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Rapid detection method and application of phomopsis nicotianae

A technology of P. pseudomycetes and a detection method, which is applied in the biological field and can solve problems such as inconvenience to distinguish diseases with similar symptoms.

Pending Publication Date: 2021-06-08
HENAN AGRICULTURAL UNIVERSITY
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

Traditional morphological classification methods are inconvenient to distinguish diseases with similar symptoms

Method used

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  • Rapid detection method and application of phomopsis nicotianae
  • Rapid detection method and application of phomopsis nicotianae
  • Rapid detection method and application of phomopsis nicotianae

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0036] 1.1 Tested strains

[0037] From 2016 to 2018, common tobacco fungal pathogens were collected in the main tobacco producing areas of Henan Province, and 11 kinds of fungal pathogens on tobacco were isolated and identified in the laboratory (Table 1). The common tobacco pathogens used in this study were all donated by the Laboratory of Tobacco Research Institute of Henan Academy of Agricultural Sciences.

[0038]

[0039] 1.2 Extraction of genomic DNA

[0040] The genomic DNA of the 11 pathogens tested was extracted using the fungal genome extraction method (CTAB method). The extracted DNA was detected by 1% agarose gel electrophoresis, and the OD of the extracted DNA was measured with an ultramicro spectrophotometer (NanoDrop 2000). 260 / 280 The values ​​are all around 1.8, the purity is high, which can meet the test requirements, and it is stored at -20°C for future use.

[0041] Optimization of PCR amplification conditions

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Abstract

The invention provides a rapid detection method for phomopsis nicotianae and application of the rapid detection method. The rapid detection method for the phomopsis nicotianae comprises the following steps of extracting genome DNA (Deoxyribose Nucleic Acid); designing a specific primer; detecting the specific primer; and detecting the sensitivity of the specific primer. According to the rapid detection method for the phomopsis nicotianae, the molecular information of the phomopsis nicotianae fungi can be rapidly, specifically and sensitively detected, and only one PCR reaction is carried out on genomic DNA of pathogenic bacteria as low as 10 pg; the method overcomes the defect that in the prior art, the time needed for morphological analysis and confirmation is too long only through strain isolated culture, disease control delay caused by the fact that symptoms are not easy to distinguish is avoided, meanwhile, pathogenic bacteria can be accurately detected in time according to the existence of trace pathogenic bacteria, disease control and early warning are facilitated, and meanwhile, a scientific basis is provided for further research of phomopsis.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a rapid detection method and application of Phomopsis tobacco. Background technique [0002] In recent years, due to a series of problems such as the expansion of tobacco cultivation area, continuous crop rotation and single variety, tobacco diseases have become more frequent or more serious. Tobacco diseases have always been an important factor restricting tobacco production. The variety of pathogens is complex, causing Fusarium wilt, black shank, root black rot, and Phomopsis stem blight caused by Phomopsis spp., causing serious harm. [0003] Traditional fungal identification methods have always been mainly based on morphology, by observing the morphological characteristics of the colony after cultivation, observing the structural characteristics with a microscope, whether spores are produced, physiological and biochemical characteristics, and so on. However, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/686C12Q1/04C12R1/645
CPCC12Q1/6895C12Q1/686
Inventor 邢国珍邱睿郑文明苏新宏程琨李博李闯孙帅李淑君李成军何雷
Owner HENAN AGRICULTURAL UNIVERSITY
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