Primer for detecting pytophthora capsici developed on basis of plygalacturonase Pcipg8 gene and method therefor

A technology of Phytophthora capsici and primer pairs, which is applied in the field of primers for detection of Phytophthora capsici, can solve the problems of weak control effect, inability to detect and early warning of pathogenic characteristics and occurrence trend of pathogens, ignoring comprehensive prevention and control and efficient control measures, etc. Achieve the effect of blocking reproduction and spread and reducing the cost of disease prevention and control

Inactive Publication Date: 2012-08-15
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 for detecting pytophthora capsici developed on basis of plygalacturonase Pcipg8 gene and method therefor
  • Primer for detecting pytophthora capsici developed on basis of plygalacturonase Pcipg8 gene and method therefor
  • Primer for detecting pytophthora capsici developed on basis of plygalacturonase Pcipg8 gene and method therefor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1. The specificity and sensitivity of the primer pair ipg8F-R for the specific detection of Phytophthora capsici

[0029] In this example, Phytophthora capsici was used as the test material, and the polygalacturonase gene of Phytophthora capsici was detected according to the qRT-PCR technology. There are Principles 2-3 (GenBank numbers: DQ415987, DQ415988), and Principles 5 (GenBank numbers: : EF558847), Principles 8-18 (the nucleotide sequences of which are respectively the sequences 3-13 in the sequence list) of these 14 gene members in the difference of RNA transcription expression levels in pepper disease fruits, compare the relevant data information, Principle 8 is Defined as the target pathogenic gene, designed and synthesized multiple pairs of specific primers for the Principle 8 gene (sequence 3), and tested the specificity of these primers to all tested bacteria and their sensitivity to Phytophthora capsici. A pair of primer pair ipg8F-R with high specific...

Embodiment 2

[0115] Example 2. The PCR detection of the specific primer pair Ipg8F-R of Principle 8 gene for different diseased tissues and soil of diseased fields

[0116] This example tested the detection effect of I pg8F-R on Phytophthora capsici in diseased soil, diseased stems, diseased fruits, and diseased leaves. The specific methods are as follows:

[0117] 1. DNA extraction

[0118] The Phytophthora capsici SD33 was transferred to a V8 solid plate, cultured in the dark at 25°C for 2-3 d, and the colony block (2mm×2mm) was taken from the edge of the colony with blown air, and the wound was inoculated to the pepper (Zhongjiao No. 6) seedling (7- 9 leaves) stem base, fruit, microwounds of the same size bacterial mass were inoculated on pepper leaves, and then different diseased tissues were cut out. Refer to the NaOH method of Wang et al. (Nucleic Acids Research, 1993, 21:4153-4154) to extract diseased tissues. DNA: Take a section of diseased stem, fruit, and diseased leaves respectively, ...

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Abstract

The invention discloses a primer for detecting pytophthora capsici developed on the basis of plygalacturonase Pcipg8 gene and a method for detecting the pytophthora capsici. The primer for detecting the pytophthora capsici developed on the basis of the plygalacturonase Pcipg8 gene consists of single-strained DNA (deoxyribonucleic acid) shown by a sequence 1 in a sequence table and single-strained DNA shown by a sequence 2 in the sequence table. The primer can be used for carrying out qualitative and quantitative molecule detection on the phytophthora capsici leonian and different phytophthoracapsici leonian materials at different periods, and detecting and easily warning the primary infection source of the phytophthora capsici leonian and the dynamic status of the growth and decline of the germs at different periods in the process of infection, so that the optimum disease preventing and controlling period can be conveniently and timely confirmed, particularly the comprehensive prevention, control and treatment can be timely carried out in the early activity period of the germs, according to the existence and the quantity for detecting the oospore in the soil, the soil can be timely treated, the breeding and spreading of source germs can be cut off, and the disease preventing and controlling cost can be reduced.

Description

Technical field [0001] The invention relates to a primer and a method for detecting Phytophthora capsici developed based on the polygalacturonase Pig8 gene. Background technique [0002] Phytopathogenic oomycetes are an important type of plant pathogens that can infect and harm a variety of plants and cause a variety of plant devastating diseases, such as Phytophthora infestans, P. capsici, and soybean disease. P. sojae, P. parasitic, P. cinnamomi and P. hibemalis cause important blights of potato, pepper, soybean, tobacco, camphor tree and citrus, respectively. Severe years and even no production. Oomycetes mainly use mycelium or oospores to survive the bad environment in the diseased body or soil and serve as the source of initial infection. Under suitable conditions, both mycelium or oospores can germinate to produce sporangia-release zoospores , The use of rain or air current to spread, infect and cause plant diseases. Therefore, timely prosecution and control of diseases ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/06C12Q1/04C12N15/11G01N21/64
Inventor 张修国
Owner SHANDONG AGRICULTURAL UNIVERSITY
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