294results about How to "Stable genetic traits" patented technology

The invention relates to a light control method for dendrobium officinale tissue culture, which comprises the following steps of: obtaining powdery seeds after dendrobium officinale fruits are sterilized, culturing the powdery seeds through a seed germination culture medium, and germinating the cultured seeds to form protocorms. The method is characterized in that: in the tissue culture process of dendrobium officinale seedlings, a light emitting diode (LED) plant growth illumination system is used as a light source, and different illumination treatment is implemented according to different stages of dendrobium officinale tissue culture. The different illumination treatment is implemented according to different growth stages so that the dendrobium officinale seedlings can furthest utilize light energy for photosynthesis and the highest growth amount is obtained. Experimental results show that: the culture period of the dendrobium officinale seedlings can be effectively shortened according to the technical scheme, the time from the seeds to the rooted finished seedlings is only 150 to 180 days, the growth period is shortened by 30 to 60 days compared with the culture under a common fluorescent lamp, and the seedlings cultured by the method have stable hereditary character and good quality.

The invention relates to acinetobacter calcoaceticus X.L.Zhu.SGD07-3-16 with free living nitrogen fixation capability. The acinetobacter calcoaceticus is preserved in China general microbiological culture collection center, and the preservation number of the acinetobacter calcoaceticus is CGMCCNo.5455. The acinetobacter calcoaceticus is high in nitrogenase activity and phosphorus and potassium dissolving capability, and has ACC (1-aminocyclopropane-1-carboxylic acid) deaminase activity, a certain amount of indoleacetic acid can be generated, and the generation of siderophore is avoided; when an acinetobacter calcoaceticus-containing liquid or solid microbial agent is used for inoculating crops such as corns, oilseed rapes and leaf mustard, the growth of the crops can be remarkably promoted, and the yield of the crops can be increased.

The present invention discloses a strain capable of producing poly-gamma-glutamic acid and a method for producing poly-gamma-glutamic acid by utilizing said strain. The described strain is X-003, a Bacillussubilis, CCTCC, NO:M202048. It adopts seed liquor preparation and liquid submerged fermentation process to produce and obtain the poly-gamma-glutamic acid product. The hereditary feature of said strain is stable, and its capacity for synthesizing poly-gamma-glutamic acid is strong, and the molecular weight of synthetic poly-gamma-glutamic acid can be up to above 10000 KD. The shake-flask fermentation level of poly-gamma-glutamic acid by utilizing said invented strain and its fermentation method can be up to 10 mg / ml, and the fermentation level of the fermentation tank with 80L is 30 mg / ml.

The invention discloses a rhamnolipid high-yielding bacterial strain, and applications thereof. The rhamnolipid high-yielding bacterial strain is named as pseudomonas aeruginosa KT1115, is preserved at China Center for Type Culture Collection, and the preservation number is CCTCC M 2016686. The rhamnolipid high-yielding bacterial strain is stable in hereditary features and high in rhamnolipid yield, and is obtained via ARTP induced mutation, blue gel plate screening, and secondary screening. According to fermentation broth of pseudomonas aeruginosa KT1115, liquid surface tension is reduced to 24.8mN / m<2>, and emulsification value is as high as 80%. Optimization of fermentation conditions including carbon source and nitrogen source single factors, cheap crude glycerine and swill-cooked dirty oil are taken as a composite carbon source, so that production cost is reduced greatly. The rhamnolipid yield of pseudomonas aeruginosa KT1115 is 50.7g / L, is increased by 18.1 times compared with that of a starting strain, conversion rate is 0.75g / g, and is relatively high. Application prospect of pseudomonas aeruginosa KT1115 in further industrialized application is promising.

The invention belongs to the fields of plant tissue culturing technologies and forest cultivation and relates to a high-efficiency lonicera edulis propagation method. The method comprises four stages of explant sterilization, inoculation, multiplication and rootage. The method solves the important technical problems of microbody quick propagation and root induction of the lonicera edulis, achieves the aims of efficiently inducing explants to separate and propagate and guaranteeing that seedlings survive with high quality, preserves all good characters of a matrix, and ensures stable hereditary character. The method can form large qualities of good test-tube seedlings within a short period and is suitable for large-scale and industrial production.

The invention relates to a culture method for efficiently obtaining adipose mesenchymal stem cells, and solves the problem that the cell viability obtained in the prior art is low. The culture method comprises the following steps: separation of the adipose mesenchymal stem cells: washing D.Hanks with fat tissues, detecting to guarantee no pollution, adding a solution mixture of I-type collagenase and trypLETM digestive enzyme with several times of volume, and stopping enzymolysis after carrying out rotary vibration digestion for dozens of minutes at the temperature of 37 DEG C, centrifuging to remove a liquid supernatant, and filtering with screen cloth to obtain an adipose mesenchymal stem cell suspension; culture of the adipose mesenchymal stem cells: inoculating a culture bottle bloodless culture medium with the adipose mesenchymal stem cell suspension, carrying out primary cell culture, observing with an inverted phase contrast microscope in the culture period, and changing a fresh culture medium, when the cells are cultured to 80% to 90% fusion, extensively inoculating a new culture bottle with the cells for culture after digestion with trypLETM, and subculturing for several generations to obtain the purified mesenchymal stem cells. The culture method has the advantages that the digestion is more sufficient, the cell yield is higher, the mesenchymal stem cells are protected from being damaged, and the viability is high.

The invention belongs to the field of plant tissue culturing technologies and forest cultivation and relates to a high-efficiency actinidia arguta propagation method. The method comprises four stages of explant sterilization, inoculation, multiplication and rootage. The method solves the important technical problems of microbody quick propagation and root induction of the actinidia arguta, achieves the aims of efficiently inducing explants to separate and propagate and guaranteeing that seedlings survive with high quality, preserves all good characters of a matrix, and ensures stable hereditary character. The method can form large qualities of good test-tube seedlings within a short period and is suitable for large-scale and industrial production.

A "Jiuxiang" strawberry stem tip rapid breeding method by tissue culture and virus removal is disclosed. The rapid breeding method includes: explant disinfection, stem tip tissue virus removal and culture, multiplication culture, rooting culture, seedling hardening and transplanting. A stem tip tissue culture method is adopted to breed "Jiuxiang" strawberry virus-free seedlings, thus ensuring that all the excellent characteristics of parents are stably inherited, and therefore influences of outside conditions can be prevented, and the rapid breeding method can be performed at all seasons, thus saving the seedling culture land, and reducing the production cost. A large quantity of good test-tube plantlets can be produced in a short period of time. Large-scale production can be performed to provide "Jiuxiang" strawberry seedlings with a large quantity and good quality for the market so as to meet market needs. The "Jiuxiang" strawberry virus-free seedlings prepared by the rapid breeding method are high in growth vigor, disease-resistant and increased in big-fruit yield. The yield can be increased by 30-50%.

The invention discloses a method for performing regeneration propagation on sterile stalks of butterfly orchids, relates to the technical field of seedling cultivation, mainly relates to a quick butterfly orchid plant reproduction technology, and in particular relates to a plant regeneration method for the butterfly orchid species. The method comprises the steps of cutting a stalk axillary bud stem with a certain length from a robust mother butterfly orchid plant, taking the stalk axillary bud stem as an explant material, wherein an axillary bud is opened and a leaf is in jade green and glossy after the sterile stalk is subjected to primary culture; after the axillary bud is cut down, culturing the stalk on a subculture medium, then continuously forming bud points at an incision of the axillary bud of the stalk to gradually form another axillary bud, continuing to cut down the axillary bud after the axillary bud is opened, and inoculating the stalk on the subculture medium again for subculture, wherein due to the culture, a large number of cluster buds can be obtained from one stalk, and part of the obtained cluster buds can be continued to be subjected to subculture by a bud-propagation-bud method. The method has the advantages that the later generations can keep a stable inheritable character of a maternal line, so that flowering phase control and industrial uniform management are facilitated, and the survival rate is up to 90 percent.

A method for separating tetradotoxin is carried out by microbe. It comprises 1) selecting fresh globefishes; 2) washing and disinfecting them to take out their ovaries, grinding the ovaries with cell dispersing liquid to obtain fresh ovary extract liquid; and 3) separately and thermally culturing in medium of marine bacteria and actinomice to separate and purify tetradotoxin. It operates easily with good effect, separates well to obtain strains with 90% achieving rate. The obtained microbe can produce tetradotoxin at high yield and breed ten more generations with stable heredity.

The invention discloses a Hyphomicrobium sp. strain and a preparation method for pyrroloquinoline quinone. The Hyphomicrobium sp. strain 1112-NTG-2318 capable of realizing high yield of pyrroloquinoline quinine is preserved in China General Microbiological Culture Collection Center with an accession number of CGMCC No. 10709. The preparation method for pyrroloquinoline quinine comprises the following steps: fermenting the Hyphomicrobium sp. strain CGMCC No. 10709 in a fermentation medium and acquiring pyrroloquinoline quinine from fermentation broth. The fermentation titer of the preparation method in a tank with a volume of 80 m<3> reaches 1783 mg / L, so the preparation method is favorable for industrial production of pyrroloquinoline quinine.

The invention discloses a tilapia fry rearing method. The tilapia fry rearing method comprises the following steps: breeding parent tilapia fries which are bigger than nine-sieve, picking parent tilapias which are disease free, injury free, strong and good in sexual maturity to rear, hybridizing the female parent tilapias and with the male parent tilapias according to ratio of stocking, picking up roes, incubating the roes with flowing incubating water in an incubating tank, cultivating the roes by marking in bold, and cultivating fries. Oviposition period is shortened by about seven days, incubating rate is as high as 67.9%, fry survival rate is 89.6%, each female tilapia lays 6700 fries each year averagely, fry production rate is improved by 30%, and the sizes of the fries are uniform. By means of the tilapia fry rearing method, fries can be reared all through a year, a large amount of tilapia fries are reared and the tilapia fries grow fast and are strong in stress resistance. The tilapia fries reared in mass are born at the same time, gonad developments are synchronous, sizes are the same, characters are unified, and heredity purity is high. Moreover, the tilapia fry rearing method has the advantages that incubating period is short, incubating efficiency is high, inheritable characters are good, fertility is strong, and fries can be reared in an overwintering mode.

The invention discloses a novel tuckahoe strain (china center for type culture collection (CCTCC) M2011072) and bag-material efficient cultivation technology. The novel tuckahoe stain is applied to cultivation by adopting the bag-material efficient cultivation technology. The novel tuckahoe stain is mainly characterized in that 1) a mycelium is strong, white, obvious in clamp connection; and hypha is spread quickly in a cultivation process; 2) the tuckahoe fruits earlier, the fruit body is even and strong, the color of pulp is white or light yellow, and the skin of tuckahoe is thin and presents purplish red color; 3)the novel tuckahoe strain has the advantages of being stable inheritable characteristics, strong in anti-bacteria infection capacity, high in biological efficiency and yield, good in quality and the like; and 4) the novel tuckahoe strain remarkably improves content of effective components of amino acid, pachymaran and the like in the tuckahoe.

The invention relates to a micrografting method for a tetraploid acacia. The method includes the steps of selecting a seedling of a diploid acacia seed as a rootstock, selecting a tissue culture seedling of the tetraploid acacia as a scion, subjecting the scion and the rootstock to cambium alignment by using a cleft grafting method, twining the interface area with silver paper slips, fixing the silver paper twining area with a grafting clamp, planting the grafted seedling into a potted tray containing matrix, then processing a routine management for 20 - 30 days, and acquiring the plant regeneration. By means of the micrografting method, an integral plant is obtained after cultivating the grafted seedling for 20 - 30 days, and accordingly the high efficiency of rapid planting is achieved. The plant regeneration cultivated by the micrografting method has the advantages of stable genetic character, healthy plant, strong growth, short seedling period, fast plant growing and high survival rate ( with a survival rate over 90% in a field planting production land).

The invention discloses a growth promotion cultivation method of fritillary bulbs. The growth promotion cultivation method mainly comprises the steps of selection and disinfection of explants, material treatment, primary culture, subculture, proliferation of bulblets, rooting culture, and management of soil for acclimatization and transplanting. Different hormones are used for adjusting at different culture phases; a formula of a culture medium is sieved accurately to realize the good effect of vigorous growth of test-tube seedlings; an effective sterilization technology is combined to achieve the aims of improving the proliferation rate, reducing variation, increasing soil fertility and reducing plant diseases and insect pests. The culture medium is replaced properly in the proliferation process of the bulblets which are subjected to subculture so as to facilitate the rapid growth of the bulblets. By comparing different parts of flakes, a best flake reproduction material is the lower part of inner-layer flakes.

The present invention discloses Agrobacteriumsp. having free-living nitrogen fixing ability, wherein a classification name of the Agrobacteriumsp. is Agrobacteriumsp. Ymu6-2, the Agrobacteriumsp. is preserved in the China General Microbiological Culture Collection Center (CGMCC), and a preservation number is CGMCC No.5007. The Agrobacteriumsp. having free-living nitrogen fixing ability in the present invention has the following advantages that: high nitrogen fixing ability is provided; high concentration Cd growth can be resisted; indole-3-acetic acid can be secreted to produce iron atoms; with the prepared microbial agent, over-accumulated plant growth can be significantly promoted, biomass of over-accumulated plants can be increased, a heavy metal enrichment coefficient and a heavy metaltransfer coefficient of over-accumulated plants can be substantially increased, soil ecology environment can be improved, and advantages of environment-friendliness and low cost are provided.

The invention relates to a tissue culture and rapid propagation method of rhododendron latoucheae. A woody plant medium (WPM) is taken as a basic culture medium, and a tissue culture and rapid propagation system of the rhododendron latoucheae is established through the isolated culture of the rhododendron latoucheae by proportioning regulation of different hormones. The method sequentially comprises the following steps of: selecting an explant; disinfecting; performing primary culturing; performing propagation culturing; performing rooting cultivation on a strong seedling; and transplanting to obtain a test tube seedling which can keep the original plant genotype, has a high aesthetic property and is better differentiated, wherein a primary induction culture medium of the explant is WPM+IBA 0.15mg / L+TDZ 0.3mg / L+cane sugar 30g / L and the pH value is 5.6; a propagation culture medium of an adventitious bud is WPM+IBA 0.15mg / L+TDZ 0.3mg / L+cane sugar 30g / L and the pH value is 5.6; and a rooting culture medium of the strong seedling is 1 / 2WPM+IBA 0.5mg / L+NAA 0.2mg / L+cane sugar 20g / L+activated carbon 0.3g / L and the pH value is 5.6. By the method, the propagation speed of the rhododendron latoucheae is greatly increased; and the method is suitable for industrialized commercial production of the rhododendron latoucheae.

The invention belongs to the field of micro-organism animal cell line and relates to a human hepatoma cell line which can emit high-intensity red or green fluorescence and has high transferring ability of lung and lymph node metastasis, and a method for establishing the same. The method comprises the following steps: using the human hepatoma cell line HCCLM3 and HCCLM6 which have high transferring ability of the lung and lymph node metastasis as mother cells, performing cotransfection on plasmid DNA of 239 cells through slow virus packaging plasmids to obtain false slow virus particles by expressing red or green fluorescent protein genes through eucaryon, and infecting liver cancer cell strains of the mother cells to obtain the chromosome integrated hepatoma cell line which has high transferring ability of the lung and lymph node metastasis and can stably expressing the red or the green fluorescence. The human hepatoma cell line which has high transferring ability of the lung and lymph node metastasis in vitro can be applied to the tracer studies on tumor cells, the molecular mechanism studies on the recurrence and transferring of liver cancer, as well as the pre-clinical drug efficacy studies on new anti-tumor drugs, thus the human hepatoma cell line has wide application prospect.

The invention relates to an agrobacterium with free living nitrogen fixation, phosphate dissolution and potassium dissolution capacity. The agrobacterium is classified and named as agrobacterium ((i)Agrobacterium( / i)(i)rsp.( / i))X.L.Zhu.SGD06-8-33 and collected in the China General Microbiological Culture Collection Center, and the collection number is CGMCC No.5457. The strain disclosed by the invention is high in nitrogenase activity, has high phosphate dissolution and potassium dissolution capacity, can generate a certain amount of indoleacetic acid and siderophores without ACC desaminase activity. If crops, such as corns, rapes and leaf mustard, are inoculated by using liquid or a solid microbial inoculum which contains the strain, the growth of the crops can be obviously promoted, and the output of the crops can be increased. The method is simple and convenient in process, rich in raw material source and suitable for industrialized production.

The invention provides a method for rapidly reproducing protruding eucalyptus through tissue culture, which comprises the steps that: excellent tree bodies are selected as explants which are then collected and sterilized; initial culture, multiplication culture and rooting culture are carried to stem sections; then the seedlings adopt seedling adaptation, transplantation from bottles and seedling management. The method of the invention can increase multiplication coefficient of 3 to 5. The inheritable characters of the germchit of the protruding eucalyptus are stable, and the germchit of the protruding eucalyptus reaches improved variety and cloning, thus increasing the wood yield of an elemental area and promoting the ecological and economic benefits of the woods of the protruding eucalyptus.

The invention belongs to the field of microorganism animal cell system, and relates to an establishment method of a fluorescence visualization human hepatoma nude mice model which can self emit high strength red or green fluorescences and has the metastatic ability. After obtaining a human high-metastatic fluorescence hepatoma cell line HCCLM3-R, HCCLM3-G, HCCLM6-R and HCCLM6-G of fluorescence genes with high chromosome conformable degree, high fluorescence intensity and stable expression by using the method of pseudotype slow virus infection, hepatoma cells expressed by the fluorescences is placed beneath nude mice skin to establish a fluorescence visualization high-metastatic human hepatoma nude mice subcutaneous tumor model or subcutaneous fluorescence expression tumor tissues is placed in nude mice hepar to establish a fluorescence visualization high-metastatic human hepatoma nude mice hepar primary tumor model. The got model can be used as a tracer of animal in vivo hepatoma cells, and can be used for molecular mechanism study of hepatoma recurrence and metastatic and prophase therapeutic effect discrimination for new anti-hepatoma therapy and new anti-hepatoma drugs, etc.

The invention provides a sexual propagation method of bletilla striata. The sexual propagation method of bletilla striata particularly comprises the steps of processing of capsules, preparation of nutrients, inoculation and cultivation, and bottle out domestication, and is characterized in that in the step of processing of capsules, an alcohol lamp drying mode replaces a filter paper sucking dry mode in the prior art; in the step of preparation of nutrients, combination of banana slurry and humus is adopted; in the step of inoculation and cultivation, only inoculation, germination cultivation and strong seedling cultivation steps are included, the conventional enlargement cultivation process in an existing tissue culturing technology is omitted, and the process is simplified and the cost is lowered while the purposes are achieved; in the step of bottle out domestication, cultivation is performed through humus in a greenhouse, domestication is finished under the condition of controlling the relative humidity, temperature and shading rate, and qualified bletilla striata seedlings are obtained. Bletilla striata seeds are used for scale seedling culturing, the survival rate is high, manual scale propagation of bletilla striata is achieved, and the obtained bletilla striata seedlings are low in contamination rate, high in differentiation rate and stable in inheritable character.

The invention provides a tissue culture quick reproduction method for dendrobium chrysotoxum lindl bud tips, which comprises the following steps of: performing induction culture of buds; performing enrichment culture of tufted buds; performing subculture of the tufted buds; and performing rooting culture to obtain a complete plant, and hardening seedling outside a bottle and transplanting. The method is easy to operate and low in production cost, environment pollution is avoided, and large-scale production can be realized. Dendrobium chrysotoxum lindl seedlings cultured by the method are stable in inheritable characters, keep the characteristics of parents and have the advantages of invariance property, small investment, high yield and short period.

The invention discloses an acinetobacter with authigenic nitrogen fixation capacity, which is classified and named as Ymu7-3 and is preserved in China general microbiological culture collection center (CGMCC) with the preservation number of CGMCC No. 5006. The free living nitrogen fixing bacteria has high nitrogen fixing capacity, is capable of resisting the growth of Cd with higher concentration, and can secrete heteroauxin so as to generate iron atom; and the prepared microbial bacterium agent can remarkably promote the growth of hyper-accumulating plants, increase the biomass liveweight of the hyper-accumulating plants, greatly improve the heavy metal enrichment factor and transfer coefficient of the hyper-accumulating plants, and improve the ecological environment of the soil, thus being environment-friendly and low in cost.

The invention discloses preparation and application of an alpha-1,6 mannose transferase gene (och1) flaw type P. pastoris X-33 bacterial strain, belonging to the technical field of biological engineering. The engineering bacterial strain P. pastoris X-33 (delta och1) is characterized in that a target gene knockout method by double-exchange homologous recombination is adopted, and a URA3 gene serves as a selective marker to knock out the alpha-1,6 mannose transferase gene (och1) of the P. pastoris X-33 (delta och1) to obtain the P. pastoris X-33 (delta och1) bacterial strain with OCH1 gene knockout. The bacterial strain provides a P. pastoris expression system which modifies protein by virtue of low N-glycosylation modification and provides a favourable basis for further glycosylation modification. The bacterial strain has the advantage of great potential application prospect.

The invention discloses a method for quickly establishing cross-bred XY holandric pelteobagrus fulvidraco. The method is characterized in that based on mating offspring of pelteobagrus fulvidraco YY physiological female fish and pelteobagrus vachelli XY male fish, cross-bred YY super-male fish is obtained by cultivation through a genetic method so as to mate with common pelteobagrus fulvidraco female fish to produce the XY holandric pelteobagrus fulvidraco. According to the method disclosed by the invention, the breeding period can be obviously shortened; compared with technologies such as Varadaraj and the like, two generations can be shortened; and compared with a method of continuously producing the holandric pelteobagrus fulvidraco by using sex reversal and gynogenesis, one generation can be shortened. Besides, the cross-bred holandric pelteobagrus fulvidraco obtained by the method disclosed by the invention is stable in genetic character, high in growth speed, large, and strong in adaptive capacity to the environment, has cross-breeding advantages and inherits advantages of the holandric fish, so that better breeding and economic benefits can be obtained.

The invention relates to a method and a substance for adjusting starch composition of root crops. The invention discloses polynucleotide constructs which is expressed by specific interference granulebound starch synthase I (GBSSI) and can be processed after being introduced into plant., and a brand-new way for changing the starch quality of the plant by interrupting the expression of GBSSI genesbased on micro-molecular RNA interface technology to achieve the aim of adjusting the starch composition in the plant.

The invention discloses a method for producing guanosine by fermentation with bacillus, which sequentially comprises the following steps: bacillus activation, seed culture, fermentation culture, guanosine extraction and refinement, and the like. The strain is Bacillus subtilis CGMCC No.4587. The invention increases the fermentation yield and conversion rate of guanosine.

The invention discloses a cultivation method for inducing scutellaria baicalensis hairy root based on agrobacterium rhizogenes infection, which uses an agrobacterium rhizogenes bacteria liquid to infect root of a scutellaria baicalensis explant and induces the scutellaria baicalensis explant to form a hairy root, the scutellaria baicalensis hairy root presents yellow hairy on a MS solid medium, so that scutelloside can be stably synthesized, an ITS sequencing is performed and the hairy root can be identified as the scutellaria baicalensis hairy root of a scutellaria. Then seed selection is performed for further in a seed selection medium and a cultivation condition can be optimized to select lines which raise the content of active ingredient scutelloside used for hairy root traditional Chinese medicines. Hairy root can be obtained through scutellaria of a scutellaria introduced by agrobacterium rhizogenes, wherein the inductivity can reach 10 - 40%, the obtained scutellaria baicalensis hairy root presents yellow hairy which is capable of stably synthesizing scutelloside. According to the invention, the scutellaria baicalensis hairy root of a scutellaria is cultivated on the optimized medium, the growth is rapid and the biomass can be increased for 23 -35 times in 30 - 35 days, and the cultivation is performed under the dark conditions. The invention has the characteristics of low energy consumption and low cost, which is suitable for industrialization production.

The invention discloses a tissue culture method of vaccinium dunalianum containing caffeoyl arbutin, relating to the technical field of tissue culture of plants. A tissue culture seedling culture system is built and the content of caffeoyl arbutin is measured by using improved WPM (woody plant medium) as basal medium at different proportion concentrations of hormone and activated carbon. The growth and root induction of aseptic seedling can be controlled by adjusting the proportion of 6-BA, NAA and the activated carbon. The induction rate of explants reaches 97 percent, the bud seedling is robust, the growth is rapid, and the rooting percentage of the bud seedling is 94%. The method has the advantages of rapid reproduction speed, short period and high content of caffeoyl arbutin, and the like, is an effective way to obtain a large number of caffeoyl arbutin, and provides reliable technical basis for large-scale production of fine rapid propagation seedlings of vaccinium dunalianum.