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以感染甜菜黑色焦枯型病毒(Beet Black Scorch Virus.BBSV)新疆及宁夏分离物的苋色藜为材料提纯病毒,提取其RNA,经琼脂糖凝胶电泳后分别回收不同的RNA组分,以不同组合接种苋色藜,证实新疆分离物RNA_1能单独侵染并复制,但RNA_2组分必须依赖RNA_1才能完成其侵染和复制过程.同时发现,宁夏分离物RNA可以代替新疆分离物RNA_1支持RNA_2的复制.通过核酸酶保护试验和对BBSV新疆及宁夏分离物cDNA标记探针与两者的RNA进行的Northern blot分析发现新疆分离物RNA_1与宁夏分离物RNA具有高度的序列同源性,新疆分离物RNA_1与RNA_2无序列同源性,根据研究结果推断,BBSV新疆分离物和宁夏分离物为同一种病毒,新疆分离物RNA_2组分可能是卫星RNA.以BBSV新疆分离物RNA_2为模板,加PolyA后,以oligo(dT)_((12)～(18))为引物合成了部分与该病毒RNA_2互补的DNA,对杂交筛选所得阳性克隆进行了核苷酸序列分析,通过与已发表的病毒核苷酸序列比较,核苷酸序列同源性均在50%以下.根据丁群对宁夏BBSV RNA序列测定及本研究结果分析,我们推测甜菜上的BBSV可能是一种未见报道的新病毒.
The virus was isolated from Begonia crigesii isolated from Beet Black Scorch Virus.BBSV Xinjiang and Ningxia isolates. The RNA was extracted and different RNA fractions were recovered by agarose gel electrophoresis. The results showed that RNA_1 could be infected and replicated independently, but the RNA_2 component must rely on RNA_1 to complete the process of infection and replication.At the same time, Ningxia isolate RNA could replace Xinjiang isolate RNA_1 to support RNA_2 .The results of Northern blot analysis of the nuclease protection test and the RNA of the BBSV Xinjiang and Ningxia cDNA tag probes and the two showed that the Xinjiang isolate RNA_1 has a high degree of sequence homology with the Ningxia isolate RNA and the Xinjiang isolate RNA_1 and RNA_2 have no sequence homology. According to the results of the study, it is concluded that the Xinjiang isolate and Ningxia isolate of BBSV are the same virus, and the RNA_2 component of Xinjiang isolate may be satellite RNA. With RNA_2 of BBSV Xinjiang isolate as template, PolyA , A partial DNA complementary to the viral RNA_2 was synthesized using oligo (dT) _ ((12) ~ (18)) as a primer. The nucleotide sequence of the positive clones obtained by hybridization screening was analyzed by According to the determination of BBSV RNA in Ningxia and the analysis of the results of this study, we speculate that the BBSV in beet may be an unseen New virus reported.