60results about How to "Increase mutagenic effect" patented technology

The present invention discloses one Penicillium citrinum strain with high nuclease P1 yield and its selective breeding process. The Penicillium citrinum strain is preserved in the preservation number of CGMCC No. 2014. Its selective breeding process includes low energy ion beam implantation on Penicillium citrinum spore suspension, transferring the mutagenized spore to plate culture medium for culturing, selecting single strain and transferring to malt wort and slant cultivating, liquid fermenting culturing to screen out strain with high nuclease P1 yield, and passage culturing for over 30 generations to obtain strain with stable high yield character. The Penicillium citrinum strain has high enzyme activity and stable high yield character, and is suitable for industrial production.

The invention relates to a polyploid inducement method of Fritillaria pallidiflora Schrenk in vitro culture, which uses fresh bulb to be induced to produce adventitious buds. Inducement treatment is applied through cultivating in MS liquid medium with different density of colchicine to produce Fritillaria pallidiflora Schrenk polyploid plant. Bulb yield and medicinal components content of the plant are improved by changing chromosome ploidy to produce plant with good characteristics. Therefore, the problem of variety degeneration of Fritillaria pallidiflora Schrenk in a long-term course of Fritillaria pallidiflora Schrenk planting is overcomed. The method has the advantages that Fritillaria pallidiflora Schrenk polyploid can be obtained quickly with effectively improved content of active components and a shorter planting cycle.

The invention relates to a protoplast mutation breeding method for improving the cordycepin content of cordyceps militaris. The method comprises the following steps of activated culture of bacterial strains, solid culture of mycelia, liquid mycelia culture, mycelia enzymolysis, protoplast nitrosoguanidine (NTG) mutation, protoplast regeneration culture, regenerated bacteria subculture, fermentation culture, filtering, further washing of mycelia, measurement of cordycepin content in the mycelia, preservation of the bacteria strain with high cordycepin content, measurement of the genetic stability and the like. Due to the adoption of the method, the cordycepin content of the cordyceps militaris can be increased.

The invention discloses a method used laser mutagenic streptomyces roseosporus to breed daptomycin high yield strain breeding. It belongs to strain mutagenesis breeding technology. The method includes the following steps: forming spore suspension; loading in aseptic quartz groove; irradiating by He-Ne laser for certain time; coating on Gao-NO.1 culture medium flat after diluting by aseptic water to culture pure streptomyces roseosporus; selecting out pure strain; and transferring it to liquid fermentation medium to culture high yield strain. The invention has the advantages of simple device, practical method, and safe operation. Its mutagenesis effect is far better than traditional physical chemistry mutagenesis method. Compared with original strain, the fermentation unit of the formed streptomyces roseosporus can rise by 0.5-3.3 times.

The invention belongs to the field of microbial fungi, and relates to a Cordyceps militaris strain capable of producing heat-resistant proteinase and cordycepin at high yield and application thereof. The strain is prepared by the following steps: selecting a Cordyceps militaris wild-type strain newly separated from the natural world as the initial strain, carrying out composite mutagenization under the actions of Cordyceps militaris protoplast nitrosoguanidine and ultraviolet light, separating and screening to obtain the strain. The strain was collected by China General Microbiological Culture Collection Center on November 11th, 2016; the collection number is CGMCC No.13179; the strain is named HYCM12; and the classification designation is Cordyceps militaris. After the strain is subjected to deep liquid ventilation fermentation, the cordycepin content reaches 2.89 g/L, and the heat-resistant proteinase activity reaches 5326U/ml. Meanwhile, the method is simple to operate and easy to implement, has the advantages of short period, abundant and accessible raw material sources, low cost, no environmental pollution, low equipment and reagent price and the like, and is convenient for large-scale production.

The invention discloses a phaffia rhodozyma strain for high yield of astaxanthin, which is a Phaffiarhodozyma LUCNOVA001 strain, is preserved in the China Center for Type Culture Collection (CCTCC), has the preservation number of CCTCC NO: M20211124, has the preservation date of September 1, 2021, and is named as PhaffiarhodozymaLUCNOVA001 in taxonomy; the strain is obtained by taking Phaffiarhodozyma as a starting production strain and carrying out ARTP mutagenesis and [beta]-ionone screening, and the breeding method comprises the following steps: (1) activating the strain; (2) preparing bacterial liquid in a logarithmic phase; (3) preparing a bacterial suspension; (4) carrying out ARTP mutagenesis; (5) screening the [beta]-violet ketone; (6) secondary screening of strains; and (7) performing continuous passage. The astaxanthin yield and content of the strain and the corresponding biomass meet the requirements of industrial production, and the strain has a great application prospect.

The invention provides a method for treating crop seeds through atmospheric pressure room temperature plasma. The method can be used for treating mung bean seeds, soybean seeds, corn seeds and wheat seeds. Compared with a control group, the bud ratio of crop seeds treated through plasma is obviously improved; the bud ratio of crop seeds subjected to germination treatment is further obviously improved, and the mutagenic effect is better, for example, the bud ratio of crop seeds subjected to mechanical seed coat breaking treatment is obviously higher than that of crop seeds subjected to seed coat complete germination, and the bud ratio of crop seeds subjected to gibberellin soaking treatment is obviously higher than that of un-soaked crop seeds.

The invention relates to a method for high-yield bacterial strains by high-flux screening of gentamycin based on an ARTP composite mutagenesis technique. The method comprises the steps of preparing aspore suspension, performing ARTP mutagenesis composite mutagenesis, performing quick detection on products, prescreening high-yield bacterial strains with gentamycin and performing screening once again to obtain high-yield bacterial strains and performing stability inspection on the high-yield bacterial strains. Compared with the prior art, the method has the advantages that the principle that OPA and the gentamycin are subjected to a reaction to form characteristic absorption peaks of about 330nm is adopted, through a microplate reader, quick determination of an OD value is realized, so thatfast detection of the gentamycin can be successfully realized. The interference problems that poor specificity of a phosphotungstic acid method and a method for forming precipitate with the gentamycin to cause reduction of the OD value, fermentation liquid color and the like, are solved. The method has the characteristics of being quick, high-flux, convenient to operate, accurate and the like, and the mutation breeding and screening efficiency of the gentamycin is improved to a great extent, so that the probability for obtaining the high-yield bacterial strain with the gentamycin is greatly improved.

The invention discloses a handling method for inducing citrus bud mutation through NaN3. The method includes the following steps that NaN3 conditioning fluid is prepared; citrus one-to-two-year-old grafted seedlings are selected, the part, which is not lignified completely, of a branch to be handled is cut off, and all leaves and germinal buds of the reserved part of the branch are removed; all leaf axils of the reserved part of the branch are incised by a sharp blade; the branch to be handled is wrapped with absorbent paper by one to two layers, and the wrapped absorbent paper is wetted by the NaN3 conditioning fluid to be saturated through a liquid-moving machine and the like; the branch is covered with a preservative film until handling is completed, so that the conditioning fluid is prevented from being evaporated; the wrapped absorbent paper is uncovered, a plant is placed under normal culture conditions such as the outside of a room to grow, it can be observed that new shoots can germinate and grow from certain leaf axils after five days, and afterwards, variation character observation and statistics of leaf morphology and the like can be conducted. In the culture growth process after handling, the buds growing at the unhandled part of the plant need to be removed in time, in accordance with experimental result statistics, the mutagenesis effect is obvious, and application value can be achieved.

The invention discloses a method for breeding an high-yield ascomycin strain by performing femtosecond laser mutagenesis on streptomyces hygroscopicus ascomycota subspecies and a culture medium preparation method; the method comprises the following steps: at room temperature, mixing mature slant spores of the streptomyces hygroscopicus ascomycota subspecies (ATCC14891) and sterile water to obtain monospore suspension containing 106-107 spores per milliliter; irradiating the spore suspension for 1-10 min by using the titanium sapphire femtosecond laser with centre wavelength of 800 nm, pulse width of 150 fs and frequency of 76 MHz under a irradiation power of 10-30m W; properly diluting the spore suspension, coating on a solid panel to culture, then screening by using a shake flask to obtain a high-yield ascomycin mutation strain. The method provided by the invention has the advantages that the used femtosecond laser mutagenesis method is feasible and safe to operate; the mutation effect is better than that of the traditional physical and chemical mutagenesis method, and the femtosecond laser mutagenesis method has great popularization value in breeding of microorganism pharmaceutical strains; by using the femtosecond laser irradiation to perform the mutagenesis, the high-yield ascomycin mutation strain can be bred. The positive mutation rate of the mutation strain is 5-30%, and fermenting unit is improved by 10-60% in comparison with that of an original strain.

The invention discloses a compound mutation method for an inosine-producing strain. Bacillus subtilis protoplast is subjected to atmospheric and room temperature plasma mutation. The compound mutation method comprises the following steps: (1) activation culture of the strain; (2) shake flask liquid culture of the strain; (3) enzymolysis and wall digestion of thallus; (4) atmospheric temperature plasma mutation; (5) regeneration culture of protoplast; (6) 24-well plate screening; and (7) shake flask fermentation and rescreening. After the strain is subjected to passage for fifty times, the genetic characters are stable. Compared with the original strain, the mutant strain has the advantages that the inosine-producing amount is increased from 60g/L to 72g/L, the conversion rate is increased by 20% and the significant effect is achieved.

The invention discloses a fungus mutagenesis method to produce, Tacrolimus. The method includes the process as follow: in the room temperature, make bevel spore into spore suspended liquid by asepsis water and enclose them into asepsis cuvette, let them be radiated in some time by He-Ne,then dilute it and besmear it to No1 cultivating basis flat to get seduced streptomycin with jarless heredity. This method has simple equipment, easily doable and safe operation.

The invention discloses a sodium azide mutagenizing method for juvenile citrus internodal stem segments, comprising: I, taking citrus young plantlets more than 1 mm in stem thickness and 5-10 cm in plantlet height, cutting to obtain internodal stem segments, soaking the internodal stem segments in a mutagenizing liquid, mutagenizing in darkness for 1-3 h, wherein the mutagenizing liquid includes sodium azide having a concentration of 0.5-2 mmol/L; II, subjecting the mutagenized internodal stem segments to tissue culture until sectional wounds of the internodal stem segments grow regenerated buds; III, using an SRAP (sequence-related amplified polymorphism) molecular markup method to detect the regenerated buds that vary, and subjecting the varied regenerated buds to rooting culture to obtain complete citrus variant plants. It is clarified that sodium azide-mutagenized juvenile citrus internodal stem segments may produce stable variant plants. The method of the invention has mutagenizing efficiency of greater than 10% and has significant mutagenizing effect and good operating convenience.

The invention provides a method for creating waterlogging-resistant wax gourd mutants by mutagenic agents EMS. The method includes (1), treating wax gourd seeds by EMS phosphate buffer liquid with thevolume ratio of 1-1.3% for 10-15hours to obtain M1-generation seeds; (2), putting the M1-generation seeds treated by the mutagenic agents EMS into a CO2 constant-temperature pregermination box, withthe temperature of 25-30 DEG C and the oxygen concentration of 10-15%, for pregermination; (3), sowing the M1-generation seeds, acquiring M2-generation seeds after selfing, sowing the M2-generation seeds, performing selfing and pollination after 15-25 days, soaking the basal parts of stems by 15-25cm for 3-4 hours in a field, selecting waterlogging-resistant wax gourd plants, harvesting and reserving seeds of the waterlogging-resistant wax gourd plants per gourd to obtain the waterlogging-resistant wax gourd mutants Mut. The method has the advantages that the wax gourd seeds are mutagenized bythe chemical mutagenic agents EMS to obtain the waterlogging-resistant wax gourd mutants, so that sources of genetic resources are widened, and new genetic resources are created.

The method discloses a method for mutating flavomycin producing bacteria by using a high-voltage corona electric field. The method is characterized in that the mutation voltage is 21kV, the needlepoint distance is 8mm, and the mutation time is 4min. A high-voltage corona electric field device adopted in the method is characterized in that the needlepoint distance is 20mm, the needle plate distance is 25mm, and the needlepoint diameter d is less than 0.01mm. As the flavomycin producing bacteria 08-28 is very sensitive to the electric field and the fatality rate of strains is increased thereupon along with the increasing of mutation dosage, and when the mutation voltage is 21kV, the needlepoint distance is 8mm and the mutation time is 4min, the mutation dosage is the best. Mutated strains No.3 are obtained through the mutation, screening and secondary culture of the electric field; and compared with the original strains, the mutated strains No.3 have the advantages that the capability of for producing the flavomycin is increased by 1.92 times, thereby showing that high-voltage corona electric field has better mutation effect on the flavomycin producing bacteria.

The invention discloses a ploidy mutagenesis method of brier grape buds covered by propyzamide. The method comprises the following steps: (1) dissolving propyzamide in dimethyl sulfoxide, and diluting with water till the mass concentration of the propyzamide is 0.005%-0.007% to obtain a propyzamide solution; and (2) preparing the propyzamide solution into propyzamide agar paste, covering the brier grape buds in the budding growth state with the propyzamide agar past for 5-7 days to complete ploidy mutagenesis of the brier grape buds. Through a covering method, the ploidy mutagenesis method induces the brier grape buds in the budding growth state to successfully obtain brier grape tetraploids; the propyzamide is used as a main reagent; when the concentration of the propyzamide solution applied to the brier grape buds is 0.005%, an optimal mutagenesis effect is achieved; the mutagenesis rate reaches 60% or above.

The invention discloses a beer yeast strain and an application thereof in beer fermentation. The invention provides a saccharomyces cerevisiae Z5 with a preservation number of CGMCC No 3218. A breeding method of the yeast strain comprises the following steps: preparation of an original parent strain, activation in a test tube, ion implantation mutagenesis, laser mutagenesis, wort agar plate primary screening, fermentation bung secondary screening, passage stability test and pilot test. The yeast strain is an excellent beer yeast strain and has a potential of large-scale production of beer breweries.

The invention discloses a method for high-throughput breeding of petroleum degradation mutant strains, which adopts Licl-ARTP combined mutagenesis of Gordon's strain to improve the ability of petroleum hydrocarbon degradation and comprises the steps: preparing bacterial suspension, preparing TSB solid culture medium containing different concentrations of lithium chloride, carrying out single chemical mutagenesis (Licl) and atmospheric pressure plasma mutagenesis (ARTP) on the strain, calculating lethality rate and determining degradation rate of petroleum hydrocarbons of the single mutagenic strain. (4) Licl-ARTP compound mutagenesis. (5) 96-well plate fermentation instead of shake flask fermentation is used to screen mutant strains by ultraviolet spectrophotometry with high throughput. The strain is Gordon's bacillus. The positive mutation rate of the mutant strain is high and the degradation ability of petroleum hydrocarbons is obviously improved. It is beneficial to industrial application and has great significance in economic, environmental and social benefits.

The invention discloses a method for inducing an astragalus sinicus tetraploid. The method comprises the following step: by using a combined mutagenic agent, performing dropping treatment on a growingpoint when a cotyledon-petiole included angle of a diploid astragalus sinicus ranges from 30 degrees to 50 degrees and a first true leaf is not released, wherein after the treatment, the optical density is reduced to 180 [mu]mol.m<-2>.s<-1>, the illumination time is controlled to be 10 h.d<-1>, the day temperature is 18 DEG C, the night temperature is 15 DEG C, and the relative humidity is 75% to90%; the combined mutagenic agent comprises 0.1 to 0.2% of colchicine, 50 to 100 mg.L<-1> of trifluralin, and 20 [mu]l.L<-1> Tween T-20; the treatment using the combined mutagenic agent is performedfor 2 to 4 times. According to the method, colchicine and trifluralin at specific concentrations are used in a mixed manner, so that the tetraploid induction rate reaches 13.7% and is improved; in addition, the concentration of colchicine is reduced, so that environmental pollution is reduced.