63 results about "Conidial suspension" patented technology

The invention discloses a rapid identification method of tobacco black shank resistance. The rapid identification method of the tobacco black shank resistance includes the following steps of (1) sterilizing the surface of tobacco seeds to be identified; (2) cultivating tobacco blank shank germs and preparing suspension liquid of conidium of the tobacco blank shank germs; (3) extracting toxin liquid of the tobacco blank shank germs; (4) germinating the seeds after soaking seed in the toxin liquid and calculating rate of emergence of the seeds and evaluating the resistance. The rapid identification method of the tobacco black shank resistance is easy to operate, low in cost, rapid, high in accuracy, and appropriate for black shank resistance identification in tobacco breeding research.

The invention discloses a method for identifying gray mold resistance of gerbera, belonging to the technical field of plant disease resistance identification. The method comprises the following steps: (1) selection of proper inoculation material: selecting freshly collected and integrated young leaves and petals as the inoculation material; (2) culture of inoculation material and preparation of conidia suspension; (3) preparation of inoculation liquid: before inoculation, diluting the conidia stock solution into an inoculation liquid containing 3 wt% of potato glucose and 1*10<6>/ml conidia, or into an inoculation liquid containing 6wt% of potato glucose and 3*10<5>/ml conidia; and (4) inoculation. The method disclosed by the invention is carried out under manually controlled stable conditions, and achieves the goal of identifying the gray mold resistance of gerbera in a simple, quick and accurate way; and the accuracy of the identification result is high.

Previous research demonstrated that aflatoxin contamination in corn is reduced by field application of wheat grains pre-inoculated with the non-aflatoxigenic Aspergillus flavus strain NRRL 30797. To facilitate field applications of the biocontrol isolate, a series of laboratory studies were conducted on the reliability and efficiency of replacing wheat grains with the novel bioplastic formulation Mater-Bi® to serve as a carrier matrix to formulate this fungus. Mater-Bi® granules were inoculated with a conidial suspension of NRRL 30797 to achieve a final cell density of ˜log 7 conidia/granule. Incubation of 20-g soil samples receiving a single Mater-Bi® granule for 60-days resulted in log 4.2 to 5.3 propagules of A. flavus/g soil for microbiologically active and sterilized soil, respectively. Increasing the number of granules had no effect on the degree of soil colonization by the biocontrol fungus. In addition to the maintenance of rapid vegetative growth and colonization of soil samples, the bioplastic formulation was highly stable, indicating that Mater-Bi® is a suitable substitute for biocontrol applications of A. flavus NRRL 30797.

The invention discloses short and dense trichoderma brevicompactum and application thereof in cucumber fusarium wilt prevention and control. According to the short and dense trichoderma brevicompactum, the strain number is BF06, and the registering and booking serial number of the short and dense trichoderma brevicompactum in the China General Microbiological Culture Collection Center is CGMCC No.11845. The short and dense trichoderma brevicompactum BF06 has the significant antagonistic action on fusarium oxysporum f.sp cucumerinum which causes the cucumber fusarium wilt; conidial suspension of the strain and ordinary cultivation soil are mixed to serve as a biocontrol matrix, therefore, the cucumber fusarium wilt can be effectively prevented and controlled, and the prevention and control effect on the cucumber fusarium wilt is 90.4%. Accordingly, an excellent strain resource is supplied to pollution-free production of cucumbers in a protected area, and the practical value on prevention and control over the cucumber fusarium wilt, especially cucumber seedling stage fusarium wilt, is achieved.

The invention aims to provide an apple tree valsa ceratosperma transformant and a preparation method thereof. The method comprises the following steps of: after mixing, co-culturing conidial suspensions of agrobacterium tumefaciens containing a binary vector and valsa ceratosperma, screening by using antibiotics to obtain the transformant. According to the method, the conidium of apple tree valsa ceratosperma is used as a receptor, agrobacterium tumefaciens is used as an amboceptor, the problems of difficulty in preparation of protoplast of the apple tree valsa ceratosperma and difficulty in genetic transformation of the valsa ceratosperma caused by low polyethylene glycol (PEG)-mediated transformation efficiency and the like are overcome; the transformation efficiency of the method can be up to 1/10<3> conidiums; simultaneously, the invention provides an apple tree valsa ceratosperma green fluorescent protein (GFP) labelled strain and a preparation method thereof; and the efficiency of the green fluorescent protein with strong expression in the transformant obtained by the method can be up to 96.7%.

The invention discloses a pesticide compounding beauveria bassiana fungus and destruxin. The pesticide is prepared by mixing the beauveria bassiana fungus, the destruxin and solvent, the concentration of live conidia of the beauveria bassiana fungus after mixing ranges from 10<7> to 10<11> conidia per liter, and the pesticidal effect is better when the concentration of the destruxin ranges from 10 mg/L to 100 mg/L. The preparation method for the pesticide comprises the following steps: preparing conidial suspension of the beauveria bassiana fungus, preparing destruxin stock solution, mixing the conidial suspension and the destruxin stock solution, and the like. The invention selects the beauveria bassiana fungus to be used with the destruxin in a mixing way for synergic action, and the pesticide has the completely new function mechanism different form the prior pesticide, has an excellent pesticidal effect particularly in poisoning and killing bemisia tabaci gennadius with serious harm, and meanwhile is environment-friendly.

The invention discloses a beauveria bassiana strain and application thereof. The beauveria bassiana strain is beauveria bassiana; the preservation name is Beauveria bassiana JXAS-02; the beauveria bassiana strain is preserved in Chinese Type Culture Collection Center at Wuhan University, Bayi Road, Hongshan District, Wuhan City, Hubei Province; the preservation date is April 25, 2019; the preservation number is CCTCC M 2019293. The beauveria bassiana strain JXAS-02 having the high-efficiency insecticidal capability to flatheaded borers is separated and identified from soil by taking flatheadedborer larvae as baits through a biological trapping method from the perspective of biological control, the bacterial agent has the strong infection prevention and control capability on larvae and adults of flatheaded borers, and the strain can be used for effectively controlling the population quantity of flatheaded borers by spraying or pouring a conidium suspension liquid of the strain into feeding tunnels of the flatheaded borers.

The invention discloses a method for obtaining a molecular marker linked with gummy stem blight resistance gene Gsb-2 and application of the molecular marker. An F2 group is constructed by PI 157082 carrying the gummy stem blight resistance gene Gsb-2 and a local susceptible variety of white skin crisp, gummy stem blight conidiospore suspension is artificially inoculated in a seedling period, and the molecular marker ISSR-57560 linked with the Gsb-2 is obtained through a simple sequence repeat (ISSR) technology. The molecular marker ISSR-57560 is used for detecting genotypes of melon varieties or strains taking PI 157082 as a parent, and can judge that whether the varieties or strains resist gummy stem blight. The invention overcomes the defects of high difficulty and longer period in breeding stable gummy stem blight resistant strains in the prior art, and provides a marker-assisted selection technology for gummy stem blight resistant breeding so as to reduce cost, improve selection efficiency, and quicken the progress of culturing good stem blight resistant melon strains.

The invention relates to a genetic transformation method for an agrobacterium tumefacien-mediated fusarium oxysporum sesame specialized strain. The genetic transformation method comprises the following steps of: (1) preparing conidiospore suspension of the fusarium oxysporum sesame specialized strain; and (2) performing the transforming-deoxyribonucleic acid (T-DNA) conversion of the fusarium oxysporum sesame specialized strain. According to the genetic transformation method, the fusarium oxysporum sesame specialized pathogenic bacteria are used as transformant, and an exogenous T-DNA sequence is guided into a genome of the fusarium oxysporum sesame specialized strain by combining an appropriate genetic transformation receptor system in an agrobacterium tumefacien-mediated method to obtain the fusarium oxysporum sesame specialized transgenic strain for the first time. The genetic transformation method is high in repeatability and high in conversion rate, and is used for constructing the following fusarium oxysporum sesame specialized mutant library; the obtained strain contains a double-tagging gene, can be used for analyzing the mechanism of sesame fusarium wilt, and is the important innovation in the study of sesame fusarium wilt pathopoiesia and sesame disease resistance.

The invention discloses a preparation method and an application of a coniothyrium minitans ZS-1SB fungicide. The preparation method comprises the following steps of: (1) first, performing PDA (Potato Dextrose Agar) slant culture for the coniothyrium minitans ZS-1SB fungicide; washing the slant with sterile water, and adjusting a conidium suspension liquid to 106 spores/ml; (2) oscillating the conidium suspension liquid of coniothyrium minitans ZS-1SB prepared by the step 1 to obtain a coniothyrium minitans ZB-1SB bacterial liquid; and (3) uniformly mixing oatmeal and husks, adding water till the water content reaches 40%, filling by an edible fungus bagging machine, and steam sterilizing at normal pressure; after cooling, inoculating the coniothyrium minitans ZB-1SB bacterial liquid, and culturing in a certain condition by a 30ml/culture bottle. The yield of coniothyrium minitans prepared by the preparation method disclosed by the invention can reach 3.2*10<10> spore/g dry compost. Under field culture condition of an antibiological inoculant, soil broadcast fertilization and overground part in blossom period realizes the preventing effect of 57% to sclerotinia rot of colza. The fungicide can be applied to large-scale disease prevention and treatment.

The invention discloses a living organism inoculation conidium generating method for quickly obtaining panax notoginseng round spot pathogenic bacterium conidia. The method comprises the following steps that 1 of pathogenic bacterium culturing; 2 conidium generating device manufacturing; 3 leaf inoculating; 4 conidium inducing, wherein after inoculating is performed for about 3 days, when obviously macroscopic disease spots are formed on hypha-infected leaves, bacterium cakes are immediately removed, spray moisturizing is continuously performed, conidia are generated at the disease spots after inoculating is performed for about 6 days, the number of the conidia generated on each leaf after inoculating is performed for 10 days reaches about 104, the conidia can be washed down with sterile water in time if the conidia need to be used after the conidia are generated, 1 mL of the sterile water is sucked up with a pipettor to wash each leaf which generates the conidia, and 1 mL of conidium suspension liquid can be obtained from each leaf. Compared with the prior art, the method has the advantages that the mycocentrospora acerina conidia for scientific experiment research can be obtained in a short time, and the stability, the operability and the repeatability of an experiment can be guaranteed.

The invention discloses a set of method for evaluating a prevention and control effect of fungicide on black spot of sweet potato seedlings. According to the method disclosed by the invention, growthconditions of the sweet potato seedlings in a large field are simulated to the greatest extent through an inoculation method comprising the steps of mixing soil or other matrix materials with a sweetpotato black spot conidiospore suspension solution and inoculating, and a cultivation method adopting a cultivation box or an artificial climate chamber, so that uniform disease infection of each treated sweet potato seedling can be ensured and conditions of evaluating the prevention and control effect of the fungicide can be met. Meanwhile, all testing conditions also can be adjustable and controllable; compared with a field experiment, the success rate and accuracy of the experiment are extremely improved and the experiment period is shortened; the disadvantage that the difference between atraditional indoor bioassay result and an actual application result is great is greatly improved. According to the method disclosed by the invention, grading of the disease infection severity degree of the sweet potato seedlings is obtained through a lot of investigation and summarization, and actual conditions of pot culture inoculation are met. Especially, disease infection of sweet potato seedling stem sections and tail wounds are innovatively and independently graded and evaluated, and the method has an important promotion effect of comprehensively evaluating the prevention and control effect of a medicament.

The method consists of placing the tomato seedlings in contact with a formulated Penicillium oxalicum conidia suspension when the plants are in the seed bed, such that the fungi generated by said conidia enhance the development or growth of the tomato seedlings.

The invention discloses isaria javanica for efficiently controlling scale insects. The first study of the isaria javanica for efficiently controlling the scale insects found that the isaria javanica has a very good disease-causing and controlling effect on the scale insects such as icerya aegyptiaca and solenopsis mealybug, an isaria javanica bacterial strain IJID003 with high pathogenicity is screened, the isaria javanica can be used as a living biological pesticide with high pathogenicity on the scale insects, and the isaria javanica for efficiently controlling the scale insects has good broad application prospects in the biological control over the scale insects. The isaria javanica is an entomopathogenic fungus, and a conidia suspension of the isaria javanica can directly act on a wormbody. Compared with current chemical control agents, the isaria javanica has the advantages of efficiency, green, environment-friendly property, environmental protection, high effectiveness and beingnot prone to producing drug resistance, has no pollution to the environment and no residue, and has good market application prospects when the isaria javanica is developed as a biocontrol agent on the biological control over the scale insects.

The invention specifically relates to a method for identifying and evaluating the level of wilt resistance of sesame, belonging to the field of timely evaluation of the disease resistance of crop varieties. The method comprises the following steps: preparation of sesame plants; preparation of a conidium suspension used for dip dyeing; dipping of roots and inoculation of bacteria; identification and evaluation of the wilt resistance of a sesame variety; etc. The method provided by the invention can realize inoculation of sesame plants in different growth stages (from a stage of two pairs of true leaves to a squaring stage) on the basis of a mature root-dipping infection method. Meanwhile, since the concentration of a wilt strain for inoculation is easy to adjust, so detailed analysis can be carried out on both the pathogen infection process and the response process of sesame seedlings, the disease resistance of different sesame germplasms and the pathogenicity of different strains can be accurately identified and evaluated, a technical base is provided for effective control of sesame wilt, and reference is provided for control of other crop diseases. The method provided by the invention has good repeatability, stable and reliable identification results and good application value in scientific research.

The invention provides a conidia coating method for producing a large number of rice blast germ sexual generations. The conidia coating method comprises the following steps: 1) preparing conidia suspensions of different mating types of rice blast germs; 2) coating conidium suspensions of different mating types on the oat culture medium according to a ratio of 1: 1; and 3) culturing for 5-15 days under the culture conditions that the culture temperature is 12-27 DEG C and the illumination is 5000lux-15000lux, so as to obtain a large number of rice blast fungus cocysts and ascospores. According to the method, the defects that sexual generation ascocysts and ascospores generated by a confrontation culture method in the prior art are small in total amount, unstable, large in sample difference, inconsistent in sexual offspring maturity and the like are overcome, a large number of rice blast bacteria sexual generations can be obtained within a short time, the formation speed of the ascocysts and the ascospores is high, the maturity is consistent, and the obtained ascocysts and ascospores can be used for quantitative comparison of sexual reproductive capacity of strains, sexual structure observation, nucleic acid extraction and gene expression quantitative analysis.

The invention discloses a pesticide prepared by mixing destruxin and paecilomyces fumosoroseus fungus, and a preparation method thereof. The concentration of the live conidium of the paecilomyces fumosoroseus fungus in the pesticide is 10<9> to 10<11> conidia per liter, and the concentration of the destruxin is 10 mg/L to 100 mg/L; and the preparation method comprises the following steps: preparing the live conidial suspension for the paecilomyces fumosoroseus fungus, preparing the destruxin stock solution, mixing the suspension and the stock solution, and the like. The invention determines the mix proportion of the destruxin and the paecilomyces fumosoroseus fungus, obtains the better synergic effect, overcomes the defects of instable use effect and low toxicity of the single reagent of the destruxin and the paecilomyces fumosoroseus fungus, and also provides a new technical proposal for biological control. The preparation method for the pesticide is simple and feasible, and is easy to be popularized.

The invention relates to application of alternaria alternata in preventing and controlling external invasion of weed solanum rostratum. Alternaria alternata is obtained by separating and purifying naturally infected plant leaves of solanum rostratum in a field natural population of Xinjiang Changji solanum rostratum, a conventional method is adopted to respectively prepare conidium suspension andcrude toxin from alternaria alternata obtained by separation and purification, the conidium suspension and the crude toxin are uniformly sprayed onto solanum rostratum plants respectively, and alternaria alternata has strong pathogenic effect and control effect on worst weed solanum rostratum. Culture operation is simple, a culture matrix material is easy to obtain, the conidium suspension and thetoxin are high in stability, the obtaining process is simple, and alternaria alternata is supportive of mass production; alternaria alternata can be used for stem-leaf spraying, is convenient in use,can effectively kill target weed solanum rostratum and has high environment safety.

The present application discloses a method for rapidly identifying the resistance of wheat to black point disease caused by Bipolaris sorokiniana, where testing takes places in an indoor, controlled environment. The test method includes surface sterilization of wheat seeds, and cultivating wheat seedlings from the sterilized wheat seeds; preparing conidial suspension of Bipolaris sorokiniana, and spraying the conidial suspension on the seedlings at one-leaf-one-shoot stage; recording the percentage of the diseased leaf area in total leaf area of the first leaf of the wheat seedling on the 10^th day of inoculation; calculating the black point incidence of the wheat according to an equation, and then evaluating the resistance of wheat to black point disease caused by Bipolaris sorokiniana through the black point incidence. Compared with existing field identification methods, the method of the present disclosure shortens identification time, simplifies identification procedure, greatly improves identification efficiency, and improves the accuracy and reliability of identification results.

The invention discloses application of Isaria javanica in control of coccoiol. It is found that Isaria javanica has a good pathogenic control effect on Icerya aegyptiaca (Douglas), Phenacoccus solenopsis Tinsley and other coccoiol pests, can be used as living coccoiol biological pesticide with a high pathogenicity of coccoiol, and has a good application prospect in biological control of coccoiol insects. Moreover, Isaria javanica is an entomopathogenic fungus, conidial suspension of Isaria javanica can directly applied to pest bodies, and when compared with current chemical control agents, high efficiency, green and environmental friendliness, environmental protection, high persistence, no easy production of chemical resistance, no pollution to the environment, no residues and other advantages are achieved; and a good market application prospect is achieved when a developed biocontrol preparation is applied to biological control of the coccoiol insects.

The invention relates to application of a G418 resistance marker in agrobacterium tumefacien-mediated genetic transformation of trichoderma. The resistance of G418 is used as a selection marker of the resistance of trichoderma. An agrobacterium tumefacien-mediated genetic transformation method of trichoderma comprises the following steps: connecting a G418 resistant gene with a carrier p1300 subjected to enzyme digestion and dephosphorylation to form a carrier p1300-neo; activating agrobacterium strains and cultivating until the OD600 value is 0.3-0.6, and using 15-25 mmol/L of CaC12 to perform resuspension on bacterial cells to prepare agrobacterium competent cells; uniformly mixing the carrier p1300-neo with the agrobacterium competent cells, keeping the temperature of 35-40 DEG C for at least 3 min, using an LB (Luria-Bertani) culture medium for cultivation at the temperature of 25-35 DEG C, and then using a YEB culture medium containing kanamycin and rifampicin for cultivation and screening out recombinant agrobacterium tumefacien; uniformly mixing an activated recombinant agrobacterium tumefacien solution in a logarithmic phase, of which the OD660 value is 0.6-0.8 with a trichoderma viride conidial suspension and performing induction cultivation, and then using an M-100 culture medium containing G418 for cultivation at the temperature of 25-30 DEG C and screening out trichoderma resisting G418. The invention provides a novel resistance selection marker for trichoderma viride and brings considerable convenience to research of the gene function of trichoderma, and the cost is low.

The invention provides an Agrobacterium-mediated leptosphaeria biglobosa genetic transformation method, which belongs to the technical field of genetic transformation of plant pathogenic fungi. The method comprises the following steps: 1) inoculating agrobacterium containing plasmid pCHs-GFP in a LB liquid medium for culture, and then transferring the agrobacterium into a CM liquid medium containing 150-250 [mu]g/mL acetosyringone to obtain agrobacterium bacterial fluid; 2) collecting the conidiospore of the leptosphaeria biglobosa, and mixing the conidiospore with sterile water to prepare a conidiospore suspension having a concentration of (0.5-9)*10<6>/mL; and 3) mixing the agrobacteriumbacteria liquid with the conidiospore suspension, co-cultivating the material for 3-4 days, transferring the material to a screening medium, and screening and culturing the material for 7-15 days to obtain a transformant; The method has high transformation efficiency, low copy and genetic stability.

The invention discloses a rapid and mass spore production method for sweet potato black rot bacteria. The method comprises the steps of collecting diseased sweet potato blocks, separating black rot bacteria by virtue of a tissue separation method, inoculating the black rot bacteria into a PS liquid culture medium, carrying out shaking culture at temperature of 31 DEG C and a rotation speed of 120rpm/min for 24 hours, carrying out gauze filtration so as to remove hyphae, carrying out microscopic examination to determine the concentration of spore suspension liquid, and adjusting the concentration of the spore suspension liquid through sterile water to meet the test requirement. The rapid and mass spore production method is scientific, efficient and simple to operate, the preparation time oftraditional black rot bacteria spore suspension liquid can be greatly shortened, the pollution of infectious microbes in a traditional spore suspension liquid preparation method is avoided, and the powerful guarantee is provided for smooth execution of the accurate resistance identification of the sweet potato black rot bacteria, the screening of indoor medicaments of the black rot bacteria, thedrug resistance risk evolution and other tests needing to be finished by virtue of conidium suspension liquid.

The invention provides a preparation method of trichoderma wettable powder, which comprises the following steps: S1, preparation of conidium suspension, S2, preparation of fermentation substrate, S3, preparation of conidium powder, and S4, preparation of the wettable powder, the composition proportion of the wettable powder is determined to be 20% of conidium powder, 1% of AB-3, 5% of dispersant NNO, 1% of ascorbic acid, and the balance of water. According to the present invention, the wetting time, the mass suspension rate and other aspects of the strain meet the related standards of the pesticide wettable powder, the important significance is provided for the development and the factory production of the T.brev T069 product, the composition of the strain wettable powder is determined, and the foundation is laid for the production development, the field application and the pre-evaluation of the biocontrol T.brev T069.

The invention provides an agrobacterium tumefaciens-mediated genetic transformation method of corynespora cassiicola. The method comprises the following steps: (1) determining the sensitivity of the corynespora cassiicola to hygromycin; (2) screening the concentration of antibiotics inhibiting the normal growth of agrobacterium; (3) preparing suspension liquid of conidia of the corynespora cassiicola; (4) preparing an agrobacterium tumefaciens bacteria solution containing a binary vector; (5) co-culturing agrobacterium tumefaciens with the conidia of the corynespora cassiicola; (6) screening transformants; and (7) performing molecular identification on the transformants, and performing insertion of a copy number and genetic stability analysis. The method provided by the invention can be used for preparing protoplast without needing removal of the fungal cell wall, and is simple to operate and high in conversion efficiency; and the obtained transformants are large in single copy ratio and can be stably inherited. The invention provides the stable and efficient agrobacterium tumefaciens-mediated genetic transformation method of the corynespora cassiicola, thus providing a potential possibility for obtaining pathogenic mutants, and laying a foundation for in-depth research on pathogenic related genes and pathogenic mechanisms.

The invention provides a genetic transformation method for an agrobacterium tumefaciens mediated fusarium oxysporum watermelon specialization strain. The method comprises the following steps: (1) preparing fusarium oxysporum watermelon specialization strain conidiophore suspension; (2) preparing an agrobacterium tumefaciens solution containing exogenous genes; (3) performing exogenous gene transformation on the fusarium oxysporum watermelon specialization strain; and (4) identifying a transformant of the fusarium oxysporum watermelon specialization strain. The exogenous genes are transferred into the genome of the fusarium oxysporum watermelon specialization strain by utilizing an agrobacterium tumefaciens mediated method, and a fusarium oxysporum watermelon specialization transgenic strain is obtained. The method is simple, rapid, high in repeatability and transformation efficiency, and can be used for the construction of a fusarium oxysporum watermelon specialization strain mutant library, host resistance screening, defense response of the host on infection of the fusarium oxysporum watermelon specialization strain and research of the infection and pathogenesis of the fusarium oxysporum watermelon specialization strain.