山东地区樱桃绿环斑驳病毒(CGRMV)的RT-PCR检测及外壳蛋白基因的克隆

DETECTION OF CHERRY GREEN RING MOTTLE VIRUS(CGRMV) AND ISOLATION OF ITS COAT PROTEIN GENES FROM SWEET CHERRY IN SHANDONG PROVINCE

为调查我省甜樱桃感染樱桃绿环斑驳病毒(Cherry Green Ring Mottle Virus,CGRMV)情况,本研究以甜樱桃(Prunus avium L.)品种"红灯"叶片总RNA为模板,根据CGRMV基因组序列设计特异引物,对山东地区37份甜樱桃"红灯"样品进行RT-PCR检测,共检测出19份阳性样品。利用CGRMV外壳蛋白基因序列引物,从阳性植物样本中分离到约800 bp的目的片段,克隆测序,序列分析显示该片段全长807 bp,编码268个氨基酸,与GenBank中已登录的CGRMV分离物的外壳蛋白基因序列一致性为87%～97%,氨基酸序列相似性为95%～99%。该结果表明山东地区甜樱桃生产园中感染CGRMV的病例较为普遍。

In order to investigate the infection of Cherry Green Ring Mottle Virus（CGRMV） from sweet cherry（Prunus avium L.） in Shandong province,total RNA was extracted from the leaves of the sweet cherry cultivar Red Lamp＇.The specific primers were designed corresponding to the CGRMV genome sequence and used for the RT-PCR detection.Thirty-seven sweet cherry samples from six orchards were analyzed in the experiment and nineteen of them were detected as positive.The gene that encodes the virus coat protein was amplified and sequenced.Sequence analysis revealed that the fragment was 807 nucleotides in length,encoding 268 amino acids.It shared 87%～97% identity to the other CGRMV isolates reported in GenBank in the nucleotide level and 95%～99% similarity in the amino acids level.These results showed CGRMV had been wide spread on sweet cherry in Shandong district.