Primer developed on the basis of pectin methylesterase Pcpme6 gene and used for detecting phytophthora capsici and method

A technology of pectin methylesterase and Phytophthora capsici, which is applied in the field of primers for detection of Phytophthora capsici, can solve the problem of detection and early warning of pathogenicity characteristics and occurrence trend of pathogenic bacteria, little control effect, and difficulty in controlling the occurrence and prevalence of diseases and other issues to achieve the effect of reducing the cost of disease prevention and control, blocking reproduction and spread

Inactive Publication Date: 2012-08-08
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional disease prevention and control strategies mainly rely on control measures such as varieties, cultivation, chemical control, and ecological regulation. Comprehensive prevention and control and high-efficiency management measures are short and get twice the result with half the effort. The control effect is very small, and it is difficult to control the occurrence and prevalence of diseases in the end.
[0003] In recent years, PCR technology and its related molecular detection technology have been widely used in the molecular detection and early warning of the growth and decline dynamics of plant pathogens. Ribosomal rDNA / ITS sequences are used to design specific primers, and targeted researches on Phytophthora soybean and Phytophthora infestans have been carried out. Molecular detection technology research on mildew, Phytophthora winteris, Phytophthora parasitica, and Phytophthora capsici, but these technical R&D achievements can only detect and warn the activity dynamics of the pathogens, but cannot detect and warn the pathogenic characteristics and occurrence trends of the pathogens

Method used

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  • Primer developed on the basis of pectin methylesterase Pcpme6 gene and used for detecting phytophthora capsici and method
  • Primer developed on the basis of pectin methylesterase Pcpme6 gene and used for detecting phytophthora capsici and method
  • Primer developed on the basis of pectin methylesterase Pcpme6 gene and used for detecting phytophthora capsici and method

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Experimental program
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Effect test

Embodiment 1

[0028]Example 1. Acquisition of the primer pair Pme6F-R for specific detection of Phytophthora capsici and its specificity and sensitivity

[0029] In this embodiment, Phytophthora capsici is used as the test material, and the difference in RNA transcription expression level of these 9 gene members of Phytophthora capsici pectin methylesterase gene Pcpme 1-9 is detected in capsicum disease fruit according to qRT-PCR technology, compares Related data information, Pcpme 6 is defined as the target disease-causing gene, designs and synthesizes many pairs of specific primers of the Pcpme 6 gene (sequence 3), tests the specificity of these primers to all the tested bacterial species and the detection of Phytophthora capsici sensitivity. A pair of primer pair Pme6F-R with higher specificity and sensitivity was selected from it as the primer pair for specific detection of Phytophthora capsici, the upstream primer is Pme6F (20bp), and the sequence is 5`-CCGAAGTTAGCTGGACCGTT-3`; the dow...

Embodiment 2

[0110] Example 2, Pcpme 6 gene-specific primer pair Pme6F-R detects different diseased tissues and diseased field soil DNA PCR

[0111] This embodiment has tested the detection effect of Pme6F-R to Phytophthora capsici in diseased field soil, diseased stem, diseased fruit, diseased leaf, specific method is as follows:

[0112] 1. DNA extraction

[0113] Transfer Phytophthora capsici SD33 to a V8 solid plate, and after culturing in the dark at 25°C for 2-3 days, take a colony piece (2 mm × 2 mm) from the edge of the colony by blowing air, and inoculate the wound on pepper (Zhongjiao No. 6) seedlings (7- 9 leaves) at the base of the stem, fruit, and micro-wounds of bacterial blocks of the same size were inoculated on pepper leaves, and then different diseased tissues were cut out, and diseased tissues were extracted with reference to the NaOH method of Wang et al. (Nucleic Acids Research, 1993, 21:4153-4154) DNA of DNA: Take a section of diseased stem, diseased fruit and diseas...

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Abstract

The invention discloses a primer developed on the basis of a pectin methylesterase Pcpme6 gene and used for detecting phytophthora capsici and a method. The primer developed on the basis of the pectin methylesterase Pcpme6 gene and used for detecting phytophthora capsici disclosed by the invention consists of a single-chain DNA (Deoxyribose Nucleic Acid) shown as a sequence 1 in a sequence table and a single-chain DNA shown as a sequence 2 in the sequence table. The primer can be used for performing qualitative and quantitative detection on phytophthora capsici and different pepper blight materials at different stages, can be used for detecting and prewarning a phytophthora capsici primary infection source and the bacterium population dynamic at every stage of an infection process, is convenient for determining an optimal disease prevention and control period in time, is particularly used for appropriately performing comprehensive prevention, control and treatment at an early bacterium activating period, appropriately treating soil according to the quantity of detected oospores in soil and blocking the propagation and spreading of source bacteria, and contributes to reducing disease prevention and control costs.

Description

technical field [0001] The invention relates to a primer and a method for detecting Phytophthora capsici developed based on the pectin methylesterase Pcpme6 gene. Background technique [0002] Phytopathogenic oomycetes are an important class of plant pathogens, which can infect and harm a variety of plants and cause a variety of plant-destructive diseases, such as Phytophthora infestans, P. capsici, soybean blight P. sojae, P. parasitic, P. cinnamomi and P. hibemalis cause important diseases of potato, pepper, soybean, tobacco, camphor tree and citrus respectively. Serious years and even extinction. Oomycetes mainly use mycelium or oospores to survive the adverse environment in the diseased residue or soil and serve as the source of primary infection. Under suitable conditions, mycelium or oospores can germinate to produce sporangia-release zoospores , spread and infect by rain or air currents and cause plant diseases. Therefore, early and timely inspection and control of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/06C12Q1/04C12N15/11
Inventor 张修国
Owner SHANDONG AGRICULTURAL UNIVERSITY
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