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Nucleotide sequence and kit for detecting Burkholderia gladioli

A technology of nucleotide sequence, rot bacteria, applied in the direction of determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., to achieve highly specific effects

Inactive Publication Date: 2013-05-01
BEIJING ENTRY EXIT INSPECTION & QUARANTINE BUREAU INSPECTION & QUARANTINE TECH CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In view of the fact that there is currently no report on the occurrence and harm of onion rot disease in my country, in order to reduce the risk of foreign onion rot pathogen introduction and protect public health, this invention researches and develops a specific onion rot pathogen detection technology

Method used

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  • Nucleotide sequence and kit for detecting Burkholderia gladioli
  • Nucleotide sequence and kit for detecting Burkholderia gladioli
  • Nucleotide sequence and kit for detecting Burkholderia gladioli

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Embodiment 1

[0032] Embodiment 1: Fluorescent quantitative PCR detection of onion rot pathogen

[0033] one. Materials and Methods

[0034] 1. Material Reagent

[0035] Burkholderia gladioli pv. alliicola ATCC20157, ATCC20159, ATCC20160, ATCC20161 were provided by Dr. Hasan Bolkan from Campbell Vegetable Research & Development Center, USA. Burkholderia andropogonis ATCC 23060, Burkholderia caryohpy ATCC 11441, Burkholderia glumae ATCC 33617 was purchased from China Culture Collection Center. Burk. Cepacia Y3, Burk. Multivorans PW99, Burk. Y10, Burk. 317, Burk.stabilis J100, Burk. vietnamiensis 419, Burk. anthina YP46, Burk. pyrrocinia 301, Burk. arboris HT1, Burk. seminaris R45, Burk. Y4 DNA was provided by Professor Xie Guanlin of Zhejiang University. Bacterial culture used NA medium, reaction solution Premix EX Taq and other reagents were purchased from Takara Company; probe primers used in PCR were synthesized from Takara Company.

[003...

Embodiment 2

[0056] Embodiment 2: the kit that detects onion rot pathogen

[0057] A method for detecting onion rot pathogen Burkholderia gladioli pv. alliicola The kit consists of the following components:

[0058] (1) PCR reaction solution: composed of 2× reaction solution Premix EX Taq, primer PBAF, primer PBAR and probe; the final concentration of reaction solution Premix EX Taq is 1×, the final concentration of primer PBAF is 0.3uM, and the final concentration of primer PBAR The final concentration is 0.3uM, and the final concentration of the probe is 0.1uM;

[0059] The 2× reaction solution Premix EX Taq was purchased from Takara Company; the primers and probes were commissioned to be synthesized by Takara Company, wherein the primer PBAF (primer 1) was the nucleotide sequence shown in the sequence table SEQ ID NO.1, and the primer PBAR (primer 2) It is the nucleotide sequence shown in the sequence table SEQ ID NO.2, the probe is the nucleotide sequence shown in the sequence tab...

Embodiment 3

[0075] Embodiment 3: the kit that detects onion rot pathogen

[0076] (1) PCR reaction solution: composed of 2× reaction solution Premix EX Taq, primer PBAF, primer PBAR and probe; the final concentration of reaction solution Premix EX Taq is 1×, the final concentration of primer PBAF is 0.3uM, and the final concentration of primer PBAR The final concentration is 0.3uM, and the final concentration of the probe is 0.1uM;

[0077] The 2× reaction solution Premix EX Taq was purchased from Takara Company; the primers and probes were commissioned to be synthesized by Takara Company, wherein the primer PBAF (primer 1) was the nucleotide sequence shown in the sequence table SEQ ID NO.1, and the primer PBAR (primer 2) It is the nucleotide sequence shown in the sequence table SEQ ID NO.2, the probe is the nucleotide sequence shown in the sequence table SEQ ID NO.3, and the 5' end of the probe is labeled with the reporter fluorescent dye 6-Carboxy fluorescein ( FAM), the 3′ end was lab...

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Abstract

The invention discloses a nucleotide sequence and kit for detecting Burkholderia gladioli and belongs to the field of plant inspection and quarantine. Primers of fluorescence quantitative PCR (polymerase chain reaction) for detecting Burkholderia gladioli has oligonucleotide sequences shown in SEQ ID No.1 and SEQ ID No.2. The invention has the advantages that: with the fluorescence quantitative PCR detection method and kit provided by the invention, the detection of Burkholderia gladioli is specific, sensitive and rapid.

Description

technical field [0001] The invention relates to a nucleotide sequence and a kit for detecting onion rot pathogen, belonging to the field of plant inspection and quarantine. Background technique [0002] Onion rot pathogen ( Burkholderia gladioli pv. alliicola ) belongs to Procaryoces, Gracilicutes, Enterobacteriaceae, Burkholderia ( Burkholderia ), Burkholderia gladiolus ( Burkholderia gladioli ). Gladiolus Burkholderia ( Burkholderia gladioli ) is a pathogen that causes gladiolus bulb rot, first described by Severini in 1913 and named as Pseudomonas gladioli . 1992, Pseudomonas gladioli and the other six belong to the same genus Pseudomonas ( Pseudomonas ) The strains of rRNA Group Ⅱ were divided into a new genus - Burkholderia ( Burkholderia ), thus being named Burkholderia gladioli . [0003] Onion rot fungus was originally recognized as a plant pathogen. The host plants are mainly alliums ( Allium cepa ) and tulips ( Tulipa spp.). The main pathog...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11G01N21/64
Inventor 高文娜王中康周琦黄魁建汪万春李建光李子龙江丽辉吕玉峰边勇王文静邓丛良梁新苗
Owner BEIJING ENTRY EXIT INSPECTION & QUARANTINE BUREAU INSPECTION & QUARANTINE TECH CENT
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