水稻橙叶病PCR检测体系的建立

利用植原体通用引物sP1和sP2,采用PCR法从发病的水稻植株中扩增植原体一段保守的16S rRNA基因的核苷酸序列. 结果表明,从发病的水稻植株中都能够稳定扩增得到1条558 bp的特异性条带. 对该条带进行克隆和序列分析表明,该片段和Genbank中公布的众多植原体的相应区域的核苷酸序列相似性都高达95%以上,表明利用植原体通用引物能有效扩增得到水稻橙叶病原的序列. 利用此引物并优化PCR反应条件,建立了水稻橙叶病的PCR检测方法. 该方法对检测水稻橙叶病具有特异、灵敏和有效性. 应用该检测体系检测从广东信宜、高州和从化采集的多份疑似标样,结果表明,在这几个地区均有水稻橙叶病的发生和危害.

The Establishment of PCR Detection System of Rice Orange Leaf Disease

It is one of the important control strategies of rice orange leaf disease by development of a rapid disease detection technique. Conserved general primer pairs sP1 and sP2 for phytoplasma were synthesized, and the 558 bp segment of the phytop!asma 16S rRNA gene was specifically amplified by PCR. The sequencing results of the segment showed that the nucleotide acid sequence of this PCR product shared above 95% identical with the corresponding segment of other kinds of phytoplasma in Genbank, which testified that the PCR product was from the pathogen of rice orange leaf disease. The PCR detection system was established by optimizing the parameters of PCR reaction system and found that this system could effectively, sensitively, and accurately detected the disease. The several samples from the different locations of Xinyi, Gaozhou and Conghua in Guangdong Province were detected by the PCR system, and the results revealed rice leaf disease existed in these locations.