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PCR detection method of sugarcane white leaf disease phytoplasma

A phytoplasma and white leaf technology, applied in the field of PCR detection of sugarcane white leaf disease phytoplasma, can solve the problems of cumbersome nested PCR experiment process, high sequencing cost, low specificity and other problems, saving detection time and reducing detection Cost, good reproducibility

Inactive Publication Date: 2019-02-22
SUGARCANE RES INST OF YUNNAN ACADEMY OF AGRI SCI
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Problems solved by technology

[0003] At present, the detection method of sugarcane white leaf disease phytoplasma is nested PCR detection using universal primers (P1 / P7 and R16F2n / R16R2) designed based on phytoplasma 16S rRNA gene. The sensitivity is not strong, and the product needs to be sequenced to determine whether it is the phytoplasma of sugarcane white leaf disease. body method

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  • PCR detection method of sugarcane white leaf disease phytoplasma

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Embodiment Construction

[0015] Specific examples of the present invention are described in detail below to facilitate a further understanding of the present invention, and those not specifically described in the examples are conventional methods.

[0016] 1. Primer design

[0017] On the basis of the SecA gene sequence of the sugarcane white leaf disease phytoplasma determined by the inventor, specific primers were designed by using the difference in the SecA gene sequence of the sugarcane white leaf disease phytoplasma and other similar phytoplasma, and the size of the expected amplified target fragment was 350bp , the designed specific primers for detecting sugarcane white leaf disease phytoplasma are made up of primer SecA-F and primer SecA-R, the nucleotide sequence of described primer SecA-F is as shown in SEQ ID NO: 1, primer SecA- The nucleotide sequence of R is shown in SEQ ID NO: 2, and the specific primer was sent to a biological company for synthesis.

[0018] 2. Total DNA extraction

[...

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Abstract

The invention discloses a PCR detection method of sugarcane white leaf disease phytoplasma. The PCR detection method of sugarcane white leaf disease phytoplasma is established by designing a specificprimer SecA-F / SecA-R according to the SecA gene sequence of the sugarcane white leaf disease phytoplasma and optimizing the reaction system and program. By adopting the specific primer and method disclosed by the invention, PCR amplification is needed only once, whether the detected sample is infected by the sugarcane white leaf disease phytoplasma can be determined according to whether the electrophoresis result has a specific band, and the amplification product does not need sequencing determination; therefore, the detection specificity and accuracy are improved, the detection time is saved,and the detection cost is lowered.

Description

technical field [0001] The invention relates to a PCR detection method for sugarcane white leaf disease phytoplasma, belonging to the technical field of plant protection. Background technique [0002] Sugarcane white leaf disease is a devastating disease of sugarcane caused by phytoplasma, which can cause huge economic losses to sugarcane and sugar industry. The disease was first discovered in Thailand in 1954, and has now widely occurred in sugarcane-growing countries such as India, Pakistan, Sri Lanka, Laos, Myanmar, Vietnam, the Philippines and Australia. In my country, the occurrence of the disease was reported in Taiwan in 1962. In 2012, the phytoplasma of sugarcane white leaf disease was detected for the first time in the Baoshan sugarcane area of ​​Yunnan Province. The main symptoms of sugarcane white leaf disease are leaf albinism, increased tillers and dwarfing. The pathogen is mainly transmitted over long distances through diseased sugarcane species, and can be t...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11C12R1/94
CPCC12Q1/686C12Q1/701C12Q2565/125
Inventor 张荣跃黄应昆李文凤单红丽王晓燕李婕罗志明尹炯仓晓燕
Owner SUGARCANE RES INST OF YUNNAN ACADEMY OF AGRI SCI
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