Multiple RT-PCR method for detecting five sugarcane viruses simultaneously as well as primer and kit thereof

Abstract

The invention discloses a multiple RT-PCR detection primer group for detecting five kinds of sugarcane viruses simultaneously. The multiple RT-PCR detection primer group for detecting five kinds of sugarcane viruses simultaneously comprises six pairs of PCR primers which aim at five sugarcane viruses such as SCBV, SSV, SCMV, ScYLV and SrMV and one sugarcane reference gene GPADH correspondingly; the six pairs of PCR primers have base sequences as shown in a sequence table SEQ ID NO.1 to SEQ ID NO.12 correspondingly. Therefore, primer concentration and annealing temperature which influence multiple RT-PCR amplification are optimized by a multiple RT-PCR system and a corresponding RT-PCR detection method is established; and by the method, the five viruses for infecting the sugarcane can be detected simultaneously from sugarcane tissues rapidly, accurately, simply and conveniently, and important significance in screening healthy seedling mother plants, monitoring healthy seedling breeding,breeding for disease resistance and predicting field disease epidemic is achieved. In order to facilitate application of the multiple RT-PCR detection primer group, a corresponding kit is designed and assembled, so that popularization and promotion in grass roots are facilitated.

Description

technical field [0001] The invention belongs to the technical field of sugarcane virus detection, and relates to a multiple RT-PCR method for simultaneously detecting five kinds of sugarcane viruses, as well as primers and a kit. Background technique [0002] Sugarcane (Saccharum officinarum L.) belongs to the grass family Saccharum and is the most important sugar crop. It is also the raw material for the production of ethanol, furfural and dextran. Sugarcane sugar accounts for more than 85% of the total sugar production in my country. Sugarcane will be harmed by various diseases, especially viral diseases, during the growth process. After long-term asexual reproduction, various viruses will accumulate in sugarcane varieties and germplasm materials, resulting in the degradation of sugarcane varieties, and the reduction of yield and sugar content. cause significant economic losses. More than 15 types of viruses have been found to infect sugarcane worldwide, including DNA vir...

AI Technical Summary

Problems solved by technology

However, single-plex RT-PCR can only detect one virus in one reaction. If the reaction template contains more than two viruses, multiple PCRs are required,

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