Primers and diagnostic methods for diagnosing mulberry bacterial wilt

A technique for bacterial wilt, which is applied in the field of primers and diagnosis of mulberry bacterial wilt by PCR method, can solve the problems of high primer specificity and long detection cycle, and achieve high primer specificity, short detection cycle and high sensitivity and high accuracy

Inactive Publication Date: 2012-02-15
ZHEJIANG UNIV
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AI Technical Summary

Problems solved by technology

[0005] The invention provides a primer for diagnosing mulberry bacterial wilt, which has high specificity and solves the problem of long detection period of traditional detection methods

Method used

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  • Primers and diagnostic methods for diagnosing mulberry bacterial wilt
  • Primers and diagnostic methods for diagnosing mulberry bacterial wilt
  • Primers and diagnostic methods for diagnosing mulberry bacterial wilt

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Embodiment Construction

[0023] Primer synthesis

[0024] By comparing the mulberry bacterial wilt pathogen (Enterobacter morus) L1, L2, L3, L4 and 7 international standard strains (the international standard numbers are respectively Enterobacter cloacae subsp.dissolvens LMG2683 T , E. cancerogenus LMG2693 T , E. pyrinus LMG22970 T , E.turicensis LMG23730 T , E. helveticus LMG23732 T , E. nimipressuralisLMG10245 T and E.agglomerans LMG2557 T ) rpoB sequence, such as Figure 4 As shown, two specific regions (gray) of the pathogen of Mulberry bacterial wilt were found. According to the two specific regions, a pair of primers were obtained by using alignment software (ClustalW) and primer synthesis software (Primer5). The upstream primer (EMrpobF) is 5'-GCCAAGCCGATTTCTGGAGCA and the downstream primer (EMrpobR) is 5'-GCATACAGGTTAGCTTTGC, through which a DNA fragment with a size of about 300bp can be specifically amplified. The above primers can be synthesized by conventional methods, and the primer...

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Abstract

The invention discloses a primer for diagnosing mulberry blight. The base sequence of the upstream primer is shown in SEQ ID NO: 1, and the base sequence of the downstream primer is shown in SEQ ID NO: 2. The invention also discloses a method for diagnosing bacterial wilt by using the primer, which comprises the following steps: peeling and chopping the mulberry roots sterilized on the surface, placing them in sterile water, and centrifugally oscillating to prepare an aqueous suspension, and The aqueous suspension is used as a template, and the primers are used to carry out PCR amplification, and the amplified product is separated by gel electrophoresis and developed with an indicator. If there is a DNA band on the gel, the mulberry tree is infected with bacterial wilt, such as If there is no DNA band, the mulberry tree is not infected with bacterial wilt. The invented primer has high specificity, and the detection period of the mulberry bacterial wilt is short by using the primer, which saves more than 90% of the time compared with the conventional method, and has high sensitivity and accuracy.

Description

technical field [0001] The invention belongs to the fields of crop disease prevention and treatment and plant quarantine, and in particular relates to a primer and a diagnosis method for diagnosing mulberry wilt by PCR method. Background technique [0002] Since 2006, a serious mulberry tree disease has been found in the main mulberry tree planting area of ​​Hangzhou, Jiahu, Zhejiang. After the summer pruning, mulberry farmers have a large area of ​​mulberry trees withered, which is called "mulberry plague". After careful observation and in-depth research on the incidence of mulberry gardens and symptoms of diseased plants, it was found that the disease was a brand-new bacterial disease of mulberry trees. , Wang Guofen, Xie Guanlin, Zhou Qin, Xu Fushou, Li Bin, Chen Jianfeng. 2009.Enterobacter spp.: New evidence of causing "mulberry blight". Chinese Science: Series C: 211-219.). [0003] In recent years, due to the continuous spread of the disease and the increasing severit...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11C12R1/01
Inventor 朱勃谢关林楼妙苗王国芬周勤余山红李斌
Owner ZHEJIANG UNIV
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