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

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...

AI Technical Summary

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 the mitochondrial cytochrome oxidase I gene (COI), and the observation and comparison of the body size of the adults of the coconut beetle and the cocoa beetle are carried out. These methods do not involve water. The precise distinction between the coconut star anise beetle and the coconut heart leaf beetle, but the traditional taxonomy method is too professional, and requires the adult to distinguish, it is difficult to distinguish from the larvae and the damage shape, and the insect body structure is required to be complete, and the characteristics of each part are fully developed. In the case of insect damage and corruption, it is difficult to identify and error-prone

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