the air exhaust uniformity and stability. When the temperature is controlled and adjusted, the air inlet hole groove on the air inlet joint is exposed, the air outside the air diffuser plate is introduced into the ventilation pipe, the air is uniformly exhausted from the air outlet on the ventilation pipe, and there are a plurality of equidistant air outlet points at the bottom of each layer of bulbs, so that the air exhaust uniformity and stability are effectively improved. When the temperature needs to be changed, the plurality of air outlets in the box body can synchronously blow out hot air or cold air, the response rate is greatly improved, and the flower bud differentiation rate is improved.

A potted Lingwu long jujube extremely-early-maturing facility cultivation temperature regulation and control method comprises the following steps: step 1, warehousing and dormancy: spraying a 5% urea solution or 5% lime sulfur before entering a refrigeration house to promote potted Lingwu long jujubes to forcibly fall leaves, manually picking off unfallen leaves, thoroughly watering, transferring the potted Lingwu long jujubes after falling leaves into a fresh-keeping refrigeration house, and storing the potted Lingwu long jujubes in the refrigeration house; the temperature of the fresh-keeping refrigeration house is set to be 2 DEG C; step 2, greenhouse temperature rise: arranging electric heating equipment in the greenhouse to adjust the air temperature and the soil temperature at the root zone; 3, temperature monitoring: setting a temperature self-recording instrument in the growth environment of the potted Lingwu long jujube, and monitoring air temperature data and soil temperature at the root zone in the flowerpot in real time. By implementing temperature regulation and control, the temperature of the environment where the potted Lingwu long jujube is located is accurately controlled, then dormancy breaking, growth and development of the potted Lingwu long jujube are promoted, and the flower bud differentiation rate and the fruit setting rate are increased.

The invention discloses a method for obtaining regenerated plants through leymus chinensis young ear tissue culture based on the same culture medium. The method comprises the following steps: 1) ear taking; 2) ear stripping and disinfection; and 3) a direct seedling method: inoculating the young ears into a culture medium, placing the culture medium in a dark 25 DEG C incubator for culturing for 20 days, then transferring the young ears into the newly-changed culture medium, placing the culture medium in a 25 DEG C incubator for culturing for 20 days under the conditions of illumination for 16 hours and darkness for 8 hours, then transferring the young ears into the newly-changed culture medium for culturing for 30 days, then transferring the young ears into a conical flask containing the newly-changed culture medium for culturing for 15 days, and taking out the young ears. The plants are transplanted into sterilized nutrient soil after being hardened; the culture medium comprises MS (Murashige and Skoog), cane sugar, agar and 2, 4-D, and the pH value of the culture medium is 5.8. According to the method, the same culture medium formula is adopted in the whole process, the use requirement of the whole operation process can be met only by preparing the same culture medium, and operation is simpler. The method is shorter in overall time consumption, lower in cost and higher in induced callus rate and differentiation rate.

This invention discloses a method for rapid propagation and seedling cultivation of *Gynostemma pentaphyllum* seedlings. Healthy, disease-free *Gynostemma pentaphyllum* plants are selected. Young leaves are washed to remove surface impurities and then disinfected. The disinfected explants are cut into pieces and inoculated into an adventitious bud induction medium to induce callus tissue. The induced callus tissue is then transferred to an adventitious bud differentiation medium to differentiate adventitious buds. The differentiated adventitious buds are then cut off of their leaves, retaining the stem segments with nodes, and inoculated into a proliferation medium. Proliferating buds with good growth and large, thick leaves are inoculated into a rooting medium. The rooted seedlings are then placed in a greenhouse for hardening off. The hardened rooted seedlings are then cleaned of the root culture medium before further cultivation. The tissue-cultured rooted seedlings produced by this method are robust, red in color, and have well-developed root systems. They exhibit high survival rates after cultivation and have a short cultivation time, providing technical support for the large-scale, standardized production of *Gynostemma pentaphyllum* and possessing broad application prospects.

The present application belongs to the technical field of organoid culture, and particularly relates to a functional bovine mammary gland organoid as well as a culture method and application thereof. The present application overexpresses BCL2 a gene in bovine embryonic stem cells, improves early cluster culture cell cluster cluster activity and / or cell cluster survival rate; bovine embryonic stem cells overexpressing BCL2 a gene are subjected to 3D culture to obtain an organoid cell cluster; 60% Matrigel is used as a supporting matrix material to provide a 3D culture environment for the organoid cell cluster, which can not only make the organoid cell cluster sprout, but also control the elongation speed of the organoid cell cluster, form a tubular structure similar to mammary ducts, and obtain a bovine mammary gland organoid with a high mammary gland differentiation rate. The obtained bovine mammary gland organoid has the function of inducing lactation, and provides a reliable model for analyzing the mammary gland development process of dairy cattle and improving lactation of dairy cattle.