46results about How to "Simplify the cultivation procedure" patented technology

The invention belongs to the field of veterinary parasites, and relates to coccidian oocyst culture preservative fluid and an application method thereof. The invention is characterized in that the culture preservative fluid takes polyhaxemethylenguanidine hydrochloride as a main ingredient. The application method of the coccidian oocyst culture preservative fluid comprises the following steps of: separating and purifying fresh coccidian oocysts from fowl dung or intestinal tract tissues containing the coccidian oocysts; adding an appropriate amount of a polyhaxemethylenguanidine hydrochloride solution; adjusting the density of the oocysts; controlling the depth of a culture medium; putting the culture medium in a shaking table to perform shaking culture; performing microscopic examination; stopping culture when the sporulation rate of the oocysts exceeds 85%; collecting the oocysts; adding an appropriate amount of the polyhaxemethylenguanidine hydrochloride solution again; and preserving in a refrigerator of which the temperature is 4 DEG C. The coccidian oocyst culture preservative fluid is efficient, long-acting and safe, pollution-free to the environment, simple to prepare and low in cost, and is ideal coccidian oocyst culture preservative fluid. By applying the application method provided by the invention, the operation is simple and convenient, the efficiency is high, the automation can be realized, and the coccidian oocyst culture preservative fluid is suitable for large-scale culture preservation of coccidian oocysts.

The invention relates to an open type simplified culture medium for taxus cuspidate. The culture medium comprises a primary culture medium and a subculture medium. The primary culture medium is prepared by adding and mixing carrageenan, white granulated sugar, indolebutyric acid IBA, active carbon AC and agricultural streptomycin into optimized B5 basic mother liquor. The subculture is prepared by adding and mixing carrageenan, white granulated sugar, indolebutyric acid IBA, 6-benzyl amino adenine BA, active carbon AC and agricultural streptomycin into optimized MS basic mother liquor. According to the culture medium provided by the invention, with the adoption of open type simplified tissue culture, the cost and the inoculating time of the culture medium are remarkably saved, the autoclaved sterilization program is cancelled, and the production flow is simplified by replacing autoclaved sterilization and strict enclosed environment operation by a bacteriostatic agent, so that the labor and equipment investment in industrialized seedling production can be reduced, and the fund is saved by 30-50%.

The invention discloses a method for directly inducing plant regeneration by using Vernicia fordii hypocotyl as an explant, comprising the following steps: using hypocotyl of a Vernicia fordii seed aseptic seedling as an explant, directly inducing an adventitious bud, proliferating and elongation culturing the adventitious bud, and performing rooting culturing, domestication and transplanting to obtain a robust regenerated plant of Vernicia fordii. Compared with the prior art, the method has the advantages such as regeneration process simplicity, hormone addition singleness, operation convenience and relatively short regeneration cycle. A new technical means to rapidly propagate Vernicia fordii is provided, and also certain technical support is provided for genetically improving Vernicia fordii by genetic engineering means in future.

The invention relates to an out-of-bottle bag rooting method for tissue culture seedlings of Europe and America hybrid aspens. The method is to the problem that the cultivation cost is high, the production phase is long and the survival rate of transplanting of tissue culture seedlings is low in a tissue culture process of the Europe and America hybrid aspens. The method comprises the following steps: 1, plastically packaging the bottom of polypropylene plastic bags with a sealing machine to manufacture rooting culture bags; 2, mixing peat soils with sands, sterilizing at high speed to obtain composts and packing the composts into the rooting culture bags; 3, burrowing in the composts in the culture bags and wetting each hole with rooting culture solution; and 4, taking out the tissue culture seedlings of the Europe and America hybrid aspens from culture mediums, planting the tissue culture seedlings into the holes of the culture bags, sealing the top of the culture bags with the sealing machine, placing in greenhouses to culture for 5-7 days, gashing the top of the culture bags and then culturing for 8-10 days to obtain the rooting seedlings of the Europe and America hybrid aspens. The method is used in the field of asexual propagation of the Europe and America hybrid aspens.

The invention relates to a one-step seedling and efficient in-vitro propagation method for Begoniaceae gynura bicolor, which is an endangered plant with homology of medicine and food. The method provided by the invention comprises the following steps of: preparing a culture medium, and adjusting the culture medium through preparing, subpackaging and sterilizing; carrying out sterile inoculating directly on gynura bicolor aseptic seedling leaves which are taken as explants; culturing the inoculated culture medium in a culture chamber for 70-80 days under appropriate conditions, wherein the number of a single explant induced seedlings is 20 above; and decapping, transplanting regenerating plants to appropriate conditions, and culturing without hardening-seedling, wherein the commodity seedling survival rate achieves 100%. According to the method provided by the invention, the gynura bicolor tissue culture seedling regeneration period is short; the original seed property is kept effectively; the vegetative propagation coefficient is improved obviously; the operation is simple and convenient; the culture program is simple; the seedling cost is lowered obviously; and the method can act as an effective technology of industrialized production of high-quality seedlings of Begoniaceae gynura bicolor.

The invention discloses a one-step plantlet formation medium and method for the isolated culture of lamiophlomis rotata. By adopting the one-step plantlet formation medium, a lamiophlomis rotata test-tube plantlet leaf forms a plantlet in one step through isolated culture. The one-step plantlet formation culture medium is characterized in that a MS (Murashige and Skoog) medium is used as a basic medium, and 6-benzylaminopurine (6-BA), naphthylacetic acid (NAA) and 2,4-dichlorphenoxyacetic acid (2,4-D) which are different in concentrations and combinations are added to the base medium. According to the one-step plantlet formation method, the test-tube plantlet leaf serving as an explant is inoculated, then is induced by a callus and is directly differentiated into an adventitious bud and an adventitious root, so that the lamiophlomis rotata regenerated plantlet is obtained by one step. By adopting the one-step plantlet formation method, the callus obtained by induction in an initial medium does not need to be transferred into a differential medium for regenerating the plantlet. Compared with an isolated culture multi-step regeneration plantlet formation method, the one-step plantlet formation method has the advantages that the steps are simplified, the culture period is short, the repeatability is good, and the planting percent is high.

The invention discloses an artificial efficient propagation method of aristolochia tuberosa. The method comprises the following steps of inoculating a disinfected and sterilized explant with a bud stem segments of the aristolochia tuberosa into a culture medium A for culture, starting propagation culture under the conditions that the illuminance is 1500-2000lx, the illumination time is 10h/d and the temperature is controlled to be 22 +/-1 DEG C, and obtaining a propagation coefficient of 3.57 after 35 days. A material is shorn into the stem segments containing 2-3 joints, the stem segments areinoculated into a culture medium B to be cultured, a clustered cluster system with base adventitious cluster buds and upper axillary buds growing is formed after 45 days, and the propagation coefficient reaches 10.0 or above. When the height of proliferated cluster buds is 3-4cm, single seedlings are inoculated into the culture medium B to be cultured for 45d to obtain rooted seedlings with strong seedlings and thick roots, and then the rooted seedlings are transplanted. The method is low in cost, short in period and high in test-tube plantlet quality and survival rate, provides technical support for protecting wild resources and breeding high-quality plantlets, and can be used for producing excellent plantlets with consistent genotype backgrounds in a short time.

The invention discloses a method and application for inducing direct somatic embryogenesis. According to the method and application, function genes RID3(AT3G49180) and YUC10(AT1G48910) capable of promoting direct somatic embryogenesis are found. Through ectopic expression of the function genes, inducing can be directly conducted on arabidopsis cotyledons to form the somatic embryo without any callus. The method for inducing direct somatic embryogenesis has the advantages of being simple in culture, short in inducing period, high in inducing frequency and the like, and can be used for study onaspects of differentiation, development and totipotency expression of botanical cells, crop variety improvement, production of artificial seeds, storage of high-quality germplasm resources, mutant screening and the like.

The invention relates to a method for establishing an efficient regeneration system by taking medical and edible plant dandelion roots as explants. The method comprises the following steps: (1) putting the dandelion roots in an MS medium to induce one-step culture; (2) putting robust cluster buds or plants into a 1/2MS rooting medium, and performing cultivation to obtain complete seedlings with roots; (3) exercising the complete seedlings with the roots, putting the complete seedlings with the roots into a substrate to grow for 20 to 30d, and then transplanting the complete seedlings with the roots to a large field. The seedlings obtained by using the method are robust and high in survival rate, and a great number of high-quality seedlings of dandelion can be provided within a short time.

The invention relates to a method for expressing an antibacterial peptide apidaecin by using Escherichia coli and for preparing the antibacterial peptide apidaecin. The method comprises the following steps: cloning AP2 gene and a polyhedron gene sequence polh into a recombinant vector in order to form a recombinant vector with a polh-AP fusion gene fragment; cloning the polh-AP fusion gene fragment into an expression vector, converting the expression vector to an expression host cell, and culturing the expression host cell to express a fusion protein with an antibacterial peptide AP2; and culturing the expression host cell to induce the expression of the recombinant protein polh-AP, centrifuging induced bacterial strains, collecting, re-suspending, carrying out ultrasonic fragmentation, centrifuging, collecting the above obtained insoluble inclusion body, purifying, and collecting a recombinant protein sample. An expression system has the characteristics of simple expression system culture program, low production cost, efficient expression of the antibacterial peptide apidaecin, and realization of AP2 and polh fusion expression, so compared with the prior art, the method has the advantages of effective reduction of toxicity of the AP2 to Escherichia coli, improvement of the stability of the antibacterial peptide, realization of high level expression of the antibacterial peptide, convenient purification of the active antibacterial peptide, and improvement of the output of the antibacterial peptide apidaecin.