66 results about "Potato virus X" patented technology

The invention relates to a ferulic acid shown in a general formula (I) and the application of derivatives thereof to pesticides. When used as novel anti-phytoviral agents, the derivatives can well inhibit tobacco mosaic viruses, pepper viruses, tomato viruses, sweet potato viruses, potato viruses, melon viruses, maize dwarf mosaic viruses and the like, and effectively prevent and control the virus diseases of various crops such as tobacco, peppers, tomatoes, melons, grains, vegetables, beans and the like, and are particularly suitable for preventing and controlling tobacco mosaic. Meanings of each group are shown in specifications.

The 19 monoclonal antibody strains of 8 monoclonal antibody for 8 plant viruses including broad bean wilt virus, cucumber mosaic virus, tomato mosaic virus, cane mosaic virus, etc are disclosed. The high-specificity high-sensitivity ACP-ELISA and TAS-ELISA methods for detecting relative viruses are disclosed on the basis of said 8 antibodies.

Disclosed are a variety of methods for achieving enhanced expression from a target nucleotide sequence in a plant e.g. comprising the step of transiently introducing into a tissue of a plant (e.g. a leaf) a first nucleic acid comprising the target nucleotide sequence and a second nucleic acid encoding a Post Transcriptional Gene Silencing (PTGS) suppressor protein (preferably of viral or plant origin), wherein the first and second nucleic acids are comprised within a single binary vector construct, or the first and second nucleic acid sequences are comprised within a first binary vector and a second binary vector construct respectively. The plant tissue may then be harvested for the protein. Such methods can give much higher levels of gene expression than are obtainable using stable transgenes, or certain replicating vectors. Also disclosed are specific PTGS suppressor proteins: potato virus X (pvx) p25 protein; african cassava mosaic virus (acmv) AC2 protein; rice yellow mottle virus (rymv) P1 protein; tomato bushy stunt virus (tbsv) 19K protein; plus variants of these. These suppressors may be used in any PTRS context, including the enhancement of transient expression systems.

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 discloses a detecting technique of potato virus and viroid molecule and atopic primer, which is characterized by the following: detecting four potato viruses (PVX, PVY, PLRV and PVS)and one potato viroid (PSTV)through once compound PCR; improving the atopy and detecting efficiency with low cost.

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 relates to a method for cultivating tobacco capable of resisting various viruses by adopting an RNAi (RNA interference) technique. The method comprises the following steps of: obtaining relatively conservative areas of four kinds of viruses within a genome range through screening by comparing full-length sequences of multiple genomes of four kinds of viruses, i.e. CMV (cucumber mosaic virus), PVY (potato virus Y), PVX (potato virus X) and TMV (tobacco mosaic virus) in a genBank; selecting virus sequences and artificially synthesizing 800bp mosaic genes; and accordingly constructing RNAi plant expression carriers of the mosaic genes and transforming common tobacco through agrobacterium to obtain transgenic plants. The method for cultivating tobacco capable of resisting various viruses by adopting the RNAi technique has the characteristics that the four kinds of major tobacco viruses in China are selected as targets, the artificially constructed hairpin structures comprising the sequences of the four kinds of viruses are transformed into the tobacco by using the plant genetic engineering technique, a hairpin double-strand RNA structure transcribed by the tobacco is cut into siRNA (small interfering RNA) by the plant self mechanism, the normal duplication and the accumulation of target virus genes in tobacco plants are specifically interfered, degraded or silenced, and new tobacco materials capable of resisting various viruses can be obtained through screening.

Disclosed are a variety of methods for achieving enhanced expression from a target nucleotide sequence in a plant e.g. comprising the step of transiently introducing into a tissue of a plant (e.g. a leaf) a first nucleic acid comprising the target nucleotide sequence and a second nucleic acid encoding a Post Transcriptional Gene Silencing (PTGS) suppressor protein (preferably of viral or plant origin), wherein the first and second nucleic acids are comprised within a single binary vector, construct, or the first and second nucleic acid sequences are comprised within a first binary vector and a second binary vector construct respectively. The plant tissue may then be harvested for the protein. Such methods can give much higher levels of gene expression than are obtainable using stable transgenes, or certain replicating vectors. Also disclosed are specific PTGS suppressor proteins: potato virus X (pvx), p25 protein; african cassava mosaic virus (acmv) AC2 protein; rice yellow mottle virus (rymv) P1 protein; tomato bushy stunt virus (tbsv) 19K protein; plus variants of these. These suppressors may be used in any PTRS context, including the enhancement of transient expression systems.

The invention discloses a duplex attenuated vaccine capable of resisting cucumber mosaic virus and potato virus X and application of the duplex attenuated vaccine. The duplex attenuated vaccine is a cucumber mosaic virus RNA2 attenuated mutant plasmid vector pCCFR2-2bPTII-PX1967-2166 inserted with a PVX gene segment; the nucleotide sequence of the PVX gene segment is shown as SEQ ID NO. 1, and thenucleotide sequence of the duplex attenuated vaccine is shown as SEQ ID NO. 2. The duplex attenuated vaccine has a cross protection effect on CMV and PVX; and a vaccine material and an effective prevention and treatment measure are provided for preventing and treating plant virus diseases caused by CMV and PVX in the field.

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 relates to the field of genetic engineering and provides construction methods and applications of two PVX (potato virus X) over-expression vectors and one BiFC (bimolecular fluorescence complementation) vector. A genome of a PVX 1985 isolate is cloned to a downstream 35S promoter through genetic recombination, obtained infectious clone can infect solanaceae crops such as tobacco, tomatoes, potatoes and the like through agrobacterium infiltration. Infectious clone is reconstructed, and the over-expression vectors capable of effectively expressing one or two types of heterologous protein in a host plant body as well as the BiFC vector capable of identifying interactions between protein are obtained.

The invention discloses a plant source antiviral agent which takes a phyllanthi fructus L. extractive as an effective component and a preparation method thereof. The preparation method comprises the following steps: crushing phyllanthi fructus L. and carrying out a certain processing process to prepare a phyllanthi fructus L. antiviral soluble liquid agent, an aqueous agent and micro-emulsion. The antiviral agent can be used for preventing and treating tobacco mosaic virus (TMV), cucumber mosaic virus (CMV), potato virus X (PVX), potato virus Y (PVY) and the like on crops including tobaccos, chilies, tomatoes, pumpkins, potatoes and the like. The preparation contains 10%-30% of the phyllanthi fructus L. extractive and a residual amount of an auxiliary agent. The plant antiviral agent not only has a good antiviral effect, but also is safe to environments, human beings, livestock and natural enemies of pests, and other beneficial organisms. The plant antiviral agent is simple in preparation process and low in cost, and is suitable for popularization and application.

The invention discloses a TMV-CMV-PVY three-virus (tobacco mosaic virus, cucumber mosaic virus and potato virus) colloidal gold rapid test strip, which is composed of a sample pad, a colloidal gold pad, an NC membrane, a piece of water-absorbent filter paper and a backing, wherein a colloidal gold labeled TMV specific antibody, a CMV specific antibody and a PVY specific antibody are enveloped on the colloidal gold pad, and the three antibodies are secreted from hybridoma cell strains BALB / c 15 50, BALBc 15 27 and BALBc 15 8; a detection line and a control line are arranged on the NC membrane; specific antigens of three viruses are enveloped on the detection line; and a colloidal gold labeled second antibody is enveloped on the control line. The test strip is especially suitable for detecting the TMV, CMV and PVY in Guangdong tobacco-growing areas, and the test strip is rapid, sensitive, accurate, low in cost and convenient to operate; the test strip is capable of simultaneously diagnosing various viruses by conducting sampling once; the test strip is quite suitable for in-situ preliminary screening of a great batch of samples; and the test strip, in actual production, has a high application value, and the test strip has a good popularization and application prospect.

The invention belongs to the field of molecular biology, and discloses a molecular marker for identifying potato virus Y (PVY) resistance of tobacco. The molecular marker contains a sequence shown in SeqIDNo.1. The invention also relates to primers for amplifying the molecular marker, and use of the molecular marker and the primers in disease-resistant resource screening and disease-resistant breeding assisted selection of tobacco. A genome DNA sequence and a tobacco PVY resistant gene are associated by the molecular marker disclosed by the invention, which is beneficial to the building of the assisted breeding selection system of the tobacco molecular marker; the molecular marker is tightly interlocked with the tobacco PVY resistant gene and the interlock distance is 0.99cM. The molecular marker and the molecular marker amplification primers disclosed by the invention can be simply and quickly applied to tobacco breeding practice at high flux.

The invention relates to a complementary DNA (cDNA) sequence of tobacco NtFT1 genes and transient expression thereof for inducing tobacco early blossoming, in particular to NtFT1 genes of tobacco FT (Flowering locus T) and application of the NtFT1 genes in inducing the tobacco early blossoming, and belongs to the field of genes in molecular biology. The invention discloses a full-length coded sequence of the NtFT1 genes derived from tobacco for controlling the plant blossom time and a protein sequence coded by using the full-length coded sequence. The NtFT1 genes are obtained by performing Blastn comparison and splice site analysis on the data searched from China tobacco genome sequencing planning data through homologous sequence searching. According to the cDNA sequence, a method for performing semi-quantitative polymerase chain reaction (PCR) or reverse transcription-polymerase chain reaction (RT-PCR) on NtFT1 gene specific primers is designed to clone the genes. According to the invention, a potato virus X (PVX) virus expression vector of the NtFT1 genes is also constructed, and the constructed virus expression vector is used for performing diafiltration and infestation on flue-cured tobacco varieties including Hongda and Yunnan tobaccos 87 and K326 through agrobacterium, and can induce the early blossoming of the varieties to prove that the genes have a function of inducing the early blossoming of cured tobaccos. When a PVX virus transient expression system of the NtFT1 genes is used for tobacco breeding, the growing time of the tobaccos can be shortened, and the fixed number of years for breeding a new tobacco variety is reduced.

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 present invention concerns a method for inducing resistance to a virus comprising a TGB3 sequence with the proviso that it is not the potato virus X, into a plant cell or plant, comprising the following steps: preparing a nucleic acid construct comprising a nucleic acid sequence corresponding to at least 70% of the nucleic acid sequence of TGB3 of said virus or its corresponding cDNA, being operably linked to one or more regulatory sequence(s) active in a plant, transforming a plant cell with the nucleic acid construct, and possibly regenerating a transgenic plant from the transformed plant cell. The present invention is also related to the plant obtained.

The present invention relates to pepper plants resistant to powdery mildew disease as well as to infection by potyviruses, particularly potato virus Y (PVY). Specifically, the present invention relates to pepper plants comprising a viral resistance allele and a powdery mildew resistant allele in cis configuration on the same chromosome, such that the two resistance alleles are in coupling phase.

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 a potato X virus attenuated vaccine and a preparation method and application thereof, and relates to the technical field of plant virus vaccine. The invention aims at the limitation of the protective varieties existing in the attenuated vaccine of potato X virus at present, Reversible mutation problem, A nucleotide sequence encoding "GGXYXDGTK" amino acid motif (where X isany amino acid) of potato virus X is completely knocked out by molecular biological technology, A modified attenuated vaccine was obtained. The recombinant virus lacking the peptide had the ability to infect plants systematically and did not cause distinguishable viral symptoms compared with healthy control plants. The invention is applicable to the development and utilization of plant attenuatedvirus vaccine.

The present invention relates to the biological function and determination method of non-structural protein gene S6 of rice dwarf virus (RDV). The non-structural protein Pns6 is encoded by the No.6 segment S6 of RDV. The infections cloning test of Potato Virus X intercellular motion defect strain and the complementary test of RDV encoded non-structural protein gene shows that non-structural protein encoded by RDV is intercellular motion protein of RDV. Intracellular location research of Pns6 shows that the S6 protein is located in the plasmodesmus of plant cell wall and has the typical characteristic of motion protein. The determination of the gene biological function of S6 in helpful to the further research of the plant infecting mechanism of RDV and obtaining more powerful RDV resisting plant.

The invention discloses preparation, detection and application of a polyclonal antibody of a virus of the potato virus X potyvirus for infecting yam virus X, (yam virus X, YVX). Based on the YVX genome, prokaryotic expression is utilized by an inventor to purify the CP (coat protein) and be used for immune a rabbit to prepare the polyclonal antibody; the polyclonal antibody can specifically recognize the CP of the YVX subjected to prokaryotic expression and the CP of the YVX virus infected by yam, and can generate a specificity immune reaction. Based on the preparation, an efficient, high-flexibility and accurate IC-RT-PCR (immunocapture-reverse transcription-polymerase chain reaction), ELISA (enzyme-linked immuno sorbent assay) and Western blot method is established by the inventor, and YVX can be detected specifically from a yam plant which is infected by the YVX. According to the invention, a material basis and a technical support can be provided for the mutual interaction research of the YVX and a host and the rapid detection, parting and molecular biology research of the virus, and a solid basis is developed for the prevalence monitoring, prevention and treatment of the virus.

The invention discloses a mutant plasmid combination resistant to both cucumber mosaic virus and potato virus X. The mutant plasmid combination is prepared from plasmids pCCFR2-2bPTII-PX1967-2066, pCCFR2-2bPTII-PX2067-2166, pCCFR2-2bPTII-PX5890-5989 and pCCFR2-2bPTII-PX5990-6089 which contain a CMVFny isolate mutant RNA (Ribonucleic Acid) 2 according to an equal volume ratio. According to the invention, four mutant plasmids capable of resisting the cucumber mosaic virus and the potato virus X are combined for use and are mixed and inoculated with plasmids of RNA (Ribonucleic Acid) 1 and RNA 3 containing CMVFny isolate, so that the control effect on the cucumber mosaic virus and the potato virus X can be obviously improved.

The present invention provides an overlapped type culture disc for potato virus-free seedlings and an application method therefor. The overlapped type culture disc comprises overlapped type culture disc bodies (1), planting holes (2), separation slide blocks (3), a thread hole (4), a separation slide block gap (5), a separation slide block rail (6), a separation handle (7), a bolt fixing hole (8), a blade rail fixing groove (9), a fixed bolt (10), a blade rail (11), a blade frame (12) and a push rod (13). The method has good controllability and strong operability; matched equipment has a simple structure and is efficient and practical; manpower is saved; and the production cost of the potato virus-free seedlings is remarkably reduced by the method and the production efficiency is improved.

The invention discloses a method for purification and rejuvenation of potato virus-free tissue culture seedlings. The method is characterized in that a culture medium suitable for growth of potato tissue culture seedlings is prepared through an MS culture medium, composite carrageenan and alpha-pimacol, the pre-cultured virus-free tissue culture seedlings are placed in the solid culture medium and cultured for 12 days, roots and stems of the virus-free potato tissue culture seedlings are strong, and the survival rate of seedling acclimatization and transplantation is high. Compared with an existing potato tissue culture seedling medium, the culture medium has the advantages of being low in cost and good in economic benefit.

This invention relates to a kind of potato virus X (PVX) yunnan outlier three antibody sandwich enzyme immune adsorbing rule (TAS-ELISA) detection kit and its preparation method. The kit comprises positive and negative contrast, PVX multi-clone antibody, PVX single clone antibody, enzyme standard antibody and other materials and medicines. The PVX multi-clone antibody is got from rabbit immune adsorbing blood serum after separately purifying the PVX yunnan outlier. The PVX single clone antibody is got from mouse immune, cell fusion, clone and purification after separately purifying the PVX yunnan outlier. The detection sensitivity of the TAS-ELISA detection kit in this invention can reach 0.01ng / ml. It coincides with the comparing accuracy rate of electrical glass detection result, and it is 100 times than indirect ELISA and DAS-ELISA detection sensitivity.

A soluble concentrate of fructus isopsoraleae chalcone relates to an agricultural soluble concentrate and comprises the following raw materials in percentage by weight: 0.1-10% of fructus isopsoraleae chalcone, 8-20% of emulsifier and a solvent which is added until the concentrate reaches 100%; the pure fructus isopsoraleae chalcone or the plant extract of fructus isopsoraleae chalcone is mixed with the solvent, stirred and completely dissolved; and then the mixture is added with the emulsifier, stirred, uniformly mixed, supplemented with the solvent to reach 100%, and then kept standing for 4-5 hours. The soluble concentrate of fructus isopsoraleae chalcone is used for preventing plant fungus diseases including rice blast, apple canker, cucumber anthracnose, tomato late blight, pepper anthracnose, cucumber botrytis and rhizoctonia solani kuhn, or used for preventing plant viral diseases including tomato mosaic virus, soybean mosaic virus, tobacco mosaic virus, tobacco virus Y and potato virus Y. The soluble concentrate is well soluble with water, so that the sterilizing effect in practical application is greatly improved; and the content of fructus isopsoraleae chalcone as an effectively component is stable.

The invention relates to the field of plant virus gene engineering, and provides screening of potato virus X (PVX) low virulent strains and application in cross protection. Based on infectious clones of PVX strong virulent strains, mutants are introduced into PVX genomes in a directed site manner through a site-directed mutagenesis technology, so as to obtain PVX low virulent strains E46A, N863A, N968A and E1001A with significantly reduced pathogenicity. The low virulent strain E1001A can effectively protect plants from being infected by the PVX strong virulent strains.

The process of extracting potato virus RNA includes grinding potato stem or leaf tissue in ice cooled sterilized homogenizer or mortar into homogeneous slurry, adding extracting buffering liquid and beta-mercaptoethanol to mixing homogeneously, adding water saturated phenol and chloroform or isoamylol, centrifugation at room temperature, adding NaAc and isopropropanol, precipitation at -20 deg.c for 3 hr, washing the precipitate with ethanol solution of 70 % concentration, dissolving with bidistilled bacteria-free water and other steps. The said process can obtain RNA with complete sequence and no degradation, and may be used in large-scale RT-PCR test of potato virus.