Primer pair, kit and PCR (Polymerase Chain Reaction) detection method for distinguishing octodonta nipae from brontispa longissima

A technology of coconut leaf beetle and water coconut star anise, applied in the field of insects, can solve problems such as difficult to distinguish, error-prone, and difficult to identify, and achieve high efficiency and high accuracy

Inactive Publication Date: 2014-01-29
FUJIAN AGRI & FORESTRY UNIV
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Problems solved by technology

However, the water coconut star anise beetle and the coconut heart leaf beetle have many of the same hosts and living habits, and there are many similarities in appearance and characteristics, which are difficult to distinguish. Especially in the import quarantine and pest census, it is necessary to accurately distinguish the two pests and take corresponding quarantine measures and emergency treatment. It is extremely important, so to carry out correct and timely prevention and control of it, the most important job is to accurately distinguish between the two
[0006] Currently, the PCR-RFLP method is used to quickly distinguish the two subspecies of the cocoa beetle through

Method used

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  • Primer pair, kit and PCR (Polymerase Chain Reaction) detection method for distinguishing octodonta nipae from brontispa longissima
  • Primer pair, kit and PCR (Polymerase Chain Reaction) detection method for distinguishing octodonta nipae from brontispa longissima

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Embodiment

[0079] Embodiment: Utilize COI-specific primer pair to distinguish water coconut star anise and coconut heart leaf beetle

[0080] Adopt the described primer pair of claim 1 and the described kit of claim 2 to distinguish the PCR detection method of water coconut star anise beetle and coconut heart leaf beetle rapidly, described PCR detection method comprises the following steps:

[0081] 1) Extract the total DNA of the water coconut star anise beetle and the coconut heart leaf beetle as DNA templates;

[0082] 2) Integrity detection of the DNA template, the DNA templates of the water coconut star anise and the coconut heart leaf beetle are formulated into a PCR system using the COI universal primer pair and the reagents contained in the kit described in claim 2, and the configured PCR system is put into Carry out amplification on the PCR machine, be used for checking the integrity of the DNA template of water coconut star anise beetle and coconut heart leaf beetle;

[0083] ...

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Abstract

The invention discloses a method for quickly distinguishing octodonta nipae from brontispa longissima through a PCR (Polymerase Chain Reaction) detection technology. The method comprises the following steps: enabling specific primers of COI (Cytochrome Oxidase Subunit I) and ITS (Internal Transcribed Spacer) gene segments of the octodonta nipae to be subjected to PCR reaction with DNA (Deoxyribose Nucleic Acid) templates of the octodonta nipae and the brontispa longissima, placing reaction products on an electrophoresis apparatus, and performing agarose gel electrophoresis, thus distinguishing the octodonta nipae from the brontispa longissima through electrophoretic bands. The method is simple, quick and high in accuracy, and plays a significant role in intensive study of the octodonta nipae and the brontispa longissima.

Description

technical field [0001] The invention relates to the field of insects, in particular to a pair of primers, a kit and a PCR detection method for distinguishing the water coconut star anise beetle and the coconut heart leaf beetle. Background technique [0002] The two pests of the water coconut star anise and the palm leaf beetle can seriously damage the palm plants, which have caused serious damage to the palm plants in recent years, causing economic losses of hundreds of millions of yuan. [0003] The water coconut star anise is a pest of the Coleoptera, and it is mainly spread by the transportation of seedlings. Its origin is Malaysia. It was discovered in Hainan Province, China in 2001. The water coconut star anise feeds on the young shoots, young stems and incompletely expanded plants of the plant. Young leaves, brown or gray-brown streaks, shrinkage, curling, etc., the young shoots will gradually wither and die, and in severe cases, the whole plant will die, which can se...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888
Inventor 张翔汤宝珍侯有明黄斌
Owner FUJIAN AGRI & FORESTRY UNIV
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