63 results about "Plasmodiophora brassicae" patented technology

This disclosure concerns a plant of the genus, Brassica, or parts thereof, which comprise one or more traits selected from the group consisting of high oleic acid content, low linolenic acid content, increased herbicide resistance, restorer of cytoplasmic male sterility, and increased clubroot disease (Plasmodiophora brassicae) resistance, compared to a wild-type plant of the same species. This disclosure further relates to wild-type and mutant alleles of genes involved in these traits, molecular markers linked thereto, and methods of their use.

The invention relates to a bacillus amyloliquefaciens for inhibiting bird rape plasmodiophora brassicae and separation, application and purposes of active substances thereof. Bacillus amyloliquefaciens HB-26 is preserved in China Center for Type Culture Collection (CCTCC), the preserving number is CCTCCNO: M2011259. A culture method is as follows: inoculating biogas slurry separated bacterial strain in an LB liquid nutrient medium, preserving the biogas slurry separated bacterial strain at 30 DEG C for one night, pouring fermentation liquor at the root of Chinese cabbage and finding the bacterial strains with bacteriostatic activity on plasmodiophora brassicae. The separated active substance is characterized in that the HB-26 is inoculated in the LB liquid nutrient medium, the fermentation liquor is dried by freezing, the frozen strain powder is added with ethyl acetate for extraction, is vibrated, rotated and evaporated, is dissolved by methyl alcohol, and then subjected to HPLC (High Performance Liquid Chromatrography) separation, the components are collected, the mass spectrum identification is conducted, and the component of the active substance can be judged according to the condition of the generation of plasmodiophora brassicae disease of the Chinese cabbage. The molecular weight of the bacteriostatic activity substance BA20 is 1494.0, and the nuclear magnetic resonance hydrogen spectrum shows that the chemical nature of the bacteriostatic activity substance is carbohydrate bacteriocin. The active substance is used for preparing a microbial agent for preventing the plasmodiophora brassicae disease of the Chinese cabbage.

Cyazofamid belongs to sulfanilamide imidazole bactericide and has high bioactivity on oomycete fungi such as phytophthora capsici, downy mildew, pseudo downy mildew and pythium and brassica plasmodiophora brassicae of plasmodiophoromycetes. A preparation method for cyazofamid aqueous emulsion has a formulation comprising 1 to 30 percent of the cyazofamid, 2 to 15 percent of emulsifying agent, 0.2 to 5 percent of co-emulsifying agent, 10 to 20 percent of solvent, 10 to 20 percent of latent solvent, 0.1 to 5 percent of anti-freezing agent, 2 to 10 percent of stabilizing agent, 0.1 to 2 percent of defoaming agent, 0.1 to 5 percent of thickening agent, 0.1 to 4 percent of pH value conditioner and the balance of deionized water, and comprises the following steps: adding the latent solvent into the deionized water according to a ratio; dispersing the latent solvent in a high-shear preparation kettle and stirring the solution evenly; adding cyazofamid raw powder and solvent as well as latent solvent mother solution into the solution; shearing the mixed solution at high speed; and finally adding the thickening agent into the mixed and stirring the resulting solution evenly to obtain the cyazofamid aqueous emulsion. The cyazofamid aqueous emulsion is a new environment-friendly water-based dosage form of agricultural bactericide, uses water as a dispersive medium to replace an expensive organic solvent, reduces environment and dust pollution, and is a new dosage form with development prospect.

The invention belongs to the technical field of agricultural sustainable development, relates to preparation of organic preparations and in particular relates to an organic preparation for controllingcruciferous vegetable club root and application thereof. The organic preparation is composed of organic raw materials and decomposition-promoting ingredients, wherein the organic materials include but not limited to one or more of crop straws, saw dust and other solid organic materials and molasses, alcoholic fermentation liquor and other liquid organic materials; the decomposition-promoting ingredients include but not limited to one or more of special microbes and organic catalysts; and a ratio (mass) of the organic raw materials to the decomposition-promoting ingredients is (95-100):(5-0).The invention further discloses a preparation method and application of the organic preparation. The organic preparation disclosed by the invention is ecologically environment-friendly and is applicable to soil with high incidence of club root after continuous cropping of various cruciferous vegetables. The quantity of plasmodiophora brassicae woron in soil and the incidence rate of the club rootof crops can be effectively reduced after treatment, the sterilization rate and the club root control rate respectively reach 60-90%, and the yield of the crops is obviously increased.

The invention discloses a method for evaluating resistance of brassica plants on club root, which belongs to the brassica plant disease resistance evaluation technical field, and is characterized in that: perlite after being soaked by acid water is mixed with crushed brassica plasmodiophora brassicae fungus root according to a ratio of 1L: 10 to 15g to form culture matrix, the culture matrix is filled inside grids of a frozen box, seeds of a specimen to be evaluated and infected specimen seeds infecting the club root are planted inside the grids of the frozen box, one seed is planted inside each grid, the frozen box is arranged inside a ceramic dish, and 1/2MS culture matrix and the crushed brassica plasmodiophora brassicae fungus root are used for being made into inoculant liquid with pH value of 5.5 and content of pathogenic bacteria spores of 1*108/ml; and the inoculant liquid is filled into the ceramic dish to be cultured inside an artificial weather box, and the resistance of the specimen to be evaluated on the club root is evaluated according to the investigated disease indexes. Due to the adoption of the method, mass evaluation of the resistance on the club root can be realized, a great amount of land can be saved, consistence of the evaluation conditions can be guaranteed, time and labor can be saved, the evaluation result is accurate and reliable, and simplicity and convenience in operation can be realized.

The invention discloses a germicide composition. The germicide composition comprises a synergic effective dose of cyprodinil and one of cyazofamid, cymoxanil, metalaxyl and hymexazol, wherein the weight part ratio of two active ingredients in the germicide composition is 40-1:1-40. The germicide composition at least comprises one surfactant which contributes to dispersing the active ingredients in water during application. The germicide composition has protection activity, treatment activity and eradicating activity, can be used for various crops and has extremely high bioactivity on oomycetes fungi such as phytophthora capsici, peronospora parasitica, pseudoperonospora parasitica, pythium and plasmodiophoromycetes plasmodiophora brassicae woron.

The invention discloses a specific PCR detection method for plasmodiophora brassicae in soil. The invention is characterized in that primers are designed as shown in SEQ ID No 1,2,3,5 according to 1-1028 sequences in a SEQ ID No 10 to form primer pairs 2,4,5 for performing the specific PCR detection. The method of the invention has the advantages of rapid detection, sensitive method, good specificity, wide application and simple operation, and can be taken as an effective technology means for detecting brassica plasmodiophora brassicae in soil and plant organization which has high practical value.

The invention provides an application method of bacillus amyloliquefaciens Bamm22 for preventing and treating plasmodiophora brassicae. The method comprises the following two methods: the method I comprises: carrying out root-irrigation treatment on a cruciferae crop by a bacillus amyloliquefaciens Bamm22 bacterial liquid with the amount of 80-100 ml per time per strain, and carrying out treatment for at least three times, wherein the treatment times are the first day, the seventh day and the fifteenth day after transplanting; the method II comprises: carrying out spraying treatment on the cruciferae crop by a bacillus amyloliquefaciens Bamm22 bacterial liquid with the amount of 150-200 ml per square meter, and carrying out treatment for at least three times, wherein the treatment times are the first day, the seventh day and the fourteenth day after seeding. The bacillus amyloliquefaciens Bamm22 provided by the invention is simple in using method, the application concentration of the bacterial liquid is less and the application dosage of the bacterial liquid is less.

The invention discloses a method for detecting plasmodiophora brassicae in Chinese cabbages. The method comprises the following steps of: making roots and leaves of the Chinese cabbages into tablets; acquiring infrared spectrograms of the tablets; and analyzing the infrared spectrograms, wherein if the tablets made from the roots have a characteristic peak in at least one part in the ranges of 1748.16+/-0.75cm<-1>, 2851.24+/-0.05cm<-1> and 3013.23+/-0.28cm<-1>, the roots of the Chinese cabbages are infected with the plasmodiophora brassicae; and if the tablets made from the leaves have a characteristic peak in at least one part in the ranges of 1252.54+/-0.50cm<-1> and 1051.98+/-4.23cm<-1>, the leaves of the Chinese cabbages are infected with the plasmodiophora brassicae. According to the method, the plasmodiophora brassicae in the Chinese cabbages can be rapidly and accurately detected in a lossless manner by virtue of a Fourier transform infrared spectroscopy technique, and the detection is convenient to implement, does not depend on skills of people and is high in security and repeatability.

The invention provides a fermentation method of bacillus amyloliquefaciens Bam22 for preventing and treating cruciferae plasmodiophora brassicae. The method comprises the following steps: (1) picking up a bacillus amyloliquefaciens Bam22 colony cultivated on a solid culture medium, and carrying out shake culture in a liquid culture medium at 30-32 DEG C for 10-12 hours, wherein the rotating speed is 200-220rpm; and the product is taken as a seed solution; (2) fermenting the seed solution obtained in the step (1) in a fermentation tank for 40-50 hours, wherein the access amount of the seed solution is 8%-10%; the temperature of the tank is 27-30 DEG C; the stirring speed of the fermentation tank is 220-250rpm; the air flow is 0.8:1 to 1:1 (V/V.m); the efficient bacillus amyloliquefaciens Bam22 for cruciferae plasmodiophora brassicae is preserved at the General Microbiology Center of the China Committee for Culture Collection of Microorganisms on September 6, 2015; and the preservation number is CGMCC No.11329. The fermentation method provided by the invention is very favorable to cultivation of the bacillus amyloliquefaciens Bam22; and multiple bacteria are obtained and have good activity.

The invention belongs to the technical field of rape breeding, and particularly relates to a breeding method for high oleic acid rape for resisting plasmodiophora brassicae and an application of the breeding method. The breeding method is characterized by comprising the following steps of enabling a high oleic acid plant and a plasmodiophora brassicae-resisting gene containing plant to be hybridized to obtain composite F1 rape seeds containing high oleic acid and CRb disease-resistant sites; planting the composite F1 rape seeds, identifying the relevant gene sites with high oleic acid and a plasmodiophora brassicae resisting molecular marker, and selecting individuals containing the sites and recurrent parents for hybridizing; performing high oleic acid and plasmodiophora brassicae resisting molecular marker analysis on backcross and selfed progeny groups, performing screening, and performing homozygosis to obtain the high oleic acid plasmodiophora brassicae resisting plant. The invention provides a simple, quick and effective method for breeding good disease-resistant high oleic acid rape kinds. The breeding method can selectively select a single plant according to the genotype atany stage in the breeding process, so that the selection efficiency is improved, and the breeding age limit is shortened.

The invention discloses an anti-plasmodiophoa-brassicae-woron inbred line breeding method which comprises the steps of seed soaking for germination hastening, vernalization, inoculation and identification, and inbred pollination. The method specifically comprises the following steps: in spring, carrying out carrying out seed soaking for germination hastening and vernalization on an F2-generation anti-plasmodiophora-brassicae breeding material, carrying out inoculationand identification by a bacterium soil process, permanently growing a disease-resistant individual plant into a flowerpot, and carrying out normal culture and management; after the plant bolts and blooms, carrying out inbred pollination, and collecting the seeds; and in next spring, continuing seed soaking for germination hastening, vernalization, inoculation and identification and inbred pollination, and after 3-5 generations of selection, breeding the anti-plasmodiophoa-brassicae Chinese cabbage inbred line with regular characters. The method can enhance the selection efficiency and accelerate the breeding progress.

The invention discloses streptomyces cuspidosporus and an application thereof in prevention and treatment of plasmodiophora brassicae. The preservation number of the streptomyces cuspidosporus WS-29246 is CCTCC (China Center For Type Culture Collection) NO: M2012413. The streptomyces cuspidosporus fermented generates a fermented liquid with a strong bacteriostatic action. The fermented liquid can be used for preparing a biocontrol suspension and a biocontrol slow release capsule for resisting plasmodiophora brassicae for biologically preventing and treating plasmodiophora brassicae.

The invention discloses a genetic transformation method of PEG/LiAc mediated plasmodiophora brassicae woronin. The genetic transformation method comprises the following specific steps: (a) preparing root secretion; (b) collecting hypnospore liquid; (c) preparing competent hypnospore liquid; (d) carrying out genetic transformation on plasmodiophora brassicae woronin; (e) carrying out resistance screening on bleomycin, repeatedly inoculating and screening for three generations, thus obtaining stable genetic transformation spores. The genetic transformation method disclosed by the invention realizes the genetic transformation of PEG/LiAc mediated plasmodiophora brassicae woronin and obtains stably genetic transformation strain capable of expressing green fluorescent proteins with a high intensity, and the transformation efficiency reaches 1000 transformants/mg of DNA (by DAN in a unit mass); in addition, a carrier used for the transformation in the genetic transformation method disclosed by the invention has enhanced green fluorescent protein genes and strong promoter RP27 in fungus, thus effectively increasing the fluorescent intensity of the transformation strain.

The invention relates to a carbon-based complex microbial inoculant and an application thereof. The carbon-based complex microbial inoculant comprises an aspergillus oryzae solid fermentation product,a zygosaccharomyces rouxii liquid fermentation product, a bacillus velezensis liquid fermentation product and biomass charcoal. According to the carbon-based complex microbial inoculant, the three fermentation products and the biomass charcoal are creatively compounded, the multiple components have a synergistic interaction effect, the carbon-based complex microbial inoculant is used in a soil remediation agent, and under the condition that no pesticide is introduced, the pH value of soil can be adjusted to be 7, so that the salt content of the soil is reduced, the content of organic mattersis increased, and the incidence rate of clubroot of green vegetables in the continuous cropping soil is reduced. The number of plasmodiophora brassicae woron in the continuous cropping soil treated with the soil remediation agent is remarkably reduced, the number of plasmodiophora brassicae woron in the root systems of the green vegetables in a harvesting period is remarkably reduced, and the diversity of microbial community species in the soil and the plant root systems is remarkably increased.

The present invention relates to the identification of clubroot resistance genes from Brassica. Clubroot resistant Brassicaceae plants are provided, as well as clubroot resistance genes and methods and means to increase clubroot resistance in Brassicaceae.

The invention relates to a bactericidal composition with synergy. The bactericidal composition comprises the following effective active ingredients of pyrimorph and cyazofamid, wherein the weight ratio of the pyrimorph to the cyazofamid is 1-80:60:1. By adding aids and excipients into the effective active ingredients to prepare wettable powder, water dispersible granules and suspending agents, the bactericidal composition has the advantage of high bioactivity on oomycetes fungi, such as phytophthora, downy mildews, pseudoperonospora, pythium and plasmodiophora brassicae in plasmodiophoramycetes.

This disclosure concerns a plant of the genus, Brassica, or parts thereof, which comprise one or more traits selected from the group consisting of high oleic acid content, low linolenic acid content, increased herbicide resistance, restorer of cytoplasmic male sterility, and increased clubroot disease (Plasmodiophora brassicae) resistance, compared to a wild-type plant of the same species. This disclosure further relates to wild-type and mutant alleles of genes involved in these traits, molecular markers linked thereto, and methods of their use.

The invention discloses a compound fertilizer special for pakchoi and application thereof. The compound fertilizer includes an organic fertilizer, an inorganic fertilizer and improved biochar in a weight ratio of 65-75:8-12:15-23, and the pH value of the compound fertilizer is 6-9, wherein the improved biochar is mainly prepared by: standing 2 parts of peanut shell charcoal and 1 part of corn straw charcoal in an LB medium of bacillus subtilis for 24-48h. The application amount of the compound fertilizer is 50-100kg/mu. The invention adopts a special compound fertilizer of appropriate dosage proportion directed at cadmium polluted pakchoi cultivated land having plasmodiophora brassicae. The compound fertilizer is not added with improved biochar, but also is introduced with bacillus subtilis having bacteriostatic effect, and is added with the organic fertilizer and inorganic fertilizer obtained by thorough decomposition of livestock and poultry excrement. Therefore, the special fertilizer not only can effectively reduce the accumulation of heavy metal cadmium in pakchoi, improve the in-situ remediation efficiency of heavy metal contaminated soil, promote the growth of pakchoi, but also can inhibit the growth ofplasmodiophora brassicae woron, and achieves the technical effect of inhibiting the plasmodiophora brassicae of pakchoi.

The invention provides a method for extracting plasmodiophora brassicae DNA of cruciferae vegetables and application of the method. A diatomite method is adopted to extract the DNA of a plasmodiophora brassicae sample of the cruciferae vegetables, the DNA of a plasmodiophora brassicae host is isolated, and a basis is laid for molecular studies of plasmodiophora brassicae. The method can be used for separating and detecting physiological strains of the plasmodiophora brassicae of the cruciferae vegetables, a specific molecular marker of the plasmodiophora brassicae is simultaneously used in a process of plant disease resistance identification, and the accuracy of specific disease resistance identification of the physiological strains can reach 100%. The method for extracting the plasmodiophora brassicae DNA of the cruciferae vegetables by using the diatomite method has good repeatability, high reliability and low detection cost and is time-saving and labor-saving.

The invention provides a quantitative detection method of plasmodiophora brassicae in different kinds of samples. The method comprises the steps of extracting DNA including plasmids, resting spores, germ carrying soil, crucifer germ carrying root tissue and seeds. ITS region gene fragments of the plasmodiophora brassicae in different samples are detected through a fluorescent quantitation PCR technique, and a regression equation between the number of the plasmodiophora brassicae in the samples and Ct value is established. The regression equations of different samples are fitted through statistics software, a universal model and an equation suitable for different samples are obtained, and quantitative detection for assessing the plasmodiophora brassicae in different samples through the universal model is realized. According to the quantitative detection method disclosed by the invention, a molecular biology technique is used, a mathematics and statistics analysis method is combined for use, the universal models for quantitative detection are fitted, the purpose of accurately assessing the quantity of the plasmodiophora brassicae in different samples is achieved, the quantitative detection method has the characteristics of saving test cost, shortening detection time and the like, and technical means and theory reference are provided for the respects of early diagnosis, epidemic early warning, prevention and control and the like of cruciferae club roots.

The invention relates to a method for inhibiting growth of plasmodiophora brassicae woronin, a method for preventing and treating club roots of cruciferae crops and application of azalomycin. Specifically, the invention provides the method for inhibiting the growth of the plasmodiophora brassicae woronin; the method comprises the steps of enabling the azalomycin to be in contact with the plasmodiophora brassicae woronin. The invention further provides the method for preventing and treating the club roots of the cruciferae crops and the method comprises the step of applying the azalomycin to the cruciferae crops. The invention further provides the application of the azalomycin to prevention and treatment of the club roots of the cruciferae crops. According to the method for inhibiting the growth of the plasmodiophora brassicae woronin, the method for preventing and treating the club roots of the cruciferae crops and the application of the azalomycin, the club roots of the cruciferae crops can be effectively prevented and treated; the azalomycin can be obtained through a microorganism manner so that the azalomycin has the advantage of safety to environment and the application prospect is wide.

The invention provides a plasmodiophora brassicae woronin DNA extraction method which comprises the following operation steps of (1) putting sterilized root nodules into an antibiotic mixed water solution to be subjected to soaking treatment for 20 to 30 hours, changing zoospores in the root nodules into resting spores, and flushing the treated root nodules by sterile water; (2) extracting plasmodiophoromycetes resting spores by using a 40-60-percent saccharose water solution by a suspension method; and (3) adding liquid nitrogen into the extracted plasmodiophoromycetes resting spores, performing sufficient grinding, then, putting the obtained material into a precooled centrifugal tube, adding a urea extracting solution, and after extraction, adding extracted plasmodiophora brassicae woronin DNA into a TE buffer solution for storage. The plasmodiophora brassicae woronin DNA extraction method provided by the invention has the advantages of short time consumption, low sample consumptionand low cost; the extracted plasmodiophora brassicae woronin DNA has high purity and few impurities; and important significance is realized for researching the genetic diversity of the plasmodiophorabrassicae woronin.