43results about How to "Culture conditions can be controlled" patented technology

The invention discloses a method for sterile seeding and rapid propagation of bougainvillea speetabilis. The method comprises the steps of conducting sterile seeding and induction germination on seeds, wherein the seeds are sterilized, then a shoot induction medium is inoculated with the sterilized seeds, and shoot induction culture is conducted at proper temperature with proper illumination time and illumination intensity; conducting seedling breeding through bud expanding propagation and rooting reinduction, wherein a sprout enrichment culture medium is inoculated with germinated buds, and bud propagation subculture is conducted at proper temperature with proper illumination time and illumination intensity; inoculating a rooting medium with the bud propagation subcultured bud seedlings, and conducting rooting breeding of seedlings at proper temperature with proper illumination time and illumination intensity. According to the method, seeds are used as breeding explant, seed treatment is easy, pollution rate is low, the germination rate of seeds reaches 100%, bud propagation rate reaches 5.22 times within 30 days, rooting rate is 95.6%, and seedling breeding is not limited by time or season so that whole-year production can be conducted.

The invention discloses a rapid algae-liquid separation method for microalgae treatment wastewater. The method includes the steps of collecting wastewater discharged by a sewage treatment plant to be put indoors for standby application, processing CODCr of wastewater through the potassium dichromate method, processing TN through the potassium persulfate oxidation ultraviolet spectrophotometry, processing TP through the antimony and molybdenum spectrophotometry, processing ammonia nitrogen through the Nessler spectrophotometry, and adjusting the pH through the glass electrode method. The method can be conducted in a cultivation device with the controllable turbulence intensity, temperature and light can be conveniently controlled, the efficiency of microalgae for processing wastewater is improved, no flocculant needs to be added under the turbulence condition, flocculating constituent is formed according to the property changes of microalgae cell surfaces so that algae and liquid can be separated, turbulence energy consumption can be sufficiently utilized, energy consumption is lowered, efficiency cost is improved, the pH of a culture solution is increased during processing, pathogenic organisms in a culture medium can be restrained, the produced flocculating constituent has high sedimentation speed, the algae-liquid separation efficiency is improved, operation is easy and convenient, cultivation conditions are easy to control, and an experimental device for the method is simple in structure, small in size, light in weight, high in practicability and repeatability, simple in process, convenient to machine, capable of saving labor, materials and financial resources, flexible to operate and convenient to clean.

The invention discloses a pluripotent stem cells directional differentiation method. The method is characterized in that pluripotent stem cells are cultured under cell culture environment of non exogenous hematopoietic cytokines, non serum, non matrix cells and clear component, after embryoid is formed, a vascular endothelial growth factor and bone morphogenetic protein 4 are added, the embryoid is attached on the surface spread with collagen IV, mesoderm differentiation is induced, and hemopoietic progenitor cells and blood corpuscles are generated. The cell culture system with determined component enables directional differentiation on the pluripotent stem cells, has the advantages of low cost, simple operation, high repeatability and ordered differentiation process, can massively generate hemopoietic progenitor cells and blood corpuscles, and has important meaning for researching a blood growth process mechanism.

The invention relates to an inducing technology of a test tube lotus root, which comprises the steps of: using terminal buds or lateral buds of the lotus root as explants; carrying out the starting of stem tips, successive proliferation and rooting; finally culturing and inducing by using a solid-liquid culture base to form the test tube lotus root. The method for inducing the test tube lotus root comprises the following steps of: cutting the stem tips of the lotus root and inoculating the stem tips on a stem tip starting culture base for culturing so as to grow up to become buds or a cluster of buds; transferring the buds or the cluster of buds to a successive culture base and carrying out successive proliferation; segmenting the cluster of buds into single bud and transferring to a rooting culture base and inducing to root to form tube seedlings; and rooting and culturing for 30d, pouring the liquid culture base into a culture container and inducing to form the test tube lotus root. In the process, the culture temperature is between 24 and 26 DEG C, the illumination intensity is 2000-25001x and the illumination is carried out for 10h every day. Through the stem tip culture and quick reproduction of the lotus root, the test tube lotus root is formed by inducing so as to quicken the reproduction speed of the lotus root, improve the transplant survival rate and industrially produce.

The invention discloses a probiotic feed additive for realizing zero discharge of manure waste. The feed additive for realizing zero discharge of manure waste is characterized by comprising Candida utilis, Bacillus lentus, Bacillus licheniformis, Lactobacillus delbrueckii, Lactobacillus plantarum, Lactobacillus reuteri, and Aspergillus oryzae. The bacterial strain matching used by the probiotic feed additive of the invention has the advantages of improving animal immunity and reducing the risk of disease caused by livestock, does not use antibiotics, and is healthy and environmentally friendly.

The invention discloses a mesenchymal stem cell and biomaterial combined microsphere and an intelligent spraying system. Firstly, sodium alginate is selected as a carrier biomaterial and subjected topartial oxidation treatment, protein components with free amino groups are immobilized through hydrazone bonds, and lithium hydroxide and salicylaldehyde are added into an immobilization process to effectively improve protein component immobilization efficiency of the material; then the hydrazone bonds are reduced by sodium borohydride to obtain C-N single bonds; finally, the protein component immobilized material and stem cells are mixed to prepare cell 3D culture microspheres, the survival rate is effectively increased after stem cell transplantation, and skin wound curative effects of stemcells are improved. Due to the fact that cell products are extremely prone to damages and loss in a transplantation process due to vulnerability, a spray dosing system matched with the microsphere isfurther provided and capable of accurately and efficiently spraying the microsphere to wounded parts, and unnecessary loss and waste are avoided.

The invention discloses a Fagus pashanica tissue culture rapid propagation method which comprises the following steps: picking Fagus pashanica branch buds as explants; peeling off all scales of the buds so as to expose the bud points; disinfecting for 5min in 0.1% of mercury bichloride; and inoculating in a culture medium with WPM+NAA (Naphthalene Acetic Acid) of 1.0mg/L, 6-BA of 2.0mg/L and carbon powder of 2.0mg/L. After the buds are inoculated for 15 days, leaves grow up, and after 45 days, the leaves expand and grow larger and the leaves are light green in color and grow exuberantly; after being continuously cultivated for 60 days, white root systems grow out from the roots, the hair roots are obvious, the root systems are developed, and complete plants are grown and can be acclimatized and transplanted. The method has the following benefits that firstly, a research material is single in resource and is accordant in clone genetic background; secondly, extremely fewer plant materials are used, so that the materials are easy to acquire with economy, convenience and high efficiency; thirdly, the cultivation condition is controllable, and anniversary test or production can be carried out; and fourthly, the Fagus pashanica grows rapidly with short cycle and strong repeatability; and fifthly, the management is convenient, and the automation control is facilitated.

The invention relates to the propagation of Sargassum thunbergii, in particular to a method for massively propagating the seedlings of the Sargassum thunbergii by a tissue culture vane method, comprising the steps of: 1) picking the complete principal branches in the sample of the Sargassum thunbergii and using disinfected seawater to disinfect; 2) then using AgCl to disinfect and process; using a bistoury to take down the wide and large vanes in the middle lower part of the principal branches and using a nipper to arrange the wide and large vanes into prepared culture liquid; the culturing temperature is between 13 and 20 DEG C; the illumination intensity is 15 to 50 Mumol/m2 per s; and the illumination: darkness in the illumination period is equal to 8 to 10: 16 to 14h; 3) needing not to ventilate 7 to 10 days before culturing; after the 7 to 10 days, beginning to ventilate and culture after the wound healing of the vanes is finished; changing the culture liquid every 35 to 45 days; after culturing for 3 months, a plurality of buds are generated in the middle lower parts of the vanes; and after culturing for 5 months, regenerated strains can be collected to carry out fixed planting and culturing. The method needs less plant materials and less culturing space, can carry out idioplasm preservation for a long period, and has large fecundity and pure seedling quality, and the like.

The invention discloses a cutting fast propagation method of aronia melanocarpa. The method comprises the following steps: selecting an aronia melanocarpa plant to be cultured as a scion mother tree,cutting a strong semi-lignified branch with no diseases or insect pests in a growing period, soaking the lower part of the branch in clear water, placing the branch in a cool environment, and keepingbranches and leaves fresh and alive; acquiring cutting slips by cutting the branch into the cutting slips with a length of 8 centimeters, carrying out disinfection, soaking the lower notches in a rooting agent solution for 2 hours, inserting the cutting slips into a soilless medium subjected to disinfection, maintaining suitable temperature, humidity and illumination conditions, and preventing andtreating diseases and insect pests; and regularly spraying water for moisture preservation, and culturing for 45 days to reach rooting rate of 80% or above, and achieve transplanting conditions, wherein the obtained cutting slips can be used for culturing seedlings for production. The materials in the method provided by the invention are simple and easy to obtain, the operation specifications ofeach link are normative, the culture conditions are controllable, the repeatability is strong, the cost is low, the promotion and the production are facilitated, the rapid propagation of excellent seedlings of aronia melanocarpa is facilitated, and the method has a promotion and application value.

The invention relates to the field of cell culture, in particular to a domestication method of a full suspension culture-type MDCK cell line. According to the domestication method, adherent MDCK cellsare domesticated into a low-serum full suspension culture-type MDCK cell line and a serum-free full suspension culture-type MDCK cell line, so that traditional production of an avian influenza virusthrough chicken embryos can be replaced, the subsequent separation and purification efficiency is improved, further upgrade of a process is promoted, a culture process is also simplified while the cost is reduced, and the domestication method is easier to industrialize.

The invention discloses a closed slope type raceway pond system used for large-scale culture and harvesting of microalgae, belonging to the field of large-scale culture of microalgae. The system mainly comprises a raceway pond subsystem used for microalgae culture, a microalgae solution adjusting subsystem and a microalgae harvesting subsystem. The culture subsystem used for microalgae culture isprovided with a raceway pond with a reserved slope, so that a microalgae solution can flow along the raceway pond under the effect of gravity, and thus the energy consumption for driving the microalgae solution to flow is greatly reduced; the raceway pond is coupled with a closed photobioreactor, so that the purposes that the manufacturing cost is low, the operation is simple and convenient, the culture conditions are easy to control, and the pollution by the environment is little are achieved; the real-time harvesting of microalgae is realized by utilizing a filtering membrane, and thus the harvesting cost is greatly reduced.

The invention belongs to the field of plant-tissue culture methods, and particularly relates to an immature-embryo culture method for red-peel peanuts. The immature-embryo culture method based on the existing peanut tissue culture comprises the following steps: disinfection and soaking of seeds, induced culture of clustered shoots, culture of strong tissue culture seedlings, rooting culture of seedlings, and transplantation after seedling hardening of complete plants. The immature-embryo culture method has the advantages that complete peanut plants can be obtained within 15 weeks by carrying out tissue culture on peanuts, so the period for tissue culture is greatly shortened, mass propagation in short time can be achieved, and the significance for propagation of rare varieties and the current peanut transgenosis is very important; secondly, culture conditions can be artificially controlled, and are not limited by regions and seasons, so that convenience is brought for stable culture production, and foundation is laid for development of tissue culture and genetic transformation of peanuts.

The invention provides an apidaecin expression vector as well as a construction method and application thereof. The construction method comprises the following steps: 1) synthesizing an expression operon gene containing encoding NTEV protease; 2) synthesizing a fusion protein expression operon gene containing encoding xylanase and apidaecin; 3) conducting double-enzyme digestion on the expression operon genes obtained in the steps 1) and 2) in a separated mode, and constructing into a vector pBluescript II SK(+) so as to obtain a recombinant plasmid; 4) extracting a genome DNA of bacillus subtilis 168, amplifying a promoter p43, cloning by virtue of a T vector so as to obtain a plasmid containing a promoter gene, and constructing into a plasmid pGJ103 so as to obtain a plasmid pGJ284; conducting reverse PCR (polymerase chain reaction) so as to obtain an efficient promoter sequence gene containing partial gene sequences of the p43 promoter, and constructing into the plasmid pGJ284 so as to obtain a plasmid pGJ288; and 5) conducting double-enzyme digestion on the recombinant plasmid and plasmid pGJ288 in a separated mode, recovering enzyme digestion products and linking so as to obtain an apidaecin expression vector. The expression vector is high in expression amount; and the apidaecin extraction method is simple and convenient, and is suitable for industrial large-scale preparation.

The invention discloses cytochalasin derived from phenylalanine and a preparation method and application thereof. The preparation method of the cytochalasin compound comprises the following steps: inoculating endophytic fungi Xyllaria sp.CFL5 in leaves of cephalotaxus fortunei Hook.f into a rice culture medium, chopping the culture medium after the endophytic fungi Xyllaria sp.CFL5 grow for 3 weeks, soaking and extracting with ethyl acetate, and concentrating under reduced pressure to obtain a crude extract; and separating the extract by macroporous resin, silica gel and a semi-prepared high performance liquid chromatography column to obtain the compound. The compound has obvious inhibitory activity on the combination of LAG3 and MHC II and the combination of LAG3 and FGL1, and further inhibits the immune escape of tumor cells so that the compound can be used as a novel anti-cancer candidate drug.

The invention discloses a pruns cistena tissue culture seedling cultivation method, relates to plant propagation and belongs to seedling production technology in forestry biotechnology. The method ischaracterized by including: fumigating an intelligent seedling culture greenhouse for disinfection, selecting pruns cistena stems, cutting the same into apical-bud-containing stems of 2-3cm after surface sterilization, putting the stems onto a reciprocating shaking seedbed, and using an adventitious bud culture medium added with 6-BA (1.0mg/l) and NAA (0.05mg/l) for culture; when plants grow to 2.5-3.5cm, transferring onto another half-reciprocating shaking seedbed, using a rooting culture medium added with IBA (1.0mg/l) and IAA (1.0mg/l) to induce rooting, hardening seedlings for 5d after 35d, and transferring the seedlings into a nutrition bowl for propagation. Conventional seedling culture and tissue culture seedling culture methods are combined together, and shallow shake culture is performed through the reciprocating shaking seedbed, the weakness that liquid culture media are prone to causing culture death due to oxygen deficiency and the weakness that solid culture media are slowin growth are avoided, and the objective of large-scale quick propagation is achieved; the method is simple in process and easy for popularization.

The invention discloses a method of culturing eurotium cristatum on a coffee bean solid medium. According to the method, a solid medium is taken as the culture medium; eurotium cristatum powder is cultured in a coffee bean solid culture medium; the culture technology is simple, the culture conditions can be more easily controlled, the manufacturing effect is more stable and better, and the cultured eurotium cristatum powder is used to prepare eurotium cristatum coffee.

The invention discloses a culture method for kombucha and application thereof. The culture method comprises the following steps: (1) preparation of culture solution, after being boiled, water is added with sugar, sweet water is prepared after the sugar is dissolved, the sweet water is then cooled to 20 DEG C to 35 DEG C, and thereby culture solution is prepared; (2) inoculated culture: an incubator is first disinfected and rinsed, mother bacteria solution and the culture solution are then poured into the incubator according to a volume ratio of 1:20 to 50, a gauze is used for sealing, the incubator is put under the conditions of a dim light environment with the illumination intensity of one thousand to eight thousand Lx, ventilation and the temperature of 30 DEG C to 35 DEG C to carry out fermentation culture for 7 to 14 days, finally, rufous floccule or lumps floating on the surface of the culture solution after the completion of culture, is separated out, and thereby the kombucha is prepared. The process of preparing the mother bacteria solution in the preparation method is simple, the materials are simple and easy to obtain, moreover, the culture conditions of the kombucha are easy to control, and the contamination rate of the cultured strain is low.

The invention discloses a kombucha metabolite used for treating toothache and a preparation method thereof. The preparation method comprises the following steps: 1) boiling water, then adding sugar, dissolving sugar to prepare syrup and then cooling the syrup to 20 to 35 DEG C to prepare a culture solution; 2) carrying out inoculated culture, i.e., subjecting an incubator to disinfection and rinsing, then adding a mixture of mother bacterium liquid and the culture solution in a volume ratio of 1: 20-50 into the incubator, sealing the incubator with gauze, and putting the incubator in a ventilation weak-light environment with illumination intensity of 1000 to 8000 Lx for fermentation culture at 30 to 35 DEG C for 7 to 14 d; and 3) subjecting the culture solution having undergone culture to solid-liquid separation and collecting liquid at a lower layer so as to obtain the kombucha metabolite. The preparation method provided by the invention is suitable for promotion and application and applicable to large-scale culture of the kombucha metabolite; and the prepared kombucha metabolite has good treatment effect on toothache.

The invention relates to the technical field of algae culture and photosynthetic bacteria culture, and discloses an anti-pollution circulating runway type culture system based on algae and photosynthetic bacteria culture. The system comprises a feeding tank, a remote main system controller, a power system, a spraying system, an oxygen supply system, a temperature control system, a PH control system, a spectrum control system, a daytime and night mode and a liquid conveying system, wherein the main control system is connected with the power system, the spraying system, the oxygen supply system, the temperature control system, the PH control system, the spectrum control system, the daytime and night mode, the spraying system, the oxygen supply system and the liquid conveying system. According to the anti-pollution circulating runway type culture system based on algae and photosynthetic bacteria culture, algae plants and photosynthetic bacteria are cultured in the mode, the culture density is high, the harvesting efficiency is remarkably improved, the culture conditions are easy to control, high-density culture is easy to realize, metabolite accumulation is facilitated, the system is not limited by regional environments, the production period is long, and production can be carried out all the year round.