77 results about "Potato virus Y" 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 invention belongs to the field of biotechnology, and provides an RNAi vector pGEM-PCTRi capable of mediating the resistance to tobacco viral diseases in China. Analyzing the genome characteristics of main virus groups in China, which comprise Potato Virus Y (PVY), Cucumber Mosaic Virus (CMV) and Tobacco Mosaic Virus (TMV) and then selecting three segments of conserved effective sequences capable of inducing RNAi in PVY CP, CMV RNA1 and TMV CP; and integrating the three segments in a tabling manner and constructing an inverted repeat sequence so as to obtain an RNAi vector pGEM-PCTRi. Massproduction of dsRNA can be realized in escherichia coli by utilizing the vector, and the resistance of tobacco to PVY, CMV and TMV can be induced to generate after the tobacco treatment. The invention has the potential of being applicable to the prevention of viral diseases for tobacco farmland production, and endowed with the advantages of broad spectrum, high efficiency, no reference to transgenosis, and the like.

The invention discloses a tobacco recessive PVY (Potato Virus Y) resistance gene eIF4E-1 and application thereof. The tobacco recessive PVY resistance gene eIF4E-1 comprises the DNA (Deoxyribonucleic Acid) fragment of an eIF4E-1 gene coding region, wherein the DNA fragment is obtained by PCR (Polymerase Chain Reaction) amplification of a specific primer to a primer 4E-1F (SEQIDNO.3) and a primer 4E-1R (SEQIDNO.4); a nucleotide sequence SEQIDNO.1 of a tobacco PVY resistance gene eIF4E-1 coding region and the amino acid sequence SEQIDNO.2 of protein coded by the tobacco PVY resistance gene are obtained first; the eIF4E-1 gene is a recessive disease resistance gene; the eIF4E-1 gene of infected tobacco is lost by a conventional breeding means, so that the non-GMO (Genetically Modified Organism) disease resistance tobacco is obtained; an interference vector plasmid is built by using the eIF4E-1 gene and converted to the infected tobacco; expression of the eIF4E-1 gene of the infected tobacco is inhibited, so that genetically-modified tobacco with obviously improved disease resistance is obtained; the gene has an important application prospect in cultivating the PVY viral disease resistance tobacco.

The invention discloses a potato virus-free seedling tissue culture propagation method which relates to the propagation technology of potatoes and belongs to the technical filed of agriculture planting. The method is characterized by comprising the following steps: performing fine selection, performing accelerating germination, performing seed airing, and inoculating aired seeds in a germination culture medium for cultivation and growth, wherein the accelerating germination and sowing are performed at the temperature being 15-20 DEG C; the method has the beneficial effects that compared with a formula based on an MS, a formula in the method is small in quantity of constitution chemical substances, low in cost, and convenient to prepare. By using the method disclosed by the invention, potato virus-free seedlings can be propagated in an opening environment, and the characteristics that the propagation speed is high, the production cost is low, the popularization and the application are easy, and the like can be realized.

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.

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 surface plasma resonance detecting method for PVY/CMV (Potato Virus Y/Cucumber Mosaic Virus) and belongs to a material detection method. The surface plasma resonance detecting method comprises the following steps: 1) preparing a PVY/CMV antigen and a monoclonal body; 2) preparing a biosensor chip; 3) preparing a standard curve and acquiring a virus computational formula; and then measuring the content of PVY/CMV by utilizing the virus computational formula. The surface plasma resonance detecting method provided by the invention has high sensitivity, can be used for qualitatively and quantitatively measuring PVY/CMV and is convenient in operation and low in cost.

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 discloses a rapid test strip for potato viruses, and a preparation method and application thereof. The rapid test strip for the potato virus comprises a sample pad, a colloidal-gold pad, a NC membrane, a test line (a T line), a control line (a C line), a water absorbing pad and a PVC base plate. The preparation method comprises the following four steps: preparation of the colloidal-gold pad; preparation of the NC membrane; preparation of the sample pad; and assembling of the test strip. Application and detection comprise the following concrete steps: treatment of a sample; dropwise addition of the sample; result interpretation; etc. The potato viruses comprise PVX, PVYNTN, PVYo, PLRV, PVS, PVM, PVA and a tomato black ring virus. The test strip is convenient to use, has good applicability, needs no laboratory, no special apparatus and no operation training of professionals, can realize testing in any place with potatoes, such as a field, a potato storage cellar and a customhouse, and can be used by any potato planter.

The invention discloses a cultivation method of a virus-free seed tuber of Solanum tuberosum and relates to the technical field of planting of Solanum tuberosum. The method comprises sowing whole tubers of virus-free seed tubers in a high-altitude aphid-free area with an elevation of 2500-2600 m, wherein each whole tuber weighs 35-45 g, and cultivating the virus-free seed tubers by means such as pre-sewing sufficient application of base fertilizers, germination accelerating and reasonable close planting. The emergence rate is high, up to 99.8%, a course of seedling emergence to maturity takes about 70 days, seedlings grow evenly during growth without disease and pest damage, harvested seed tubers are good in quality, total virus (PVY (Potato Virus Y) and PLRV (Potato Leaf Roll Virus)) rate is 4%, bacterial wilt rate is 0.35%, indexes comply with related specifications in seed tubers of Solanum tuberosum GB 18133-2012, the yield per mu can be increased by 35%, and the yield of small potatoes is up to 70%.

The invention discloses a hydroponic bed for potato virus-free seedlings and relates to the technical field of hydroponics for potato virus-free seedlings. The hydroponic bed comprises a bed body, a fixing plate is arranged in the bed body, and multiple seedling holes used for storing nutrition solutions for seedling are formed in the fixing plate; water serving as an insulation medium is filled between the fixing plate and the bed body, a heating device and a temperature controlling device are further arranged on the bed body, and the heating device and the temperature control device are cooperated with each other to have water temperature regulated; the temperature control device comprises a display and a heat sensor, and the heating device and the heat sensor are immersed in water. By the arrangement, the temperature of the hydroponic bed can be regulated to promote rapid growth of the hydroponic seedlings, time for entering mist culture is shortened, quality of stock seeds of potatoes is improved, and yield is increased; when the seedling viruses occur, infection sources cannot be spread with the nutrition solutions, and the viruses are easily controlled.

The invention belongs to the technical field of crop disease diagnosis and molecular biology and discloses a multi-PCR (Polymerase Chain Reaction) method for synchronously detecting four chilli viruses. A PCR system contains four pairs of specific primers (SEQ ID NO: 1 to NO: 8); In one PCR system, the four viruses are synchronously amplified to obtain 1406bp, 1202bp, 600bp and 376bp specific bands; a multi-RT-PCR reaction system capable of synchronously detecting BBWV2 (Broad Bean Wilt Virus 2), PVY (Potato Virus Y), ChiVMV (Chili Veinal Mottle Virus) and CMV (Cucumber Mosaic Virus) is established and optimized. A detection result shows that the optimized multi-RT-PCR reaction system can be used for rapidly, efficiently and accurately detecting field samples. The multi-PCR method has very important meanings on predication of field occurrence condition and disease epidemics of four common virus diseases including PVY and the like on chilies.

The invention discloses a soil cultivation method for potato virus-free basic seeds. The method comprises: 1) production of a tissue culture seedling; 2) rooting and strengthening; 3) soil cultivation; and 4) harvesting and storage, wherein the step 2) rooting and strengthening comprise: taking out the tissue culture seedling suitable for fixed planting out of a culture bottle; removing root systems and cleaning a culture medium; soaking the roots for 10-15 minutes by using a NAA50 mg/L+2-4 D 5 mg/L rooting agent; then fixedly planting the roots in a soilless culture medium, and keeping the temperature and humidity by using a plastic arch-shaped shed to accelerate the tissue culture seedling to take root, sprout and quickly grow; 12 days after fixed planting, starting spraying a nutritional liquid, spraying the nutritional liquid once at an interval of five days, and spraying the nutritional liquid three times; in 29-32 days after fixed planting, uncovering plastic of the arch-shaped shed, and selecting robust tissue culture seedlings, wherein the height of the seedlings reaches 10 cm and the blades are longer than 1 cm, and transplanting the tissue culture seedlings in pre-treated field soil. By adopting the soil which replaces the medium to culture the potato virus-free basic seeds, the production cost is greatly lowered, so that the market supply price is lowered, and the method plays a positive role in generalizing and popularizing virus-free potatoes.

The invention discloses a method for cultivating potato virus-free seedlings, relates to the virus elimination and tissue culture techniques of plants, and belongs to the field of agricultural seedling breeding. The method is characterized by comprising the following steps of: transferring virus-free potato tissue culture seedlings into a MS culture medium to culture, wherein a bacteriostatic agent and sugar are added into the MS culture medium; and after the culturing is completed, soaking the seedlings into a CIP improved rooting agent, and carrying out cuttage planting and nutrient solution applying on the seedlings, and after the seedlings are mature, harvesting the seedlings. The method disclosed by the invention has the beneficial effects that through cultivating virus-free seedlings in a special culture medium, the contamination rate in the culture medium is reduced; then, the plant seedlings cultivated in the culture medium are subjected to cuttage, and before cuttage, the cuttage seedlings are placed in the CIP improved rooting agent, and nutrient solutions N, P and K are applied, so that the survival of virus-free seedlings is ensured, the potato growth amount and the expansion of tubers are promoted, thereby improving the yield; and the single-plant potato growth amount of breeder seeds is improved, so that the production cost of breeder seeds is reduced by 25%, thereby providing a reliable technical support for the industrialized development of potatoes.

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 discloses a method for rapidly removing a potato virus. The method comprises the steps that a plant resistance inducer for inhibiting the virus is sprayed on a tissue culture seedling at 3-5 days before stem tip peeling based on conventional stem tip detoxication, wherein the plant resistance inducer includes but is not limited to amino-oligosaccharin, ningnanmycin, 3-hydroxyl-3-acetonyl ketoindole, and the like; a stem tip is taken and separately cultured for 7-28 days; the plant resistance inducer for inhibiting the virus is sprayed for one time; 0.1-0.7mm stem tip is taken and separately cultured for 7-28 days; and the step is repeated for three times to obtain a virus-free tissue culture seedling. A virus-free rate of the obtained tissue culture seedling is higher than 80%. According to the method, the time for obtaining the virus-free tissue culture seedling is short; a virus rate of the obtained tissue culture seedling is low; operation is simple; and production popularization is facilitated.

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.

The invention discloses a RT-PCR detection kit for GMBFV, PVY and GCLV viruses of garlic, which is characterized in that RNA of garlic tissue is used as a template, cDNA is obtained through reverse transcription, then three pairs of specific primers are adopted, one-time RT-PCR is carried out to detect the main garlic viruses simultaneously, such as the Garlic mite borne filamentous virus, the Potato virus Y and the Garlic common latent virus, and the three pairs of primers are shown in SEQ ID NO.1-SEQ ID NO.6. The detection kit provided by the invention has high detection efficiency, low detection cost and is suitable for virus detection of large-scale garlic materials.

The invention discloses a detection method for poison of CMV (Cytomegalovirus) and PVY (Potato Virus Y) of single-head tobacco aphids. The detection method mainly adopts reagents and materials including CMV and PVY primary antibodies, a corresponding enzyme label second antibody, a sample diluting solution, a washing solution and a developing solution. A detection plate is a nitrocellulose membrane (NCM) coated with CMV and PVY virus protein antigens carried by the tobacco aphids; an enzyme conjugate working solution is a goat anti-rabbit antibody coated with the CMV and PVY primary antibodies (goat anti-rabbit) and horse radish peroxidase. According to the detection method, the single-head tobacco aphids are subjected to a series of treatment and an antigen and antibody reaction basic principle is utilized to detect that whether the tobacco aphids have poison or not in time. The detection method has the active effects of strong specificity, high sensitivity, simplicity in operation and low detection cost; the detection method is suitable for being popularized and applied in fields in a large scale and has a wide application prospect.

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.