1508 results about "Specific primers" patented technology

The invention discloses a primer combination for detecting monkey pox viruses and identifying monkey pox viruses Ia, Ib and II, and belongs to the technical field of molecular biology, the nucleotide sequences of specific primers and probes are shown as SEQ ID NO.1-SEQ ID NO.12, and the primer combination aims at solving the problem that in the prior art, new evolutionary branch Ib types of monkey pox viruses cannot be accurately typed, and the detection sensitivity is high. According to the present invention, the defects that the monkey pox virus and the orthopox virus have the cross reaction are overcome, the genome difference of different evolutionary branches is analyzed through the bioinformatics system, the primer probe composition capable of simultaneously detecting the monkey pox virus and carrying out accurate typing is analyzed and screened, and the primer combination can detect 200 copies / mL of the monkey pox virus with high specificity and high sensitivity. Meanwhile, detection of the monkey pox virus and identification of the types Ia, Ib and II of the monkey pox virus are realized in a reaction system, so that different monkey pox branches can be distinguished, and treatment and prevention can be performed in a more targeted manner.

The invention discloses a double-dominant SCAR marker for identifying allium fistulosum and allium fistulosum polluted by onion pollen and application of the double-dominant SCAR marker. The specific fragment length of the SCAR marker of an abnormal plant polluted by onion pollen is 1481 bp and 851 bp, the nucleotide sequences of the SCAR marker are shown as SEQ ID NO.1 and SEQ ID NO.2 respectively, the specific fragment length of the SCAR marker of the allium fistulosum is 840 bp, and the nucleotide sequence of the SCAR marker is shown as SEQ ID NO.3; the invention also discloses a specific primer of the SCAR marker. When the marker determined by the method is used for verifying different varieties of green Chinese onions and abnormal plants of the green Chinese onions polluted by the onion pollen, the green Chinese onions and the abnormal plants of the green Chinese onions polluted by the onion pollen can be quickly and stably identified, and compared with a traditional seed purity identification method, the method is not influenced by natural environment and human factors, can be used for batch experiments, improves the identification efficiency, and has a wide application prospect. The invention provides a rapid and feasible method for solving the problem that allium fistulosum varieties are mixed due to the fact that allium fistulosum pollen is polluted by the allium fistulosum pollen cannot be identified in production.

The invention discloses SSR (simple sequence repeat) multiple PCR (polymerase chain reaction) primers for paternity test of grass carp and application of the SSR multiple PCR primers. The primers comprise 16 pairs of specific primers, and base sequences of the 16 pairs of specific primers are respectively shown as SEQ ID NO.1-32 in sequence. The invention further discloses a fish paternity test kit comprising the SSR multiple PCR primer, an SSR multiple fluorescent PCR method for grass carp paternity test and application of the SSR multiple PCR primer, the kit or the method in grass carp paternity test. According to the kit, 16 pairs of specific SSR primers, multiple fluorescent PCR and a universal amplification primer technology are combined, 16 sites can be detected at a time, universal primers can be repeatedly used, and compared with simple single-site detection, the efficiency is improved, and the cost is reduced; the microsatellite loci contained in the invention are 3-6 basic groups, the allele size interpretation is more accurate, and the accuracy of genotype data is improved.

The invention discloses an SCAR marker for identifying onion pollen pollution and application thereof, the specific fragment length of the SCAR marker is 811bp, the nucleotide sequence of the SCAR marker is as shown in SEQ ID NO.1, and the SCAR marker of the onion has no target band; the invention also discloses a specific primer of the SCAR marker. When the marker determined by the method is used for verifying the allium fistulosum of different varieties and the allium fistulosum YZD polluted by the onion pollen, the allium fistulosum and the allium fistulosum YZD polluted by the onion pollen can be rapidly and stably identified, compared with a traditional seed purity identification method, the method is not influenced by natural environment and human factors, batch experiments can be carried out, the identification efficiency is improved, and the method is suitable for large-scale popularization and application. The invention provides a rapid and feasible method for solving the problem that allium fistulosum varieties are mixed due to the fact that allium fistulosum pollen is polluted by onion pollen in production.

The invention discloses a molecular marker closely linked with wheat flag leaf width major QTL and application of the molecular marker in prediction of wheat flag leaf width, and belongs to the technical field of wheat molecular biotechnology and breeding. Wherein the molecular marker is 4BFLW-290, the nucleotide sequences of the molecular marker are as shown in SEQ ID NO: 3 and SEQ ID NO: 4, the specific primer pair of the molecular marker is 4BFLW-290-F / R, and the nucleotide sequences of the specific primer pair are as shown in SEQ ID NO: 1 and SEQ ID NO: 2. The molecular marker 4BFLW-290 provided by the invention fully embodies the flag leaf width major QTL of the wheat variety (line), can quickly and accurately predict whether the wheat variety (line) has the flag leaf width major QTL or not, and provides excellent gene resources and selection tools for wheat yield trait molecular breeding.

The invention discloses protease for improving the resistance of potatoes to late blight as well as a coding gene and application thereof, and belongs to the technical field of genetic engineering and plant disease resistance breeding. The amino acid sequence of the protease StRD21 is as shown in SEQ ID NO.2, or the protease StRD21 has a derivative sequence with the same disease-resistant function. The nucleotide sequence of the coding gene is as shown in SEQ ID NO. 1. The invention also provides a specific primer pair for cloning the gene, a potato recombinant overexpression vector containing the gene and a transgenic host cell. The StRD21 gene is introduced into potatoes and overexpressed, so that the resistance of the potatoes to late blight can be remarkably enhanced. The invention provides an effective gene resource and a biotechnological means for cultivating a new variety of disease-resistant potatoes.

The invention relates to the technical field of crop molecular genetic markers, and discloses an InDel molecular marker of avena nuda and application thereof.The molecular marker is a group of 314bp deletion variation and 2bp insertion variation in the 610439484-610440026 interval of the 5C chromosome position of a reference genome of an avena nuda variety OT3098, the nucleotide sequence of the deletion variation is shown as SEQ ID NO.1, the nucleotide sequence of the insertion variation is shown as SEQ ID NO.2, and the nucleotide sequence of the insertion variation is shown as SEQ ID NO.3. The nucleotide sequence of the insertion variation is 5 '-AA-3'. The molecular marker provided by the invention can realize stable detection in agarose gel electrophoresis by combining a specific primer pair (SEQ ID NO.2-3) with an optimized touchdown PCR (Polymerase Chain Reaction) program, has the advantage of being not influenced by a tissue part and a growth period, and provides an efficient and reliable technical means for oat germplasm identification, seed purity detection and molecular breeding.

The invention discloses a specific primer for distinguishing rice and barnyard grass and application of the specific primer, and belongs to the technical field of biological species identification. The sequences of the specific primers are as shown in SEQ ID NO. 1 and SEQ ID NO. 2. The specific primer is designed according to the nuclear gene ITS sequence of the rice, the specific primer can accurately identify the rice through PCR, the identification method is stable, the detection time is short, the specificity is high, the detection limit is low, the identification method is not affected by material characters, rapid and accurate identification of the rice and barnyard grass can be achieved, and the identification efficiency is high. The problem that a method for effectively and rapidly distinguishing rice and barnyard grass does not exist at present is solved.

The invention provides an RPA-CRISPR-Cas12a system-based detection reagent for the drug resistance of a chilo suppressalis organophosphorus insecticide, a detection method and application, and belongs to the technical field of molecular detection. The invention provides a detection reagent for chilo suppressalis organophosphorus insecticide resistance, the reagent comprises a specific RPA2 primer pair designed for chilo suppressalis acetylcholin esterase ace-1 gene A314S resistance mutation site and crRNA, a double screening mechanism of RPA amplification and CRISPR-Cas12a is combined, wild type and mutant type genes can be accurately distinguished, and the detection reagent can be used for detecting the insecticide resistance of the chilo suppressalis organophosphorus insecticide resistance to the chilo suppressalis organophosphorus insecticide resistance to the chilo suppressalis organophosphorus insecticide resistance to the chilo suppressalis organophosphorus insecticide resistance to the chilo suppressalis. The rapid, sensitive and visual nucleic acid detection of the chilo suppressalis ace-1 gene A314S resistance mutation site is realized, and a visual detection means which is high in specificity, simple and convenient to operate, economical and efficient is provided for field on-site determination of chilo suppressalis resistance populations.

The invention relates to an IGF2BP3 gene molecular marker primer related to pigeon weight traits and application of the IGF2BP3 gene molecular marker primer, and belongs to the technical field of biology. According to the invention, genome DNA of a to-be-detected pigeon is taken as a template, a specific primer is adopted to carry out PCR amplification on the genome DNA, then a PCR amplification product is subjected to Sanger sequencing and SNP molecular marker genotyping, and then pigeon body weight character selection is carried out. The weight of G / G genotype pigeons is higher than that of A / G and A / A genotype individuals, and the weight of A / G genotype pigeons is higher than that of A / A genotype individuals. By eliminating A / G and A / A genotype individuals and retaining G / G genotype individuals, the molecular marker has the beneficial effects that the molecular marker is used as a genetic marker for breeding pigeons, and the pigeons with large weight and uniformity are bred; the method can efficiently and quickly identify the weight traits of the pigeons, improves the uniformity of the weight of pigeons, provides a scientific basis for the early breeding of the pigeons, and better serves the breeding of the pigeons.

A triple droplet type digital PCR (ddPCR) detection method for the severe fever with thrombocytopenia syndrome virus (SFTSV), spotted heat group rickettsia (SFGR) and tsutsugamushi Ot is established. According to the method, specific primers and probes are designed based on an SFTSV L gene, an SFGR 17Kd gene and an Ot 47Kd htrA gene, the reaction temperature and the concentration of the primers and probes are optimized to establish an SFTSV, SFGR and Ot triple ddPCR reaction system, and the specificity, sensitivity and repeatability of the triple ddPCR reaction system are analyzed. According to results, the optimal primer concentration and probe concentration of triple digital PCR are 600nmol / L and 250nmol / L respectively, and the optimal annealing temperature is 60 DEG C; the correlation coefficients of the triple ddPCR SFTSV, the triple ddPCR SFGR and the triple ddPCR Ot standard curve are 0.98, 0.99 and 0.98 respectively, and the lowest detection lower limits are 0.59 copies / mu L, 0.46 copies / mu L and 0.36 copies / mu L respectively. According to the invention, the SFTSV, SFGR and Ot triple digital PCR detection method is established, the sensitivity is high, the specificity is strong, and quantitative detection of three pathogens in SFTS suspected cases can be realized.

The invention discloses a primer group, a kit and a method for detecting brucella, and belongs to the technical field of gene detection. The primer group comprises two pairs of specific RPA (recombinase polymerase amplification) primers: a primer pair B1F1 (SEQ ID No.3) and a primer pair B1R1 (SEQ ID No.6) targeting a B1 sequence, and a primer pair B2F1 (SEQ ID No.8) and a primer pair B2R1 (SEQ ID No.11) targeting a B2 sequence. The invention also provides a kit containing the primer group, and a detection method based on the RPA-CRISPR-Cas12a. The invention also provides a kit containing the primer group and a detection method based on the RPA-CRISPR-Cas12a. According to the application, the high efficiency of RPA amplification is combined with the ultrahigh specificity of CRISPR-Cas12a detection, the defects of long time consumption, low sensitivity and easy generation of false positive in the traditional method are overcome, rapid, sensitive and specific detection of Brucella is realized, and the application has important application value in early screening and monitoring of Brucella.

The invention provides a composition, a kit and a detection method for detecting staphylococcus aureus or a drug-resistant gene thereof based on RPA-CRISPR / Cas12a and application of the composition, the kit and the detection method, and belongs to the technical field of gene detection. A CRISPR / Cas12a system is combined with recombinase polymerase amplification (RPA), an RPA-CRISPR / Cas12a detection method for rapidly detecting the staphylococcus aureus in the wastewater and the drug-resistant gene of the staphylococcus aureus is established, a specific primer and crRNA are designed, the high-specificity nucleic acid cutting capability of a gene editing enzyme CRISPR / CAS system and the isothermal rapid amplification advantage of a nucleic acid isothermal amplification technology are fully fused, and the detection method has the advantages of high specificity, high specificity, high specificity, high specificity, high specificity, high specificity, high specificity, high specificity, high specificity, high specificity, high specificity, high specificity, high specificity, high specificity, high specificity, high specificity, high specificity, high specificity, high specificity, high specificity, high specificity, high specificity, high specificity, high specificity The fluorescent probe has high-specificity sequence recognition capability, realizes efficient detection of target nucleic acid, is strong in detection specificity, high in sensitivity and simple and convenient to operate, does not need complex equipment and professionals, and is suitable for rapid detection in an on-site environment with limited resources.

The invention discloses a gene, a primer and a method for specific molecular detection of alternaria ginseng and application, and belongs to the technical field of molecular detection. Aiming at the problems of long time consumption, low accuracy and lack of specific molecular markers of a traditional detection method for black spot pathogenic bacteria in ginseng planting, a nucleotide sequence as shown in SEQ ID NO: 1 is provided as a specific molecular detection target. The sequence lays a foundation for designing high-specificity primers and supports establishment of a detection system based on conventional PCR and real-time fluorescent quantitative PCR, and the lowest detection concentration reaches 0.5 fg / mu L. The technology is mainly used for rapid qualitative and quantitative detection of ginseng alternaria in ginseng plants, seeds and soil, disease early warning is realized through early pathogen monitoring, and technical guarantee is provided for safety production of ginseng.

The invention discloses a nucleic acid product, a kit and a method for detecting various pathogens. The nucleic acid product comprises a primer pair and a probe, wherein the nucleotide sequences of the primer pair and the probe are respectively shown as SEQ ID NO: 1-24. Specific primer probes are respectively designed for conserved regions of eight respiratory tract pathogenic bacteria, synchronous detection of multiple pathogens is realized through a one-tube eight-color fluorescent PCR, freeze-drying and ultrasonic direct diffusion method technology, experimental operation steps are remarkably reduced, the detection time is shortened, the result interpretation complexity is reduced, and in addition, the detection efficiency is greatly improved. The anti-interference capability is strong, and the sensitivity is high.

The invention relates to the technical field of microbiological detection and molecular biology, in particular to a specific detection primer for bifidobacterium longum subsp. Longum BBMN68 and application of the specific detection primer. The specific primer for detecting the bifidobacterium longum subsp.longum BBMN68, provided by the invention, comprises an upstream primer and a downstream primer, and nucleotide sequences are shown as SEQ ID NO.7-8. The invention also provides a kit for detecting the bifidobacterium longum subsp.longum BBMN68. The specific DNA fragment provided by the invention is a specific molecular marker of BBMN68, and the specific primer is designed aiming at the fragment sequence, so that the specific amplification of the strain can be realized, no cross reaction is caused to other related strains, the detection sensitivity is higher, and the detection requirements of samples with different concentrations can be met; the method can be used for specific detection and accurate quantification of the BBMN68 strain in yoghourt and other products, and has good practical application value.

The invention relates to the technical field of phytopathogen detection, and particularly discloses a primer, a kit and a method for detecting crop root rot fusarium solani and application, the primer comprises a specific RPA primer RPA-F and a specific RPA primer RPA-R; the specific RPA primer RPA-F is as shown in SEQ ID NO. 1, and the specific RPA primer RPA-R is as shown in SEQ ID NO. 2; the kit further comprises a guide RNA primer crRNA, and the guide RNA primer crRNA is as shown in SEQ ID NO. 3. The kit comprises a specific RPA primer RPA-F, a specific RPA primer RPA-R, a Cas12 protein, a T7 transcriptase, a crRNA DNA template, a DNA polymerase and a Cas12 / 13 nucleic acid detection test strip. The kit has the characteristics of good sensitivity, relatively good specificity and enough reliability.

The invention belongs to the technical field of biological detection, and particularly discloses a targeted sequencing primer group and a kit for detecting common pathogens of children and drug-resistant genes thereof. The primer group comprises 145 primer pairs of common children pathogen primer groups and 67 primer pairs of drug-resistant gene primer groups of the common children pathogen primer groups. According to the kit provided by the invention, nucleic acid in a clinical sample is directly extracted, 212 pairs of specific primers are adopted to realize multi-targeted amplification of a pathogen target area, a high-throughput sequencing platform is adopted to accurately identify pathogens and drug-resistant mutant genes thereof, 72 common children pathogens and drug-resistant genes thereof can be detected at the same time, and the kit has a wide application prospect. More accurate diagnosis information is provided for clinic, a key technical support is provided for early-stage accurate treatment of children infectious diseases, and the technical problems that in the prior art, in children pathogen detection, the detection speed is low, the number of missed detection is large, and the drug-resistant blind area is large are solved.

The invention relates to the technical field of biological detection, and discloses a mycoplasma pneumoniae nucleic acid rapid detection method, which comprises the following steps: mixing a sample to be detected with a multifunctional pretreatment buffer solution containing tris (hydroxymethyl) aminomethane, potassium acetate, polyethylene glycol, saponin, spermine tetrahydrochloride and tris (2-carboxyethyl) phosphine hydrochloride, and carrying out cracking treatment; a cracking mixed solution is obtained; redissolving the enzymatic isothermal amplification basic reagent, the specific primer and the probe to obtain a redissolving amplification system; sucking the supernatant of the cracking mixed solution, adding the supernatant into a redissolving amplification system, and adding a magnesium acetate solution to obtain a reaction mixed solution to be detected; and carrying out isothermal amplification and signal acquisition to obtain a detection conclusion. The multifunctional pretreatment buffer solution is used for cracking the to-be-detected sample, sample liquefaction, cracking and nucleic acid release are completed in a single system, the nucleic acid extraction and purification step of centrifugal column adsorption or magnetic bead elution is avoided, and the operation process is simplified.

The invention belongs to the technical field of plant fungus molecular biology detection, and discloses an LAMP primer group, a kit and a detection method for detecting oat smut pathogenic bacteria. A group of LAMP (loop-mediated isothermal amplification) specific primers are designed according to a specific sequence on a whole genome of the oat smut pathogenic bacteria Ustilago hordei, results are judged through a real-time fluorescence quantification method, an agarose gel electrophoresis method and an SYBR Green I fluorescent dye developing method, and the oat smut pathogenic bacteria carried by oat seeds are detected. The LAMP detection method for the oat smut pathogenic bacteria, established by the invention, is strong in specificity, high in sensitivity, high in speed and low in cost, provides a new technical means for detection of the oat smut pathogenic bacteria carried by the oat seeds, and has relatively high practical application value.

The invention discloses a molecular marker linked with yellow peel of bitter gourd and application of the molecular marker. It is found through analysis that mutation of a base G at the 199th site of SEQ ID NO.1 into A is a reason for formation of bitter gourd yellow peel. A specific primer is designed aiming at the mutation site, a bitter gourd genome is amplified by utilizing the primer, a PCR amplification product is subjected to incision enzyme XhoI enzyme digestion, and the bitter gourd variety with yellow peel can be identified according to the size of an enzyme digestion product. The method can accurately identify the peel color in the early growth stage of the bitter gourd, improves the breeding efficiency of the yellow peel variety of the bitter gourd, and has the advantages of high accuracy, simplicity and convenience in operation and the like.

The invention belongs to the technical field of lake and reservoir water source risk prediction, and provides a lake and reservoir water source algae bloom risk early warning system based on a 16S / 18SrRNA gene expression quantity threshold, and the method comprises the following steps: S1, collecting a sample from a lake and reservoir water source surface layer water body; s2, using 7-gate water bloom algae specific primers for blue-green algae, green algae, diatom, euglena, dinoflagellate, chrysophyta and cryptoalga; s3, based on the qPCR standard curve, calculating the copy number of the 16SrRNA / 18SrRNA gene of the seven water bloom algae; s4, when the gene expression quantity of a certain algal bloom algae continuously reaches 106-7 copy number / mL for 7-12 days, determining that the algal bloom algae is in a window phase; according to the threshold value of the gene expression quantity of the water bloom algae 16SrRNA / 18SrRNA, the window period of water bloom algae cells can be accurately recognized, so that early warning of the algae bloom risk in the lake and reservoir water source is achieved, and early warning of algae bloom outbreak in the lake and reservoir water source is achieved one week or above in advance.

The invention relates to the technical field of crop molecular breeding and detection, in particular to a sweet waxy corn molecular detection system based on an sh1 gene InDel marker and application of the sweet waxy corn molecular detection system, the sweet waxy corn molecular detection system is composed of a forward primer, a reverse primer and a transposon specific primer, and the sequences are shown as SEQ ID NO: 1-3. The sh1 gene InDel marker-based sweet waxy corn molecular detection system provided by the invention can be used for corn sh1 gene Mu insertion rapid detection and / or corn molecular marker-assisted sweet waxy corn breeding. By adopting a sweet waxy corn molecular detection system based on the sh1 gene InDel marker, the accuracy rate is 98.7%. And the detection cost of each sample is 0.15 dollars, and compared with the traditional SSR marking technology, the cost is reduced to 3% of that of the traditional SSR marking technology. From DNA extraction to final result interpretation, the whole detection process takes 3 hours.

The invention belongs to the technical field of molecular diagnosis, and particularly discloses a high-risk HPV16 rapid detection system based on RPA-CRISPR / Cas12a, which is prepared by the following steps: (1) mixing a nucleic acid lysis solution containing Tris (pH = 8.0), NaHCO3, polyvinylpyrrolidone and Tween-20 with Proteinase K to treat a sample, and performing pyrolysis at room temperature for 5 minutes to obtain a nucleic acid template; (2) an RPA reaction system contains an RNA inhibitor, a reverse transcriptase and an RPA reaction essential enzyme, and after the template and the starting agent in the step (1) are added, RPA amplification is carried out at 37 DEG C; (3) dropping the reaction liquid on a colloidal gold test strip subjected to position exchange and transformation of a sample pad and a water absorption pad, and carrying out result interpretation; and (4) adding the RPA product into a CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) reaction system containing the LbCas12a protein, the specific gRNA and the probe. According to the scheme, a rapid, high-sensitivity and cross-reaction-free detection system of the HPV16 is realized by optimizing a nucleic acid lysis solution formula, designing a specific primer and gRNA and modifying a test strip for detection.

The invention discloses a kit for simultaneously detecting listeria monocytogenes and a drug-resistant gene fosX, and belongs to the technical field of bacterial molecular diagnosis. In order to solve the technical problems that in the prior art, a traditional method for detecting the listeria monocytogenes and the drug-resistant genes fosX of the listeria monocytogenes is low in efficiency and high in equipment dependence degree, the invention provides a kit for simultaneously detecting the listeria monocytogenes and the drug-resistant genes fosX of the listeria monocytogenes, and the kit comprises an EmDEA specific primer pair and an RNA probe, and a nucleic acid amplification group enzyme system and a signal amplification group enzyme system. The kit for simultaneously detecting the listeria monocytogenes and the drug-resistant gene fosX of the listeria monocytogenes, provided by the invention, has relatively strong specificity, repeatability, stability and reliability, and the sensitivity can be as low as 500 copies / mu L. The kit provided by the invention is simple in operation process, can be carried to rural sites, remote areas or any other scenes needing instant detection for detection, and has a wide application prospect.

The invention discloses a gastric signet-ring cell carcinoma biomarker and a detection kit thereof, belongs to the technical field of biological medicines, and particularly relates to the detection kit of the gastric signet-ring cell carcinoma biomarker, which comprises a total RNA (Ribonucleic Acid) extraction reagent, an RNA purification reagent, a cDNA (Complementary Deoxyribonucleic Acid) first strand synthesis premixing reagent, a rapid fluorescent quantitative premixing reagent (SYBR Green), nuclease-free double distilled water and a specific primer. The RNA purification reagent at least comprises isopropanol, chloroform and absolute ethyl alcohol; the specific primer comprises an upstream specific primer and a downstream specific primer. The detection kit for the gastric signet-ring cell carcinoma biomarker is high in RNA purification and recovery efficiency and excellent in specificity.

The invention discloses a method for detecting drug resistance of botrytis cinerea to succinate dehydrogenase (SDHI) bactericides. On the basis of an RPA-CRISPR / Cas12a system, a specific primer pair amplification target fragment and crRNA for activating Cas12a trans-cleavage activity are designed aiming at key mutation sites related to drug resistance in a bactericide action target SdhB gene, so that fluorescent light visible to naked eyes can be generated only when a resistant strain sequence is matched with the crRNA, and the fluorescent light is opposite to the fluorescent light generated when a sensitive strain sequence is matched with the crRNA. The method provided by the invention overcomes the dependence on laboratories and large instruments in the traditional drug resistance detection method, provides key technical support for accurate prevention and control and drug resistance management of grape gray mold, and provides direct drug use basis for scientific prevention and control in the field.

The invention discloses a targeted sequencing method for various pathogenic microorganisms. The targeted sequencing method specifically comprises the following steps: collecting a biological detection material containing the pathogenic microorganisms; extracting nucleic acid in the pathogenic microorganisms to be detected; designing specific primers and probes according to the target pathogenic microorganisms; amplifying a nucleic acid sequence through a PCR (Polymerase Chain Reaction) technology; carrying out deep sequencing based on a high-throughput sequencer; according to the present invention, the specific primers and the probe are designed, such that the nucleic acid sequence of the target pathogenic microorganism can be efficiently enriched, and the high-throughput sequencing technology is adopted to perform the deep sequencing so as to significantly improve the sensitivity and the accuracy of the detection of the plurality of pathogenic microorganisms; the method can be used for simultaneously detecting various different types of pathogenic microorganisms such as bacteria, viruses, fungi and the like, is simple and convenient to operate, short in time consumption and low in cost, and can provide powerful support for quickly and accurately identifying the pathogenic microorganisms and timely taking effective prevention and control measures.

The invention relates to a wheat stem rot regulatory gene Tatrx-m, and an encoding protein, a recombinant vector and an application of the wheat stem rot regulatory gene Tatrx-m. The genomic sequence of the gene is SEQ ID NO.1, the CDS sequence of the gene is SEQ ID NO.2, and thioredoxin shown as SEQ ID NO.3 is encoded. The invention further provides a specific primer pair used for gene identification, a silent vector (Tatrx-m-VIGS) and an overexpression vector (LGY-OE3-Tatrx-m) are constructed, and the function of the primer pair is verified through a virus-induced gene silencing (VIGS) technology and agrobacterium-mediated transformation of wheat. Experiments show that silence of Tatrx-m results in significant reduction of wheat stem rot resistance, and overexpression of the gene can improve resistance. Based on hypha quantification, DAB staining, H2O2 content and cell death analysis, Tatrx-m is revealed to enhance wheat resistance by regulating active oxygen removal and cell wall strengthening pathways. The gene TaTrx-m has the positive regulation effect in wheat stem rot for the first time, can be applied to disease-resistant molecular marker development, gene editing and transgenic breeding, provides core genetic resources for disease-resistant variety breeding, and has theoretical and application values.

The invention relates to the technical field of sweet potato virus detection, and provides a sweet potato virus G detection method. The invention relates to a sweet potato virus G specific primer pair. The nucleotide sequence of a forward primer SPVG-F of the specific primer pair is as shown in SEQ ID NO: 1; and the nucleotide sequence of the reverse primer SPVG-R is as shown in SEQ ID NO: 2. According to the technical scheme, the problems of low detection sensitivity and low detection efficiency when a sweet potato G virus amplification primer and a sweet potato G virus same family virus compositely infect a sample in the prior art are solved.