甘蔗黄叶病毒的RT-PCR检测技术

Detection of sugarcane yellow leaf virus by RT-PCR

以甘蔗黄叶病毒(ScYLV)特异性引物YLSF111和YLSR462为引物,对福建蔗区8个罹病品种的疑似病株进行RT-PCR检测,扩增出352 bp特异性片段.将PCR产物克隆后测序,序列分析表明,该片段是ScYLV外壳蛋白(CP)基因的一部分,核苷酸和氨基酸序列同源性达95%以上,与美国、印度、巴西等国家ScYLV分离物(GeneBank登录号分别为AF157029、AY236971、AF141385)CP基因同源性达100%,证实我国福建地区甘蔗黄叶综合症的病原体为ScYLV.本研究同时建立了ScYLV的RT-PCR检测技术.

Using primers YLSF111 and YLSR462, a 352 bp fragment was amplified by RT-PCR in total RNA extracted from leaves of eight infected sugarcane varieties in Fujian, P. R. China. The PCR product was cloned and sequenced. Nucleotide and deduced amino acid sequences were greater than 95% homologus with ScYLV isolates from other countries and were 100% homologus with ScYLV isolates from America, India and Brazil （ GenBank accession numbers AF157029, AY236971 and AF141385 respectively）. The results confirmed that the causal organism of YLS of sugarcane in China was ScYLV. The protocol for the use of RT-PCR to detect ScYLV was described in the paper.