110results about How to "Maintain genetic stability" patented technology

The invention discloses a relay-intercropping cultivation method of polygonatum sibiricum. The relay-intercropping cultivation method is characterized in that an uncaria relay-intercropping mode is adopted and imitated wild plantation is performed at borderlands of mountain forests. The cultivation method comprises following steps of: firstly, selecting moisture and completely-shaded land parcels; secondly, performing fixed planting operation on uncaria seedlings in accordance with 1.0m*1.5m distance between plants; thirdly, making a compartment surface, 50cm wide and 30cm high, between lines of uncarias; fourthly, planting two rows of polygonatum sibiricum on compartment surfaces in the shapes of regular triangles or inverted triangles; fifthly, managing uncaria and polygonatum sibiricum daily and preventing and controlling plant diseases and insect pests; and lastly, timely harvesting and processing polygonatum sibiricum.The relay-intercropping cultivation method of polygonatum sibiricum has following beneficial effects: a shed may not be required for shading since a growth condition is provided for imitated wild growth of polygonatum sibiricum, thereby solving the problem that manually-planted medicinal materials have poor quality; and high yield of polygonatum sibiricum is realized while an additional amount of uncaria is harvested such that land resources are fully utilized and comprehensive income is increased.

The invention relates to using a mouse able to generate human IgGl heavy chain- k light chain to prepare humanized monoclonal antibody and its application. The characteristic: respectively knocking in huma k-chain and gl-chain constant gene at the sites of gene in mouse k- light chain constant region and heavy chain gl constant region; the humanized monoclonal antibody: the constant region is fully the same as that of human antibody and the variable region is mouse sourece, and the preparing steps: obtaining mouse k-chain and gl-chain gene groups; constructing a knoncking-in carrier; making homologous recombination in ES cell; producing the mouse with light-heave conversion; further mating to obtain a mouse with complete chimeric gene; the application: taking spleen and marrow cells of an immunized chemeric mouse to make fusion and culture; screening hybridomas of humanized monoclonal antibody generating antigenic specificity; preparing the humanized monoclonal antibody with antigenic specificity. The advantages is easy to obtain high-affinity human-mouse chimeric antibody.

The invention discloses a method for planting similarly-wild dendrobium. The method for planting the similarly-wild dendrobium comprises the seedling screening step, the host selection step, the planting geographical climate condition selection step, the planting environment selection step, the planting environment optimization step, the colonization cultivation step and the planting management step. The ecological environment for planting of the dendrobium is accurately selected and optimized, three-dimensional large-scale planting of the dendrobium in a tea garden is achieved, chemical fertilizer or pesticides are not applied, independent occupation of land resources is not caused by planting, quality of dendrobium products can be greatly improved, and the planting technology is easy to learn, small in investment, low in cost and applicable to wide popularization and application.

The invention provides an ultralow temperature storage and regeneration culture method of an in vitro stem tip of Jerusalem artichoke. The method comprises the following steps: obtaining materials; peeling to obtain the stem tip; pre-culturing saccharose; loading a loading liquid; carrying out dehydration treatment by using PVS2 (Plant Vitrification Solution) vitrification protective liquid; carrying out ultralow temperature cryopreservation; unfreezing and washing; and recovering regenerative culture. By adopting the ultralow temperature storage technology by a droplet vitrification method, the vitro stem tip of a test-tube plantlet of Jerusalem artichoke stored at the ultralow temperature is defrosted and recovered in cultivation and is directly regenerated to form a seedling. The method is a long-term safe, stable, reliable and simple and effective method for storing germplasm resources of Jerusalem artichoke, wherein the survival rate can reach over 90%, the regeneration rate can reach over 80% and the regenerated plant is good in recovery growth.

The invention discloses a method for culturing tissue of 'Zhongzhen No.1'. The method comprises the following steps of: performing sterile pre-culture on 'Zhongzhen No.1' seeds to obtain sterile seedlings; and then performing primary culture on stem tips of the sterile seedlings to obtain regenerated seedlings. Experiments show that by performing pre-culture before producing the 'Zhongzhen No.1' regenerated seedlings, the differentiation rate of the stem tip of the pre-cultured seedling reaches 87.77 percent while the differentiation rate of the stem tip of the seedling cultured in nutrient soils is only 21.07 percent. In the method of the invention, the seeds are used as explant materials, and by adopting a mode of preculture of the sterile seedlings, the genetic stability of excellent characters is guaranteed; moreover, the method of the invention has the advantages that: the antiseptic processing method is improved; and the types of suitable anti-browning materials and hormones and the concentrations of the materials are screened out, the pollution and browning problems in present researches are solved, and a 'Zhongzhen No.1' sterile cultivation system is established successfully.

The invention discloses a tissue culture method of pitayas, belonging to the technical field of plant tissue culture. The method comprises the following steps: sterilizing a ripe stem section of a pitaya, inoculating the sterilized stem section on an axillary bud induction culture medium containing MS+0.1-1.5mg / l of paclobutrazol, cutting generated axillary bud and placing on the other culture medium containing MS+6-1.0-6.0mg / l of BA+0.1-1.5mg / l of chlormequat chloride, taking root simultaneously for 42% of test tube seedling which can be transplanted directly with above 95% of transplant survival rate. The method in the invention can be applied to industrialization production, and has the good effect of formula and fast budding. The pitaya germchit produced by the method has the advantages of high genetic stability, strong operability and high application value. By using the method, fast factorial growing seedlings can be realized and pitaya is developed and utilized more effectively.

The invention provides a rapid propagating method of Bletilla. Bletilla seedlings are obtained through the steps of treatment of seedling culturing beds, selection and pretreatment of explants, cluster bud induction, Bletilla seedling root system culturing, Bletilla division culturing, final-period management and the like. The method is very suitable for industrial production, and has the characteristics of excellent formula effect, high bud ratio, short culture time, vigorous plant growth, high transplanting survival rate, strong operability, high application value, low investment, high yield and the like.

The invention provides a method for rapidly breeding seedlings by using bletilla striata tubers. According to the method, rapid breeding is realized by using bletilla striata tubers, and the technical route of synchronization of bud increase and rooting is adopted, complete plants can be formed after about 50 days, and a large number of complete plants can be obtained within 4-6 months, so that large-scale industrial seedling breeding of bletilla striata is realized. The method for rapidly breeding seedlings by using bletilla striata tubers is simple, and the seedling breeding cost is reduced greatly. By adopting the method, quick budding and high reproduction rate are realized, the method is simple, tissue culture equipment is not required to be added, the production cost is low, batch production is realized and the application value is high.

The invention discloses a quick propagation method for humulus scandens. The method comprises the following steps of: culturing tissues, such as stem section of the humulus scandens, on an LS culture medium; accurately cutting and taking the stem section with 1-2 axillary buds as an explant; disinfecting and then inoculating in an LS+6-BA 1.2mg / L+NAA 0.2mg / L culture medium for culturing; wherein the explant buds after being inoculated for about 6-8 days and grows into a plantlet with leaves after being inoculated for about 10 days; cutting off the plantlet and transferring to an LS+6-BA 0.5mg / L+IAA 0.4mg / L culture medium, so as to keep high-speed increasing and a propagation coefficient above 5; selecting LS+6-BA 0.5mg / L+IAA 0.4mg / L+NAA 0.5mg / L as a rooting culture medium, wherein the rooting rate is above 90% and the complete plant can be obtained only after about 25 days; and transplanting a test-tube plantlet into sandy soil, carefully keeping water and humidity, and establishing a set of high-frequency stable regeneration system after the transplanting survival rate is above 90% after transplanting for 1 month. The method provided by the invention has the advantages of excellent stability, convenience in operation, high propagation speed and low production cost, achieves industrial level, and the like.

The invention relates to a quick adventitious buds inducing method for the panax tuber section. The method uses the panax tuber section as the explants, and induces the adventitious buds on the MS substrate of the additional hormone, and to proceed with the multiplication of the adventitious buds. The invention can directly induce the buds through the path produced by the direct organs, the speed is high, the sprouting rate is high, the cost is low, and the inducing process of the adventitious buds can be completed after 25 days about, the reproducing frequency is high, the variation rate is low, the period is relatively short, and the un-differentiated cells can directly differentiate into buds, so that the clone variation of the body cells is small. The invention can better keep the genetic stability of the host plant, and can ensure the transformed exogenous gene to transmit stably, the limit from the genotype is low, the adventitious buds obtaining rate is relatively high, and thereby is an effective measurement for obtaining the artificially cultivated panax seedlings in large scale.

The invention discloses a seedling method for paris polyphylla. A large number of complete plants can be obtained by breaking dormancy of latent buds at a low temperature and then carrying out shoot-inducing and root-inducing treatment, so that large-scale industrial breeding of the paris polyphylla is achieved. According to the seedling method, the inhibiting effect of various latent buds on a single rootstock of the paris polyphylla is completely avoided; the propagation coefficient is increased to the maximal extent; the hereditary stability of the paris polyphylla variety is fully ensured; and meanwhile, the phenomenon that variation of seeds and tissue culture seedlings is easily generated is avoided. Compared with other propagation methods employing the rootstocks, the seedling method has the advantages that completely separate plants can be formed; and seedlings with a plurality of buds can also be formed.

The invention provides a construction method of porcine reproductive and respiratory syndrome virus (PRRSV) recombinant plasmid for expressing ASFV (African swine fever virus) p12 protein or p17 protein. The invention also provides a genetic engineering vaccine constructed according to the recombinant plasmid and a construction method. Indirect immunofluorescence is carried out on rPRRSV-p12 and rPRRSV-p17 infection holes, and it is found that specific fluorescence for resisting PRRSV N protein and specific fluorescence for resisting ASFV p12 protein or p17 protein appear in the visual field.It shows that the recombinant PRRSV expressing African swine fever virus p12 protein or p17 protein can ensure that the p12 or p17 protein of exogenous protein ASFV can be well, efficiently and stablyexpressed. Researches show that p12 and p17 are important antigen proteins and can be used for ASF vaccine development.

The invention discloses a rapid propagation method of triarrhena sacchariflora. The rapid propagation method comprises the following steps of: selecting an excellent single plant of the triarrhena sacchariflora; cutting to take a stem section with 1-2 axillary buds as an explant; after sterilization, inoculating the explant in MS+6-BA0-2.0 mg / L culture medium to culture for 3-5 days, wherein the axillary buds start to sprout and a plantlet is formed around 7-9 days; cutting the plantlet and inoculating the plantlet in MS+6-BA 2.0-6.0 mg / L +IAA0.1-0.5 mg / L+NAA0-0.5 mg / L enrichment culture medium, and keeping high-speed increasing by degrees, wherein enrichment coefficient is above 4; synchronously inducing buds and roots on the enrichment culture medium which is added with NAA, wherein rootage rate is above 85 percent, and a complete plant can be formed only within about 35 days; and transplanting a test tube plantlet to natural soil, wherein transplanting survival rate reaches above 90 percent; thus a set of high-frequency and stable regeneration system is established. The method greatly simplifies the operation process, shortens the culturing time, saves the production cost, and provides a seedling growing method with short cycle, high propagation rate and low cost for large scale planting of the triarrhena sacchariflora.

The invention discloses a method for rapidly propagating wild ginseng. The method mainly comprises the following steps of: inducing callus, differentiating buds, performing successive transfer culture, performing multiplication culture, performing rooting culture, and hardening seedlings and transplanting. Good effects of high propagation speed, stable hereditary character, high propagation coefficient and eugenic test-tube seedlings are achieved; and through culture, the germination rate of the callus of the wild ginseng is 95 percent, the proliferation times is 4.8 times, the rooting percentage is 96 percent, the final survival rate is 98 percent and the quality is well ensured.

The invention belongs to the plant tissue culturing technical field, discloses a method for isolated culturing hypericum regenerated plant, the steps comprising: A: inoculating non-bacterial seeds of pre-cultivating hypericum different varieties in an MS minimal medium to cultivate for generation; B: formulating regeneration culture medium: MS minimal medium+TDZ0.08-0.8 mg / L+Alpha-fruitone 0.01-0.05 mg / L+filter parer as filter parer bridge; C: under sterilized condition, inoculating the blade of hypericum non-bacterial seeds of hypericum cultured for 40-45 days successively as explants in the MS regeneration culture medium, enabling the paraxial surface to be toward the regeneration culture medium, converting to cultivate under light irradiation after dark cultivating 15-25 days, after 24-26 days, obtaining the regeneration bud; D: inoculating the cutting regeneration bud of the step C in the MS minimal medium to cultivate successively to obtain the complete regenerated plant. The invention is simple, the operation is convenient, the regeneration frequency is high, the preparations of genetic transforming of the cutting flower hypericum and the molecule breeding materials can be quickened.

The invention relates to a method for seed selection of an excellent honeysuckle flower Lonicera japonica Thunb II variety and is suitable for seed selection of a novel honeysuckle flower variety. The method comprises the following steps of collecting branches of a large chicken feet-type farm-variety plant in the main honeysuckle flower producing area in the Pingyi county of the Shandong province, carrying out irradiation mutagenesis by 15 to 25Gy of <60>Co-gamma rays, carrying out cottage and transplant, carrying out single-plant seed selection of variant individuals, carrying out cottage and transplant to obtain a plant system, and carrying out variety comparison and production test. The novel Lonicera japonica Thunb II plant has a short, thick and straight limb, a large amount of branches, long circle-shaped leaves, sparse and short pubescence at leaf veins and leaf edges, internode length of 2.5 to 7cm, average flower bud length of 4.16cm, a large amount of flowers, a long bud period, compact pier piles, concentrated buds, chicken feet-shaped flowers and an early flowering phase, can be picked easily, and is suitable for close planting. Compared with the original large chicken feet-type farm-variety, the honeysuckle flower Lonicera japonica Thunb II obtained by the method has a honeysuckle flower medicinal material yield improved by 20.2%, chlorogenic acid content improved by 17.95% and galuteolin content improved by 15.20%.

The invention discloses a tissue culture quick propagation method of Polygonatum kingianum. The method comprises the following steps: explant selection and treatment, induced culture, successive transfer culture, enrichment culture, rooting culture, and acclimatization and transplant. The induced culture medium is composed of MS, 1.0-3.0mg / L6-BA, 0.5-1.0mg / L 2,4-D, 1.0-2.0mg / L gibberellin, 5.0-10.0g / L agar and 20.0-30.0g / L sucrose. The rooting culture medium is composed of 1 / 2MS, 0.2-1.0mg / L NAA, 0.2-0.6mg / L IBA, 0.2-0.5% of activated carbon, 5-20% of banana, 6.0-8.0g / L agar and 20.0-25.0g / L sucrose. The method effectively reduces the seed / seedling culture time and cost, increases the quick propagation speed, effectively enhances the Polygonatum sibiricum propagation coefficient, and can well keep the hereditary stability of Polygonatum sibiricum; and the survival percent after transplant can reach 98% or above, thereby creating favorable social benefits and economic benefits.

The invention relates to a rapid proliferation method of hosta undulata bailey. The rapid proliferation method comprises the following main steps of selecting and disinfecting explants, carrying out subculture enrichment culture on the explants, carrying out rooting culture on the explants, and carrying out acclimatization and transplant on the explants. The rapid proliferation method provided by the invention has the advantages that in a subculture process, the illumination intensity is improved, the photoperiod is prolonged, the culture temperature is reduced, meanwhile, a technological method for transforming the subculture culture medium is adopted, and the good effects that the proliferation speed is rapid, the inheritable character is stable, and the proliferation coefficient is high are achieved.

The present invention provides novel methods of maintaining genetic stability of non-human animal inbred strains. In the methods, pedigree-tracked cryopreserved embryos or gametes or pre-gametes derived from a foundation colony are produced and used to re-establish the foundation colony at appropriate intervals.

The invention relates to a breeding method for an improved variety of Lonicera japonica Thunb No.1, and is applicable to breeding of new variety of Lonicera japonica Thunb. Farmyard germplasm type Lonicera japonica Thunb branches from main producing area of pingyi county, ShanDong province, are collected; 15-25 Gy60Co-Gamma is taken for radiation induced mutation, and then cuttage and transplant are carried out; seed selection, cottage and transplant for variation individual plant are performed so as to form a strain; the Lonicera japonica Thunb No.1 obtained through variety comparison and production testing has thick and upright branches, oblong, linear oblong and oblong-ovate leaves, has dark green and thick leaves, and has more blossom buds; and flower buds are 4.5 cm in length, bracts are oval and oblong-ovate, the flower season comes earlier, saline-alkaline tolerance and barren resistance and strong adaptability are achieved, and plant diseases and insect pests are less. Compared with common Lonicera japonica Thunb, the Lonicera japonica Thunb No.1 has the advantages that the flower season starts two to three days earlier, the flower bud yield is improved by 16.20 percent, the content of galuteolin of flower bud is 0.106 percent, and the content of chlorogenic acid accounts for 1.75 percent, which is higher than the quality standard of Pharmacopoeia of the People's Republic of China (2010).

The invention discloses a Hibiscus aridicola Anthony purple flower excellent single plant tissue culture rapid propagation method. Hibiscus aridicola Anthony purple flower excellent single plants areadopted as explants, and through a series of culture steps of explant sterilixation, induction, differentiation, proliferation and rooting, the bottle neck problems of tissue culture rapid propagation, introduction and domestication and commercial development and application of Hibiscus aridicola Anthony are effectively solved, the situations that natural plants are reduced and natural vegetationregions are damaged because of over-use of wild resources are effectively avoided, and particularly the method has the beneficial effect of maintaining stability of special shapes of excellent singleplants. The method has a very good development and application prospect. By adopting the method, the induction differentiation rate is 80%, the breeding cycle is 30 days, the growth coefficient is 4,the rooting rate is 95%, the transplanting survival rate is 92%, the reproduction number and the growth velocity of the Hibiscus aridicola Anthony are greatly increased, and technical supports for introduction and domestication, preservation and on-scale production of the specimen are provided.

The invention discloses a construction method of an in vitro regeneration system of a tea tree. According to the construction method, the construction of the in vitro regeneration system takes a stem with an axillary bud as an explant, the stem is cultured, the axillary bud is germinated to grow leaves upwards and roots downwards, and then a seedling is directly formed without callus induction. According to the invention, the stem of Fuding Dabaicha is used as the explant and the construction method of the regeneration system is researched; the hereditary stability of tea tree varieties is kept and the storage of germplasm resources of tea trees can also be promoted.

The invention provides a construction method of porcine reproductive and respiratory syndrome virus (PRRSV) recombinant plasmid expressing African swine fever virus (ASFV) p30 protein, and a genetically engineering vaccine constructed based on the recombinant plasmid and a construction method thereof. Indirect immunofluorescence is carried out on rPRRSV-p30 infected wells, it is found that the specific fluorescence of anti-PRRSV N protein and anti-ASFV p30 protein appears in the field of vision. Protein samples of rPRRSV-p30 infected wells are collected and analyzed by Western blot, results show that p30 bands of the expected size are found. It is suggested that the recombinant PRRSV expressing African swine fever virus p30 protein can ensure good, efficient and stable expression of foreign protein ASFV p30 protein. However, some studies show that p30 is an important antigen protein, and can be used in the development of ASF vaccine.

The invention relates to a rapid propagation method of nothapodytes pittosporoides, and the method is characterized in that tissues such as tender shoot sections are cultivated on a murashige and skoog (MS) culture medium, a tender shoot stem section with one to two sprouts is cut as explants, the explants are grafted in MS+6-butyl acrylate (BA) 1.2mg / L+ indole butyric acid (IBA)0.2mg / L culture medium after being sterilized, the bud is germinated in 10 to 15 days after the grafting, seedlings with leaves are formed in 30 days, the seedlings are cut off and transferred onto the MS+6-BA1.2mg / L+IBA0.4mg / L culture medium to keep the high-speed increase, and the propagation coefficient is more than 5. The MS+6-BA0.5mg / L+ naphthyl acetic acid (NAA)1.5mg / L is selected as rooting culture medium, the rooting rate reaches more than 95 percent, and a complete plant can be formed in 30 days. Test-tube plantlets are transplanted in sandy soil, attention is paid on the moisture preservation, the transplanting survival rate can reach up to 93 percent after the test-tube plantlets are transplanted for 45 days, and a high-frequency stable renewable system is established. The method has the advantages of good stability, simplicity and convenience in operation, fast propagation speed, low production cost, industrialization and the like.

The invention discloses a mutant vector pCCFR2-2bPTIII containing cucumber mosaic virus Fny isolate RNA2. The CMVFny mutant RNA2 nucleotide sequence of the vector is shown as SEQ ID NO. 1. The mutant is mixed and inoculated with CMVFnyRNA1 and CMVFnyRNA3, and has a cross protection effect on CMV; and the mutant vector pCCFR2-2bPTIII at least can accommodate 350bp exogenous fragments and maintain the genetic stability, provides a more optimized basic vector for researching and developing multivalent attenuated vaccines, and provides vaccine materials and effective prevention and treatment measures for preventing and treating CMV and other viruses in the field.

The invention provides a method for conserving radix ginseng germplasm resources and propagating seeds and seedlings. Excellent radix ginseng varieties or the germplasm resources can be planted in sunlight greenhouses in an isolation manner by the aid of the method. The technical method includes (1), selecting locations; (2), constructing the sunlight greenhouses; (3), preparing seedbed soil substrates; (4), turning, disinfecting and laying soil; (5), directionally arranging beds; (6), carrying out germination accelerating treatment on the seeds; (7), establishing germplasm resource propagation gardens; (8), carrying out seed sowing modes; (9), carrying out management in growth periods; (10), carrying out processes for propagating improved varieties; (11), controlling diseases and insect pests; (12), harvesting fruits and storing the seeds; (13), carrying out processes for lifting, collecting and storing the seedlings, and the like. The method has the advantages that the scientific method for protecting the radix ginseng germplasm resources and propagating the seeds and the seedlings is provided according to biological characteristics of radix ginseng, and material guarantee and technical support can be provided for conserving and innovatively utilizing the radix ginseng germplasm resources.

The invention discloses a rapid propagation method of tissue culture of female and male plants of Zhejiang hemsleya amabilis. The stem segments of the female and male plants of Zhejiang hemsleya amabilis of which the sex is known are utilized as explants, and the method further comprises the following steps: disinfection of the stem segments, starting cultivation of axillary buds, enrichment culture / rapid augmentation of the stem segments of aseptic seedlings, induction of blade callus, and differentiation culture, rooting culture and transplantation of adventitious buds. The method successively realizes the rapid propagation of the tissue culture of the female and male plants of Zhejiang hemsleya amabilis, can rapidly cultivate a large quantity of test-tube plantlets, of which the sex isknown, within a short period of time, provides a feasible propagation mode for the recovery of an extremely small population, and further provides a feasible development project for planting traditional Chinese medicine GAP simultaneously.

The invention relates to the biologically technical field, in particular to a mass differentiation and rooting method of protocorms of dendrobium candidum. The invention adopts the main technical scheme that the mass differentiation and rooting method comprises the following steps: sterile explant obtaining: soaking dendrobium candidum capsules with a cleaning powder for 3 minutes, then fishing out, washing with tap water, sterilizing the surface with 70 weight percent of alcohol for 1.5 minutes under aseptic conditions, cleaning with sterile water for 1 minute, then soaking with 2 weight percent of sodium hypochlorite solution for 10-15 minutes, finally cleaning with sterile water for 4-5 times for 1 minute at a time and after the end of washing, absorbing moisture on the surfaces of thedendrobium candidum capsules with sterile filter paper. The mass differentiation and rooting method provided by the invention has the benefits that based on the study on rapid propagation of mass differentiation and rooting of the protocorms of the dendrobium candidum, the rapid test tube propagation can be performed, so that a reliable technical support is provided for the oriented large-scale industrial production of the dendrobium candidum; the method is simple and practical, efficient and fast, and suitable for large-scale production.

The invention relates to the technical field of plant tissue culture and provides a tissue-culture rapid propagation method for craigia yunnanensis. According to the tissue-culture rapid propagation method, induction and differentiation culture are integrated, multiplication and subculture are integrated, and seedling strengthening and rooting culture are integrated, so that the culture method issimplified. The tissue-culture rapid propagation method has advantages that the problem of difficulty in short-time mass propagation, storage and sustainable utilization due to less natural resourcesof craigia yunnanensis can be solved, a biotechnological research gap of craigia yunnanensis is filled, and a foundation is laid for comprehensive development and sustainable utilization of craigia yunnanensis.