Method for authenticating Peking pantoea through primer pair AHF-AHR

A specific primer pair, pantoea technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve problems such as difficulty in control and loss of industrial production of Pleurotus eryngii

Active Publication Date: 2016-04-20
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

Once bacterial soft rot of Pleurotus eryngii occurs in the production workshop, it is difficult to control, causing serious losses to the industrial production of Pleurotus eryngii

Method used

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  • Method for authenticating Peking pantoea through primer pair AHF-AHR
  • Method for authenticating Peking pantoea through primer pair AHF-AHR
  • Method for authenticating Peking pantoea through primer pair AHF-AHR

Examples

Experimental program
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Effect test

Embodiment 1

[0028] Example 1. Identification of Pantoea beijing with PCR primer pair AHF-AHR

[0029] 1. PCR reagents for identification of Pantoea beijing

[0030] The PCR reagent for identifying Pantoea beijing in this example consists of the specific primer pair AHF-AHR, 2×TaqPCRMix (Bioeasy) and ddH. 2 O composition.

[0031] Among them, the Beijing Pantoea specific primer pair AHF-AHR consists of AHF (upstream primer) and AHR (downstream primer). The nucleotide sequence of AHF is 5'GTCGTTGCTAAAGGCGGGAGC3' (SEQIDNo.1), and the nucleotide sequence of AHR is 5. 'TGTGGGTGATGGTGCGGGTAT3' (SEQ ID No. 2).

[0032] 2. Identification of Pantoea Beijing

[0033] 2.1. Extract the DNA of the strain

[0034] Strains stored from -80℃ (Pantoeabeijingensis sp.nov.) JZB2120001 T ;PantoeagaviniaeDSM22758;PantoeaeucalyptiDSM23077;PantoeaanthophilaDSM23080;PantoeaagglomeransDSM30072;PantoeadispersaDSM30073;PantoeaagglomeransJCM1236;PantoeaagglomeransJCM20143 or Pantoeaplenonov.Jtis Bantoea T ) Streak in solid LB ...

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Abstract

The invention discloses a primer pair AHF-AHR for authenticating Peking pantoea. The primer pair AHF-AHR is composed of a primer AHF and a primer AHR. The primer AHF is a single-chain NDA molecule shown in SEQ ID No.1, and the primer AHR is a single-chain DNA molecule shown in SEQ ID No.2. According to the Peking pantoea specific primer pair AHF-AHR, only a specific stripe is amplified in the genome DNA of Peking pantoea, amplification products are not obtained in other strains, the Peking pantoea specific primer pair AHF-AHR can be used for rapidly authenticating Peking pantoea. The technical support is provided for early preventing infection sources of pleurotus eryngii soft rot and pleurotus nebrodensis soft rot caused by Peking pantoea and timely cutting off the transmission route, and pleurotus eryngii soft rot and pleurotus nebrodensis soft rot caused by Peking pantoea can be early prevented and controlled.

Description

Technical field [0001] The invention relates to a method for identifying Pantoea beijing by using primer pair AHF-AHR in the field of biotechnology. Background technique [0002] my country began to grow Pleurotus eryngii on a large scale in the 1990s. Pantoea includes several species as plant growth promoting bacteria, biocontrol bacteria or plant pathogens (Pantoea: insightsintoa highly versatile and diverse genus within the Enterobacteriaceae, FEMS Microbiology Reviews (2015) 39:968-984). The bacterial soft rot of Pleurotus eryngii caused by Pantoea is a serious disease that has occurred in the industrial production of Pleurotus eryngii at home and abroad in recent years. The pathogen of Pleurotus eryngii soft rot reported in Beijing was identified and named Pantoea Beijing (Pantoeabeijingensis sp.nov., isolated from the fruiting body of Pleurotuseryngii. Liuetal. Antonievan Leeuwenhoek (2013) 104: 1039-1047). Once the bacterial soft rot disease of Pleurotus eryngii occurs i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/689
Inventor 荣成博刘宇王守现马元伟许峰赵爽宋婧祎李瑞杰黄斌王晶王兰青宋爽牛玉蓉
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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