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Primer pair and detection method used for detecting citrus branch tumor pathogenic bacteria

A pathogen detection and pathogen technology, applied in the field of PCR amplification detection, can solve the problems of PCR primer design and other problems, and achieve the effects of high sensitivity, good repeatability and rapid customs clearance

Active Publication Date: 2013-04-03
CHINA CERTIFICATION & INSPECTION GRP HUBEI CO LTD
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  • Abstract
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  • Claims
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Problems solved by technology

Searching from the NCBI database, there are only two ribosomal sequences, and the design of PCR primers has become a problem

Method used

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  • Primer pair and detection method used for detecting citrus branch tumor pathogenic bacteria
  • Primer pair and detection method used for detecting citrus branch tumor pathogenic bacteria

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Embodiment Construction

[0012] The present invention will be described in detail below in conjunction with specific embodiments.

[0013] In this example, the strain Sphaeropsis tumefaciens Hedges used was purchased from the American ATCC Culture Collection Center, No. 20908 TM , and operate and use strictly in accordance with the quarantine dangerous strains. Verified by DNA cloning and sequencing before use.

[0014] In the primer pair provided by the present invention, the nucleotide sequence of the sense primer SPT1 is shown in SEQ ID NO: 1: 5'-GAACGCAGCGAAATGCGATAAG-3'; the nucleotide sequence of the antisense primer SPT2 is shown in SEQ ID NO: 2 Shown: 5′-ATGCTNAAGTTCAGCGGGTAT-3′.

[0015] The development method of the above primer pair is as follows: search for homologous sequences in GenBank with the 437bp ribosomal sequence of Sphaeropsis tumefaciens strain ATCC 20908 (GenBank accession number: JX431882), according to the different taxonomic status and sequence coverage of the strains (le...

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Abstract

The invention provides a primer pair used for detecting citrus branch tumor pathogenic bacteria, wherein the nucleotide sequence of the sense primer SPT1 is represented by the SEQ ID No:1: 5'-GAACGCAGCGAAATGCGATAAG-3', the nucleotide sequence of the antisense primer SPT2 is represented by the SEQ ID No: 2: 5'-ATGCTNAAGTTCAGCGGGTAT-3'. The invention synchronously provides a method for detecting the citrus branch tumor pathogenic bacteria by using the above primer pair, which comprises the following steps of: (1) extracting target object DNA (deoxyribonucleic acid); (2) performing PCR (polymerase chain reaction) amplification aiming at the target object DNA by using the primer pair according to claim 1, so as to obtain a PCR reaction product; and (3) detecting the PCR reaction product by using a gel electrophoresis method. The technical scheme provided by the invention can exactly and rapidly diagnose out whether the citrus carries citrus branch tumor pathogenic bacteria or not; and meanwhile, the method is excellent in repeatability and high in sensitivity, and the type of the pathogenic fungus carried by the citrus is likely to identified out by only 100ng / mu L target object DNA.

Description

technical field [0001] The invention provides a pair of primers for detecting the citrus branch pathogen, and also provides a method for using the primer pair to perform PCR amplification detection. Background technique [0002] Sphaeropsis tumefaciens Hedges, the pathogen of citrus branch tumors, mainly affects Mexican limes, limes and most citrus plants. There are two types of symptoms, branch tumors and arbuscular branches, but branch tumors and branch tumor ulcers are the main symptoms. Diseased plant tissues can transmit pathogenic bacteria. Pathogens can be spread in short distances by wind, rain, etc. Diseased plant tissues such as branches, diseased leaves and reproductive materials are the way to spread pathogens over long distances. Many citrus cultivars are susceptible to this disease and the effects are severe. On May 22, 2008, the EU Food Safety Authority released the risk assessment report on citrus pests completed by the French authorities, which included t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11C12R1/645
Inventor 王振华张建坤叶芸李凤新
Owner CHINA CERTIFICATION & INSPECTION GRP HUBEI CO LTD