50 results about "In vitro tissue culture" patented technology

The invention discloses a method for inducing and multiplying a prothallium of Huperzia serrata. The method comprises the following steps of taking a sporangium or a spore as an explant, performing in vitro tissue culturing on the explant; establishing an induced system of the prothallium of the Huperzia serrata, wherein the induction frequency of the prothallium reaches 15 percent; and under an artificial regulation, performing multiplication culturing for the prothallium until the month multiplication time reaches 5 times. The method is easy, has high efficiency, and can be used for establishing an import technical base for artificial propagation for the Huperzia serrata.

The invention relates to a method of excysting and growing protozoal oocysts by in vitro tissue culture resulting in production of a continuous culture of merozoites. The invention also provides an economical and reliable supply of cultured Eimeria sp. for vaccine production, assays and research. Domesticated avians that have been vaccinated using the provied Eimeria sp. are also provided.

The invention relates to a fast propagation method for aquatic plant lotus flowers. According to the method, in vitro tissue culture is firstly adopted for obtaining tissue culture seedlings, and the in vitro tissue culture comprises the following steps that: (1) lotus flower seeds are used as initial raw materials, after the lotus flower seeds are subjected to sterilization and disinfection treatment, plumules are taken out under the aseptic condition and are inoculated into a basic culture medium to be cultured for 2 to 3 weeks, and aseptic seedlings are obtained; (2) the hypocotyl upper parts and cotyledons of the aseptic seedlings containing growth point parts are cut off and are inoculated into a propagation culture medium, the light illumination culture is carried out for 3 to 4 weeks, and tissue culture calluses are obtained; (3) buds with the length being 2 to 3cm in the tissue culture calluses obtained in the second step are cut off and are inoculated into a strong seedling culture medium to be cultured; and (4) after the seedlings in the third step grow to 4 to 6cm, the seedlings are transplanted onto a rooting culture medium for carrying out rooting culture, and the tissue culture seedlings are obtained. The fast propagation method provided by the invention solves the problems of few lotus root resources and long propagation period in the prior art, and a new path is provided for meeting the market requirements.

The invention discloses a method for platycerium wallichii tissue culture and rapid propagation and relates to a method for obtaining the platycerium wallichii test-tube plantlet in a short time through a plant tissue culture technology. The in-vitro tissue culture and rapid propagation method of the platycerium wallichii is built by taking the platycerium wallichii tender sporophyl as an explant, disinfecting the explant, and performing GGB induction, proliferating, differentiating and rooting. The growth cycle is shortened, the shortage of the platycerium wallichii commercialized produced seedlings can be relieved and a foundation is laid for innovating the germplasm resources of the pteridophyte in future.

A porous chamber for tissue culture in vitro. The porous chamber includes a main body made of porous biologically absorbable polymer material, comprising an inner surface surrounding a hollow cavity, the hollow cavity having an aperture opened at an upper surface of the main body, wherein the aperture is communicated with outside of the main body for seeding tissue blocks in the hollow cavity, and an upper cover passing through the upper surface of the main body to seal the aperture to prevent the tissue blocks from outflowing.

An in vitro tissue culture process of rooting Hoodia explants by using an inventive combination of phytohormones.

A fast reproduction technology for the Freesia by in vitro tissue culture and soilless seedball reproduction includes such steps as using the young stachys, rhachis, stem, leaf, floral base, mature bud, root, etc of Freesia X hybrida as explant, preparing culture medium, inducing regeneration, and special treating. A freeze storage method for the calli is also disclosed. Its advantages are high reproduction coefficient, short reproduction period, and no need of soil.

An external microtuber reproduction method for Dioscorea nipponica includes such steps as using the tender stem segment of Dioscorea nipponica, or the stem segment with bud or small tube with bud of its test-tube plantlet as explant, preparing culture medium, inducing test-tube plant, special treating to generate microtuber, and outdoor culture. The method for outdoor culture is also disclosed. Its advantages are short period, high survival rate and no need of ground.

The invention relates to a method for culturing a wild jujube grafted by a jujube in-vitro tissue culture seedling, which is characterized by comprising the following steps of: (1) carrying out bud forcing treatment on a wild jujube seed, and seeding the wild jujube seed in a nutrition bowl filled with medium when the seed begins to bud; (2) hardening a tissue culture seedling growing to the length of 5-7cm and being cultured for 25-35 days in an opened bottle under hard light for 5-10 days, shearing the tissue culture seedling into a stem with two buds, wherein the stem with the two buds is used as a scion; and cutting the lower part of the scion to be wedged; (3) after the main leaf of a wild jujube gives off, cutting off a growing point, remaining two seed leaves, cutting from the middle of the two seed leaves by using a blade through a cleft graft method, meanwhile, rapidly inserting the wedged part of the scion into a cut, and aligning the scion to two ends of a stock stem and twining by the sealing film; and (4) cutting off sprout tillers of the wild jujube in time after one week since grafting, and growing for 15-30 days to obtain a grafted seedling. The method has the beneficial effects that: 1, the problem that the jujube tissue culture seedling is difficult or unstable in rooting can be solved; and 2, the grafted seedling is directly obtained without being limited by seasons and can be produced all the year round.

The invention belongs to the technical field of plant propagation and particularly relates to a method for rapid seedling of red-core hylocereus undatus through in vitro tissue culture. The method comprises the following steps: by taking stems with areole of the red-core hylocereus undatus as an explant, disinfecting, then picking off small thorns, respectively carrying out induction culture, enrichment culture, rooting culture and transplanting culture. The improved areole is immersed into a culture medium, the sprouting of gemmule can be obviously accelerated, and the quantity of the femmuleis increased; in the rooting culture process of the gemmule, through primary sprouting of adventitious roots, namely transplanting culture, the production period of seedlings of the red-core hylocereus undatus is greatly shortened, the growth coefficient is 3-5 times, the rooting percentage is 100%, and the rate of survival after transplanting is 96% or above, the production period of the seedlings is about 90d. the method is a new technology for rapid propagation of the red-core hylocereus undatus, is suitable for industrialized, large-scale and intensive production of the seedlings of the red-core hylocereus undatus.

The invention provides a tissue engineering material construction method based on amniotic mesenchymal stem cells, and relates to a stem cell tissue engineering material. The construction method comprises: 1) obtaining and separating fresh human amnion, 2) carrying out in situ amplification on the obtained amniotic mesenchymal stem cells, 3) adopting different cell differentiation culture mediums to induce differentiation of the amniotic mesenchymal stem cells toward cells with specific types, and 4) constructing tissue engineering materials with different forms. According to the present invention, the in vitro tissue culture technology is adopted to carry out in situ amplification on the amniotic mesenchymal stem cells, the differentiation of the mesenchymal stem cells toward nerve cells, liver cells and other types of the cells is induced, and the tissue engineering material having the corresponding cell type is constructed, and can be used for corresponding disease treatments.

The invention discloses an in-vitro tissue culture and rapid propagation method for viburnum burejaeticum and belongs to the technical field of plant tissue culture. The problems that the pollution rate is high and the survival rate is low in the conventional viburnum burejaeticum tissue culture are solved. The method disclosed by the invention comprises the following steps: adopting a plant tissue culture technology, taking semi-lignified stems of viburnum burejaeticum as explants, obtaining explants with high survival rate and relatively low death rate and pollution rate by using an explantsterilization method, taking MS as a minimal medium, adding different plant hormones of appropriate types and concentrations to induce germination of axillary buds, and performing enrichment culture,rooting culture and transplanting under appropriate culture conditions, thereby obtaining lots of high-quality regenerated plants in a short time. A certain foundation is laid for tissue culture and rapid propagation of the viburnum burejaeticum, and necessary technical support is provided. The method disclosed by the invention can provide technical reference for large-scale propagation of high-quality seedlings of the viburnum burejaeticum, and has significances on comprehensive development and utilization of the viburnum burejaeticum.

The invention provides a method for in vitro tissue culture and nuisance-free planting by taking maternal tissue leaves of Ypsilandra thibetica as explant materials. Large quantities of tissue culture seedlings can be obtained by the method, which meets the planting requirement. In the method, plantation in a natural forest is adopted to overcome pollution caused by pesticides and heavy metals, the survival rate reaches 95.5%, new medicinal materials can be supplied within 3 years, weight of an individual plant is twice more than that of wild medicinal materials and non-proliferated medicinal materials at the same year, the content of effective ingredients which can induce uterus contraction and arrest bleeding is higher than that of the wild medicinal materials at the same year, thus meeting the requirement of pharmaceutical enterprises for raw medicinal materials. The method helps culture the tissue culture seedlings in batches and plant the tissue culture seedlings at a large area, and obtain the nuisance-free medicinal materials 2 years earlier, therefore, the method has popularization value.

The invention discloses a breeding method for bilinear ultra-small embryo hybrid rice and belongs to the technical field of special rice seedling. Bilinear sterile line and restoring line immature embryos to be mutagenized are subjected to in vitro tissue culture induction to form callus, 60 Co-gamma rays with the dosage being 11 to 15Gy are utilized for irradiation treatment, ultra-small embryo property mutants are obtained, then, the ultra-small embryo bilinear sterile line and restoring line are breaded through selecting properties of agriculture, yield, flowering and the like, and further, the bilinear ultra-small embryo hybrid rice combination is formed through matching. The content of the embryo is reduced through the breeding method, and the endosperm specific gravity improvement is reached, so the goals of improving the grain thousand seed weight, the yield and the polished rice rate are reached. The breeding method provided by the invention has the advantages that the ultra-small embryo property mutation frequency reaches 0.01 to 0.02 percent, the embryo weight of the breaded bilinear ultra-small embryo hybrid rice only accounts for 1 to 1.2 percent of the coarse rice grain weight, the size is only 1 / 3 to 1 / 2 of that of the ordinary rice embryo, and the polished rice rate is improved by 1.5 to 2.5 percent through being compared with that of the ordinary rice.

The invention discloses a method for endangered Chinese azalea in vitro tissue culture propagation and storage. The best rapid propagation system and storage system are built in the Chinese azalea tissue culture process. According to the method, an optimum explant selection method, an efficient multiple sterilization method, optimum culture medium components, adventive bud rooting, acclimatizating and transplanting condition control for Chinese azalea in vitro growth propagation and storage and other key technological problems are included, and finally large-quantity, rapid and high-quality propagation of Chinese azalea in a short time is achieved. The Chinese azalea test tube plantlet differentiation rate can reach 100%, the adventive bud growth coefficient month proliferation rate can reach 8-10.5, and the rooting rate and the acclimatizating survival rate are 85% or above after 40 d, so that the increasing requirement of people for Chinese azalea can be effectively met, then, damage by people to wild Chinese azalea is reduced, protection of the endangered wild Chinese azalea is facilitated, and huge scientific research value, economic value and ecologic value are achieved.

The invention discloses a semiliquid in-vitro tissue culture and rapid propagation method for MuSa basjoo. According to the method, a semiliquid induction medium comprises an MS culture solution, 6-BA with the concentration of 4 mg / L, NAA with the concentration of 0.15 mg / L, sucrose with the concentration of 30 g / L, activated carbon with the concentration of 0.5 g / L and agar with the concentration of 3 g / L, explants are cultured in a constant-temperature shaking incubator with the shaking frequency of 30 times per minute under the conditions that pH is 5.6-5.8, the temperature is 28-30 DEG C, the illumination intensity is 1,600-2,000 lx and the illumination time of 8 h / d; a multiplication subculture medium mainly comprises the MS culture solution, 6-BA with the concentration of 3 mg / L, NAA with the concentration of 0.15 mg / L and the activated carbon with the concentration of 0.5 g / L; a rooting medium mainly comprises the MS culture solution, NAA with the concentration of 0.5 mg / L, IBA with the concentration of 0.1 mg / L and the activated carbon with the concentration of 0.5 g / L. The browning ratio of the MuSa basjoo is reduced, the induced differentiation ratio and the multiplication coefficient are increased remarkably, and the culture period is shortened.

The invention provides a Terminalia mantaly Tricolor tissue culture and propagation method. The Terminalia mantaly Tricolor tissue culture and propagation method comprises the following steps of: carrying out tissue culture and rapid propagation under different hormone ingredients and concentration levels by using MS as basic culture medium, adding a natural organic mixture, and using the leaves or twigs of the Terminalia mantaly Tricolor as explants; carrying out disinfection treatment and inoculation induction to form a callus tissue; carrying out callus tissue differentiation, bud multiplication and rooting culture to form a complete plant seedling; transplanting the complete seedling in a raising substrate for growing into a normal Terminalia mantaly Tricolor plant, wherein the basic culture medium is based on MS culture media and supplemented with the ingredients including 6-benzylaminoadenine, 2,4-dichlorphenoxyacetic acid, naphthylacetic acid, sucrose, active carbon and the like. According to the in-vitro issue culture and rapid propagation method of Terminalia mantaly Tricolor disclosed by the invention, the tissue culture is applied to overcome the problems that the Terminalia mantaly Tricolor is long in seedling raising period and low in propagation coefficient, so that rapid industrialized seedling can be carried out for meeting the requirements of the market on the Terminalia mantaly Tricolor seedling.

This invention characterizes the specific peptide fragment derived from specially prepared zinc charged fetuin and a method of preparation thereof, wherein the fragment was found to contain an apoptosis-inducing activity. Specifically, the amino acid sequence of this peptide is His Thr Phe Ser Gly Val Ala Ser Val Glu and correlates to amino acid no. 300-309 of fetuin, referred to herein as Fetuin Peptide Fragment (FPF 300-09). FPF 300-09 strongly induced apoptosis in LNCaP (prostate cancer) and HT-29 (colon cancer) cells without affecting CCD 18 Co (normal colon) cells. The in vitro tissue culture study demonstrated that the FPF 300-09 is more potent than the parent molecule (full-length zinc charged fetuin) in inducing apoptosis. FPF 300-09 has a LD50 of 0.3-0.4 μM, while the LD50 for zinc-charged fetuin is 3-10 μM.

The invention discloses a tissue culture method of raspberries. The method comprises the following steps: explant selection and disinfection, induction culture, enrichment culture, rooting culture, acclimatization and transplanting, and the like. The invention provides a raspberry in-vitro tissue culture rapid propagation packaged technology. Different hormones are utilized for adjustment in each phase to achieve the goals of enhancing the multiplication rate and reducing variation; the survived seedlings are convenient for transportation; and the method has the advantages of low requirements on the female parent material, low cost, short time, high survival rate and favorable product quality, and implements large-scale standardized production on raspberry seedlings.

The invention belongs to the technical field of plant genetic engineering transformation and discloses a genetic transformation method for stab-vacuum infiltration-assisted agrobacterium tumefaciens-mediated castor seeds. The method comprises the steps of pre-culturing castor seeds, stabbing epicotyls of the pre-cultured castor seeds, performing vacuum infiltration for enabling an agrobacterium tumefaciens solution containing target genes to infect the castor seeds, performing co-culture, removing agrobacterium tumefaciens, and performing transplantation, wherein according to the vacuum infiltration, the stabbed seeds are treated in the agrobacterium tumefaciens solution for 5-20 minutes under the vacuum negative pressure of -30kPa to -70kPa, air is discharged for 2-5 minutes, and the stabbed seeds are treated for 2-5 minutes under the negative pressure of -30kPa to -70kPa again. According to the method, the pre-cultured seeds are directly taken as receptors, so that in vitro tissue culture is not required; and a transgenic plant can be obtained in 15 days from castor seed pre-culture to extracted DNA detection, so that the transformation cycle is short, the transformation rate is high, the operation is relatively simple, and the repeatability is high.

The invention provides a culture method for proliferation and rejuvenation of subculture buds in tissue culture of masson pine. The culture method comprises the following steps of selection of subculture buds, induction of axillary buds, and rejuvenation culture; selecting the subculture single bud of the masson pine after conventional disinfection, inoculation and subculture, transferring into asubculture bottle of a solid culture medium I, and performing induction of the axillary buds; adding a liquid culture medium II into the original subculture bottle by a double-layer and double-phase culture method, so as to obtain the subculture clumpy bud of the masson pine with strong growth vigor. The culture method has the advantages that the transferring step in the traditional subculture method of tissue culture of masson pine is simplified, and the subculture clumpy bud of the masson pine with high subculture proliferation coefficient, strong bud and seedling vigor and difficulty in aging is obtained; the quick propagation and utilization of scale in-vitro tissue culture of high-quality variety of masson pine are realized; the better economic, social and ecological benefits are realized.

The invention belongs to the field of tissue engineering, and particularly relates to a high-compliance tissue engineering blood vessel preparation template. The high-compliance tissue engineering blood vessel preparation template comprises a framework main body (1) and an inner core (2), wherein the inner core (2) is arranged inside the framework main body (1); the outer diameter of the inner core (2) is matched with the inner diameter of the framework main body (1); the main body of the framework main body (1) comprises support layers (11) and degradation layers (12) in alternate arrangement; the inner surface and the outer surface of the framework main body are the support layers (11); and the degradation layers (12) are made of degradable materials. The high-compliance tissue engineering blood vessel preparation template is used for being implanted under the skin of an animal or being used for in-vitro tissue culture; then, the inner core (2) is pulled away; and after the decellularization treatment, the high-compliance tissue engineering blood vessel can be prepared. The high-compliance tissue engineering blood vessel preparation template has the beneficial effects that the tissue engineering blood vessel prepared by using the high-compliance tissue engineering blood vessel preparation template has good compliance; the intimal hyperplasia can be inhibited.

The invention discloses a method for in vitro tissue culture, reproduction and preservation of Huperzia serrata. An optimal rapid reproduction system in the tissue culture process of Huperzia serratais established, comprising a most suitable selection method of explants, and an efficient multi-sterilization method. The key technical problems of in vitro growth, reproduction and preservation of different genotypes of Huperzia serrata are solved, and the large-scale, rapid and high-quality reproduction and preservation of Huperzia serrata within a short time are finally realized. The Huperzia serrata from different provinces and different genotypes can be reproduced in vitro, and the in vitro Huperzia serrata tissues can produce huperzine A. The method can effectively meet the growing demand for the Huperzia serrata and further reduce the damage to the wild Huperzia serrata, is conducive to the protection of the wild Huperzia serrata, and has huge scientific research value, economic value and ecological value.

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 relates to a micro-needle based in vitro culture method for testicular tissue and applications thereof, and belongs to the technical field of biology. The method for in vitro culturing testicular tissue to promote proliferation of spermatogonial stem cells is provided. The method includes: soaking PEGDA micro-needles in a culture medium for replacement, so as to obtain a culture system including replaced PEGDA micro-needles and the culture medium; and adding isolated testicular tissue in the culture system to culture, so as to proliferate the spermatogonial stem cells in the isolated testicular tissue. The method improves situations of death of tissue centers during tissue culture in vitro, so that large tissue culture can be realized. Therefore, a physiological structure of the tissue can be maintained to the greatest extent, which is more in line with in vivo situations. In addition, the method can promote the proliferation of the spermatogonial stem cells in vitro, the self-renewal of the spermatogonial stem cells (SSCs) is supported, and the dryness of the spermatogonial stem cells can be maintained.

The invention relates to tissue culture media of Glossostigma elatinoides. The tissue culture media comprise an initial culture medium, a subculture medium and a rooting culture medium which are used for different stages. The initial culture medium prepared by adding potassium iodate, 2,4-D and paclobutrazol into an ER culture medium. The subculture medium is prepared by using an MS culture medium as the basis, regulating the concentration of ammonium nitrate to be 1200-1300mg / L and then adding urea, ABA, indole acetic acid, glutamine and activated carbon. The rooting culture medium is prepared by adding ammonium molybdate, TDZ and amylopectin into an N6 culture medium and using sodium hydroxide to regulate pH to be 7.0-7.2. When the in-vitro tissue culture media are used to perform tissue culture on the Glossostigma elatinoides, the Glossostigma elatinoides is high in survival rate, the obtained Glossostigma elatinoides seedlings are robust and has tenacious vitality, and the Glossostigma elatinoides can fast adapt to natural growth conditions after transplanting.

The invention discloses a method for in vitro tissue culture, reproduction and preservation of endangered sheep roe deer, establishes the best rapid propagation system and preservation system in the tissue culture process of sheep roe deer, including the most suitable selection method for explants, and is highly efficient. The multi-sterilization method, the most suitable medium composition for the in vitro growth and preservation of the sheep hemp , High-quality reproduction. The invention can make the differentiation rate of the test-tube seedlings of the goat deer reach up to 100%, the multiplication factor of the adventitious buds reaches 8-10.5 per month, and the rooting rate and domestication survival rate after 40 days are both over 85%. This can effectively meet people's increasing demand for sheep roe, thereby reducing people's damage to wild sheep roe, which is conducive to the protection of endangered wild sheep roe, and has great scientific research value, economic value and ecological value.