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Rapid detection method of pathogens of corn southern leaf blight

A technology of corn spot disease and detection method, which is applied in the field of specific primers for rapid detection of corn spot spot and its detection field, which can solve the problems of difficult pathogenic bacteria, long time-consuming, strong experience, etc.

Inactive Publication Date: 2016-05-04
HENAN AGRICULTURAL UNIVERSITY
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  • Claims
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Problems solved by technology

Traditional pathogen identification methods are time-consuming, low-sensitivity, and highly empirical. It takes several days to isolate and identify pathogens from diseased plants, making it difficult to monitor the occurrence of diseases in a timely manner and effectively control the spread of pathogens and epidemics

Method used

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  • Rapid detection method of pathogens of corn southern leaf blight
  • Rapid detection method of pathogens of corn southern leaf blight
  • Rapid detection method of pathogens of corn southern leaf blight

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Embodiment Construction

[0017] A kind of rapid detection method of corn leaf spot pathogen, described detection method comprises the following steps:

[0018] (1) Collection of fungal hyphae;

[0019] (2) Extraction of fungal DNA;

[0020] (3) PCR amplification reaction.

[0021] The sequence of the forward primer X-EF is shown in SEQ ID NO: 1, and the sequence of the reverse primer X-ER is shown in SEQ ID NO: 2.

[0022] A kind of rapid detection method of corn leaf spot is used as the application of early monitoring of the small spot pathogen in corn leaves, and the specific steps are as follows:

[0023] 1. Collection of fungal hyphae

[0024] Take the preserved bacterial strain (see attached table 1) and inoculate it on a PDA plate, grow it at 25°C for 3 days, pick fresh mycelia, and inoculate it in a 250mL Erlenmeyer flask filled with 1 / 3 volume of PDB. 25°C, 120r / min, culture for 3-4 days, until a large number of mycelium clusters are formed. Filter with double gauze, rinse continuously wi...

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Abstract

The invention belongs to the technical field of gene engineering, relates to a method for rapidly detecting pathogens by a specific primer, and in particular relates to the specific primer for rapidly detecting corn southern leaf blight and a detection method of the corn southern leaf blight. A rapid detection method of pathogens of the corn southern leaf blight comprises the following steps: (1) collecting fungal hyphae; (2) extracting fungal DNAs; (3) carrying out PCR (Polymerase Chain Reaction) amplification, wherein a PCR system is as follows: 1 microliter of a forward primer X-EF, 1 microliter of a reverse primer X-ER, 1 microliter of a DNA template, 12.5 microliters of Taq PCR Master Mix, and the balance of ddH2O which is added until the volume is 25 microliters; a PCR procedure: pre-denaturing for 3 minutes at 94 DEG C; denaturing for 30 seconds at 94 DEG C, annealing for 30 seconds at 52.9 DEG C, and extending for 30 seconds at 72 DEG C, circulating for 32 times; extending for 10 minutes at 72 DEG C and preserving at 4 DEG C. In order to provide evidences for rapidly and accurately monitoring whether corn leaves are latently infected by bipolaris maydis or not, prevention and control measures can be easily and effectively adopted as soon as possible.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and relates to a method for rapidly detecting pathogens with specific primers, in particular to a specific primer for rapidly detecting maize leaf spot and a detection method thereof. Background technique [0002] Corn small spot bacterium is half-knowledge fungus, Helminthosporium genus, Helminthospora maize, formal name used at school: Bipolarismaydis (NisikadoeTMiyake) Shoem, synonym: HelminthosporiummaydisNisikadoeTMiyake, Drechsleramaydis (NisikadoetMiyake) Subram&Jain. The sex is Cochliobolus Heterostrophus (Drechsler) Drechsler, and its synonym is: Ophiobolus Heterostrophus Drechsler. The fungal ascus is black, nearly spherical, 357-642×276-443 (μm), the apex of the ascus is blunt, and the base has a short stalk, the size is 124-183×23-29 (μm). The number of ascospores in each ascus varies, generally about three, and the ascospores are long linear, about 7.3×6.3-8.8 (μm) in si...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04
CPCC12Q1/686C12Q1/6895C12Q2531/113
Inventor 张猛马庆周耿月华武海燕于思勤
Owner HENAN AGRICULTURAL UNIVERSITY
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