57 results about "Tomato yellow leaf curl virus TYLCV" patented technology

The invention discloses application of guaiacol to tomato yellow leaf curl virus. The application concentration of guaiacol chemical liquid is 50-500 micrograms/mL, and an emulsifier is added into the guaiacol chemical liquid to serve as a surfactant. The guaiacol is used in prevention and treat of the tomato yellow leaf curl virus, has a prevention and treat effect of 50 percent for the tomato yellow leaf curl virus, and has an effect of treating both diseases and insects when used together with chemical pesticide for preventing and treating a virus transmitting mediator bemisia tabaci. Experiments prove that guaiacol can be used for effectively preventing and treating the tomato yellow leaf curl virus.

The invention belongs to the technical field of plant molecular marker technology, and particularly relates to a double-SNP marker of a gene Ty-1 related to resistance to tomato yellow leaf curl virus disease, cDNA sequences of disease-resistant gene Ty-1 and disease-infected gene (namely allele gene) ty-1 and positions thereof on a whole genome can be obtained from the reported WO2012125025 document. Through sequence comparison, the disease-infected and disease-resistant specific primers NL2 and NL3 can be designed respectively by selecting two multielement SNP-locus GT/CG and GTG/ACA, and are mixed to form a NL2-3 double-SNP marker primer. The three types of Ty-1 and ty-1 can be detected by the NL2-3SNP marker through once PCR and gel electrophoresis display.

The invention relates to a method for identifying tomato yellow leaf curl virus resistance, which enables infestive clonal construction of TYLCV in Tianjin region by aiming at a typical tomato yellow leaf curl virus sequence in Tianjin region. The resistibility of different tomato materials on the virus can be identified through inoculating tomato yellow leaf curl virus by injecting in stalks. The method avoids culturing Bemisia tabaci (Gennadius), ensures simple operation, high efficiency, high repeatability and high controllability, provides a relatively consistent inoculation pressure, obtains accurate result and is a simple and effective inoculation identification method.

The invention discloses a hybridoma cell strain capable of secreting a tomato yellow leaf curl virus (TYLCV) monoclonal antibody and application of the monoclonal antibody. A coat protein gene of a tomato yellow leaf curl virus separator (TYLCV-SH2) is cloned; the coat protein of the virus is expressed by a prokaryotic expression system; BALB/c mice are immunized by using expression and purification protein as antigen; a hybridoma cell strain D10 capable of performing passage stably and secreting the TYLCV monoclonal antibody is obtained through cell fusion, screening and cloning; and the collection number is CGMCC No.5538. The D10 monoclonal antibody ascites indirect ELISA valence reaches more than 10<-6>; and the antibody type and subclass are IgG1 and kappa chains. A dot-ELISA detection method for detecting the TYLCV of the tomatoes is established by the D10 monoclonal antibody; and when the leaf suffering from the disease is diluted according to the ratio of 1:320 (w/v and g/mL), the virus can still be detected. By the dot-ELISA and Tissue-blot ELISA method, the TYLCV of tomato samples in the field can be detected accurately, specifically and sensitively. Due to establishment of a preparation method for the TYLCV monoclonal antibody and a detection method, technological and material support is provided for diagnosis, prediction and scientific prevention and control of the tomato virus disease.

The invention relates to a seedling growth matrix capable of reducing plant virus harm and a preparation method and application thereof. The seedling growth matrix is composed of mushroom residue fermentation rotten matters, grass carbon and vermiculite. The disease index of the seedling growth matrix is more than 40 percent lower than that of a grass carbon matrix on average. The average prevention efficiency of the seedling growth matrix on cucumber mosaic viruses, tomato yellow leaf curl viruses and tobacco mosaic viruses is respectively 40 percent, 40 percent and 50 percent. The seedling growth matrix is simple in production process, low in cost and capable of reducing plant virus harm and usage amount of the grass carbon, and has remarkable economic benefit and economic benefit.

The invention relates to primers and a probe used for carrying out qualitative and quantitative detections upon a virus through a biological method, and especially relates to primers, a TaqMan probe and a kit used for carrying out qualitative and quantitative detections upon tomato yellow leaf curl virus (TYLCV) through a real-time fluorescence quantitative PCR technology. The base sequence of an upstream primer of the primers is represented by SEQ ID NO: 1, and the base sequence of a downstream primer of the primers is represented by SEQ ID NO: 2. The DNA sequence of the probe is represented by SEQ ID NO: 3. A 5' terminal of the probe is labeled with reporting fluorophores, and a 3' terminal of the probe is labeled with quenching fluorophores. The kit comprises the primers and the probe. The primers and the probe provided by the invention can be used in quantitative and qualitative analysis of TYLCV. The primers and the probe have important significances in research fields such as the determinations of a permanent planting regularity and a replication level of the virus in a plant, and the determinations of whether the virus is removed after medication. The primers, the probe and the kit can play important roles in large-scale quarantine, epidemiological study and early diagnosis of TYLCV.

The invention discloses a kit for rapidly detecting tomato yellow leaf curl virus and application thereof. A pair of specific primers is designed according to the nucleotide sequence of a dominant strain of tomato yellow leaf curl virus in our country, and also an extraction buffer and a washing buffer for processing a sample are designed. By using the kit for rapid detection on tomato yellow leaf curl virus, a virus-inflected tomato sample does not need extraction of DNA, only a small amount of a disease leaf (about 10 mg) is needed, and the disease leaf only needs once PCR reaction after being processed by the extraction buffer and the washing buffer, so that identification on tomato yellow leaf curl virus is realized. Experiments prove that the method has relatively high sensitivity and the sensitivity reaches 1E4. Also, the method is capable of performing virus containing identification on virus-transmission vectors Q-biotype bemisia tabaci and B-biotype bemisia tabaci of tomato yellow leaf curl virus. Compared with traditionally biology and serology, the method is a simple, convenient, rapid, economic and sensitive method for detecting tomato yellow leaf curl virus.

The invention discloses a compound microbial agent and application thereof in preventing and treating various plant diseases. The compound microbial agent has a remarkable inhibition effect on variousmain soil-borne diseases such as bacterial wilt, root rot, soft rot, black shank, gibberellic disease and root black rot caused by bacteria, fungi and oomycetes; meanwhile, the biomass of solanaceaecrop tomatoes, brassicaceous crop brassica campestris and cucurbitaceae crop cucumbers is remarkably improved, and a remarkable inhibiting effect is also achieved on tomato yellow leaf curl viruses which are plant virus diseases. The compound microbial agent effectively solves the problems of environmental pollution and food quality caused by the fact that the conventional microbial agent only aims at single pathogenic bacteria and chemical pesticide prevention and treatment, and has a good application prospect in green and environment-friendly and agricultural sustainable development.

The invention discloses a method and a kit for synchronously detecting four viruses inducing a tomato yellow leaf curl disease and belongs to the field of plant protection. The four viruses include tomato yellow leaf curl virus, Taiwan tomato leaf curl virus, Guangdong tomato yellow leaf curl virus and Guangdong tomato leaf curl virus. Four pairs of specific primers are designed; total DNA of a sample which is to be detected is extracted; the extracted DNA is taken as a template and the four pairs of specific primers are taken as primers to perform PCR (polymerase chain reaction) in the same system; the PCR product is detected; and according to the fragment size of the PCR product, the virus which invades the sample can be identified. The method and the kit are quick, simple, convenient, accurate and low in cost, can be used for synchronously detecting the four viruses which induce the tomato yellow leaf curl disease, and can be applied to the actual production to quickly, accurately and effectively detect the plant disease sample.

The invention relates to the technical field of molecular biology and crop breeding, in particular to KASP primers for detecting typing of a tomato yellow leaf curl virus disease-resistant gene Ty-1 and an application of the KASP primers. The tomato yellow leaf curl virus disease-resistant gene Ty-1 has an SNP site at the position of the 3461st < base, and the mutation of the base of the SNP sitecauses the change of the resistance of the Ty-1 gene. A group of KASP primers capable of being used for genotyping and a kit containing the KASP primers are designed on the basis of the SNP site. TheKASP primers and the kit can be used for identifying whether tomatoes contain the tomato yellow leaf curl virus disease-resistant gene Ty-1 or not, and distinguishing tomatoes which contain Ty-1 and resist tomato yellow leaf curl virus diseases from tomatoes which do not contain the Ty-1 and are infected with the tomato yellow leaf curl virus diseases. Compared with the prior art, rapid, accurateand high-throughput disease resistance identification can be performed on tomato plants in the seedling stage, the workload of manual inoculation in the seedling stage and field disease resistance identification is reduced, the tomato breeding efficiency can be improved through the application of the KASP primers, the breeding cost is saved, and the breeding process is accelerated.

The invention discloses an SSR (simple sequence repeat) marker for tomato yellow leaf curl disease resisting character co-segregation and application thereof. The SSR marker includes primers: upstream primer T5-F: 5'-CAATCAATCGATGCACAAAACACC-3'; downstream primer T5-R: 5'-GACTGACTGCATTGGATTTGGCTT-3'. The marker can be amplified in CLN32120a-23 into a 580bp strip, can be amplified in Money marker into a 640bp strip and can be amplified in F1 into the above two strips, marking results are substantially consistent to that of field identification, it is proved that the marker can distinguish anti-disease materials, disease-sensitive materials and hybrid anti-disease materials and is a co-dominant marker in tight linkage with tomato yellow leaf curl disease virus resisting gene ty-5. Molecular marker detection is 90% matching with field detection in terms of detection results. The marker can be used in PCR (polymerase chain reaction) quick detection for tomato yellow leaf curl virus resisting ty-5 gene, and certain theoretical and practical basis is provided for the application of ty-5 molecular marker assisted selective breeding.

The invention discloses a method for synchronously detecting four whitefly transmitted gemini-viruses infecting tomatoes. The method can be used for simultaneously identifying four viruses by utilizing four specific primers ToL-F/ToL-R, TY-F/TY-R, PaL-F/PaL-R, AY-F/AY-R especially for four viruses, namely, tomato leaf curl China virus, tomato yellow curlleaf China virus, papaya leaf curl China virus and ageratum yellow vein China virus by combining a PCR (polymerase chain reaction) detection technology. According to the method, after a total DNA (deoxyribonucleic acid) of a sample is extracted, the four whitefly transmitted gemini-viruses can be detected by only once PCR detection, and a simple method is provided for investigation and identification. Compared with the traditional method which can be used for detecting one virus by once PCR, the method has the advantages of reducing cost, saving time, guaranteeing sensitivity and specificity and simplifying operation, and is a specific, sensitive, convenient, economical and effective detection method.

The invention discloses a method for rapidly detecting Chinese tomato yellow leaf curl virus (TYLCCNV) by using a loop-mediated isothermal amplification (LAMP) technology. The method comprises a reaction system and a specific primer group contained in the reaction system. The method comprises the steps of extracting DNA (Deoxyribonucleic Acid), carrying out LAMP, carrying out electrophoresis detection or fluorescent staining and the like. The method has the advantages of strong specificity, high sensitivity, simplicity, convenience, rapidness, low cost and the like, and special instruments such as a PCR (Polymerase Chain Reaction) instrument and equipment such as gel electrophoresis equipment are not required, so that the rapid detection on virus disease samples of laboratories and fields is facilitated.

WRKY transcription factor family widely exists in plants. In accordance with the quantity of WRKY structural domains and the characteristics of a zinc finger structure, the WRKY transcription factor family can be divided into three sub-families (I, II and III). An WRKY transcription factor gene Soly WRKY41 is analyzed and identified from a tomato variety which is infected by tomato yellow leaf curl virus. The transcription factor has one WRKY structural domain and a zinc finger structure of C2HC, belonging to members of the WRKY III sub-family. Based upon subcellular localization analysis, itis indicated that the SolyWRKY41 transcription factor belongs to nucleoprotein. A virus-induced gene silencing result shows that the SolyWRKY41 gene can implement negative regulation of the tomato yellow leaf curl virus. Through functional researches of the SolyWRKY41 transcription factor, the molecular mechanism of tomatoes in response to the tomato yellow leaf curl virus is enriched; and the SolyWRKY41 transcription factor is applicable to tobacco disease-resistance breeding.

The invention discloses a microbial antiviral composition containing a glycoprotein GP-1, a preparation and an application. The microbial antiviral composition containing the glycoprotein GP-1 comprises berberine and further comprises the glycoprotein GP-1 and/a metabolite of Streptomyces kanasensis ZX01). The antiviral composition has a good effect on preventing and controlling various plant virus diseases such as Tobacco mosaic virus, Cucumber mosaic virus, Tomato yellow leaf curl virus, Potato virus X/Y, Rice dwarf virus and is novel in acting way and capable of effectively relieving the problem of resistance brought by long-term use of conventional pesticides and improving the disease resistance of plants. The antiviral preparation disclosed by the invention has the characteristics such as high efficiency, wide spectrum and low cost, and a conventional pesticide application method can be adopted, so that the environment is not polluted.

The invention belongs to the field of agricultural microbial technology, and relates to a bio-control strain DX3 used for preventing and controlling tomatos yellows leaf curl virus and applications thereof. It is identified that DX3 belongs to Bacillus sp.; DX3 is preserved in China General Microbiological Culture Collection Center from January, 31th, 2013; and the strain preservation number is CGMCC NO.7239. The results of a greenhouse experiment show that: the biocontrol strain DX3 is capable of reducing the frequency of toamtos yellows leaf curl virus disease significantly. A biocontrol agent is prepared by using DX3, and is diluted to 100-fold for spray processing; and bio-control effect of toamtos yellows leaf curl virus disease at room temperature can reach 57.38% by spray processing. In addition the bio-control effect under field conditions can reach 51.70 to 53.35%. Therefore, the strain and the bio-control agent prepared by using the strain can be used for preventing and controlling toamtos yellows leaf curl virus and yield increase.

The invention discloses a high-yield and high-quality tomato breeding and planting method which comprises the following steps: breeding a strain which is excellent in commodity character, simultaneously contains Cf-5, Mi and Ty-13 resistance genes and can resist tomato leaf mold, tomato root knot nematode disease and tomato yellow leaf curl virus disease, and carrying out planting to obtain seeds; soaking the obtained seeds with a 0.1% forchlorfenuron solution, stirring and soaking the seeds with warm water, accelerating germination until the seeds germinate, planting the seeds in planting soil obtained by mixing and stirring planting land soil, amino acid long-acting slow-release nitrogen, monopotassium phosphate and glycan polypeptide biological potassium, and performing conventional seedling raising; after tomato seedlings are 15-20 days old, transplanting the tomato seedlings into the planting area where soil preparation is completed, during transplanting, transplanting the tomato seedlings and the planting soil together, and conducting reasonable field management and fertilizer and water management till tomatoes are harvested. On the premise that high yield, low morbidity and high variety of the tomatoes are guaranteed, rapid breeding of good varieties of the tomatoes can be achieved.

The invention relates to the technical field of plant biology, and particularly relates to an SNP locus combination for detecting resistance of a tomato yellow leaf curl virus disease and application thereof. Based on this, the invention develops a primer combination, a kit and a detection method capable of quickly, intuitively and effectively identifying the genotype state of a target SNP locus. The SNP locus combination can realize quick, accurate and high-throughput detection of haplotypes of sections Ty-1/Ty-3 and Ty-2 of resistance genes of the tomato yellow leaf curl virus disease, has the advantages of simple operation, low cost, automation, high throughput efficiency, stable marking, safety, no toxicity, harmlessness and the like, can quickly and accurately identify the resistance of the tomato yellow leaf curl virus disease at high throughput in the seedling stage of tomatoes, reduces the workload of artificial inoculation identification and field transplantation, improves the breeding efficiency, reduces the breeding cost, accelerates the breeding process, and is very suitable for modern commercial breeding application and large-scale genetic improvement research.

The invention discloses a reagent kit for detecting tomato yellow leaf curl virus carried by single-head whiteflies. A primer group includes a primer 1, a primer 2, a primer 3 and a primer 4, wherein nucleotide sequences of the primer 1, the primer 2, the primer 3 and the primer 4 are respectively a sequence 1, a sequence 2, a sequence 3 and a sequence 4 in a sequence table. An experiment of the reagent kit proves that the reagent kit aims for the single-head whiteflies, a LAMP primer is designed according to genome sequences of TYLCV preponderant strains occurred in our country, toothpicks or suckers are directly utilized to puncture the single-head whiteflies, deoxyribonucleic acid (DNA) is not required to be extracted, the LAMP reaction is directly performed, and the reagent kit has the advantages of being simple, quick, sensitive and the like and overcomes the shortcomings including time consumption, nucleic acid extraction, high instrument requirements, complicated steps and the like in traditional technologies.

The invention relates to the technical field of prevention and control of tomato virus and discloses a method for preventing and controlling tomato yellow leaf curl virus. The method comprises the following steps: step 1, seedling culture and selection of tomatoes; step 3, treatment of a solar greenhouse before tomato planting; step 4, installation of an exhaust system of the solar greenhouse; step 5, installation of a sprinkling irrigation system of the solar greenhouse; and step 6, management after tomato planting. Compared with the prior art, the method has the beneficial benefits as follows: 1, the method can effectively reduce the incidence of the tomato yellow leaf curl virus and effectively solve the problem of tomato cultivation and planting in the solar greenhouse after autumn; 2,by high-temperature greenhouse sealing, pests are prevented and controlled under the condition of no pesticides are used, and the environmental and ecological safety is facilitated; 3, through simulated rainfall, bemisia tabaci is killed and the use quantity of pesticides is reduced; and 4, through simulated rainfall and exhaust, the cooling effect of the solar greenhouse is good, the environmental condition for growth of tomatoes is improved, and the yield of the tomatoes is increased.

The invention discloses a cultivation method for controlling tomato yellow leaf curl virus, which comprises the following steps: using tomato seedlings which are not resistant to yellow leaf curl virus, performing field planting when four leaves and one core exist, and irrigating roots with 0.01-0.02% (mass concentration) Silatrane aqueous solution on the same day after field planting; delaying seedling hanging and conducting watering once every 2-6 days before seedling hanging; and when the trunk of each plant has 16 leaves and 1 core, starting to hang seedlings, and spraying 60 mg/kg chitosan oligosaccharide aqueous solution on the leaf surfaces on the seedling hanging day. According to the cultivation method, the morbidity of the yellow leaf curl virus of infected tomato varieties is reduced from 56.8%-61.6% to 3.5%-4.1%; the soluble solid content of tomatoes can reach 4.5%-5.2%; and the single plant yield of the tomatoes can reach 4.81-5.28 kg.