48 results about "Soybean mosaic virus" patented technology

The invention belongs to the technical field of biology, and relates to a disease resistance detection of a plant antiviral gene and an application thereof. According to the invention, a soybean endogenous antiviral gene Gm-RSMV3 is used to construct a transformation vector which is used for transforming sensitive varieties of soybean mosaic virus; and the transformed plants are subjected to herbicide resistance screening, and then subjected to genome DNA PCR, bar gene expression detection, RT-PCR, Southernblot and inverse PCR, thereby verifying that the obtained plants are indeed transgenic and exogenous genes are correctly expressed. T0 generation plants are selfed to obtain T1 generation plants which are also selfed to obtain T2 generation homozygous plants, and the obtained T2 generation homozygous plants are inoculated with the soybean mosaic virus. A disease resistance testing is carried out and results show that transgenic plants of varieties being originally not resistant to the soybean mosaic virus are relatively high in disease resistance. Conventional cross-breeding and transgenic technologies can be used for enhancing disease-resistance performance of plants and increasing production.

The invention discloses molecular markers of soybean mosaic virus resistant gene (RSC8) and applications thereof. The invention discloses two pairs of novel primers for screening resistant gene (RSC8) related with soybean anti-soybean mosaic virus in the soybean molecular breeding field, a preparation method of molecular markers and applications. The analysis results show that the provided molecular markers (ZL-39 and ZL-52) can precisely trace the resistant gene (RSC8) of a soybean disease-resistant species (Kefeng No.1), and can predict the resistance of soybean to soybean mosaic virus, so that researchers can more conveniently identify and screen anti-soybean mosaic virus materials under the laboratory conditions. The molecular marker assisted breeding technology can avoid the influences of environmental factors and artificial factors on phenotype; and the provided two pairs of molecular markers (ZL-39 and ZL-52), which are tightly interlocked with the resistant gene (RSC8), can improve the accuracy in selection of anti-soybean mosaic virus breeding materials so as to achieve the goals of breeding soybean species which can resist soybean mosaic virus.

The invention relates to a method for culturing anti-soybean-mosaic-virus transgenic plants, and belongs to the technical field of plant biology. According to the method provided by the invention, a sequence of an RNA (ribonucleic acid) fragment shown by SEQ-1 or the sequence complementary with the segment is used. The RNA fragment can be used for interfering the copying and the movement of the soybean mosaic virus so as to inhibit the development of the soybean mosaic virus symptom. The method for culturing the anti-soybean-mosaic-virus transgenic plants provided by the invention comprises the following steps of expressing the RNA molecules in the target plant; obtaining the transgenic plants with the obviously improved resistance on the soybean mosaic virus. The method has an important application value on the culturing of wide-spectrum anti-soybean-mosaic-virus plants.

The invention discloses a one-step loop-mediated reverse transcription isothermal amplification detection method for soybean mosaic virus. The primer group is used to carry out RT-LAMP (Reverse Transcription Loop-Mediated Isothermal Amplification) reaction; the reaction product is placed to 1% of TBT (Tetrabromoethane) agar gel for electrophoresis or visualization is carried out to the amplification product by using a fluorescent dye SYBR Green I; if the reaction is positive, the soybean mosaic virus exists; if the reaction is negative, the to-be-detected sample does not contain soybean mosaic virus. According to the one-step loop-mediated reverse transcription isothermal amplification detection method for soybean mosaic virus disclosed by the invention, the RNA (Ribonucleic Acid) quick crude extracting method is combined with the RT-LAMP detection, so that the total RNA nucleic acid of the quickly and crudely extracted to-be-detected sample is used as a reaction template of the RT-LAMP, therefore, the redundancy time for the conventional RNA extracting method of the RT-LAMP is saved, the operation process is simplified, the detection speed is improved, and the one-step detection for the soybean mosaic virus is realized.

The invention provides a soybean mosaic virus resistant gene GmNN1 and a susceptible allelic gene thereof. Nucleotide sequences of the soybean mosaic virus resistant gene GmNN1 and the allelic gene are respectively shown as SEQ ID No.1 and SEQ ID No.2. The soybean mosaic virus resistant gene GmNN1 and the allelic gene have the advantages that difference of expression of the GmNN1 among different soybean resistant and susceptible varieties is analyzed, participation of the GmNN1 to SMV (soybean mosaic virus) resistant reaction is clarified, difference between GmNN1 gene sequences of the different soybean SMV resistant and susceptible varieties is studied, functional markers GmNN1-AC/GmNN1-GT are developed according to the difference between the sequences, association between the markers and the disease resistance of soybean is verified by the aid of SMV resistant and susceptible RIL (recombinant inbred line) populations, and the different SMV resistant and susceptible soybean varieties can be differentiated by the aid of the markers; functional genes and selection markers can be provided for SMV resistant soybean breeding by study results. The invention further provides a method for identifying the functional markers of the soybean mosaic virus resistant gene GmNN1 and application of the soybean mosaic virus resistant gene GmNN1 to breeding selection.

The invention discloses a soybean mosaic virus major gene locus and application thereof, and in particular relates to the soybean mosaic virus major gene locus and the application of the soybean mosaic virus major gene locus in molecule marker assistant selective breeding. The soybean mosaic virus major gene locus is qRSC.13-1 with a contribution rate of 32.4%; the molecule which is tightly interlocked with the mosaic virus major gene locus is marked as GmSSR13-23. The tightly interlocked molecule marker is utilized to detect whether the breeding population system contains the major gene locus or not, so that the mosaic virus performance can be predicated, and the selection rate of the soybean mosaic virus breeding is greatly improved.

The invention provides an SMV (soybean mosaic virus)-resistant gene GmST1, a cultivation method of GmST1 transgenic soybeans and application, and belongs to the technical field of soybean genetic breeding. Aiming at solving the problems of polyploidization and low conversion rate of cultivation of disease-resistant varieties with a traditional method, the cultivation method of the SMV-resistant transgenic soybeans is provided and comprises steps as follows: the SMV-resistant transgenic soybean gene GmST1 with the nucleotide sequence represented as SEQ ID NO:1 is constructed to a plant expression vector, and a soybean gene GmST1 recombinant vector is obtained; the recombinant vector is introduced into agrobacterium and infects the soybeans, and after cultivation, a transgenic positive plantis obtained by identification. The SMV-resistant gene GmST1 and the cultivation method can be applied to SMV-resistant crop breeding work.

The invention discloses a soybean mosaic virus sequence capable of inhibiting growth of agrobacteria, and proves that the sequence can be used for directly screening recombinant clones in agrobacteria. The agrobacteria are important host bacteria in a plant transformation system, but the constructed recombinant clones need to be screened in Escherichia coli before being transferred to the agrobacteria. The soybean mosaic virus strain SC15 has the characteristic of inhibiting growth of the agrobacteria, but strains SC3 and SC7 do not have the same effect; by virus sequence truncation experiments, the sequence SC15P associated with growth inhibition of the agrobacteria in SC15 is identified; a vector compatible with a gateway cloning system is constructed by the sequence, LR reaction and transformation experiments are conducted, and finally, the assumption of recombinant clone in the agrobacteria by SC15P is achieved.

The invention discloses a preservation method of soybean virus. The preservation method provided by the invention comprises the following steps: a virus preserving fluid is adopted for preserving soybean virus; the virus preserving fluid comprises a polymer in the formula (I) shown in the specification and ascorbic acid; and n represents a natural number ranging from 20 to 70. Experiments prove that the soybean virus preserved by using the preservation method provided by the invention can completely maintain activities within three years under an extra low temperature of -80 DEG C, and can be taken as a drug source for being used directly at any time without need of excitation, the efficiency for identifying the soybean virus is improved greatly, unnecessary subcultures during the multiplication process of the mosaic virus are reduced, mutual cross infection of virus seeds can be avoided, the probability of virus variation is lowered, and the genetic stability of viruses can be kept. Therefore, the preservation method provided by the invention has a significant application value.

The invention provides a soybean GmST1 gene mutant plant and a preparation method thereof and belongs to the technical field of plant biology. The method for preparing the soybean GmST1 gene mutant plant to define the disease resistance function of genes related to anti-mosaic virus of soybeans comprises the following steps: designing a gRNA single target primer according to a soybean GmST1 gene;preparing a gRNA single target primer dimer; inserting the primer dimer into a Cas9/gRNA vector to construct a soybean GmST1 gene knock-down vector; transforming soybeans with the GmST1 gene knock-down vector constructed in the step (3), performing screening to obtain the soybean GmST1 gene mutant plant. The method determines that the GmST1 can participate in the resistance reaction of the anti-mosaic virus of soybeans and provides theoretical basis to researches on anti-mosaic virus related genes of soybeans.

The invention discloses a soybean mosaic virus SC18 line resistance gene GmNIK and an application thereof. A nucleotide sequence of the gene GmNIK is shown as SEQ ID NO. 1, and an amino acid sequenceof the encoded protein is shown as SEQ ID NO. 2. A bean pod mottle virus-induced gene silencing technology (VIGS) is used to silence the GmNIK gene in a diseased variety Kefeng No.1 and inoculate theSC18 line, the disease-resistant variety Kefeng No. 1 generates symptoms of necrosis and floral leaf disease sensitivity, after GmNIK is silenced, the resistance of the disease-resistant variety Kefeng No.1 to the soybean mosaic virus SC18 is changed, and the disease resistance is susceptible. The GmNIK gene of the Kefeng No.1 determines the resistance to the SC18 line. The introduction of the GmNIK gene by a transgenic technology or molecular marker-assisted selection is expected to increase soybean resistance to the soybean mosaic virus.

The invention discloses a method for assisting in authenticating a soybean mosaic virus resistant gene and application thereof, and belongs to the field of the soybean genetic technology. According to the method, DNA is separated and extracted from soybeans to be tested, PCR amplification is carried out by marking primers of SSR 64 and primers of SG1 through molecules, whether marked genetic locus bands exist in amplification products of the primers of the SSR 64 or not is detected through electrophoresis and/or the gene types of the amplification products of the primers of the SG1 are analyzed by utilizing a genetic typing analysis system, and whether soybean germplasm resources to be detected have the mosaic virus resistant gene or not is determined. By the adoption of the method, high efficient selection of resistance materials of soybean mosaic virus N1 strain can be achieved, select breeding of new strain of anti-SMV soybeans can be quickened, a novel soybean cross parent selection system is established, waste of manpower and material resources can be reduced, seed breeding efficiency and accuracy are improved.

The invention discloses a primer, a probe, a kit and application for detecting soybean mosaic virus based on an RPA technology. A base sequence of an upstream primer is shown as SEQ ID NO.1, the basesequence of a downstream primer is shown as SEQ ID NO.2, and the base sequence of the probe is shown as SEQ ID NO.3. Besides the primers and the probes, the kit also comprises one or more of a flow measurement chromatography test strip, a flow measurement chromatography test strip buffer solution, an RPA enzyme, an RPA buffer solution, a magnesium acetate solution, ddH2O and a reference substance.The invention also discloses a method for detecting the soybean mosaic virus. Specific primers and the probes are designed based on a soybean mosaic virus CP gene, and the kit is prepared by a recombinase polymerase amplification-flow chromatography method. The sensitivity is high, the specificity is strong, the diagnosis is accurate, the whole detection process only needs 30 minutes, and the judgment of the result is extremely simple and convenient.

The invention discloses a soybean mosaic virus infectious clone vector capable of infecting tobacco, an agrobacterium strain and application of the s vector and the agrobacterium strain. The sequenceof the soybean mosaic virus infectious clone vector capable of infecting tobacco is shown as SEQ ID NO.1. The soybean mosaic virus infectious clone vector pCB301-SMV (SC7) is applied to tobacco infection. According to the invention, a soybean mosaic virus strain (SC7) capable of infecting tobacco is selected to construct an infectious clone vector, agrobacterium tumefaciens is transformed and injected into the tobacco, and then systematic infection is successfully formed on the tobacco. Therefore, a solid and reliable foundation is laid for the application of tobacco in the aspect of soybean mosaic virus research.

The invention discloses application of anti-soybean mosaic virus gene GmRDBM. A gene GmRDBM for encoding a CC-NBS-LRR functional domain is cloned from big white gastrodia of an anti-soybean mosaic virus variety. The gene is sequenced, and the amino acid sequence encoded by the gene is obtained by means of software prediction. A Crispr/Cas9 technology is used for editing the GmRDBM sequence of thebig white gastrodia, and plants knocked out of GmRDBM are inoculated with a soybean mosaic virus for carrying out an experiment; the gene is proved to be the anti-soybean mosaic virus gene of the bigwhite gastrodia variety. The obtained gene and a protein sequence thereof can be used for the improvement of resistance of soybeans to the soybean mosaic virus.

The invention provides a primer pair and an internal standard amplification sequence. The primer pair has a nucleotide sequence represented by SEQ ID NO: 1 and a nucleotide sequence represented by SEQ ID NO: 2, and the internal standard amplification sequence has a nucleotide sequence represented by SEQ ID NO: 3. The invention further provides an application of the primer pair and the internal standard amplification sequence in the preparation of a product for screening or auxiliary screening of soybean mosaic viruses and a method for screening the soybean mosaic viruses based on an RPA-IAC technique. Experiments prove that the screening method of the soybean mosaic viruses established by combining with the internal standard amplification sequence based on an RPA-IAC primer is good in specificity, high in sensitivity, short in screening time, and wide in application range, has a good quality control effect, does not require a special instrument and has guiding significance to the screening of imported and exported relevant plants and products.

The invention discloses an SSR marker CH0211 for identifying the resistance of soybeans to an SMV (Soybean Mosaic Virus) epidemic strain SC3 as well as a detection method and application of the SSR marker CH0211. The SSR marker CH0211 which is newly developed and closely linked with the gene of the anti-soybean mosaic virus SC3 strain is used for carrying out disease resistance selection on a soybean variety so as to obtain a new soybean germplasm of the anti-SMV SC3 strain. According to the method, the defects of long period, large workload, low selection efficiency, susceptibility to environmental influence on selection of disease-resistant phenotypes and the like of conventional soybean mosaic virus-resistant breeding are overcome, the selection efficiency of disease-resistant breedingcan be greatly improved, possibility is provided for direct selection of genotypes, and the disease-resistant breeding process of soybeans is greatly promoted.

The invention discloses a variety breeding method. According to the variety breeding method, a drought-resistant soybean variety, a saline-alkaline-tolerant soybean variety, a budworm-resistant variety, a cyst nematode disease-resistant variety, a soybean downy mildew-resistant variety, a soybean grey spot-resistant variety and a soybean mosaic virus disease-resistant variety are used as cross parents, and a variety is bred through hybridization and multiple-cross polymerization and through multi-generation stress resistance and disease and pest resistance identification and screening. The high-stress-resistance soybean variety which can highly resist seven kinds of stress and diseases and pests and is excellent in agronomic and yield traits is bred. Compared with a common soybean variety, the yield of the bred soybean variety is increased by 15-18% per mu, the pesticide spraying cost is reduced by 80-90 yuan per mu, the economic benefit can be increased by 300-450 yuan per mu, meanwhile, the variety can be planted in arid areas and saline-alkali areas, the economic income of farmers in the arid areas and the saline-alkali areas can be greatly increased, and the life of the farmers is improved; and through a seed screening machine, seeds are selected, and the seeds which cannot well develop are removed, so that the variety excellencyof the final seeds is ensured, and the survival rate is increased.

Provided are a bavachalcone soluble concentrate and a preparing method and application thereof. The bavachalcone soluble concentrate relates to an agricultural soluble concentrate and is composed of raw materials according to weight percentage: 0.01-5% of bavachalcone, 8-20% of emulsifiers and solvents which complement the solution to be 100%. The preparing method of the bavachalcone soluble concentrate is that pure bavachalcone or bavachalcone plant extracts are mixed with the solvents, dissolved, added with the emulsifiers, stirred and uniformized, the insufficient portion is complemented by the solvents to be 100%, and the 100% solution stands for 4-5 hours. The bavachalcone soluble concentrate is used for preventing plant fungus diseases such as rice blast, apple rotting, cucumber anthracnose, tomato late blight, hot pepper anthracnose, cucumber gray mold and rice sheath blight disease or plant viruses such as tomato mosaic virus, soybean mosaic virus, tobacco mosaic virus and tobacco and potato Y virus. The bavachalcone soluble concentrate has good mutual solubility with water, and accordingly the sterilizing effect in practical application is greatly improved and the bavachalcone soluble concentrate is stable in active constituent content.

The invention discloses a mosaic-virus resistance GsCAD1 gene separated from wild soybeans. The nucleotide sequence of the GsCAD1 gene is shown in SEQ No.1 and closely related to mosaic virus resistance. The invention further discloses a protein encoded by the GsCAD1 gene. The amino acid sequence of the protein is shown in SEQ No.2. The invention also discloses application of the GsCAD1 gene. Whenin use, the GsCAD1 gene is introduced into a target plant to obtain a transgenic plant of which the plant mosaic-virus resistance is higher than that of the target plant. Overexpression of the GsCAD1gene can significantly improve the mosaic-virus disease resistance of the soybeans, a method for permanently and effectively preventing and treating soybean mosaic-virus diseases is provided, and thus the yield of the soybeans and economic benefits of farmer households are ensured.