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Primer, probe and method for detecting tilletia walkeri

A technology of ryegrass and smut, applied in the detection of T. ryegrass, primers and probes, can solve the problems of limited practicality, large amount of spores, difficulty in obtaining a large number of spores, etc., and achieve shortened detection time and high sensitivity Improved effect

Inactive Publication Date: 2007-10-17
上海出入境检验检疫局
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AI Technical Summary

Problems solved by technology

Conventional PCR technology requires teliospores to germinate, culture and reproduce to obtain hyphae. The whole process of treatment and identification lasts for 2-3 weeks, and the dormancy and non-viability of teliospores also affect the identification of pathogens
If the spores are detected directly, and DNA is extracted from the spores for PCR detection, the amount of spores required is relatively large, and it is difficult to obtain a large number of spores in the actual detection process, and there may be several pathogenic spores in the actual sample
Therefore, the practicability of the above two identification methods is limited to a certain extent.

Method used

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  • Primer, probe and method for detecting tilletia walkeri
  • Primer, probe and method for detecting tilletia walkeri
  • Primer, probe and method for detecting tilletia walkeri

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Embodiment Construction

[0022] The present invention will be described in further detail below in conjunction with the accompanying drawings and embodiments.

[0023] The following examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental method that does not indicate specific condition in the embodiment, generally according to conventional conditions, such as people such as Sambrook, molecular cloning: the condition described in the laboratory handbook (New York: Cold Spring Harbor Laboratory Press, 1989), or according to manufacturer's suggested conditions.

[0024] A TaqMan-MGB (Minor Groove Binder) probe TWAL was designed for the detection of T. rye for the specific site difference (G / A) of the ITS (internal transcribed spacer, ribosomal gene transcription spacer) sequence of the pathogen , established a real time PCR detection technique for a single teliospore, and Figure 1 is a technical flow chart for the dete...

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Abstract

The present invention discloses specific primer and probe for detecting Tilletia walkeri and method for detecting Tilletia walkeri using the primer and probe. The inventive method can shorten greatly the detection time and promote the sensitivity. Single teliospore can be detected. Two kinds of pathogen teliospore can be simultaneously detected in the invention so detection efficiency is promoted.

Description

technical field [0001] The invention relates to agriculture and plant quarantine, in particular to a detection method for Tilletia rye smut, and primers and probes used in the detection method. Background technique [0002] Tilletia indica Mitra is a dangerous pest of concern to the world, and it is also a type of dangerous pest announced by my country. It is mainly distributed in India, Pakistan, Iran, Iraq, Nepal, Afghanistan, Brazil, and Mexico. , the United States and South Africa. Tilletia walkeri is a new species similar to Tilletia indica. In recent years, with the opening of the grain market and the increasing quantity of imported grain, the chances and risks of Tilletia walkeri being introduced are also increasing. Increased, especially the main grain exporting country, the United States, also has the occurrence of Tilletia rye smut, which brings new problems in the detection of germs for my country's port quarantine. Currently commonly used detection and identifica...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11C12Q1/04
Inventor 易建平周国梁印丽萍
Owner 上海出入境检验检疫局
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