Molecular detection primer and method for potato rhizoctonia solani
A technology for R. solani and molecular detection, which is applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the time-consuming and laborious detection of R. solani potato disease and various fungi Combined infection and other problems, to achieve the effect of easy satisfaction of detection conditions, convenient storage and transportation, and easy access
Active Publication Date: 2015-01-07
黑龙江省农业科学院植物脱毒苗木研究所
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Problems solved by technology
[0003] The present invention aims to solve the practical problem that the traditional method is time-consuming and laborious to detect Rhizoctonia solani disease and is difficult to distinguish and identify when multiple fungi are infecting together, and provides molecular detection primers and detection methods for Rhizoctonia solani
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specific Embodiment approach 1
[0014] Embodiment 1: The molecular detection primers for Rhizoctonia solani of this embodiment, the primer sequences are shown in Seq ID No: 1 and Seq ID No: 2 in the sequence table.
specific Embodiment approach 2
[0015] Specific embodiment two: the molecular detection method of the Rhizoctonia solani of the present embodiment, it is carried out according to the following steps:
[0016] One, take the potato genomic DNA to be detected as template, adopt the primer of claim 1, carry out PCR amplification;
[0017] 2. Get the PCR product for gel electrophoresis detection. If a fragment with a size of 242 bp can be detected, it can be determined that there is Rhizoctonia solani in the potato.
specific Embodiment approach 3
[0018] Embodiment 3: This embodiment is different from Embodiment 2 in that: the extraction site for extracting the potato genomic DNA to be detected is potato rhizome or tuber. Others are the same as in the second embodiment.
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The invention relates to a molecular detection primer and method for potato rhizoctonia solani, and solves the actual problems of time and labor waste during detection of potato rhizoctonia solani disease and difficult distinguishing and identification of various fungi during complex infection in a conventional method. The molecular detection method comprises the following steps: by taking the standard strains of an anastomosis group capable of causing the rhizoctonia solani disease of potato tissues as positive materials, the potato tissues as negative materials, and phytophthora infestans causing potato late blight, alternaria dauci causing potato early blight and three main fusaria which cause potato dry rot as identification materials, extracting total DNA (deoxyribonucleic acid) respectively, and performing PCR (polymerase chain reaction) detection by specific primers, wherein the positive materials can be amplified to obtain specific fragments and the stripes cannot be generated on the negative materials and the identification materials. The molecular detection method is simple, accurate in result, high in sensitivity and good in repeatability, and can be applied to detection of a large amount of samples.
Description
technical field [0001] The invention relates to a molecular detection primer for Rhizoctonia solani and a detection method thereof. Background technique [0002] Rhizoctonia solani is an important soil-borne disease that threatens potato production in my country. The potato canker (seedling stage) and black mole (tuber) caused by it can not only greatly reduce the potato yield but also seriously affect its commerciality. In view of the prevalence of the disease in potato-producing areas in my country, rapid and accurate detection techniques and methods are crucial for the scientific control of Rhizoctonia solani disease. At present, the detection of Rhizoctonia solani is still mainly based on the traditional isolation and culture method, that is, the results of morphological identification are used as the basis for discrimination. However, this method has many steps and a long cycle. Once the sample is not properly separated, it will lead to corresponding information. lost...
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IPC IPC(8): C12Q1/68C12Q1/04C12N15/11
CPCC12Q1/6895
Inventor 杨帅闵凡祥高云飞王文重郭梅李学湛吕典秋王晓丹
Owner 黑龙江省农业科学院植物脱毒苗木研究所
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