57 results about "Potato virus Y PVY" patented technology

The invention relates to a method for detecting tobacco viruses by a reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) technology, which comprises the steps of tobacco virus total ribonucleic acid (RNA) extraction, RT-LAMP reaction, electrophoresis detection and virus type determination, wherein tobacco viruses are selected from two or more than two kinds of viruses from cucumber mosaic viruses (CMV), potato viruses Y (PVY), tobacco etch viruses (TEV), tobacco mosaic viruses (TMV) and tobacco vein banding mosaic viruses (TVBMV). LAMP primers are designed according to a case protein gene conserved region of the tobacco viruses, and a fast, sensitive and high-specificity method is built for detecting various main tobacco viruses by the one-step RT-LAMP technology. A newmeasure is provided for the fast detection of the tobacco viruses.

The invention discloses a abrasive paper rubbing method of tobacco virus disease, including steps: (1) bruising leaf tissue with typical symptom of tobacco virus disease in mortar, adding water, filtering by gauze and extruding juice, then constant volume by distilled water to obtain inoculation liquor, compouded when need; (2) dividing a abrasive paper into equal portions and folding as arris; (3) dipping arris of the abrasive paper with inoculation liquor, sliding from one end to the other end of the arris on inoculation leaf, finishing inoculation. The method gets material easily, has simple, quick operation with little inaccuracy, only has three steps, success ratio of inoculation is 100%. The method suits for tobacco virus disease infected by machinery juice such as TMV, CMV, PVY, PVX and TEV and so on, especially for selecting and identifying fastness of tobacco species and experiment of prevention and cure tobacco virus disease.

The invention relates to the field of plant antiviral genetic engineering, and provides a plant antiviral new target. On the basis of tobacco vein banding mosaic virus infection cloning, photosystem II oxygen-evolving complex protein PsbO1 is screened through immunoprecipitation and mass spectrography, and a PbsO1 gene is obtained through reverse transcription PCR amplification. Immunoprecipitation and bimolecular fluorescence complementation prove that the PsbO1 can interact with the tobacco vein banding mosaic virus 6K2. Reduction in expression of the PsbO1 gene enables an infected plant to generate resistance to the tobacco vein banding mosaic virus and potato virus Y.

The invention discloses salt-tolerance bacillus E40207a2 and application thereof. The salt-tolerance bacillus E40207a2 is preserved in Guangdong Microbiological Culture Collection Center on January third, 2019, and the preservation number is GDMCC NO:60498. A fermentation solution of a secondary metabolite product of a strain has an obvious antagonistic function on viruses and diseases caused by Potatovirus Y (PVY), and the material is wide in source, low in cost, and safe to the environment.

The invention relates to clone, reconstruction and transformation of a precursor (pre-miR159a) of arabidopsis thaliana microRNA159a and antivirus analysis of a transgenic plant and belongs to the technical field of molecular biology and biology. The technology clones a nucleotide sequence of pre-miR159a from the arabidopsis thaliana and carries out site-directed mutagenesis on the sequence producing mature miR159a. 126kDa protein of tobacco mosaic virus, 25kDa protein of potato virus X and HC-Pro protein of potato virus Y are subjected to genetic mutation to obtain three sections of the mutation sequences. The three sections of the sequences are in series to construct the plant expression vector; and an agrobacterium-mediated method is adopted to transform nicotiana benthamiana to obtain transgenic tobacco. The transgenic plant has high disease resistance on three viruses. The antivirus strategy has the advantages of strong disease resistance, durable disease resistance, biological safety and the like and has wide application prospect in the breeding field of plant antivirus genetic engineering.

The invention discloses a preparation method of an anti-tobacco potato Y virus vaccine. The method includes the following steps that 1, pCaPVX440-eIF4E-6 plasmids are prepared; 2, the pCaPVX440-eIF4E-6 is transferred to agrobacterium to prepare the anti-tobacco potato Y virus vaccine. The anti-tobacco potato Y virus vaccine prepared by means of the preparation method is suitable for prevention and control of tobacco PVY viruses and has an important practical value for prevention and control of diseases caused by the PVY viruses in the tobacco leaf production process. Compared with the prior art, the virus vaccine is good in specificity, good in safety, simple, convenient and quick to operate and low in cost.

The invention discloses a method large-scale breeding non-toxic myzus persicae. A tobacco material resistant to TMV (Tobacco Mosaic Virus) and PVY (Potato Virus Y) is used as a breeding host of aphids. After seedling growing or potting or transplanting, the aphids are artificially bred in a net house or a greenhouse, and a double-resistant strain with a high aphid breeding efficiency is selected to be used for breeding non-toxic aphids. The method adopts the tobacco material resistant to the TMV and the PVY to breed the aphids and can effectively reduce the potential risk of the aphids carrying virus diseases under the condition that the current aphid parasitoid breeding process and the cost are not increased. The method has the characteristics of simplicity, convenience and easiness in operation, economy, reliability and the like and can promote popularization and application of aphid parasitoid control crop aphids.

The invention relates to a method for compositely detecting tobacco common mosaic virus and potato virus Y in one step, which is characterized by comprising the following steps of: screening and comparing conservative domains in virus genome, and artificially screening and synthesizing two composite RT(Reverse Transcription)-PCR (Polymerase Chain Reaction) specific primers suitable for viral diseases; analyzing to obtain TMV (Tobacco Mosaic Virus) and PVY (potato virus Y) composite tobacco-infected samples, and integrating enzymes and buffer solution required for reverse transcription and PCR; and then establishing the method for compositely detecting the TMV and the PVY in one step by reaction condition optimization. The method has the advantages that 1) the pathogens of the TMV and the PVY are more convenient to detect, test pollution and influencing factors in the reaction are reduced, and the detection time is greatly shortened; 2) the provided TMY and PVY specific primers have high viral pathogen specificity and can effectively and independently or compositely identify the TMV and the PVY, and ensure the accuracy of the detection result; and 3) when in use, the method saves time and labor, needs low requirement on sample materials, and has less limitation on experimental conditions.

The invention discloses a hybridoma cell strain secreting potato-virus-Y(PVY)-resistant monoclonal antibodies and monoclonal antibody application thereof. Pure PVY virion purified through a differential centrifugation method serves as antigens to immune BALB/c mice, cell fusion, screening and cloning are performed, one hybridoma cell strain 3B2 which is stable in passage and can secrete PVY-resistant monoclonal antibodies is obtained, and the preservation number of the hybridoma cell strain 3B2 is CGMCC No. 12001. The ascites indirect ELISA titer of the monoclonal antibodies secreted by the cell strain reaches 10<-7>, according to the antibody type and the subclass, the monoclonal antibodies are composed of IgG1 and kappa light chains, and the monoclonal antibodies and PVY coat protein of 30 kDa have a specific reaction. The 3B2 monoclonal antibodies are used for establishing ACP-ELISA, dot-ELISA and Tissue blot-ELISA detecting methods of PVY on crops, wherein the diseased leaf detecting sensitivity of the ACP-ELISA method and the diseased leaf detecting sensitivity of the dot-ELISA method reach 1:81920 and 1:10240 fold dilution (w/v, g/mL) respectively. Preparation of the PVY-resistant monoclonal antibodies and establishment of the detection methods of the PVY-resistant monoclonal antibodies provide matter and technologic support for detection and diagnosis, epidemiological analysis and scientific prevention and control of potato virus diseases.

The invention discloses a quadruple RT-PCR method simultaneously detecting multiple kinds of pepper viruses and an application of the method.The method comprises the steps that firstly, tender pepper blades with mosaic symptoms in a filed are selected, saved on ice and brought back to a laboratory, and RNA is extracted through a Trizol method; secondly, the RNA obtained in the first step serves as a template, 6-base Random Primers serve as reverse primers, and by means of inverse transcription, cDNA is obtained; thirdly, the cDNA obtained in the second step serves as a template, and PCR amplification is conducted on the tender pepper blades through four pairs of primers; fourthly, PCR products are subjected to gel electrophoresis, a gel-imaging system takes pictures, and the virus infection situation is judged according to the size of a target fragment and positive control.According to the detection method, one RT-PCR is used for simultaneously detecting main pepper viruses including a cucumber mosaic virus (CMV), a tobacco mosaic virus (TMV), a potato virus Y (PVY) and a broad bean wild virus-2 (BBWV-2).The kinds of detected virus are increased, the detection efficiency is improved, and the detection cost is low.

The invention respectively designs five pairs of primers for detecting potato viruses according to conserved regions of coat protein sequences of five types of viruses comprising a potato virus X, a potato virus M, a potato virus Y, a potato virus A and a potato virus S, and simultaneously discloses a method for detecting the potato viruses by utilizing the five pairs of primers. Specifically, the five types of viruses in potato leaves are detected in an amplification manner by virtue of the five pairs of specific primers via a multiplex RT-PCR method, and the verification is carried out by cloning, sequencing and comparing amplified products, so that the detection method capable of simultaneously and reliably detecting the five types of potato viruses via an one-step reaction is established. The method has the characteristics of high detection efficiency, high sensitivity and low cost.

The present invention relates to two-step synchronous double RT-PCR process of detecting Y virus and leaf roll virus of potato, synchronous triple RT-PCR process of detecting X virus, A virus and S virus of potato, and the detecting kit. The present invention can detect five main potato viruses within 4 hr in high sensitivity, high specificity, high stability and high reliability. The present invention is suitable for the early diagnosis on potato seedling and potato seeds, and is significant on the quality monitoring of potato seed and virus disease prevention.

The invention discloses a preparation method of test strip for rapid field detection of tobacco potato virus Y, which comprises the following steps of: preparing an antibody, preparing colloidal gold, preparing a colloidal gold-antibody conjugate, preparing a nitrocellulose filter, and preparing a colloidal gold chromatographic test strip. The prepared test strip for rapid field detection of tobacco potato virus Y according to the technical proposal has an optimum detection temperature of 15-35 DEG C, sample detection amount of 0.1-0.3g and tobacco sample area of 3-5cm<2>. The detection method comprises charging tobacco leaf in a plastic bag containing buffer, grinding, immersing the test strip in the buffer, reacting for 30min, and observing the result. The false positive rate is less than 1%. The invention has the advantages of convenient application and high reliability; and is especially suitable for rapid field detection.

The invention discloses a dithioacetal derivative containing methoxyacrylate, a preparation method and an application thereof. The general formula is as shown in (I), wherein, Y is N or CH; R1 is a substituted halogen atom, a methyl, a methoxy, a ethoxy or a 3, 5 -dimethoxy, the halogen atom may be fluorine, chlorine, bromine; R2 is a substituted aromatic ring, a heterocyclic ring, a 1- propyl alcohol, a ethyl, a propyl, a isopropyl. The corresponding position of the aromatic ring contains a fluorine or chlorine, and the heterocyclic ring is a morphinane ring. The derivative has better activity on plant viruses such as potato Y virus, cucumber mosaic virus, tobacco mosaic virus, tomato chlorosis virus and the like, and has simple structure, simple preparation process and low production cost.

The invention discloses a method for potato Y virus, potato X virus, potato A virus and potato leaf roll virus in Yunnan potato main culture species Li potato No.6, Yun potato 401 and Xuan potato No.2 infected plants. The specific mixed antibody is utilized to entrap the potato Y virus, potato X virus, potato A virus and potato leaf roll virus in Yunnan potato main culture species Li potato No.6, Yun potato 401 and Xuan potato No.2 infected plants, so that the total RNA (ribonucleic acid) of the sample does not need to be extracted to perform random primer reverse transcription and specific virus primer compound PCR (polymerase chain reaction) reaction. The method combines quick specific immune precipitation and sensitive compound RT-PCR (reverse transcription-polymerase chain reaction), sets healthy potato random primer cDNA (complementary deoxyribonucleic acid) product and corresponding virus PCR specific segment positive plasmid as negative and positive controls, and thus, is quickly, accurately and sensitively used in detecting field potato plant samples, antiviral materials, field environment suspect infection sources and the like.

The invention relates to the field of virus detection, in particular to a kit for rapidly detecting potato viruses Y (PVY for short) and a detection method thereof. The rapid detection kit comprises a sampling tube, a rinsing tube, a nucleic acid denaturing tube, an amplifying and detecting tube, a negative control tube, a positive control tube, a rapid drying solution as well as a FTA (Fault Tree Analysis) membrane, a grinding bar, toothpicks, a suction tube and the like which are respectively packaged in a sterile bag. The formed detection method simplifies operation steps, reduces experimental cost and has stronger practicality in fields or rooms.

The invention discloses a method for detecting potato virus Y by using digital PCR and a special complete set of reagents thereof. The complete set of reagents is prepared from a primer pair X1 and a probe PVY-p, wherein the primer X1 consists of a primer PVY-f and a primer PVY-r; the primer PVY-f is a single-chain DNA molecule expressed by a sequence 1 in a sequence table; the primer PVY-r is a single-chain DNA molecule expressed by a sequence 2 in the sequence table; the sequence of the probe PVY-p is a sequence 3 in the sequence table; an FAM fluorescent mark is arranged at a 5'end of the probe PVY-p; a TAMRA fluorescent mark is arranged at a 3'end of the probe PVY-p. Experiments show that the method for detecting the potato virus Y by using the digital PCR is high in specificity and high in sensitivity, is applicable to detection of latent diseases and the PVY in samples with low virus content, and has an important application value.

The invention belongs to the field of pesticides, and provides a bactericidal composition containing zhongshengmycin. The main active ingredients of the bactericidal composition containing zhongshengmycin are chloroindole hydrazide and zhongshengmycin, wherein the weight ratio of chloroindole hydrazide to zhongshengmycin is 0.5: 5-40: 1, the content of chloroindole hydrazide in the bactericidal composition is 0.5 to 40%; the content of Zhongshengmycin in the bactericidal composition is 0.5 to 10%; the bactericidal composition can be used for prevention and treatment of various virus diseases of vegetables, melons, fruits, crops and the like, particularly, common cucumber bacterial angular leaf spot, Chineses cabbage soft rot, potato virus Y, camellia flower leaf virus, and peanut stunt virus, and possesses effect on Toamtos Yellows Leaf Curl Virus, rice black-streaked dwarf disease, and the like which are difficult to prevent and treat, possesses both prevention and treatment effect, is capable of preventing and controlling infection and development of plant virus diseases; and using of the bactericidal composition before or after disease infection is capable of achieving obvious prevention and treatment effect, and reducing pesticide using amount.

The invention discloses bacillus subtilis JQ2.11-12 and application thereof. The bacillus subtilis JQ2.11-12 is preserved in the Guangdong Microbial Culture Collection Center on June 17 th, 2019, andthe preservation number is GDMCC No:6096. The strain is separated from distilled grains of highland barley baijiu, and it is proved through experiments that a fermentation product of the strain can effectively inhibit potato dry rot pathogens and potato viruses Y, has a potential and application prospect as a biocontrol bacterium preparation for preventing and controlling the potato dry rot and potato viruses Y.

The invention relates to a method of breeding a tobacco plant having broad-spectrum resistance to potyviruses. The method comprises the following steps: polymerizing an eIFiso4E-S gene mutant, an eIFiso4E-T gene mutant and an eIF14E1 gene mutant in the tobacco plant to achieve the resistance of the tobacco to four potyviruses comprising a potato virus Y (PVY), a chilli veinal mottle virus (ChiVMV)and a tobacco vein banding mosaic virus (TVBMV). The method has great application prospect for cultivating the anti-Potyviruses tobacco.

The invention discloses a detection kit for new potyviridae virus in nelumbo nucifera. The detection kit comprises two specific primers, and also comprises M-MLV reverse transcriptase, an M-MLV enzyme reaction buffering solution, dNTPs, Taq, DNA polymerase, Mg<2+> and a PCR reaction buffering solution. The invention also provides a detecting method for the new potyviridae virus in the nelumbo nucifera. The detecting method comprises the following steps: (1) extracting total RNA of a sample to be detected; (2) designing and synthesizing primers; (3) performing RT-PCR; (4) performing electrophoresis detection. The invention provides the detection kit and the detecting method for the new potyviridae virus in the nelumbo nucifera; the new potyviridae virus in the nelumbo nucifera is detected by adopting an RT-PCR method; cumbersome steps for separating nucleic acid of a virus of the nelumbo nucifera are avoided; time is shortened; the operability, the flexibility and the repeatability of the detection are greatly improved; an effective method is provided for the prevention and the treatment of the new potyviridae virus and the detection of a virus-free seedling.

The invention discloses a method for rapidly and sensitively detecting a tobacco potato virus Y. A tobacco extracting solution is used as a reaction template, a polyvinylidene fluoride film is used as a reaction film, and an TR-PCR is combined to detect the potato virus Y, a tobacco mosaic virus and a cucumber mosaic virus. By adopting the method, the potato virus Y, the tobacco mosaic virus and the cucumber mosaic virus can be accurately and effectively detected, the operation is simple and convenient, the detecting sensitivity is improved, the detection time is shortened, and the detection cost is saved.