64 results about "Rice dwarf virus" patented technology

The invention provides a method for rapidly identifying the rice black-streaked dwarf virus and southern rice black-streaked dwarf virus, belonging to the field of plant protection. Three primers are designed according to the nucleotide sequence of the rice black-streaked dwarf virus and southern rice black-streaked dwarf virus, which are respectively [A] RB1_S9_F, [B] RB2_S9_F and [C] RB_S9_R, wherein the combination of A and C detects the rice black-streaked dwarf virus and the combination of B and C detects the southern rice black-streaked dwarf virus. After the total RNA is extracted fromthe samples, the two viruses of the rice black-streaked dwarf virus and the southern rice black-streaked dwarf virus can be detected by only one RT-PCR, thus realizing the identification of two viruses. Compared with the traditional method in which one RT-PCR can only detect one virus, the invention greatly reduces cost, reduces time, and is a detection method which is special, sensitive, economic and convenient.

The invention provides a pesticide composition, and in particular relates to an active pesticide composition, compounding one or more of toxic fluoride phosphate, the weight ratio of toxic fluoride phosphate to other active substances is 30:0.15 to 3.3:30, and the content of effective components by weight is 1%-90%. The compound composition can be processed into water dispersible powder, suspending agent, microemulsion, aqueous emulsion, water dispersible granule, dry suspension emulsion and seed coating agent and the like, after accessory ingredient or carrying agent is added. The composition can be used to prevent and treat virus diseases caused by rice stripe virus, rice dwarf virus, maze rough dwarf virus, rice black-streaked dwarf virus, tomato virus, cucumber mosaic virus and tobacco mosaic virus, and has the beneficial effects of enhanced effect, concurrent treatment, postponed pesticide resistance, decreased dosage of toxic fluoride phosphate and reduced cost.

The invention discloses a method for obtaining Laodelphax striatellus specially for inoculating black streaked dwarf virus of rice and application thereof in agriculture. The principle of the invention is to obtain Laodelphax striatellus without carrying stripe virus of rice by manual screening, feed the Laodelphax striatellus on rice carrying the black streaked dwarf virus without the stripe virus to obtain virus, transfer the Laodelphax striatellus to healthy rice sprouts without carrying viruses to feed for living through cycle period of the viruses, and further obtain the Laodelphax striatellus carrying the black streaked dwarf virus of the rice without the stripe virus of the rice. The established method for identifying the resistance of a rice variety with the black streaked dwarf virus can suppress interference due to that the Laodelphax striatellus simultaneously transmits the stripe virus of the rice, so that the identifying result is more objective and accurate, and the evaluation of resistance is more scientific and reasonable. The identifying method can be applied in discovery, identification, the evaluation of variety resistance, the breeding of disease-resistant variety, the research on resistance genetic development for rice variety resources and other fields.

The invention discloses a method for obtaining sogatella furcifera carrying SRBSDV (southern rice black-streaked dwarf virus) and an application sogatella furcifera. The method for obtaining the sogatella furcifera carrying the SRBSDV, comprises the following steps of: feeding the sogatella furcifera by utilizing rice infecting the SRBSDV or cryopreserved rice diseased plant so that the sogatella furcifera obtains the SRBSDV, transferring the infected sogatella furcifera to a healthy rice to be fed till passing through a circulation period of virus, and detecting the sogatella furcifera by utilizing an RT-PCR (reverse transcription-polymerase chain reaction) technology and a serology method to obtain the sogatella furcifera carrying the SRBSDV, wherein the carrier rate of the sogatella furcifera can reach more than 50% through a manual feeding manner based on the experiment. Through utilizing the method for obtaining the sogatella furcifera carrying the SRBSDV, the obtained sogatella furcifera carrying the SRBSDV can be applied to screening of SRBSDV-resisting monoclonal antibody to carry out a rice inoculation experiment to identify the resistance of the rice, screen disease-resistant variety and provide a service of the breeding for disease resistance. The invention can be further applied to researching mutual relation between the SRBSDV and a vector insect sogatella furcifera and provides powerful theoretical evidence for prevention and treatment on the viruses.

The invention provides a method for laodelphax striatellus to obtain a rice black-streaked dwarf virus (RBSDV) from refrigerated diseased leaves. The principle of the invention is as follows: storing plant leaves carrying RBSDV by using a refrigerated method, obtaining RBSDV for laodelphax striatellus from refrigerated diseased leaves by a feeding method, and obtaining laodelphax striatellus with RBSDV applied in inoculating and identification after laodelphax striatellus passes a circulative period of virus. The varietalresistance identification method established by the invention can overcome the technical bottleneck of difficult long-term storage of diseased plants by adopting the method of obtaining the virus which feeds laodelphax striatellus with diseased plant better, broadens the varietalresistance identification time, and is hopeful for quickening the screening speed of disease-resistant resources and breeding process of disease-resistant varieties.

The invention discloses molecular markers closely linked to an RBSDV (rice black-streaked dwarf virus) major QTL (quantitative trait locus). The essence of the invention is that according to the linkage and segregation rules, the F2 population of the hybrid generation of Tetep and RBSDV susceptible varieties is taken as the mapping population and the SSR (simple sequence repeat) markers RM5626 and RM7097 on the No.3 chromosome, which are closely linked to the RBSDV resistant major QTL are obtained through analysis of linkage between the results of evaluation of disease resistance and the SSR marker data. Whether a plant has resistance to RBSDV can be judged by detecting the SSR marker banding pattern of the single rice plant of the hybrid combination generation of Tetep and derived varieties (lines) thereof and the RBSDV susceptible varieties. After the SSR markers are applied to assistant selection breeding of the RBSDV resistant varieties of Tetep and derived varieties (lines) thereof / RBSDV susceptible varieties, the defects of poor stability and repeatability of RBSDV resistance evaluation, time and labor waste, high technical threshold and the like in conventional breeding can be overcome and the results can conduce to simplifying the selection method and improving the breeding efficiency and then accelerating the breeding process of RBSDV resistant varieties.

The invention provides a q SRBSDV6 (Southern rice black-streaked dwarf virus 6) and a molecular marker method thereof. BC3F7 is obtained through hybridization, backcross and selfing of Guangxi common wild rice material y11 and the susceptible variety Guanghui 998 and then is subjected to association analysis and genetic linkage analysis to obtain the q SRBSDV6, and the q SRBSDV6 is positioned between RM20148 and RM 20297; when the molecular marker RM20148 primer has q SRBSDV6, the amplification band is 129 bp band; when the molecular marker RM 20297 primer has q SRBSDV6, the amplification band is 172 bp band. According to the invention, the molecular markers of q SRBSDV6 genes can be adopted to detect whether the resistant material y11 and the derived varieties (series) contain the q SRBSDV6, the resistance level of the SRBSDV6 can be predicted, and the selective efficiency of the q SRBSDV6 rice can be greatly improved.

The invention provides a rapid detection technology for rice black-streaked dwarf viruses in plants such as rice, wheat and corn and the like and an application method thereof in the diagnosis of diseases. By the method, the interference of southern rice black-streaked dwarf disease can be eliminated, and plant samples of the rice black-streaked dwarf viruses can be rapidly identified, so that targeted preventive strategies can be used, and the loss brought by the diseases is reduced.

The invention discloses application of OsAGO18 protein or an encoding gene of the OsAGO18 protein or a recombinant vector containing the coding gene to the improvement of the resistance of rice on virus diseases caused by an RDV (Rice Dwarf Virus) or a virus in the same family as the RDV. The amino acid sequence of the OsAGO18 protein is concretely a sequence 2 or a sequence 4 in a sequence table. Experiments prove that the plant disease resistance in the rice with overexpression of the OsAGO18 gene is enhanced; otherwise, the disease resistance of an ago18 mutant is reduced; and the result shows that the OsAGO18 protein coded by the gene achieves an important effect in the anti-RDV reaction of the rice.

This invention discloses a method of improving the dwarf virus resistance of rice. The rice with dwarf virus resistance is obtained by transferring the coding gene of interfering RNA of S2 gene (RDV-S2) of the rice dwarf virus into the cell or tissue infected with dwarf virus. This invention uses RNA interfering technology inhibiting the expression of RDV-S2 gene in rice infected with RDV, which made ent-kaurene oxidation enzyme gibberellin playing normal function, and then the level of gibberellin returns to normal, so the dwarfing symptoms of the rice infected with RDV will disappear and it will return to normal growth. The invention has extensive prospective in RDV prevention and cure field, and it also has positive effect in researching pathogenic mechanism of other Reovirus members.

The invention discloses application of RING1 protein for improving RDV (rice dwarf virus resistance) of plants, and particularly provides application of protein RING1 or a coding gene of the protein RING1 or a recombinant vector containing the coding gene for regulating RDV resistance of plants. The amino acid sequence of the protein RING1 is the sequence one in the sequence table. The experiment disclosed by the invention proves that the RING1 is E3 protein-ubiquitin ligase and capable of interacting with RDV coat protein P2 to promote the P2 to be degraded by 26S proteosome, and the cumulant of the P2 can be reduced by excessively expressing the RING1 protein in rice, thereby inhibiting the amplification of RDV and improving the anti-virus capability of rice.

The invention relates to a rice black-streaked dwarf virus (RBSDV) resistant locus qRBSDV11 of a rice variety 9194 and a molecular marker method thereof. Four RBSDV resistant genetic loci of the RBSDV resistant variety 9194 are obtained by carrying out genetic linkage analysis on the genotype of a single plant F2 obtained by hybridizing the RBSDV resistant rice variety 9194 (male) with a susceptible variety Suyunuo (female) and the RBSDV resistance grade of a corresponding F2:3 family, wherein qRBSDV11 is between a marker RM26062 and a marker RM536. The RBSDV resistance levels of the resistant variety 9194 and derived varieties (lines) thereof can be predicated, and the selection efficiency of the RBSDV resistant rice can be greatly improved by detecting whether the resistant variety 9194 and the derived varieties (lines) thereof contain the RBSDV resistance genetic loci via the molecular markers of the RBSDV resistant genes.

The invention provides a method for SRBSV (southern rice black-streaked dwarf virus) detection on rice plants and accurate quantification of RNA (ribose nucleic acid) copy number. The method can be used for eliminating the interference of the SRBSV, the copy number of the virus RNA in the SRBSV plants is fast obtained, and further, the basis is laid for studying the SRBSV pathogenic mechanism, the molecular biology study and the like.

The invention discloses combinations of reference genes for gene function analysis on an RBSDV (rice black-streaked dwarf virus) infected plant. The combinations are obtained according to the following steps: analyzing the expression stability of a fluorescent quantitative PCR candidate reference gene in the RBSDV infected plant by utilizing geNorm and NormFinder, then the paired difference value V of a normalized factor is calculated after a new reference gene is introduced according to a geNorm program, and determining optimal reference gene combinations according to the variation of V, so as to obtain the optimal reference gene combinations for gene function analysis on the gene function analysis, wherein the optimal reference gene combinations are UBC and Actinl, and when the RBSDV infected plant is taken as a target, the optimal reference gene combinations (UBC and Actinl) can be used for obtaining more accurate and reliable gene expression results as compared with a single reference gene. The combinations can be used for researching the expression level and function of related genes of the RBSDV infected plant and the infecting mode of RBSDV.

The invention provides a method for quickly and synchronously detecting rice stripe virus (RSV) and rice black-streaked dwarf virus (RBSDV) in an insect vector laodelphax striatellus, and belongs to the field of plant protection. According to nucleotide sequences of the rice stripe virus RNA4 and the rice black-streaked dwarf virus S2, three primers are designed as follows: [A] RS-F, [B] RB-F, and [C] RSRB-R, wherein A and C are combined to detect the rice stripe virus; the B and C are combined to detect the rice black-streaked dwarf virus. After extracting total RNA of single laodelphax striatellus, only one RT-PCR is needed, so that the rice stripe virus and the rice black-streaked dwarf virus can be detected, and thus, and quick and synchronous detection for the two viruses in the laodelphax striatellus is realized. Compared with the conventional method for detecting one virus by virtue of one RT-PCR, the cost is greatly saved and the time is saved, and thus, the method is a quick, simple, flexible and specific detecting method. The method disclosed by the invention can be applied to monitoring work of a virus carried rate of a field laodelphax striatellus colony, and can also be applied to related research work of a virus and vector interaction mechanism.

The invention discloses a resistance identification method for southern rice black-streaked dwarf virus variety. The method comprises the steps of (a) acquiring sogatella furcifera, and separating and purifying a large amount of uniform and consistent 5-age sogatella furcifera nymphs; (b) breeding, separating and screening a large amount of uniform and consistent 2-age sogatella furcifera nymphs; (c) feeding virus to the obtained 2-age sogatella furcifera nymphs; (d) calculating the sogatella furcifera nymphs; (e) quantitatively co-breeding rice to be tested and the 2-age sogatella furcifera nymphs fed with virus, and spreading virus; (f) counting land symptomatic appearance, namely, transplanting the rice to be tested to a farm sogatella furcifera separating net chamber, investigating and counting the ratio of infected holes of southern rice black-streaked dwarf virus at the peak tillering phase, so as to accurately identify the resistance of the rice variety to be tested. With the adoption of the resistance identification method, the problems of the interference of antixenosis and the mass acquiring of sogatella furcifera can be overcome; the quantitative and reinforced inoculation identification of southern rice black-streaked dwarf virus can be achieved; the resistance level of the rice variety to southern rice black-streaked dwarf virus can be accurately evaluated.

The invention discloses a hybridoma cell strain excreting a monoclonal antibody (MAb) resisting a rice blackstreaked dwarf virus (RBSDV) and the application of the MAb. A coat protein (CP) of the RBSDV is expressed into antigen immune BALB / c mouse by a prokaryotic expression method, and the hybridoma cell strain 5G1 which can stably passage and excrete the MAb resisting the RBSDV is obtained through cell fusion, selection and cloning, wherein the preserving number is CGMCC No. 5537. Ascitic indirect enzyme-linked immuno sorbent assay (ELISA) valence of the 5G1 MAb is over 10<-6>, and the antibody type and the subtype are IgG1 and kappa chain. The antibody excreted by the cell strain and the coat protein of the RBSDV have specific immunobinding reaction. By the dot-ELISA detection method for detecting rice planthopper and the RBSDV in rice and established by using the 5G1 MAb as a core, the virus can be still detected when a single-head small brown rice planthopper is diluted to 1,600 microlitres and sick leaves are diluted (w / v, g / mL) in the proportion of 1:160. The preparation of the MAb resisting the RBSDV and the establishment of the detection method thereof provide technical and material support for the diagnosis, forecast and scientific prevention and control of rice viruses.

The invention relates to rice black-streaked dwarf virus (RBSDV) RNA interference vectors, a construction method and application, and belongs to the technical field of biology. RBSDV RNA interference vectors, namely pMCG161+ / -B and pCMBIA1301+ / -B, are constructed by using a sequence shown in SEQ ID NO:3 as positive-sense and antisense strands. The invention provides a virus gene interference expression vector and a construction method and application thereof for plant disease-resistant genetic engineering, and provides a novel way of thinking for application of transgenic technology and RNA interference in biological resistance breeding. By transferring the vector into rice, antiviral plants are successfully obtained, the expected goal for obtaining new anti-RBSDV materials of rice is achieved by using an RNA interference principle, successful examples for RNA interference-mediated plant resistance breeding and genetic engineering are provided and a gap of RNA interference-mediated RBSDV resistance in the research of the field is filled.

The invention relates to an RT-PCR detection method of a south rice black-streaked dwarf virus (SRBSDV). In the detection method, SRBSDV specific primers are used for carrying out RT-PCR amplification on a sample to be detected and then carrying out electrophoretic identification. The specific primers are obtained by the method that by utilizing the conservatism of the evolution of viral nucleotide sequences, the tenth section of RNA of SRBSDV and nucleotide sequences of the tenth section of RNA of similar viruses are searched and downloaded from NCBI / GenBank, the sequences are compared by utilizing MEGA4.0 software, and then a pair of primers which are shared by all the SRBSDV nucleotide sequences reported by different researchers and are different from other viral sequences are found out from a file with an extension name of mas. Proved by a BLAST result, sequences which 100 percent cover the primers and 100 percent same as the primers are all the SRBSDV nucleotide sequences. The primers can be used for accurately separating SRBSDV diseased plants from ordinary RBSDV diseased plants without a nested RT-PCR detection.

The invention discloses a primer and a reagent kit capable of being used for detecting southern rice black-streaked dwarf virus (SRBSDV) and also discloses a method for quantitatively detecting the SRBSDV by the reagent kit. According to the method, firstly, the primer is synthesized; then, an RNA (ribonucleic acid) extraction reagent kit is utilized for extracting RNA of the primer from samples; the RNA is used as a template, in addition, a cDNA (complementary deoxyribonucleic acid) synthetic reaction system is utilized for synthesizing cDNA, then, positive standard samples are prepared and are subjected to real-time fluorescence quantification PCR (polymerase chain reaction) procedure drawing, and a standard curve of circulation number-initial concentration logarithmic values with fluorescence signals reaching the threshold values is obtained; and finally, a real-time fluorescence quantification RT-PCR (reverse transcription-polymerase chain reaction) system is prepared by using the synthesized cDNA as the template, the amplified reaction is carried out through the real-time fluorescence quantification RT-PCR procedure, and the initial concentration value of the SRBSDV in the samples is measured according to the collected fluorescence signals and the standard curve. The method has the advantages that the precision is high, the stability is good, the repeatability is good, the specificity is high, the sensitivity is high, and the like.

The invention provides a rapid detection technology for rice black-streaked dwarf viruses in plant hopper and a method for applying the rapid detection technology to disease diagnosis and prediction. By the method, the interference of southern rice black-streaked dwarf viruses can be eliminated, and whether the rice black-streaked dwarf viruses are carried in the plant hopper is rapidly identified, so that a pertinent control strategy is adopted to reduce the loss caused by diseases.

The invention provides RT-PCR used for detecting rice black streaked dwarf virus (RBSDV) and southern rice black-streaked dwarf virus (SRBSDV) and an application of the RT-PCR and belongs to the technical field of PCR detection. Through comparison of genomic sequences of RBSDV with SRBSDV, a conservative-area target sequence is determined, a specific primer pair used for detecting the RBSDV and the SRBSDV is designed, and the nucleotide sequences are as shown in SEQ ID NO. 1-4. The invention further provides a method for detecting the RBSDV and the SRBSDV and a detection kit. The method has the advantages of accurate detection, high flexibility, high specificity, convenience and rapidness, crop infection of the RBSDV or the SRBSDV can be distinguished, low cost is achieved, and good economic value and application prospect are realized.

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 provides a rapid detection technology for southern rice black-streaked dwarf viruses in plant hopper and a method for applying the rapid detection technology to disease diagnosis and prediction. By the method, the interference of rice black-streaked dwarf viruses can be eliminated, and whether the southern rice black-streaked dwarf viruses are carried in the plant hopper is rapidly identified, so that a pertinent control strategy is adopted to reduce the loss caused by diseases.

The invention relates to a rice variety IR24 black-streaked dwarf virus resistant site and a molecular marker method thereof. According to the rice variety IR24 black-streaked dwarf virus resistant site and the molecular marker method, the filial generation F2 population of a rice black-streaked dwarf virus resistant variety IR24 and a susceptible variety is adopted as a mapping population; the rice black-streaked dwarf virus resistance is identified by using a combined method of natural field identification and improved laodelphax striatellus antixenosis identification; linkage analysis is carried out on virus resistance identification results and SSR marker data to obtain a SSR marker, closely linked to rice black-streaked dwarf virus resistance major QTL, on the chromosome 8; the resistance of a plant to the rice black-streaked dwarf virus can be predicated by detecting banding pattern data of the primer marker on the chromosome 8. Therefore, the selecting efficiency of black-streaked dwarf virus resistant rice is greatly enhanced and the rice breeding progress is accelerated.

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 provides application of reducing expression of OsSAMS1 protein and an encoding gene thereof in improving plant resistance to rice dwarf virus. The amino acid sequence of the protein OsSAMS1 is a sequence 2 in a sequence table. The experiments prove that RDV coat protein Pns11 can interact with the rice OsSAMS1 protein, so that the OsSAMS1 enzyme activity is increased, the OsSAMS1 protein excessively-accumulated rice is more susceptible to diseases, and the transgenic rice with the OsSAMS1 eliminated by a CRISPR / Cas9 pathway has enhanced resistance to the RDV.