三系杂交水稻真伪及纯度实时PCR技术鉴定的研究

杂交水稻的研究和推广为国民经济的发展起到了重要的作用，并已经在东南亚地区推广。如何建立一套快速、准确的杂交水稻真伪及纯度检测体系一直是困扰着口岸检疫人员的技术难题。实时荧光PCR技术是近年来新兴的检验技术。我们通过对已报道的雄性不育特异片段R2-630WA进行PCR验证，发现其可以作为鉴定杂交稻不育胞质的分子标记。依据此片段设计并合成一对引物和一条TaqMan荧光探针，对17个水稻品种进行实时荧光PCR检测。结果表明：此探针可以特异扩增三系杂交稻F1及不育系，并可初步用于三系杂交稻的纯度检测，由此建立了一套准确、快速的三系杂交稻鉴定体系。

A preliminary research on application of Real-time Fluorescent PCR assay to identify three-line hybrid rice

The research and development of the hybrid rice had paid great contribution to the development of the national economy, and had been popularized in the Southeast asia. How to build up a set of fast, accurate ture-false and purity test system of hybrid rice has been technique hard nut to crack which perplexed quarantine researchers for long time. The real-time fluorescent PCR technique is a newly arisen detect technique in recent years. Using PCR amplification, The reliability of DNA fragment R2-630 was confirmed which related to male sterity reported in 1995.Based on this fragment, one TaqMan probe and one pair of primers were designed, which can detect 17 hybrid rice samples. The result shows: this probe can with peculiar to amplify, hybrid rice and sterile lines peculiarly, and can be used in the purity identification basically. Thus the first step of one time- saving, accurate and quick real-time fluorescent PCR test system was built up.