44results about How to "Reduce the chance of mutation" patented technology

The invention discloses a method for producing enramycin by using microbial fermentation. The method comprises the following steps of: preparing spore liquor of streptomyces fungicidicus SDSL1205 CGMCC No. 3933, and preserving the spore liquor under the environment of liquid nitrogen; inoculating the frozen and preserved spore liquor into a seed culture medium to culture seeds so as to obtain seed liquor; inoculating the seed liquor into a fermentation medium for fermenting and culturing according to the inoculum size of between 4 and 6 weight percent; and further processing fermented fermentation liquor to obtain enramycin crystals. The method has the advantages of omitting complicated seed making process, reducing microbiological contamination rate, along with simpleness and practicability, easy control of inoculum size, stable fermentation result and high repeatability. Compared with the conventional process, the method improves the fermentation result to some extent; and the maximum yield of the enramycin is over 8,500 mu g / ml (HPLC).

The invention belongs to the technical field of microorganisms, relates to a method for preserving and separating a toadstool strain through sporocarp dried in the shade, and aims to solve the technical problems that a conventional toadstool strain preservation method is limited by season, complex to operate, easy in causing pollution and relatively high in cost. According to the technical scheme, the method for preserving and separating toadstool strain through sporocarp dried in the shade comprises the following steps: a, preserving the strain; b, separating the strain. The method provided by the invention is low in cost, simple to operate, unlimited by season, capable of effectively preserving activity of the strain, and convenient in transporting and preserving the strain.

The invention relates to a method for cultivating jade needle organism, wherein it comprises that: using fresh bud, via initial cultivation to form germ-free seed, without cure organism to induce the bud to increment, to root and cultivate. The invention has simple process and high yield.

The invention relates to a high-yield planting method of salix integra. The method comprises the steps of (1) treating cutting planting and seedling soil; (2) selecting cutting branches; (3) treating the cutting branches; (4) cutting; (5) managing after cutting; (6) transplanting; (7) daily managing. With the adoption of the method, the survive rate of the treated salix integra is high; diseases and insect pests are reduced; a large amount of seedlings per unit area can be cultured; the land can be saved; the economic benefit is high.

The invention relates to a high-survival-rate azalea seedling cultivating method. The method comprises the step one of cuttage planting soil treatment, the step two of cuttage branch pretreatment, the step three of cuttage branch treatment, the step four of cuttage and the step five of management after cuttage. Before planting, liquid traditional Chinese medicine is used for treatment, the disease resistance ability of azalea seedlings is improved, and branches can be promoted for germinating and surviving; as the length of the cuttage branches is reduced, the number of bacteria on the seed branches is reduced, the mutant probability of the seedlings is reduced, the strain genuineness of azalea can be kept, the applicability of the azalea is enhanced, and the survival rate of the seedlings is increased.

The invention discloses a method for producing avilamycin by fermenting. The method includes steps of preparing streptomycesolivaceoviridis with inclined surface with spores, connecting spores on the inclined surface to a seed bottle to culture to obtain seed solution, preserving the seed solution in hydraulic hydrogen; inoculating the frozen seed solution to a seed culture medium to culture to obtain the seed solution; inoculating the seed solution to a fermenting culture medium to be subjected to fermenting culture by the inoculation quantity of 5wt%; and further treating the fermented liquid obtained after fermentation to obtain crystals of avilamycin. The method is easy to carry out, completed seed making process is omitted, degeneration is reduced, contamination risk is lowered, inoculation quantity is easy to control, repeatability is good, fermenting results are good, and production cost is low. Further, the fermenting effect is higher than that of the prior art, and the outputof avilamycin can be over 6500 microgram / microliter.

The invention discloses a method for prolonging the subculturing interval time of tissue culture seedlings of apple. A culture medium comprises the following components: 0.5-0.75 mg / L of MS+BA, 0.05 mg / L of NAA, 40.0 g / L of white granulated sugar and 6.0 g / L of agar; the packing quantity is 40-50% more than that in conventional breeding, and a bottleneck is sealed by using a plastic sealing film; tissue culture seedlings of apple are transferred to the culture medium and slowly grow about 2 weeks under the culture conditions that the temperature is 22-28 DEG C, the illumination intensity is 2000 lx, and the illumination time is 10-14 h / d, and then the tissue culture seedlings of apple are placed in a chemical agent storage box with a temperature of 2-8 DEG C so that the growth is slowed down; and when the culture medium is approximately dried, the tissue culture seedlings of apple are taken out of the refrigerated container to grow normally for 1-2 weeks or are directly transferred to a new culture medium. In such a way, the subculturing interval time of tissue culture seedlings of apple can be successfully prolonged from 2-3 months to 15-18 months, the survival rate of stem tips is more than 90%, the workload of subinoculation is greatly reduced, and the mutation probability is also reduced. The technique is applied to the germplasm preservation of plantlets of apples and other parts of orchardfruits and flowers, is simple, convenient, saving, and high in efficiency, and provides a new way for the preservation of germplasm resources.

The invention relates to a culture and cryopreservation method of amniotic mesenchymal stem cells. The culture method comprises amniotic tissue separation, amniotic mesenchymal stem cell separation, P0 generation amniotic mesenchymal stem cell culture and amplification culture. According to the culture method of the amniotic mesenchymal stem cells, at a separating stage of the amniotic mesenchymalstem cells, a special mixed enzyme digestive juice system (the final concentrations of components in the digestive juice are as follows: 1.5-2U / mL of neutral protease, 0.5mg / mL of deoxyribonuclease Iand 1mg / mL of collagenase IV) is adopted for digestion, so that the amniotic mesenchymal stem cells can be more effectively separated from amniotic tissues, and then the yield of P0 generation cellsis obviously improved. Compared with conventional two-step digestion, the culture method provided by the invention has the advantages that a one-step digestion method is adopted, operation is more convenient, and the cultured amniotic mesenchymal stem cells have good activity, high yield, high purity and good repeatability.

The invention relates to a microbial strain preserving method, in particular to a method for preserving microbial strains for a long time. The method comprises the main steps of: inoculating the strains onto a slant culture medium; cultivating the strains for 20-30 hours at a temperature of 20-40 DEG C; adding a little calcium carbonate onto a slope, wherein the calcium carbonate is subjected to dry heat sterilization; filling a liquid tubular culture medium into a test tube; plugging the test tube by using a sterile rubber plug; and putting the test tube in a refrigerator for preservation at a temperature of 2-4 DEG C. When the method is adopted to preserve acetic acid strains, reproducing times of the acetic acid strains can be reduced, namely mutation probability can be reduced. The method is simple to operation; the preservation period can reach 2 years; and the microbial strains can be put into production after being activated by using the slant culture medium for two times when used, without complex operation processes. In addition, the purpose of preservation can be achieved without any expensive instrument; the method is suitable for mass production; and according to the method, cost can be lowered, and energy consumption can be reduced.

The invention belongs to the technical field of plant cultivation and provides an efficient cultivation method of osmanthus fragrans. The efficient cultivation method includes the steps of 1), pretreatment of soil; 2), selection and treatment of cutting branches; 3), treatment of ends of the cutting branches; 4), cutting; 5), management after cutting. With the method, the survival rate in cultivation of the osmanthus fragrans is increased, disease resistance of the osmanthus fragrans is enhanced, and adaptability is improved.

The invention discloses a preservation method of soybean virus. The preservation method provided by the invention comprises the following steps: a virus preserving fluid is adopted for preserving soybean virus; the virus preserving fluid comprises a polymer in the formula (I) shown in the specification and ascorbic acid; and n represents a natural number ranging from 20 to 70. Experiments prove that the soybean virus preserved by using the preservation method provided by the invention can completely maintain activities within three years under an extra low temperature of -80 DEG C, and can be taken as a drug source for being used directly at any time without need of excitation, the efficiency for identifying the soybean virus is improved greatly, unnecessary subcultures during the multiplication process of the mosaic virus are reduced, mutual cross infection of virus seeds can be avoided, the probability of virus variation is lowered, and the genetic stability of viruses can be kept. Therefore, the preservation method provided by the invention has a significant application value.

The invention discloses a method of prolonging the preservation time of a grape tissue culture seedling with mineral oil coverage. The method includes the steps of removing leaves from tender shoots of the grape tissue culture seedling, cutting the grape tissue culture seedling into single-bud stem segments in about 1.5 cm, planting the single-bud stem segments into a subculture culture medium, covering sterilized mineral oil on the culture medium with the thickness being about 5-6 cm, sealing the culture medium with a sealing film, and culturing the single-bud stem segments under common culture conditions (temperature: 25 + / - 3 DEG C, illumination intensity: about 2000 lx, and illumination time: 10-14 h / d). By means of the method, the preservation time of the grape tissue culture seedling can be prolonged from 4 months to longer than 15 months, thereby greatly reducing subculture planting work load and reducing mutation rate. After the mineral oil is removed, the growth of the preserved tissue culture seedlings can be recovered without washing the tissue culture seedling. The method is used for germplasm preservation of the grape tissue culture seedling, is wide in available varieties, is long in preservation time, is low in cost since the mineral oil can be reused, is simple in operation, allows growth to be recovered easily and is an excellent method of in-vitro preservation of grape germplasm.

The invention relates to a novel cutting seedling method for grape. The novel cutting seedling method comprises the steps that 1 the oil for cutting planting is processed; 2 a cutting branch is selected and processed; 3 the end of the cutting branch is processed; 4 cutting is conducted; 5 management after cutting is conducted. According to the novel cutting seedling method for grape, the cutting technology is improved, an original cutting branch is shortened to be about 10 cm, the length of the cutting branch is shortened, the usage amount of a mother branch is reduced, the cost is saved, the number of cutting seedlings in unit area can be improved greatly, the seedling density is increased, the gowning way of grape seedling is balanced, and the survival rate is high.

The invention provides a fast cultivation method for strawberry biochemical callus. The method comprises the following steps: 1) cleaning and sterilizing raw materials; 2) primarily culturing: using a cutter for cutting 0.5mm stem tip growing point, inoculating into a 0.1mg / L 6-BA MS culture medium for culturing and growing clumpy buds; 3) sub-culture: adapting the clumpy buds into the MS culture medium containing 0.2mg / L 6-BA and 0.01mg / L NAA and sub-culturing; 4) rooting culture: cutting big buds from the clumpy buds, adapting to the MS culture medium containing 0.5mg / L IBA and 0.1% activated carbon for inducing rooting, adapting the residual callus into the culture medium in the step 3) and culturing; 5) transplanting domestication: taking out the rooting seedlings, cleaning the root culture medium, transferring into a hole plate filled with the medium in greenhouse and domesticating. According to the method provided by the invention, the seedling culturing time is short, the subculture speed is high and the cultivating cost is saved.

The invention discloses a method for producing avilamycin by fermenting. The method includes steps of preparing streptomycesolivaceoviridis with inclined surface with spores, connecting spores on the inclined surface to a seed bottle to culture to obtain seed solution, preserving the seed solution in hydraulic hydrogen; inoculating the frozen seed solution to a seed culture medium to culture to obtain the seed solution; inoculating the seed solution to a fermenting culture medium to be subjected to fermenting culture by the inoculation quantity of 5wt%; and further treating the fermented liquid obtained after fermentation to obtain crystals of avilamycin. The method is easy to carry out, completed seed making process is omitted, degeneration is reduced, contamination risk is lowered, inoculation quantity is easy to control, repeatability is good, fermenting results are good, and production cost is low. Further, the fermenting effect is higher than that of the prior art, and the outputof avilamycin can be over 6500 microgram / microliter.

The invention discloses a method for preserving and taking potato germplasm resources. The method comprises the following steps that S1, single sections of potato tissue culture seedlings are shearedand cut on a culture medium in a culture bottle, and a bottle opening is sealed by using a sealing film; S2, subculture is performed on the potato tissue culture seedlings treated in step S1 at the temperature of 20-22 DEG C, the illumination intensity of 3,200-3,500 lx and the illumination time of 15-18 hours per day; after two months, the illumination intensity is reduced to 2,300-2,500 lx, andculture is continued; after two months, the illumination intensity is increased to 2,700-3,000 lx; and after two months, microtuber is naturally fruited, the illumination intensity is reduced to 0, and continuous culturing is carried out until the microtuber passes away dormancy and starts to germinate; and S3, the microtuber starting to germinate obtained in step S2 is inoculated to a culture medium in a culture bottle, a bottle opening is sealed by using a sealing film, and a new round of germplasm resource preservation is carried out according to the operation in step S2 when the microtubergrows into a new test-tube plantlet. The method for preserving and taking potato germplasm resources is simple, convenient, easy to operate, low in cost and good in effect.

The invention relates to a method for culturing and freezing amniotic mesenchymal stem cells. The culturing method includes amniotic membrane tissue separation, amniotic mesenchymal stem cell separation, P0 generation amniotic mesenchymal stem cell culture and expansion culture. The culture method of amniotic mesenchymal stem cells of the present invention, in the separation stage of amniotic mesenchymal stem cells, adopts a special mixed enzyme digestive solution system (final concentration of each component in the digestive solution: 1.5-2U / mL neutral protease, 0.5mg / mL deoxyribonuclease I and 1mg / mL collagenase IV) for digestion, can more effectively separate amniotic mesenchymal stem cells from amniotic tissue, and then significantly improve the yield of P0 generation cells. The present invention adopts one-step digestion Compared with the traditional two-step digestion method, the operation is more convenient, and the cultured amniotic mesenchymal stem cells have good activity, high yield, high purity, and good repeatability.

The invention provides a rapid reproduction method of schlumbergera seedlings. The method comprises the following steps: inoculating a selected, disinfected and sterilized schlumbergera explant onto a culture medium for induction and carrying out synchronous induction and proliferation culture until the explant sprouts and differentiates out a bud; sub-culturing until being proliferated to 3-4 folds, and then transferring to a rooting medium for rooting culture until 3-5 fine roots are grown at the base of a plant; conventionally hardening and then transplanting to a mixed substrate of humus soil, red soil and pearlite; and watering and fertilizing conventionally for growing to obtain the schlumbergera seedlings. In the method, key links in the whole production technical flow of the schlumbergera seedlings are optimized and integrated, the culture medium for induction is blended and bud induction and proliferation culture are carried out synchronously, thus simplifying tissue culture procedure and shortening breeding cycle; and only by adopting two types of the culture media, the bud can be directly formed without callus formation, thus reducing probability of variation occurred on an adventitious bud, effectively controlling generation of a variant strain, being capable of fixing and maintaining fine variety of the schlumbergera seedlings rapidly, improving seedling quality and solving the problems of complicated parental source and unstable quality of the seedlings produced by the conventional breeding system.

The invention relates to a high-surviving-rate salix integra seedling culturing method. The high-surviving rate salix integra seedling culturing method comprises the steps of 1, cutting planting soil treatment, 2, cutting branches selection, 3, cutting branch treatment, 4, cutting and 5, management after the cutting. After salix integra seedlings are treated by the method, soil insects in soil are killed firstly, and the surviving-rate of the salix integra branch cutting is high.

The invention relates to a method for cultivating jade needle organism, wherein it comprises that: using fresh bud, via initial cultivation to form germ-free seed, without cure organism to induce the bud to increment, to root and cultivate. The invention has simple process and high yield.

The invention relates to a mixed protease system used for gene cloning and assembling, a kit containing the mixed protease system and a use method of the kit. By utilizing the mixed protease system, specificity recombinant reaction of two or more DNA can be completed at the high temperature of 40-70 DEG C. The invention also relates to the kit containing the mixed protease system. Compared with enzyme or enzyme system with the same function, the mixed protease system keeps high activity at the high temperature of 40-70 DEG C, then the specificity of recombinant reaction is improved, generation of meaningless mutation is effectively avoided, high fidelity cloning and assembling of two or more DNA can be realized, the stability is good, the reaction precision is high, and operation is simple, convenient and efficient.

The invention discloses a full-ecological vertical cultivation method for dendrobium officinale. The cultivation method comprises the following steps of: selecting planting materials; selecting arenga engleri ropes for binding arenga engleri sheets; selecting the dendrobium officinale; selecting spring water; a planting process: moving pines arranged in matrixes into pits in a leveled greenhouse at intervals of 1.5 m, filling and leveling up the pits with soil, wrapping two-year-old dendrobium officinale domestication seedlings in a bottom-up manner from the bottoms of the pines, and fixing the seedlings on the pines; exposing stems, planting a second circle of the dendrobium officinale domestication seedlings at vertical intervals of 20 cm in a bottom-up manner, and then planting the rest seedlings in the same manner. The full-ecological vertical cultivation method has the beneficial effects that a full-ecological vertical cultivation mode is adopted, the original wild growth strength of the dendrobium officinale is fully released, under the artificial large-area cultivation conditions by modern facilities, the dendrobium officinale can grow mostly close to the wild state, organic production is realized in the whole process and the organic standards are wholly met; in addition, the stability of variety inheritance of the dendrobium officinale can be ensured and the probability of mutation is reduced so as to guarantee pure and good-quality varieties; furthermore, the greenhouse space can be fully utilized for realizing vertical space high-density cultivation and increasing the yield per mu.

The invention discloses a tissue culture and rapid propagation process of Chinese Pistache. The tissue culture and rapid propagation process of Chinese Pistache includes the steps of A, collecting ripe seeds of Chinese Pistache, and removing episperm by sulfuric acid digestion; sterilizing; inoculating the seeds to MS (Murashige and Skoog) minimal medium in which no hormone is added; B, cutting aseptic seedlings into stem segments with buds, removing part of leaves, inoculating to rooting medium or inoculating to enrichment medium prior to transferring to the rooting medium; and C, exercisingand transplanting the tissue culture seedlings. The tissue culture and rapid propagation process of Chinese Pistache has the advantages that the tissue culture method is modified, a medium formula isoptimized, and aseptic seedlings can be obtained easily and can be propagated rapidly and grow healthy and vigorous; mass high-quality test-tube seedlings can be obtained rapidly from the Chinese Pistache which is difficult or slow to propagate under natural conditions and can be applied to culture and production, and variation rate of offspring of the Chinese Pistache obtained by the process is low.

The invention discloses a rice disease-resistant gene RGA5-HMA5 and application thereof in rice breeding. On the basis of an RGA5 gene, a heavy metal-associated (HMA) structural domain at a carbon endof protein encoded by the RGA5 is subjected to point mutation to obtain the RGA5 gene mutant RGA5-HMA5. Experiments indicate that the protein encoded by the RGA5-HMA5 can specifically recognize effector protein AVR-Pik of magnaporthe oryzae to relieve the inhabitation function of the effector protein AVR-Pik on an RGA4 gene and activate immunoreactions of rice, and recognition of AVR-Pik effectorprotein is changed, so that rice containing the disease resistant gene obtains resistance to magnaporthe oryzae containing the AVR-Pik effector protein. The rice disease-resistant gene RGA5-HMA5 hasimportant significance in rice blast resistant genetic improvement and disease-resistant variety breeding of rice.

The invention provides a rapid reproduction method of schlumbergera seedlings. The method comprises the following steps: inoculating a selected, disinfected and sterilized schlumbergera explant onto a culture medium for induction and carrying out synchronous induction and proliferation culture until the explant sprouts and differentiates out a bud; sub-culturing until being proliferated to 3-4 folds, and then transferring to a rooting medium for rooting culture until 3-5 fine roots are grown at the base of a plant; conventionally hardening and then transplanting to a mixed substrate of humus soil, red soil and pearlite; and watering and fertilizing conventionally for growing to obtain the schlumbergera seedlings. In the method, key links in the whole production technical flow of the schlumbergera seedlings are optimized and integrated, the culture medium for induction is blended and bud induction and proliferation culture are carried out synchronously, thus simplifying tissue culture procedure and shortening breeding cycle; and only by adopting two types of the culture media, the bud can be directly formed without callus formation, thus reducing probability of variation occurred on an adventitious bud, effectively controlling generation of a variant strain, being capable of fixing and maintaining fine variety of the schlumbergera seedlings rapidly, improving seedling quality and solving the problems of complicated parental source and unstable quality of the seedlings produced by the conventional breeding system.

The invention belongs to the technical field of medicines, and particularly relates to application of glutamine in preparation of a drug for inhibiting escherichia coli from generating gene mutation.According to the application, the glutamine and ampicillin are combined for use, and after multiple passages, the escherichia coli can be inhibited from generating drug-resistant gene mutation, so that mutation probability of drug-resistant genes is remarkably reduced, the increase times of bacteria on the lowest inhibitory concentration of the ampicillin after passage is reduced, and the survivalrate is reduced. Animal experiments also prove that passage bacteria added with the glutamine and the ampicillin are easier to remove by the body, so that the effect of slowing down the drug-resistant gene mutation of the escherichia coli on the ampicillin is achieved.