56 results about "Ustilaginoidea virens" patented technology

The invention relates to a Bacillus subtilis T-500. The Bacillus subtilis T-500 has been preserved in CGMCC (China General Microbiological Culture Collection), and the preservation number is CGMCC NO:6058. The Bacillus subtilis T-500 can be used as a biological pesticide for preventing and treating multiple plant diseases, and can also be used as a biological fertilizer for promoting the yield increase of crops. The Bacillus subtilis T-500 has higher antibacterial activity on Magnaporthe oryzae, Pellicularia sasakii, Xanthomonas oryzae, Ustilaginoidea virens, Xanthomonas oryzae pv. oryzicola, Fusarium graminearum, Aspergillus flavus, Aspergillus niger and other pathogenic bacteria, and has better prevention and control effects on diseases caused by the pathogenic bacteria. The Bacillus subtilis T-500 can prevent and treat rice leaf diseases when being diluted 200-500 times on the leaves of rice, can prevent and treat bakanae diseases of rice after seed soaking, and can prevent and treat soil-borne diseases of vegetables and the like after root irrigation.

The invention relates to a simple and convenient method for quickly separating microorganisms, in particular to a method for quickly separating ustilaginoidea virens, which belongs to the technical field of the microorganisms. The method comprises the following steps of: scratching and washing rice curve balls in sterilized water to remove contaminated microbes on a surface layer of the rice curve balls; sterilizing the rice curve balls for 30 to 60 seconds by using 75-percent ethanol solution, and then sterilizing the rice curve balls for 2 to 4 minutes by soaking the rice curve balls in 0.1-percent corrosive sublimate aqueous solution; washing the rice curve balls in the sterilized water, and then adding the rice curve balls into the sterilized water to smash the rice curve balls; using smashing solution to flush a separation Wakimoto culture medium; air-drying the water on the surface of the culture medium, and then putting a culture dish in a thermostat for dark culture at 28 DEG C; after the sixth day of the culture, picking yellow or white small colonies and white mycelia by using a sharp end picking needle, and moving the yellow or white small colonies and the white mycelia into a culture Wakimoto culture medium to perform purification culture for 10 days; and moving the ustilaginoidea virens to a PSA slant culture medium and culturing the ustilaginoidea virens for 10 days to obtain corresponding ustilaginoidea virens. The method can quickly and effectively separate a large amount of the ustilaginoidea virens from the rice curve balls in a short period, and can effectively the problems such as pollution, time waste, poor reliability, high purification difficulty and the like in other methods for separating the ustilaginoidea virens.

The invention discloses a method for separating ustilaginoidea virens. According to the method, a special spore dispersion filament is matched with a spore dispersion plate, single chlamydospore is directly separated from rice grains suffering from false smut, and a false smut bacterial strain with single genetic composition is formed through culture. The implementation and operation steps of the isolated culture are as follows: 1) collecting and preparing a standard sample of false smut; (2) preparing spore dispersion filaments; (3) preparing a spore dispersion plate; (4) dispersing the chlamydospores; (5) cutting out discrete single chlamydospores; and (6) culturing the monospores and forming monospore strains. The method has the following advantages: 1) a key appliance for separating spores is simple and easy to manufacture, and the separation operation is simple and easy; 2) single spore strains can be separated and obtained at one time; 3) the separation work consumes less time and labor; and 4) the working efficiency of Ustilaginoidea virens separation and the reliability of the separation result can be obviously improved.

The invention discloses a plate culture and preparation method of ustilaginoidea virens thin-wall conidium. The core of the method is that the thin-wall conidium is directly utilized for culturing and breeding new thin-wall conidium through a plate culture medium. The method comprises the following steps: 1) preparing an initial thin-wall conidium solution; 3) preparing a sporulation culture medium plate; 3) performing sporulation cultivation; 4) collecting new thin-wall conidium; and 5) purifying conidium. The method has the advantages that the cultivation and preparation can be performed within a shorter time which is generally 4 to 6 days; the operation is simple, easy and convenient; oscillating culture equipment is not required; the thin-wall conidium in an obtained culture product is mainly the conidium of the same generation; the conidium solution is relatively high in quality; no purifying treatment is required; and the method is applied to a plurality of research works.

The invention relates to an effective Ustilaginoidea virens separation method, and belongs to the technical field of microbe. The method comprises the steps of: scraping a small amount of chlamydospore powder on surfaces of false smut balls with a pricking needle; placing the powder in a sterile petri dish; adding 0.5mL of sterile water suspension chlamydospores; then diluting the chlamydospore suspension by 2-3 gradients, with the ratio of the chlamydospore suspension to sterile water being 1:5 in a 1.8mL Eppendorf tube through a gradient dilution method; transferring 0.2mL of the diluted chlamydospore suspension to surface of a Wakimoto toceshi solid culture medium for separation (containing 50 mug / ml chloramphenicol); smearing evenly; inversely placing the petri dish into an incubator at 28 DEG C for dark culture; and effectively separating out target pathogens six days later. The method provided by the invention obviously weakens interference effect of infectious microbe in chlamydospores of rice false smut balls, has very simple operation procedure, is suitable for separation of rice false smut balls stored at normal temperature or 4 DEG C in a refrigerator within 3 months, and has the best separation effect on rice false smut ball bacteria just acquired in the field.

The invention relates to the technical field of separating and preserving ustilaginoidea virens, in particular to a simple method for separating and preserving the ustilaginoidea virens. The separation method comprises the following steps that fresh yellow, green and black rice false smut ball samples are collected from an attacked rice field, drying is conducted at the room temperature for 7-10 d for standby application; rice false smut balls of which the covering depth of chlamydospore powder is larger than 1 mm are selected in the collected samples and placed on an ultra-clean working table, and sterilization is conducted through an ultraviolet lamp for 30-40 min; single-grain rice false smut balls are taken through tweezers under the aseptic condition, and a small amount of chlamydospore powder on the surfaces of the rice false smut balls is shaken off slightly on a PSA culture medium; 150-200 microliters of sterile water is taken through a pipette and drips on a PSA flat plate containing chlamydospore. By means of the simple method for separating and preserving the ustilaginoidea virens, the ustilaginoidea virens can be separated to the objective strains rapidly and preserved, the problems that time is consumed, and the efficiency is low during separation of the ustilaginoidea virens are solved, and meanwhile the problems that in the preservation process of the strains, the pathogenicity is deteriorated, and the space is occupied and consumed are solved.

The invention relates to a pesticide combination, in particular to a pesticide combination containing effective dose of synergic action propiconazole and validamycin A, wherein the weight ratio of the propiconazole to the validamycin A is preferably 10:1 to 1:10, more preferably 10:1 to 1:6, further preferably 1:1 to 1:6, and most preferably 1:2 to 1:6. The pesticide combination is particularly suitable for preventing and treating the holing illness and the ustilaginoidea virens of the paddy rice.

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 separation method of Ustilaginoidea virens (Cooke) Takahashi, and especially an effective separation method of Ustilaginoidea virens (Cooke) Takahashi capable of producing five ustiloxins. The effective separation method comprises a step of false smut ball collection and a step of strain isolation. According to the effective separation method, collected false smut balls are immersed in 75% alcohol for 10s, and are subjected to spontaneous combustion for 5s using an alcohol lamp; water on the surface is removed using sterilized filter paper; surface tissue is cleared under sterile operation conditions; the false smut balls are cut using a sterile scalpel; middle layers are collected, and inoculation on sterile PSA plating medium containing 0.05g / L chloromycetin is performed; after inoculation, obtained culture dishes are subjected to incubation at a constant temperature of 28 DEG C in an incubator; and bacterial strain preservation is carried out after strain purification. Isolation success rate of the effective separation method on collected fresh single yellow, yellow green, or green false smut balls is about 100%; isolation success rate on single yellow, yellow green, or green false smut balls stored at 4 DEG C for 4 months or shorter is 90% or higher, and isolation success rate on single yellow, yellow green, or green false smut balls stored at -70 DEG C for 1 month or shorter is 80% or higher.

The invention discloses a potato culture medium suitable for ustilaginoidea virens spore production and an application thereof. Potatoes are prepared into a plate culture medium as a spore production culture medium so as to culture and prepare ustilaginoidea virens thin-wall conidiospore. Preparation for the culture medium and an application method for the culture medium comprise the following steps: 1) preparing the culture medium; 2) inverting a culture flat plate; 3) implanting ustilaginoidea virens; 4) carrying out constant-temperature culture; and 5) collecting a new generation of spores. The potato culture medium has the characteristics and the advantages that: the culture medium is simple in composition; the operation is simple, easy and convenient, and shaking culture equipment is not needed; time of obtaining a great number of spores is short; a high spore production state lasts for a long time; and a spore liquid is relatively pure, is less in mixed mycelia, and is generally suitable for being used in a lot of research work without purifying treatment.

A kind of Trichoderma atroviride TA‑9, which has been preserved in the General Microorganism Center (CGMCC) of the China Committee for the Collection of Microorganisms, and the preservation number is CGMCC No.9698. The Trichoderma atroviride TA-9 of the present invention can be used as a biological pesticide to prevent and control various rice diseases. Trichoderma dark green TA‑9 showed strong hyperparasitic ability against Rhizoctonia solani and Ustilaginoidea virens. The present invention also relates to the high-yield chlamydospore and conidia liquid fermentation technology of Trichoderma dark viridans TA-9, after its fermented liquid is diluted 50 times, it can control rice sheath blight caused by Rhizoctonia solani and rice sheath blight caused by Chlorosclerotinum oryzae. Aspergillus has a good control effect.

The invention discloses a method for quantitatively detecting ustilaginoidea virens from seed-borne and soil-borne media. The method comprises the steps of (1) pre-treating a detected sample, and quickly extracting DNA of the detected sample; (2) constructing a standard curve; and (3) performing reaction by using a fluorescent nucleic acid thermostatic amplification detection technology. The method can be used for quantitatively detecting the ustilaginoidea virens, overcomes the defects of the conventional first generation LAMP (loop-mediated isothermal amplification) detection method, and has the advantages of high sensitivity, high specificity, low pollution, stable reaction and the like. By adopting the strategy of sensitively detecting the reaction product by adopting a closed-tube detection technique, the method disclosed by the invention is unlikely to be affected by pollutants and can be used for detecting a soil sample on site and analyzing and judging the reaction product, so that the method is extremely simple and suitable for being widely popularized and applied.

The invention provides a kit for quantitatively detecting Ustilaginoidea virens. The kit comprises an upstream primer UvP292, a downstream primer UvP397, a fluorescence probe UvP333, 10*PCR buffer solution, 2mmol / L of dNTPs, 25mmol / L of Mg<2+> and TaqDNA polymerase, wherein eh sequence of the upstream primer UvP292 is 5'-GAACCAAGAGATCCGTTGTTGAA-3', and the sequence of the downstream primer UvP397 is 5'-GCCGGAGGATACAACCAAAA-3' The kit can highly sensitively detect Ustilaginoidea virens spores surrounding the rice field, especially spores in air surrounding the rice filed, and well facilitates the epidemiology research of the Ustilaginoidea virens.

The invention discloses atransferring and dispersing method for thin-wall conidia of ustilaginoidea virens. Spores are picked up from a spore-forming colony and are directly transferred and dispersed to an observation plate. The method comprises the following steps: 1) preparation of materials and appliances; 2) preparation of a spore source; 3) preparation of a spore observation plate; 4) picking of spores and diluting of dense spores; 5) transferring and releasing of the spores; and 6) culturing and observing of the spores. The method has the following advantages: 1) a key working material (thin-wall conidia) does not need to be prepared into a spore suspension, so that the pollution risk is reduced; and 2) the germination and growth of thin-wall conidia can be dispersed and observed on the long-strip-shaped culture plate, so that the efficiency and effect of observing spore germination activity are improved, and multi-factor action design and batch test comparison in spore germination biological research are facilitated.

The invention discloses a LAMP-specific primer composition used in a q-LAMP rapid detection method for rice smut spores. The LAMP-specific primer composition is composed of four primers: upstream and downstream outer primers and upstream and downstream inner primers. The present invention also discloses a loop-mediated isothermal amplification kit for the quantitative detection of rice smut spores, which comprises the above-mentioned LAMP-specific primer composition, and also contains betaine, 10×SYBR Green I, 8U / μL Bst DNA polymerase, etc. The invention also correspondingly discloses a loop-mediated isothermal amplification method for quantitatively detecting the spores of the rice false smut fungus.

The invention discloses a primer composition for LAMP (loop-mediated isothermal amplification) rapid detection of ustilaginoidea virens and application of the primer composition. The primer composition disclosed by the invention is prepared from the following primers: F3, B3, BIP, FIP, Loop F and Loop B. The primer composition is used for carrying out the LAMP rapid detection on the ustilaginoideavirens and has strong specificity and high sensitivity, so that the primer composition disclosed by the invention can be used for detecting and / or identifying the ustilaginoidea virens; a simple, rapid and low-cost detection technology is provided for scientific researches and production practice and theoretical foundation and technical guidance are also provided for early warning and reasonableapplication of ustilaginoidea oryzae; the primer composition has realistic and profound significances of improving ecological social and economic benefits.

The invention relates to an Ustilaginoidea virens (Cke.) Tak., especially an Ustilaginoidea virens (Cke.) Tak. producing five Ustiloxins and an acquisition method thereof. The Ustilaginoidea virens (Cke.) Tak. is obtained through separation and purification of a collection source. The Ustilaginoidea virens (Cke.) Tak. obtained by the method can produce five Ustiloxins. The Ustilaginoidea virens ZZY-2 (Ustilaginoidea virens ZZY-2) is sent to China Center for Type Culture Collection (CCTCC) in Wuhan University of Wuhan, Hubei province on April 26, 2016 for preservation, and the preservation number is CCTCC NO:M2016230.

The invention relates to the technical field of ustilaginoidea virens prevention and control, in particular to bacillus pumilus ZHQN-6 for preventing and controlling ustilaginoidea virens. According to the bacillus pumilus ZHQN-6 for preventing and controlling the ustilaginoidea virens, the preservation number is 16565. The bacillus pumilus has a good prevention and control effect on the ustilaginoidea virens, an experiment is carried out through a three-point confrontation culture method, and the result shows that the inhibition zone of the bacillus pumilus ZHQN-6 on the ustilaginoidea virensis 31.67 mm.

The invention discloses a pathogenic factor for negatively regulating ustilaginoidea virens spore production, a gene and an application thereof. An amino acid sequence is shown as SEQ ID No. 1, and the gene sequence is shown as SEQ ID No. 2. The researches find that the fungus pathogenic gene ustilaginoidea virens UvPSR1 derived from ustilaginoidea virens plays an important role in fungus vegetative growth, spore yield, infected hypha formation and pathopoiesis process, and the gene or protein encoded by the gene can be used as a target for designing and screening antifungal drugs.

The invention relates to a composite germicide composition for crops. The composite germicide composition comprises 10-30 parts of difenoconazole, and 70-90 parts of prochloraz-manganese chloride complex; the sum of parts by weight of the difenoconazole and the prochloraz-manganese chloride complex is 100 parts; besides, the composite germicide composition further comprises 2.0-6.0 parts of organic silicon synergistic agent. A composite germicide for crops prepared by use of the composite germicide composition comprises 71-85% of composite germicide composition, 8-20% of carrier and 7-9% of auxiliaries. The form of the composite germicide is wettable powder; the composite germicide is used for preventing and treating ustilaginoidea virens, rice blast, sheath blight and dendrobium candidum anthracnose. The germicide composition is high in efficacy, high in microbicidal efficacy and relatively low in harm to crops.

The invention provides a method for detecting a gene nucleotide mutation site of rice ustilaginoidea virens cyp51, belongs to the technical field of molecular biology and relates to a primer for detecting the gene nucleotide mutation site of the rice ustilaginoidea virens cyp51. Sequences of the primer are shown as SEQID No.1 and SEQID No.2. The method for detecting a tebuconazole resistance mutation site of rice ustilaginoidea virens has the advantages of being high in sensitivity, simple, rapid, good in stability and wide in applicability, can effectively distinguish sensitive strains and drug-resistant strains and is used for detecting tebuconazole resistance occurrence and development trend of the rice ustilaginoidea virens. The molecular detection method has the important significance on early warning of drug resistance of the rice ustilaginoidea virens, formulation of a reasonable disease management scheme and effective control of drug resistance development of diseases.

The invention discloses a preparation method of an ustilaginoidea virens (Cooke) Takahashi inhibitor prepared from potato-xylose. The preparation method comprises following steps: 1) preparation of a potato-xylose mixture; 2) high temperature treatment; 3) cooling; and 4) antibacterial effect examination of the high efficiency inhibitor, wherein a culture medium is prepared from the obtained high efficiency inhibitor, and is inoculated with ustilaginoidea virens (Cooke) Takahashi, and cultivation is carried out at an appropriate temperature. It is shown by the results of antibacterial effect examination that the high temperature treatment product of the potato-xylose mixture is capable of inhibiting growth of ustilaginoidea virens (Cooke) Takahashi with high efficiency. Advantages of the preparation method are that: the inhibitor capable of inhibiting growth of ustilaginoidea virens (Cooke) Takahashi with high efficiency can be prepared from the potato-xylose mixture via simple high temperature treatment.

The invention discloses an ustilaginoidea virens inhibitor derived from a potato-xylose mixture. The inhibitor is derived from a mixture of potato and a compound xylose, and is generated by high-temperature processing the potato-xylose mixture, and the inhibitor has a function of efficiently inhibiting ustilaginoidea virens growth and development. The ustilaginoidea virens inhibitor derived from the potato-xylose mixture has the beneficial effect of providing an effective and feasible technological beginning and technological means for developing ustilaginoidea virens prevention and cure drugs.

The invention discloses a culture medium suitable for growth of Ustilaginoidea virens. The main body of the culture medium comprises, by weight, 0.1 to 5% of cane sugar, 0.1 to 5% of peptone, 0.1 to 1% of gellan gum and 0.02 to 0.1% of Mg<2+>, with the balance being water. Trace amounts of inorganic salts consisting of potassium hydrogen diphosphate, sodium chloride, ferrous sulfate, sodium molybdate, manganese sulfate and calcium carbonate are added into the main body of the culture medium so as to obtain the culture medium suitable for growth of Ustilaginoidea virens.

The invention discloses ustilaginoidea virens elicitor protein SGP1, oligopeptide and application thereof. The elicitor protein and the elicitor oligopeptide can obviously activate plant defense response, improve plant disease resistance, enhance plant resistance to pathogenic bacteria infection and effectively prevent or relieve diseases. The elicitor protein and the elicitor oligopeptide provide a new way for improving the disease resistance of plants, provide resources for development and application of biopesticides in the future, can be used as a novel microbial protein pesticide for defending the invasion of diseases and pests, also provide a basis for transgenic plants with the gene in the future, and have a wide application prospect in agricultural production.

The invention discloses pseudomonas aeruginosa 9 # and application thereof, and belongs to the technical field of microorganisms. The invention discloses a pseudomonas aeruginosa strain 9 #, and the preservation number of the pseudomonas aeruginosa strain 9 # is CCTCC NO: M2021178. The pseudomonas aeruginosa strain 9 # disclosed by the invention can inhibit the growth of a Ustilaginoidea virens strain YY7850, which is an example that the pseudomonas aeruginosa strain is found to have antagonism to the Ustilaginoidea virens pathogenic bacteria for the first time, and due to the fact that the strain 9 # has the capacity of inhibiting the Ustilaginoidea virens pathogenic bacteria, the strain has the potential of being developed into a Ustilaginoidea virens biocontrol bacterium.

The invention discloses a xylose medium suitable for ustilaginoidea virens spore production and an application method thereof. A potato xylose medium is prepared from xylose after filter sterilization and serves as a spore producing medium for culturing thin-wall conidia of ustilaginoidea virens. The preparation and application method of the medium comprises the following steps: 1) preparing a basic medium; 2) performing filter sterilization of xylose; 3) preparing a xylose medium plate; 4) implanting ustilaginoidea virens; 5) performing constant-temperature culture; and 6) collecting new-generation spores. The xylose medium has the characteristics and advantages that the time for obtaining a large quantity of spores is short; the continuous spore producing time is long; the operation is simple and convenient, and shaking culture equipment is not needed; and the new-generation spores are relatively pure.

The invention relates to a molecular marker of a new anti-ustilaginoidea virens gene of rice, belonging to the field of rice breeding for disease resistance. According to the invention, each strain of Teqing background backcross introgression lines obtained by the cross and backcross between a susceptible parent Teqing and a disease-resistant parent Lemont is subjected to evaluation of disease resistance and molecular genetic linkage analysis and positioned to the anti-ustilaginoidea virens gene qFSR10.5 on the 10th chromosome of rice; and the molecular marker linked with the qFSR10.5 is RM258. By checking the RM258 banding pattern data and screening the ustilaginoidea virens resistance controlled by the qFSR10.5 resistance allele, the breeding efficiency of the anti-ustilaginoidea virens rice variety can be improved.

The invention discloses an efficient and poisonless agent fertilizer special for preventing and treating ustilaginoidea virens and a preparing method thereof. The agent fertilizer is prepared from, byweight, 30-50 parts of urea, 15-30 parts of monopotassium phosphate, 3-8 parts of microelement fertilizer, 8-15 parts of sodium humate, 3-8 parts of potassium fulvic acid, 2-6 parts of KOM, 0.5-2 parts of chitosan oligosaccharide, 1-5 parts of vinasse extracting solution, 6-12 parts of potassium hydroxide, 3-6 parts of citric acid, 6-10 parts of EDTA-disodium, 1-3 parts of sodium gluconate, 0.2-0.6 part of xanthan gum, 0.1-0.3 part of sodium carboxymethylcellulose and 800-1,000 parts of water. The efficient and poisonless agent fertilizer can still achieve the effect of preventing and treating ustilaginoidea virens while traditional pesticide components are completely avoided from being used, and also has the effects of adjusting the growth of rice and providing nutrients.