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Primers and method for detecting phytophthora capsici based on development of pectate lyase Pcpe l15 gene

A technology of Phytophthora capsicum and primer pairs, which is applied in the field of primers for detecting Phytophthora capsicum, can solve the problems of little control effect, inability to detect and early warning of pathogenic characteristics and occurrence trends of pathogens, neglect of comprehensive prevention and control measures and efficient treatment measures, etc. Achieve the effect of blocking reproduction and spread and reducing the cost of disease prevention and control

Inactive Publication Date: 2012-09-19
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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  • Primers and method for detecting phytophthora capsici based on development of pectate lyase Pcpe l15 gene
  • Primers and method for detecting phytophthora capsici based on development of pectate lyase Pcpe l15 gene
  • Primers and method for detecting phytophthora capsici based on development of pectate lyase Pcpe l15 gene

Examples

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Effect test

Embodiment 1

[0028] Example 1. Acquisition of the primer pair pel15F-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 Pcpel1-2 (GenBank number is respectively: FJ213434-5), Pcpel4 (GenBank number is: FJ213436), Pcpel9 ( GenBank number is: FJ213442), Pcpel11 (GenBank number is: FJ213444) and Pcpel15 (its nucleotide sequence is the sequence 3 in the sequence table) gene these 6 gene members in the difference of RNA transcript expression level in capsicum disease fruit, compare Related data information, Pcpel15 is defined as the target disease-causing gene, designs and synthesizes many pairs of specific primers of the Pcpel15 gene (sequence 3), tests the specificity of these primers to all the tested bacterial species and the sensitivity to the detection of Phytophthora capsici sex. A pair of primer pair pel15F-R with high specificity and sensitivity was selected from it ...

Embodiment 2

[0111] Example 2, Pcpel15 gene-specific primer pair Pme6F-R detects different diseased tissues and diseased field soil DNAPCR

[0112] In this example, the detection effect of pel15F-R on the diseased field soil, diseased stems, diseased fruits and diseased leaves of Phytophthora capsici was tested. The specific method is as follows:

[0113] 1. DNA extraction

[0114] 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 the DNA of the diseased tissues was extracted with reference to the NaOH method of Wang et al. (NucleicAcidsResearch, 1993, 21:4153-4154) : Take a section of diseased stem, disease...

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Abstract

The invention discloses primers and a method for detecting phytophthora capsici based on the development of a pectate lyase Pcpe l15 gene. The primers consist of single-stranded deoxyribonucleic acid (DNA) shown as a sequence 1 in a sequence table and single-stranded DNA shown as a sequence 2 in the sequence table. The primers can perform qualitative and quantitative molecular detection on phytophthora capsici in different periods and different pepper phytophthora blight materials, detect and warn a primary infection source of phytophthora capsici and the regression trends of pathogenic bacteria in each period in the infection process of phytophthora capsici, determine the optimal control period in time, and particularly perform comprehensive control and treatment in the active period of the early stage of the pathogenic bacteria appropriately, for example, soil is treated appropriately according to presence and number of detected oospores in the soil, so that the reproduction and propagation of source pathogenic bacteria are blocked, and the cost of controlling diseases is reduced.

Description

technical field [0001] The invention relates to a primer and a method for detecting Phytophthora capsici developed based on the pectin lyase Pcpel15 gene. Background technique [0002] Phytopathogenic oomycetes are an important class of plant pathogens that can infect and harm a variety of plants and cause devastating plant diseases, such as Phytophthora infestans, P. capsici, and Phytophthora soybean (P.sojae), Phytophthora parasitic (P.parasitic), Phytophthora camphora (P.cinnamomi) and Phytophthora winteriformis (P.hibemalis) respectively cause important diseases of potato, pepper, soybean, tobacco, camphor tree and citrus. Years and even extinct. 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. T...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/06C12Q1/04C12N15/11
Inventor 张修国
Owner SHANDONG AGRICULTURAL UNIVERSITY
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