124results about How to "Short conversion cycle" patented technology

The invention discloses a method for preparing levodopa by virtue of an enzymic method. The method comprises the following steps: picking up an annular stenotrophomonas maltophilia strain slant for inoculation into a seed culture medium for culturing, thereby obtaining a primary seed liquid; inoculating a fermentation culture medium with the primary seed liquid at the inoculum size of 3-10% for fermentation culturing, centrifuging after the fermentation is ended and collecting bacterial cells; and adding tyrosine and the bacterial cells to a buffer solution to have an enzymatic reaction under the conditions of 18-30 DEG C and the pH of 5.0-6.0, thereby converting the tyrosine into the levodopa. According to the method, due to the resting cell transformation technology, and the preferred intermittent weak ventilation technology, adopted in the transformation process, the catalytic efficiency of the enzyme is improved; the concentration of the levodopa is 27g/L and the molar transformation rate of the tyrosine is above 99%; and besides, the method is mild in conditions, high in enantio-selectivity and free from racemization effect, is more suitable for industrial production, and has remarkable economic benefit and industrial application value.

The invention discloses a nonglutinous rice callus culture medium and a genetic transformation method mediated by agrobacterium tumfaciens. The culture medium contains major elements, minor elements, organic elements and hormone in different culture stages which are required for the growth of nonglutinous rice calluses. The genetic transformation method of nonglutinous rice based on the culture medium comprises the steps of induction, subculture, pre-culture, co-culture, sieving, aftersieving, differentiation, rooting of the calluses and the like. The method is widely suitable for nonglutinous rice genotypes, all genetic transformation of the tested 35 nonglutinous rice genotypes are successful, and the transformation rate is between 20% and 50%. The genetic transformation method solves the technical problem of low genotype reliance and transformation frequency existing in the nonglutinous rice callus culture and mediation of agrobacterium tumfaciens for a long time, and is widely applied to the research in the molecular biology, the functional genomics of the nonglutinous rice and the genetic improvement of varieties. The planting area of the nonglutinous rice accounts for more than 80% of the total production area of the nonglutinous rice, and the genetic transformation method has an important function on the research of the basic biology, the improvement of new varieties and the safety guarantee of future grain.

The invention belongs to the technical field of plant biology, and particularly relates to a method for agrobacterium tumefaciens-mediated genetic transformation of dried radix rehmanniae. A leave disc formed in a shearing mode after blades of aseptic dried radix rehmanniae seedlings are unfolded initially and veins are removed serves as the infection receptor of agrobacterium tumefaciens, and a transgenetic seedling of the radix rehmanniae is finally acquired through the processes of agrobacterium infection, co-culture, resistant callus induction, resistance bud initiation, and resistance bud rooting. The method is short in culture period and high in transformation efficiency, successfully achieves genetic transformation of the dried radix rehmanniae, lays the foundation of stable expression of an exogenous gene in the dried radix rehmanniae, and has significance in radix rehmanniae genetic improvement and the molecular biology study.

The invention provides a method for quickly building a genetic transformation regeneration system of wine grape Riesling, and belongs to the technical field of transgenic breeding of grapes. The method is characterized by comprising the following steps: performing tissue culture to induce grape anther to obtain callus; subculturing and screening to obtain embryonic callus; with embryonic callus as a receptor, screening through a selective medium by an agrobacterium tumefaciens-mediated method to obtain resistant callus; and then regenerating a grape plant. Whether the gene gfp is expressed can be observed and reported by PCR (Polymerase Chain Reaction) verification and through a stereo fluorescence microscope, and a transformant also can be identified quickly, therefore, the grape transgenosis efficiency can be improved. Compared with the prior art, the method has the advantages that the culture medium has a good effect on culturing and screening, and achieves quick regeneration of a Riesling transgenic grape plant, thus the grape transgenosis efficiency is greatly improved, and the regeneration rate reaches 81%; the transformation cycle is short, the transgenic plant can be obtained within seven months; the transformant is greatly increased; the transformation rate can reach 63%.

The invention discloses a method for agrobacterium-induced genetic transformation of Ustilaginoidea virens. The method comprises the following steps of carrying out pre-induction culture on agrobacterium containing a hygromycin B gene and a marker gene, mixing an Ustilaginoidea virens spore liquid and the pre-induced agrobacterium liquid according to a volume ratio of 1: 1 to obtain a mixed bacterium liquid, uniformly coating the mixed bacterium liquid on an acetosyringone-containing modified AIM solid medium flat plate of which the surface is coated with a filter paper, a PVDF film or a nitrocellulose film, carrying out co-culture in the dark at a temperature of 20-24 DEG C for 36-48h, shearing the co-cultured film into small slices, paving the small slices on a modified selective medium CM, and carrying out culture at a temperature of 26 DEG C for 4-5 days to obtain hygromycin B gene-containing Ustilaginoidea virens. The method shortens converter time by 4-5d and has a conversion rate of 91.3%. The transgenic Ustilaginoidea virens has stable offspring, a low gene loss rate and converter stability of 95%.

The invention relates to a soybean conversion method which comprises the following steps: cotyledon and the first euphylla of germinated soybean seeds are removed, and naked meristem is obtained; the naked meristem is inoculated to a pretreatment culture medium containing cytokinin for pretreatment; meristem blocks after the pretreatment are infected by agrobacteria. According to the soybean conversion method, the soybean meristem blocks after the pretreatment are used as explant materials for the first time; the explant materials are subjected to high-concentration cytokinin treatment before agrobacterium infection so that the meristem of soybeans can be in an active state and exogenous genes can be inserted easily; meanwhile, the meristem is multiplied and enlarged, so that the infection process is more convenient to operate; in the process of agrobacterium infection, ultrasonic treatment is added, so that the efficiency of insertion of the exogenous genes in a plant genome is greatly improved, and the conversion rate of the soybeans can reach 10%; meanwhile, the conversion period is short, the number of chimera is small, and conversion cost is low.

A provided method for improving cotton transgenosis efficiency comprises the following steps: a, employing an agrobacterium-mediated method to infect cotton aseptic-seedling shoot tips with a plant expression carrier containing a target gene, and placing the infected cotton shoot tips in a co-culture medium for culture; and b, after co-culture is performed, transferring the cotton shoot tips to a selective medium for culture. The co-culture medium and the selective medium both contain KT with a concentration of 0.3 mg/L and IAA with a concentration of 0.2 mg/L. By utilizing the method, the aseptic seedling culture time is 2-3 days, the co-culture time is 2 days, the selective culture time is 15-20 days, the rooting culture time is 20 days, the conversion period does not exceed 2 months, and the positive conversion rate reaches 8% after screening is performed.

The invention discloses a vacuum-infiltration assisted exogenous gene transforming method of a soybean germinating embryo, which comprises the following steps of: 1. carrying out disinfection and pre-culture processing on seeds; 2. after the expansion and the germination of seed embryos, cutting, exposing or damaging germinating embryo parts; 3. infecting the germinating embryos by agrobacteria containing target genes; 4. carrying out co-culture; 5. regenerating and screening clustered buds as well as rooting and screening plantlets; and 6. regenerating and screening the clustered buds as well as rooting and screening the plantlets. The method is applicable to multiple genotypes, has high regenerating speed and is a reliable system for soybean transgenic breeding.

The invention discloses an adventitious bud transformation method for sweet potatoes mediated by agrobacterium. The method comprises the following steps: (1) acquisition and culture of aseptic seedlings; (2) pre-culture of stem segments of the sweet potato; (3) preparation of activating agent bacteria solutions of strains; (4) infection and co-culture of the agrobacterium; (5) bacteria-elimination of the agrobacterium and adventitious bud regeneration; (6) segmentation and rootage of gemmules of the sweet potato; (7) selection of transgenic plants; and (8) detection and transplantation. The invention provides a new method of genetic transformation of the sweet potatoes, which aims to use the stem segments of the sweet potato as an explant and obtain the transgenic plants through infecting and inducing the adventitious bud, therefore, the invention is suitable for various kinds of gene types with fast regeneration speed, and is a reliable system for sweet potato transgenic breeding.

The invention discloses a rapid and high-efficiency transgenic method for indica rice. The method comprises the following steps of: exposing indica rice mature seeds to Agrobacterium tumefaciens 5 to 15 days after callus induction, co-culturing, performing resistant callus screening for 10 to 20 days, performing differentiation culture for 10 to 20 days, performing rooting culture for 5 to 10 days, and performing resistance screening again. The transgenic plants of indica rice can be produced within 40 to 50 days, and the conversion efficiency reaches 20-30%. In the conversion process, the culture conditions are as follows: the temperature ranges from 30 DEG C to 33 DEG C and the light is continuously supplied with the intensity of 80 to 120 mu mole/m<-2>s<-1>, except that the co-culture stage is carried out at 25 DEG C in the dark. The method provided by the invention greatly shortens the in vitro culture time of calli, increases the differentiation efficiency, reduces the mutation frequency of somatic cells, simplifies the operation procedure and saves a large amount of labor and material resources.

The invention discloses a genetic transformation system of a new strawberry variety 'Shenqi'. An agrobacterium tumefaciens-mediated 'Shenqi' leaf and petiole genetic transformation system is established through a leaf and petiole in-vitro regeneration method, and the method comprises the following steps: preparing a sterile leaf; screening the optimal exogenous hormone concentration to induce the leaves and petioles of the strawberries' Shenqi 'to regenerate adventitious buds; analyzing the sensitivity of 'Shenqi' leaf regeneration on antibiotics; activating and culturing agrobacterium tumefaciens; inducing and culturing adventitious buds of the leaves and the petioles; soaking leaves with agrobacterium and performing co-culturing; screening positive strains with antibiotics; and detecting a transformed strain and the like. According to the method disclosed by the invention, adventitious buds are induced by utilizing strawberry leaves and petioles, the leaves are impregnated by agrobacterium, and transformed strains are identified by methods such as antibiotic screening, fluorescence screening and PCR (Polymerase Chain Reaction) detection after co-culture, so that the method is suitable for tissue culture efficient regeneration of the strawberry 'Shenqi' leaves and petioles and agrobacterium-mediated genetic transformation application; the method has important significance on genetic engineering of a new variety of 'Shenqi' strawberries.

A process for quickly culturing transgenic plant of potato includes such steps as agroinfecting the potato explant cultured in test tube, culturing to introduce the target gene to potato receptor, direct culturing in solid differential culture medium to induce resistant buds, culturing in selective rooting culture medium to induco normal root, and verifying by PCR, PCR-southern, or Northern hybridizing method.

This invention relates to Gluconobacter oxydans mutant (CGMCC No. 1184) and its application in preparing 6-(2-hydroxyethyl) amino-6-deoxy-alpha-L-sorbfuranose by microbe conversion. The Gluconobacter oxydans mutant has such advantages as high stability, mild and controllable culture conditions, rapid propagation and short conversion time.

The invention discloses an agrobacterium tumefaciens-medicated genetic transformation method of wheat. The agrobacterium tumefaciens-medicated genetic transformation method comprises the following steps of: firstly, inoculating recombinant Agrobacterium tumefaciens containing a target DNA (Deoxyribonucleic Acid) into wheat immature embryos; and then co-culturing the inoculated wheat immature embryos for enabling the T-DNA of the recombinant Agrobacterium tumefaciens to be integrated with a chromosome of the wheat; and secondly, directly inoculating the wheat immature embryos co-cultured in the first step in a long-seedling culture medium for culturing to obtain transgenic wheat seedlings with roots, stems and leaves. The method disclosed by the invention has the advantages that the applicability is wide, so that the method is suitable for transforming various wheat species; the method is simple in operation, and thus a transgenic plant can be obtained without inducing a tissue culture and regenerating the plant; and the transformation period is short, i.e. the process from the infection to the obtaining of the transgenic wheat seedlings with the roots, the stems and the leaves only requires 11-14 days. The method has very important significance for genetic improvement on the wheat species and the obtaining of new species of the transgenic wheat.

The invention discloses a method for preparing gamma-decalactone. The method comprises the steps of preparing seed culture fluid with yeast CICC 32859 serving as bacteria, then inoculating the seed culture fluid to a transform culture medium, adding L-carnitine into the transform culture medium, adding a small quantity of gamma-decalactone compound into fermentation liquor in the transformation process, conducting biotransformation with ricinoleic acid serving as substrate, and preparing the gamma-decalactone. According to the method for preparing the gamma-decalactone, the product formation rate is promoted, the fermentation period is shortened, and more than that, a new method for restraining saccharomycetes to conduct open circle degradation on the product, namely the gamma-decalactone, is provided, so that the one-time cumulant of the gamma-decalactone is greatly increased.

The invention relates to a method for treating corn straws by using two kinds of insects. The method is characterized by comprising the following steps: smashing fresh corn straws for feeding the locusta migratoria manilensis; crushing dry corn straws, mixing the crushed dry corn straws, locusta migratoria manilensis feces generated after feeding of the locusta migratoria manilensis and remainingcorn straw residues after feeding of the locusta migratoria manilensis, and laying the mixture; stacking and composting the laid mixture; turning the compost pile; carrying out soil preparation disinfection treatment on a protaetia brevitarsis culture shed to manufacture a culture purse net; inoculating protaetia brevitarsis larvae in a breeding shed; and harvesting adult of the locusta migratoriamanilensis and the aged larvae of the protaetia brevitarsis for breeding. The method has the beneficial effects that the corn straws can be completely transformed by utilizing the combination of thetwo kinds of environmental insects, the transformation period is short, the synergistic effect of the locusta migratoria manilensis and the protaetia brevitarsis is fully achieved, and high speed, high efficiency and high value transformation of the corn straws are guaranteed on the premise that auxiliary materials are not added, the process energy consumption is low, the transformation period isshort, and secondary pollution is avoided.