44 results about "Resting spore" patented technology

The invention discloses bacillus velezensis ZF481 and application thereof. The strain number of the bacillus velezensis is ZF481, the bacillus velezensis is preserved in the China General Microbiological Culture Collection Center on July 8, 2020, and the registration number of the bacillus velezensis is CGMCC No.20320. The bacillus velezensis ZF481 or / and the metabolite of the bacillus velezensis ZF481 can be used for preventing and / or treating the plant clubroot, or preparing a product for preventing and / or treating the plant clubroot. The strain has an inhibiting effect on the activity and germination of plasmodiophora brassicae resting spores, has a good growth promoting effect on brassicaceous crops such as Chinese cabbage and the like, and also has a good antibacterial effect on phytophthora capsici, stemphylium solani, fusarium oxysporum, alternaria solani, corynespora, Botryis cinerea, rhizoctonia solani and the like.

The invention discloses a rapeseed endogenous trichoderma atroviride ReTv2 strain and a preparation method and application thereof. The trichoderma atroviride ReTv2 strain is separated from a rapeseed root tissue by the applicant, and the number CCTCC NO is M2014407. The ReTv2 strain is activated and cultured in a PDA culture medium at the temperature of 28 DEG C for 2-4d to obtain conidiospores, and the spores are inoculated into a PDB culture medium for enlarged culture for 6d, wherein the content of the conidiospores can achieve 1.0*10<9> / mL. The trichoderma atroviride ReTv2 strain disclosed by the invention is the rapeseed endogenous strain, can be successfully colonized at the root of rapeseed and has an effect on promoting growth of the rapeseed, a fermentation broth has an obvious effect on inhibiting germination of resting spores of pathogenic bacteria of rapeseed clubroot, and the control effect of the strain against the rapeseed clubroot achieves 66.4%; furthermore, the ReTv2 strain can be directly applied during seeding, is simple in an application method, can save labor force and further has the property of being safe to people and animals of biological pesticides.

The invention discloses a Bacillus velezensis F85 for preventing and treating clubroot disease. The Bacillus velezensis F85 is preserved in China Center for Type Culture Collection, and the preservation number is CCTCC NO: M2019105. The strain has inhibitory effect on both the vitality and germination of resting spores of plasmodiophoromycetes, and can significantly inhibit the infection of plasmodiophoromycetes to rape root hairs. Pot experiments show that root irrigation treatment with the Bacillus velezensis F85 fermentation broth can effectively prevent and treat the clubroot disease of rapes, the comprehensive preventing effect reaches 83.4%, the treatment is at a higher level at present, and the strain also has the advantages of stable effect, wide environmental adaptability and etc.

The invention relates to the technical field of biology, in particular to a downy mildew bacterium culture and observation method. The method comprises the following steps: collecting downy mildew bacteria; preparing sporangium suspension liquid; and culturing the sporangium suspensions. Through repeated tests, a technology capable of inducing sporangium of cucumber downy mildew to germinate andrelease zoospore is obtained, the zoospore forms static spores through movement and morphological changes, the static spores germinate to form appressorium, and the appressorium germinates to generateinfection filaments. The technology can provide an effective technique for researchers to deeply research downy mildew biological basis, disease resistance identification, bactericide screening and the like. By means of the method, researchers can achieve quantitative statistics and monitoring of the number of zoospore of downy mildew, morphological changes and the number of formed infectors, andresearch of the action mechanism of the bactericide can also be achieved. Several development stages which must be experienced before downy mildew bacteria invade a host are clearly observed. The systematic observation method can provide first-hand data for prevention and treatment of the pathogenic bacteria on symptomatic drugs.

The invention discloses reference genes in growth, development and infection stages of peronophythora litchi, and primers and application thereof. Four peronophythora litchi genes PlACTIN, PLGAPDH, PlrpS5 and Pl006400 are selected, the expression quantity changes of the genes in the germination and infection stages of peronophythora litchi hyphae, sporangia, zoospores, resting spores and resting spores are measured by a real-time fluorescence quantitative PCR method, the stability of the expression quantity of the selected genes is compared, and the result shows that the Pl006400 and PlACTIN can be used as the reference genes with stable expression quantity. The specific primer primers of the reference genes Pl006400 and PlACTIN can be widely applied to the gene expression study in the development and infection stages of peronophythora litchi, the blank of lack of reliable reference genes of peronophythora litchi is filled, and important application values are realized.

The invention provides a resting spore inoculated biological organic fertilizer. The resting spore inoculated biological organic fertilizer comprises the following components in parts by weight: 300-600 parts of an organic matrix, 30-200 parts of diatomite, 200-500 parts of ammonium sulphate, 30-200 parts of phosphorite powder, 30-250 parts of potassium feldspar powder and 0.5-5 parts of fungal spore powder agents. The organic fertilizers disclosed by the invention are rich in nutrient elements and have the advantages of preserving moisture and fertility, improving soil, being low in cost andthe like, the yield and the quality of crops are greatly increased, and soil acidification, soil alkalization and soil salinization are improved.

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 discloses a method of enabling exogenous DNA to penetrate cell barriers to enter penicillium notatum resting spores. The method includes three steps: penicillium notatum culturing and spore collection, penicillium notatum spore pretreatment and using of a HDEN method to electrically shock penicillium notatum spores to obtain the penicillium notatum spores with to-be transformed plasmids guided in. Ungerminated spores are used as a starting material for guiding in exogenous molecules, and HDEN electric transformation technology is applied to guide exogenous DNA into the penicillium notatum resting spores, so that a complex step of spore germination can be omitted, steps of preparing protoplast or agrobacterium-mediated transformation in conventional methods are omitted, transformation rate is high, and each transformation reaction system at least can realize effect of not less than 6000 positive transformants.

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.

The invention relates to the fields of microorganisms and phytopathology, and particularly discloses a method for counting plant pathogenic oomycetes zoospores. According to the invention, the cameraphase parameter is set, so that the two morphological characteristics of the zoospores, namely blue spores and orange spores, can be clearly observed, thereby realizing the counting of the zoospores.According to the method, the zoospores and the resting spores can be counted simultaneously, so that the related experiments of the oomycetes zoospores are facilitated, and the accuracy and the precision of experimental results are improved. The method disclosed by the invention is simple and reliable, and all microscopes with imaging equipment can be suitable for counting oomycetes zoospores, sothat an evaluation index is provided for evaluating the severity of plant diseases and the pesticide effect of the bactericide.

The invention provides a method for separating plasmodiophora brassicae monospore on the basis of a methylene blue agarose method. The method comprises the following steps: firstly, preparing a plasmodiophora brassicae resting spore suspension with the concentration of 5*10<3> spore.mL<-1>; adding 0.01% methylene blue staining solution to a 1.3% agarose solution to prepare a mehtylene blue agaroseglass slide coagulation layer; uniformly coating the surface of the coagulation layer with 2 mu L of plasmodiophora brassicae resting spore suspension, performing observation under a microscope to determine that only one spore exists in the visual field, separating the coagulation layer containing the monospore, placing the coagulation layer at root hair of cruciferae seedlings upside down, and performing cultivation to obtain a plasmodiophora brassicae monospore system. The plasmodiophora brassicae monospore is separated by the methylene blue agarose method, the background is slightly blue,no impurities or bubbles are produced, and the plasmodiophora brassicae monospore can be clearly distinguished. The method is simple in process and easy to operate, and the separation efficiency and accuracy of the plasmodiophora brassicae monospore are greatly improved.

The present invention discloses A METHOD TO PRODUCE PROTEIN IN ASPERGILLUS NIGER'S SLEEPING SPORES USING single stranded RNA. The method includes three steps of culture of Aspergillus niger and collection of spores, pretreatment of Aspergillus niger spores, and electroporation of Aspergillus niger spores by using HDEN method. In the present invention, non-germinated spores are used as a starting material for introduction of exogenous molecules. The exogenous protein coding single stranded RNA is introduced into the resting spores of Aspergillus niger by employing the HDEN electrotransformation technique to express protein. The method of this invention is simple and fast, the effect is excellent, and the transformation rate reaches more than 90%.

The present invention discloses a method for direct transformation of exogenous DNA into resting spores of Penicillium amagasakiense. The method includes three steps of culture of Penicillium amagasakiense and collection of spores, pretreatment of Penicillium amagasakiense spores, and electroporation of Penicillium amagasakiense spores by using HDEN method, to obtain Penicillium amagasakiense spores with introduction of plasmids to be transformed. In the present invention, non-germinated spores are used as a starting material for introduction of an exogenous molecule, and exogenous DNA is introduced into the resting spores of Penicillium amagasakiense by employing the HDEN electrotransformation technique, whereby the complex step of spore germination is omitted, and steps of protoplast preparation or Agrobacterium-mediated transformation in conventional methods etc. are omitted. Moreover, the transformation efficiency is high, and at least an effect of no less than 6000 positive transformants per transformation reaction system can be achieved.

The present invention provides a Bacillus velezensis strain HMQAU19044, which was deposited in the General Microbiology Center of the China Committee for the Collection of Microbial Cultures on January 19, 2020, with a deposit registration number of CGMCC No.19420. Bacillus velezensis (Bacillus velezensis) strain HMQAU19044 provided by the present invention has significant control effect to cucumber downy mildew, and the research of embodiment shows, bacterial strain HMQAU19044 fermentation filtrate has obvious inhibition to the release of cucumber downy mildew zoospore Effect; the fermentation liquid of strain HMQAU19044 has obvious inhibitory effect on the release of zoospores of cucumber downy mildew; To a certain extent, the lesion expansion of the infected cucumber leaves is inhibited; the fermentation filtrate and fermentation liquid of the strain HMQAU19044 of the present invention have a good control effect on cucumber downy mildew.

The invention discloses a method for expressing protein in the resting spores of rhizopus stolonifer. The method comprises the following three steps: rhizopus stolonifer culture and spore collection, pretreatment of rhizopus stolonifer spores and electric shock of rhizopus stolonifer spores by using a HDEN method. According to the method, the step of fungal spore germination is avoided, the HDEN electrotransformation technology is adopted, the extracorporeal single-chain coding protein RNA is delivered to penetrate through the cell wall and cell membrane, and the protein is expressed in the resting spores of rhizopus stolonifer. The method disclosed by the invention has simple and quick steps and realizes a perfect effect while the transformation rate exceeds 90%.

The invention discloses a method for expressing protein in geotrichum candidum resting spores. The method comprises the following three steps: geotrichum candidum culture and spore collection, geotrichum candidum spore pretreatment and electric shock of geotrichum candidum spores using a HDEN process. According to the method, without a step of fungal spore germination and by adopting the HDEN electrotransformation technology, the in-vitro single-stranded coded protein RNA is delivered to pass the cell wall and cell membrane, and the protein is expressed in the geotrichum candidum resting spores. The method disclosed by the invention has the advantages of simple and fast steps and perfect effect while the transformation rate reaches 90% or above.

The invention provides a Streptomyces griseochromogenes YN-6 separated from germ-carrying soil of cruciferae crops infected with club root, and the CCTCC No is M2013187. The strain YN-6 has good control effect on the club root of the cruciferae crops through verification of club root germ resting spore germination inhabitation and greenhouse pot experiments. The strain is high in adaptability and colonization capacity under field natural conditions, not prone to antagonistic reaction with complicated microorganism species in field soil, the disease control effect is stable, defects of chemical control, disease-resistant variety and agricultural control methods are effectively overcome, and the strain is an ideal anti-biological inoculant applicable to field club root control.

The invention discloses a method for directly transforming exogenous DNA to enter aspergillus nidulans resting spores through non-medium dependency. The method comprises three steps: aspergillus nidulans culture and spore collection, aspergillus nidulans spore pretreatment and electric shock of aspergillus nidulans spores using a HDEN process to obtain aspergillus nidulans spores with introduced to-be-transformed plasmid. In the invention, the non-terminated spores are used as a start material for introducing exogenous molecules, the exogenous DNA is introduced into the aspergillus nidulans resting spores by the HDEN electrotransformation technology, and a complicated step of spore germination can be saved; the steps such as preparation of protoplast and agrobacterium-mediated transformation in traditional method are saved, and the transformation rate is high; and in each transformation reaction system, an effect of at least 6000 positive transformants can be realized.

The invention discloses an internal reference gene in the growth and development and infection stages of Pythophthora litchi, primers and application thereof. The present invention chooses 4 litchi downy mildew genes PlACTIN, PLGAPDH, PlrpS5 and P1006400, has measured these genes in litchi downy mildew hyphae, sporangia, zoospores, restenia, restenia germination and The expression levels of the infection stages were changed, and the stability of the expression levels of the selected genes was compared, and it was found that the expression levels of Pl006400 and PlACTIN were stable and could be used as internal reference genes. The specific primers of the internal reference genes P1006400 and PlACTIN of the present invention can be widely used in the gene expression research of the development of Lychee Pyrosophthora and its infecting litchi stages, and fill the gap that Lychee Pyramids lacks reliable internal reference genes, and has important Value.

The invention discloses a method for medium-independent direct transforming of exogenous DNA into Rhizopus oryzae resting spore, wherein the method comprises three steps such as Rhizopus oryzae culturing and spore collecting, Rhizopus oryzae spore pre-treating, and electric shocking of the Rhizopus oryzae spore with a HDEN method so as to obtain the Rhizopus oryzae spore introduced into plasmid to be transformed. According to the present invention, the non-germinating spore is adopted as the starting material for introducing exogenous molecules and the exogenous DNA is introduced into the Rhizopus oryzae resting spore by using the HDEN electrotransformation technology, such that the complex step spore germination can be eliminated, the steps such as protoplast preparation or Agrobacterium transformation, and the like in the traditional method can be eliminated, and the transformation rate is high, wherein at least not less than 6000 positive transformants can be achieved in each transformation reaction system.

The invention discloses a method for directly delivering exogenous DNA into monascus resting spores. The method includes three steps: performing monascus culture and spore collection; pre-treating monascus spores; using a HDEN method to apply electric shock to the monascus sports to obtain monascus spores with to-be-converted plasmids guided in. Non-germinal spores are used as a starting material for guiding in exogenous molecules, and HDEN electrotransformation technology is applied to guide the exogenous DNA into the monascus resting spores, so that a complex step of spore germination can be omitted, and steps of protoplast preparation or argrobacterium-mediated transformation in conventional methods are omitted; transformation rate is high, and each transformation reaction system can realize effect of at least 6000 positive transformants at least.

The present invention discloses A METHOD TO PRODUCE PROTEIN IN PENICILLIUM AMAGASAKIENSE'S SLEEPING SPORES BY TRANSFORMATION OF SSRNA. The method includes three steps of culture of Penicillium amagasakiense and collection of spores, pretreatment of Penicillium anagasakiense spores, and electroporation of Penicillium anagasakiense spores by using HDEN method. In the present invention, non-germinated spores are used as a starting material for introduction of exogenous molecules. The exogenous protein coding single stranded RNA is introduced into the resting spores of Penicillium amagasakiense by employing the HDEN electrotransformation technique to express protein. The method of this invention is simple and fast, the effect is excellent, and the transformation rate reaches more than 90%.

A method to produce protein in Aspergillus niger's sleeping spores using single-stranded RNA is provided. The method includes three steps: culture of Aspergillus niger and collection of spores, pretreatment of Aspergillus niger spores, and electroporation of Aspergillus niger spores using HDEN method. Non-germinated spores are used as a starting material for introduction of exogenous molecules. The exogenous protein coding single-stranded RNA is introduced into the resting spores of Aspergillus niger by employing the HDEN electrotransformation technique to express protein. This method is simple and fast, the effect is excellent, and the transformation rate reaches more than 90%.

The present invention discloses a method for direct transformation of exogenous DNA into resting spores of Penicillium amagasakiense. The method includes three steps of culture of Penicillium amagasakiense and collection of spores, pretreatment of Penicillium amagasakiense spores, and electroporation of Penicillium amagasakiense spores by using HDEN method, to obtain Penicillium amagasakiense spores with introduction of plasmids to be transformed. In the present invention, non-germinated spores are used as a starting material for introduction of an exogenous molecule, and exogenous DNA is introduced into the resting spores of Penicillium amagasakiense by employing the HDEN electrotransformation technique, whereby the complex step of spore germination is omitted, and steps of protoplast preparation or Agrobacterium-mediated transformation in conventional methods etc. are omitted. Moreover, the transformation efficiency is high, and at least an effect of no less than 6000 positive transformants per transformation reaction system can be achieved.

The invention discloses a method for directly introducing exogenous deoxyribonucleic acid (DNA) into resting spores of Pyricularia oryzae. The method comprises the three steps, i.e., carrying out Pyricularia oryzae culture and spore collection, pretreating Pyricularia oryzae spores and carrying out electroshock on the Pyricularia oryzae spores by using an HDEN (High-density Distributed Electrode Network) method, thereby obtaining the Pyricularia oryzae spores which are used for being introduced into plasmid to be transformed. According to the method, un-germinated spores serve as a starting material for introducing exogenous molecules, and the exogenous DNA is introduced into the resting spores of the Pyricularia oryzae by using an HDEN electrotransformation technology, so that a complicated step, i.e., carrying out spore germination can be avoided, and a step of preparing protoplast or transforming Agrobacterium tumefaciens and the like in the traditional methods can be avoided; the transformation ratio is high, and each transformation reaction system at least can achieve the effect of no less than 6,000 positive transformants.

The invention discloses a method for allowing transformed-external-source desoxyribonucleic acid to enter rhizopus stolonifer resting spores. The method includes the steps that rhizopus stolonifer is cultivated, and spores are collected; the rhizopus stolonifer spores are pretreated; the rhizopus stolonifer spores are subjected to electric shock with the HDEN method, and the rhizopus stolonifer spores with led-in to-be-transformed plasmids are obtained. According to the method, the non-germinated spores serve as initiative materials for leading in external source molecules, external source DNA is led into the rhizopus stolonifer resting spores with the HDEN electrotransformation technology, the complex spore germination step can be omitted, the protoplast preparing step or the agrobacterium-mediated transformation step and the like in the traditional method is omitted, the transformation rate is high, and the effect that 6,000 positive transformants or more exist in each transformation reaction system is achieved.

The invention discloses a method for directly importing foreign DNA into fusarium moniliforme resting spores. The method comprises the following three steps: performing fusarium moniliforme culture and spore collection, pretreating fusarium moniliforme spores and performing electric shock on the fusarium moniliforme spores by using a high-density distributed electrode network (HDEN) method, so that the fusarium moniliforme spores imported with to-be-transformed plasmids are obtained. Non-germinated spores serve as starting materials of importing foreign molecules, and the foreign DNA is imported into the fusarium moniliforme resting spores by employing the HDEN electroporation technology, so that a complicated step of germinating the spores can be saved, and the steps of preparing protoplast or performing agrobacterium-mediated transformation and the like in the traditional method are saved. Moreover, the transformation rate is high, and an effect of obtaining not less than 6000 positive transformants can be achieved at least in each transformation reaction system.

The invention relates to a water quality regulator for fancy carp culture and a preparation method thereof. The water quality regulation mechanism of the quick lime is as follows: hydroxyl ions dissociated from calcium hydroxide generated by the action of the quick lime and water can enable a water body to be strongly alkaline, and harmful species, moss and other aquatic plants and partial microorganisms, parasites and other pathogens and resting spores remaining in a pond can be killed. The selected chitosan, sodium alginate and Na-montmorillonite have certain adsorption and flocculation effects and can adsorb, flocculate and precipitate various impurities in the water body and can improve the water quality, and the chitosan and sodium alginate are biodegradable and cannot cause secondarypollution to the water body, so that the environment is protected. Besides, the modified montmorillonite and the tourmaline ceramsite are matched with each other, so that negative ions can be released, trace elements can be supplemented, water can be electrolyzed, water molecular groups can be reduced, impure substances in water can be removed conveniently, and the water purification performanceof the water purifier is further improved.

The invention relates to a new application of methyl propyl trisulfide, a sulfide compound, for inhibiting Phytophthora spp., and the compound can effectively inhibit Phytophthora capsici. The compound of the present invention has remarkable effects on inhibiting the release of zoospores of Phytophthora, swimming of zoospores, germination of restospores, growth of hyphae and promoting the cracking of zoospores. The compound of the invention can be used for the prevention and control of Phytophthora in soil, preventing and inhibiting the infection of Phytophthora to plants, especially crops from the source. The compound described in the invention is a biological source compound, which can reduce the generation of Phytophthora drug resistance and reduce the environmental pollution caused by pesticides.