97 results about "Tomato yellow leaf curl virus" patented technology

The invention discloses a method for preventing and treating tomato yellow leaf curl virus diseases. The method includes cultivating tomatoes in sunlight greenhouses in large rows and small rows; transplanting abutilon theophrasti 15-20 days before the tomatoes are planted in fields; planting one row of abutilon theophrasti at the planting distances of 20cm along the inner side of the root of a north wall of each greenhouse; planting a row of abutilon theophrasti at the planting distances of 2.0m between each two adjacent small field planting rows; pulling out the abutilon theophrasti among the small field planting rows after fruits of fifth panicles of the tomatoes are set. The row spacing of the large rows of the tomatoes planted in the sunlight greenhouses is 80cm, and the row spacing of the small rows of the tomatoes planted in the sunlight greenhouses is 60cm. The leaf age of the abutilon theophrasti is 4 leaves and 1 core when the abutilon theophrasti is transplanted. The method has the advantages that the incidence of the tomato yellow leaf virus diseases can be effectively reduced, and the incidence of tomato yellow leaf virus disease infected tomato varieties can be within the range of 10% under the control generally.

The invention provides bacillus velezensis for inhibiting virus and promoting plant growth and application of a bacillus velezensis SNB55 strain in prevention and treatment of plant virus diseases andplant fungal diseases and promotion of plant growth. The bacillus velezensis for inhibiting virus and promoting plant growth and the application of the bacillus velezensis have the beneficial effectsthat the bacillus velezensis has no toxicity and pathogenicity, is safe to human and livestock, is environment-friendly, and has high stress resistance and stable antibacterial activity; the bacillusvelezensis can inhibit phytopathogens such as bacillus cereus, tobacco mosaic virus, tomato yellow leaf curl virus, fusarium wilt of cucumbers, tobacco target spot pathogens, phytophthora capsici, rice sheath blight disease and botrytis cinerea, can be secrete indoleacetic acid to promote seed germination and plant growth; and the strain is a multifunctional biocontrol bacterium and has a wide application prospect.

The invention disclose a dot- ELISA (dot Enzyme-Linked Immunosorbent Assay) method and tissue printing ELISA method for detecting the presence of the tomato yellow leaf curl virus in a plant as well as a reagent kit and an application thereof. A monoclonal antibody prepared for tomato yellow leaf curl virus coat protein is utilized to build the dotenzyme-Linked immunosorbent assay (dot-ELISA) method and the tissue printing ELISA method with optimal proportions, and the rapid-detection reagent kit is developed. The dot- ELISA (dot Enzyme-Linked Immunosorbent Assay) method and tissue printing ELISA method for detecting the presence of the tomato yellow leaf curl virus in a plant as well as the reagent kit and the application thereof are suitable for such solanaceae plants as tomato, capsicum, eggplant, tobacco, night shade and jimson weed; the situation of the presence of tomato yellow leaf curl virus in field vegetable samples is examined to detect the incidence of the virus disease and evaluate the occurrence and distribution and the prevalence trend of the tomato yellow leaf curl virus under the field condition; and the detection method has high sensitivity and good specificity, needs short time, is low in cost, and provides a technical support for rapid and large-scale detection of 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 discloses a molecular marker auxiliary selection method for tomato yellow leaf curl viral disease and knot nematode. The method is characterized by comprising the following steps of: (1) extracting DNA from a tomato material F1 and a selfed separation offspring F2 individual; (2) performing multiple PCR amplification on the tomato material F1 and the selfed separation offspring F2; and (3) performing resistance evaluation on the breeding separation generation material according to the detection result. The multiple PCR amplification method can amplify Ty-1 genes and Mi genes in the same reaction system at the same time, so the method greatly simplifies experiment operation and experiment expense, is suitable for identifying and analyzing a large amount of sample material, greatly shortens the time and reduces the workload.

The invention discloses a quick, simple and effective tomato yellow leaf curl virus resistance determination method and belongs to the field of plant protection. Traditionally, the determination of the tomato yellow leaf curl virus resistance is performed by a Bemisia tabaci (Gennadius) inoculation determination method, while the Bemisia tabaci (Gennadius) inoculation determination method has difficulty in providing uniform and consistent inoculation pressure due to the influences (such as the number of the inoculated Bemisia tabaci (Gennadius), virus rate and feeding habit) of the Bemisia tabaci (Gennadius) and at the same time, suffers interference from other Bemisia tabaci (Gennadius) transmitted viruses, so the accuracy of the determination result is low. The agrobacterium rhizogenes injection inoculation method well solves the problem. The resistance of a tomato material to the tomato yellow leaf curl viruses can be determined by using constructed infectious clones and directly inoculating the tomato yellow leaf curl viruses by injection in stalks. The method avoids culturing Bemisia tabaci (Gennadius), ensures simple operation, high efficiency, high repeatability and high controllability, provides a relatively consistent pressure, obtains accurate result and is a simple and effective inoculation determination method.

A Lycopersicon esculentum plant comprising within its genome at least one tomato yellow leaf curl virus (TYLCV) resistance allele and at least one root knot nematode resistance allele, characterized in that the resistance alleles are present in coupling phase at different loci on one chromosome and in that the plant is highly resistant both against TYLCV and to at least one root knot nematode species selected from the group consisting of Meloidogyne arenaria, Meloidogyne incognita and Meloidogyne javanica.

The invention relates to a plant-virus resisting composition which contains active constituents. The plant-virus resisting composition is characterized in that the active constituents contain amino-oligosaccharins and ribavirin; the weight ratio of the amino-oligosaccharins to the ribavirin is 1: (0.03-20). The invention also relates to a plant-virus resisting medicament prepared from the plant-virus resisting composition. The dosage form of the plant-virus resisting medicament can be soluble liquid, soluble powder, water dispersible granules, granules or water aqua. The invention also relates to an application of the plant-virus resisting medicament in prevention and treatment of one or multiple plant viruses comprising tobacco mosaic viruses, cucumber mosaic viruses, potato Y viruses, tomato mosaic viruses, tomato yellow leaf curl viruses and tomato spotted wilf viruses. The plant-virus resisting composition and the plant-virus resisting medicament are good in prevention and treatment effects, and therefore, the medicament using amount can be reduced, the cost is reduced, and the pollution to the environment is relieved; the plant-virus resisting medicament does not only contain a single active constituent, so that the plant viruses difficultly produce drug resistance; the plant-virus resisting medicament can prevent and treat various different types of plant-virus diseases.

The invention relates to a method for cultivating an anti-TYLCV (Tomato Yellow Leaf Curl Virus) tomato plant by using an RNAi (RNA interference) technology, belonging to the field of biotechnology. According to the method, the conserved sequences in genes of V2 and C1 of TYLCV are inosculated, an RNAi carrier with inverted repeat sequences is constructed, the aseptic seedling cotyledons or hypocotyl explants of the tomato are transformed through an agrobacterium-mediated transformation method, the transgenic tomato expresses resistance to the TYLCV after the inoculation of the agrobacterium infectious clone injection of the TYLCV and the TYLCV-carrying Bemisia tabaci virus, and no obvious disease symptoms are observed. According to the method, the anti-TYLCV transgenic tomato plant can be cultivated by using the RNAi technology, so that the technical support for the control of the TYLCV is provided.

The invention discloses identification and an application of a tomato SolyWRKY54 transcription factor in regulating and controlling tomato yellow leaf curl virus. According to the invention, a WRKY transcription factor gene SolyWRKY54, which responds to the tomato yellow leaf curl virus, is identified from tomatoes; protein coded by the gene belongs to III sub-family in a WRKY transcription factorfamily. A semiquantitative PCR result shows that a susceptible variety, namely 'Jinpeng No.1', is higher than a disease-resistant variety, namely 'Zheza 301', in virus accumulation. Based upon fluorescent quantitative PCR analysis, it is indicated that the expression amount of the SolyWRKY54 gene is reduced in the disease-resistant tomato variety, namely 'Zheza 301'. Through a virus-induced genesilencing experiment, it further provides a negative regulation function of the SolyWRKY54 gene in a process of responding to the tomato yellow leaf curl virus. The SolyWRKY54 transcription factor gene, which is identified by the invention, is beneficial to analyzing and researching a resistance mechanism of tomatoes responding to the tomato yellow leaf curl virus; and the SolyWRKY41 transcriptionfactor is applicable to tobacco disease-resistance breeding.

The invention relates to an InDel molecular marker based on a TYLCV (tomato yellow leaf curl virus) resistance gene Ty-3. The nucleotide sequence of the molecular marker is represented as SEQ ID No.1. The invention further relates to a primer pair for amplifying the InDel molecular marker based on the TYLCV resistance gene Ty-3. Sequences of the primer pair are as follows: a forward primer sequence: 5'-TGCAGGAACAGAATGATAGAAAA-3', and a reverse primer sequence: 5'-GCTCAGCATCACCTGAGACA-3'. The molecular marker is totally based on the TYLCV resistance gene Ty-3. According to the developed InDel molecular marker based on the TYLCV resistance gene Ty-3, the marker is totally based on the resistance gene Ty-3, is completely linked with the resistance gene Ty-3 and is a codominant molecular marker. When the marker is applied to molecular marker-assisted selection of the TYLCV resistance gene Ty-3, the accuracy of selection can be greatly improved, the breeding cycle can be shortened, and accordingly, the breeding process is accelerated.

The invention provides a method for quickly and synchronously detecting two virus components, including the tomato yellow leaf curl virus and the accompanying China tomato yellow leaf curl virus satellite, on tomatoes, and belongs to the field of plant protection. According to the nucleotide sequence of the tomato yellow leaf curl virus and the nucleotide sequence of the China tomato yellow leaf curl virus satellite, two pairs of primers are designed, namely TYmF / TYmR and TycnbF / TycnbR, wherein the TYmF / TYmR combination is used for detecting the tomato yellow leaf curl virus, and the TycnbF / TycnbR combination is sued for detecting the China tomato yellow leaf curl virus satellite. After total DNAs are extracted from a sample, only one time of PCR is needed to achieve detection of the two components including the tomato yellow leaf curl virus and the accompanying China tomato yellow leaf curl virus satellite. The method solves the problems existing in traditional biology, serology, electron microscopy observation and other methods. Compared with the traditional method that a virus is detected through one-time PCR, the detection method reduces cost, saves time and is a special, sensitive, economical and convenient 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 provides a fast, simple and effective method for identifying resistance to tomato yellow leaf curl virus, belonging to the field of plant protection. The tomato yellow leaf curl virus is a virus spread by bemisa babaci, the multi-sampling bemisa babaci inoculation identification method is applied to the traditional virus resistance identification; however, as affected by the bemisa babaci (such as inoculation quantity, virus-carrying rate and chow characteristic of the bemisa babaci) the bemisa babaci inoculation identification method is very difficult to provide uniform inoculation pressure and at the same time is interfered by other virus spread by the bemisa babaci, which leads to poor accuracy and repeatability of the identification result. However, the toothpick inoculation method well solves these problems, uses the constructed infective cloning, inoculates the tomato yellow leaf curl virus by puncturing the phloem of a tomato by a toothpick and identifies the resistance of the tomato material to the tomato yellow leaf curl virus. The method of the invention has the characteristics of no need to raise the bemisa babaci, simple operation, high efficiency, good repeatability, good controllability, accurate identification result and capability of providing relatively uniform inoculation pressure, thus being a simple and effective inoculation identification method.

The invention relates to a duplex PCR (polymerase chain reaction) primer and a method for identifying tomato chlorosis viruses and tomato yellow leaf curl viruses. The method comprises the following steps that (1) plant total RNA (ribose nucleic acid) is extracted; (2) the plant total RNA is used as a template for synthesizing cDNA (complementary deoxyribonucleic acid) through reverse transcription; (3) the cDNA is used as a template for performing PCR amplification on a nucleotide sequence coded by ToCV and TYLCV capsid protein; (4) a PCR product prepared in the step (3) is subjected to agarose gel electrophoresis analysis. The invention relates to a PCR primer combination. A duplex PCR method is used for performing ToCV and TYLCV identification; the result is accurate; the method is fast, simple and convenient; the detection cost is reduced; the virus detection efficiency is greatly improved; good application prospects are realized.

The invention discloses a molecular marker method for identifying a tomato yellow leaf curl virus resistance gene Ty-1. The method comprises the following steps: extracting a tomato gene group DNA; performing PCR amplification on the extracted tomato gene group DNA by using a molecular marker primer and performing electrophoretic analysis on the obtained PCR amplification product, representing that the yellow leaf curl virus resistance gene Ty-1 in a plant is a disease-resistant plant if a stripe with the length of 690bp is generated in an electrophoretogram, and representing that the plant is a plant system containing ty-1 gene site and is not a disease-resistant plant if the stripe with the length of 409bp is generated in the electrophoretogram. The electrophoretogram in the marker method is simple in stripe form, easy to identify, can be directly applied to marker assistant selection and can improve the identification accuracy; and moreover, the time is saving, and the material resources and the labor force are reduced.

The invention provides a method for rapidly identifying two similar viruses (a tomato yellow leaf curl virus and a papaya leaf curl China virus) on a tomato, belonging to the field of plant protection. The method comprises the following steps: according to the nucleotide sequences of the tomato yellow leaf curl virus and the papaya leaf curl China virus, designing three primers, i.e. PA_F362, TY F833 and PATY_R, wherein the PA _ F362 and the PATY_ R are combined to detect the papaya leaf curl China virus, and the TY _ F833 and PATY _ R are combined to detect the tomato yellow leaf curl virus;and extracting total DNA of samples, and carrying out PCR only once, thus realizing identifying the tomato yellow leaf curl virus and the papaya leaf curl China virus. The method of the invention overcomes the problems of the traditional methods in biology, serology, electron microscope observation and the like; and compared with the method for detecting a virus by PCP once, the method of the invention reduces cost and saves time, thus being a specific, sensitive, economical and convenient detection method.

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 provides a method for fast inoculating a tomato yellow leaf curl virus and belongs to the field of plant protection. The tomato yellow leaf curl virus is fast inoculated by a stem injection method by using built infectious clone PTYj01. The tomato yellow leaf curl virus is frequently inoculated by the conventional bemisia tabaci gennadius inoculation method in which the bemisia tabaci gennadius needs to be specially fed, and the inoculation efficiency is influenced by the virus transmitting efficiency of the desmone insect and is interfered by the virus transmitting of other bemisia tabaci. The method effectively overcomes the defects of the conventional bemisia tabaci gennadius inoculation method, and has the advantages of no need of feeding the bemisia tabaci gennadius, simple operation, high efficiency, high repeatability, and high controllability, and is a simple and convenient and effective inoculation method.

Molecular markers are described for determining the presence or absence of a gene conferring resistance to tomato yellow leaf curl virus from S. chilense (Ty-1) in a host plant. Also described, are methods for producing a host plant comprising a gene conferring resistance to tomato yellow leaf curl virus from S. chilense (Ty-1), including the analysis of the presence or absence of the molecular markers. A plant, and parts thereof, obtained by such a method are also described.

The invention relates to a CAPS molecular marker based on tomato yellow leaf curl virus disease resistance gene Ty-3. The nucleotide sequence of the molecular marker is disclosed as SEQ ID No.1. The invention also relates to a primer pair for amplifying the CAPS marker based on tomato yellow leaf curl virus disease resistance gene Ty-3. The sequences of the primer pair are as follows: the sequence of the forward primer is 5'TTGCCACATTAAGCAGAACG-3'; and the sequence of the reverse primer is 5'-TGACGGTCATTGAATGTGCT-3'. The marker is a codominant marker based on PCR (polymerase chain reaction) technique, and can obviously lower the cost and save the labor as compared with the RFLP, AFLP, and other markers. Besides, the marker is completely based on the disease-resistant gene Ty-3, is coseparated from the Ty-3 gene, and thus, does not have the problem of false positive when being used for detection. When being used for seedling stage selection, the marker can reduce the workload, and can avoid the difficulty in accurately screening out the individual plant with the disease-resistant gene due to the insufficient inoculation, thereby accelerating the breeding progress.

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 discloses a method of detecting tomato yellow leaf curl virus based on RPA (recombinase polymerase amplification). The invention provides a primer pair for detecting tomato yellow leaf curl virus; the primer pair includes a primer pair (a) composed of two single-stranded DNA molecules shown by sequence 1 and sequence 2 in a sequence table, and a primer (b) as functional as the primer pair (a) and composed of two single-stranded DNA molecules shown by sequences formed by subjecting the sequence 1 and sequence 2 in the sequence table to substitution and / or deletion and / or addition of one or more nucleotides. The RPA primers herein can effectively amplify target genes, the specificity reaches 100%, and sensitivity reaches 6*105 c / mu L; the primers show no cross-reaction with common tomato viruses. The method acts as an available quick detection method for screening plantlets of nurseries, and is significant to preventing spreading of tomato yellow leaf curve virus.

The invention discloses a method for breeding a novel tomato line rich in lycopene and resisting against a yellow leaf curl virus. The method comprises the following steps: screening a recessive mutant gene dg specificity Caps molecular marker according to tomato genome DNA (Deoxyribonucleic Acid) sequence signatures; screening a pair of Caps molecular markers which are closely linked with the anti-TYLCV (Tomato Yellow Leaf Curl Virus) gene Ty-3; hybridizing a tomato material containing the TYLCV resistance gene and the tomato mutant dg material with a common tomato good selfing line; conducting multi-generation selfing and taking the dg and Ty-3 molecular markers as basis to screen single plants; at last, obtaining the excellent selfing line aggregating the dg gene and the Ty-3 gene. The method can also be applied to a hybrid which is obtained by utilizing the selfing line to be hybridized with the other tomato selfing lines.

The present invention relates to the field of tomato plants having an intense phenotype and Tomato Yellow Leaf Curl Virus (TYLCV) resistance.

The invention discloses a preventing and treating method of the tomato yellow leaf curl virus. Tomatoes are cultured in a solar greenhouse according to size, wherein the large line spacing is 80 cm, and the small line spacing is 60 cm; gathering inducing plants are transplanted 15-20 days before tomatoes are planted, wherein a line of gathering inducing plants are planted between two lines of planted tomatoes, and the row spacing is 25 cm; substrate seedling trays containing carbonized rice husks are adopted for culturing seedlings of the gathering inducing plants, carbonized rice husks are additionally applied in planting pits, and seedling tray dipping and root irrigating are conducted through liquid thiamethoxam; before the gathering inducing plants are transplanted, a vent hole of thesolar greenhouse is closed by a 40-mesh sophocarpidine pest-killing protection net. On the premise of greatly reducing the chemical pesticide consumption and achieving main crop non-contact chemical prevention and treatment, the morbidity of the tomato yellow leaf curl virus can be effectively reduced, and the morbidity of the tomato yellow leaf curl virus of the infected tomato species can generally be controlled to be 4% or lower.

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.