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菊花矮化病由菊花矮化类病毒(Chrysanthemum stunt viroid,CSVd)引起。从北京、海南万宁、合肥和哈尔滨共采集了122个野生及商业种植的菊花样品,使用改进的CTAB法提取菊花总RNA后,通过斑点杂交检测CSVd,应用RT-PCR对阳性样品进行克隆、测序。斑点杂交结果表明,在检测的122个样品中有14个样品感染了CSVd,感染率为11.5%;除哈尔滨外的其余3个地区均有CSVd的发生。序列比较分析结果表明,中国的CSVd分离物与韩国和日本分离物的序列最为接近,对其二级结构进行分析发现,与CSVd的参考序列(NC002015)相比,所获得的中国CSVd分离物的序列中虽然有21个碱基发生了突变(大多位于二级结构上的致病区内),但并未影响其折叠成稳定的拟棒状结构。
Chrysanthemum dwarf disease is caused by Chrysanthemum stunt viroid (CSVd). A total of 122 wild and commercially grown chrysanthemum samples were collected from Beijing, Hainan Wanning, Hefei and Harbin. Total RNA was extracted from chrysanthemum using improved CTAB method. CSVd was detected by dot blot hybridization and positive samples were cloned by RT-PCR. Sequencing. Dot blot results showed that 14 of the 122 samples tested were infected with CSVd and the infection rate was 11.5%. CSVd occurred in the remaining three regions except Harbin. Sequence comparison analysis showed that the Chinese CSVd isolates were the closest to the sequences of Korean and Japanese isolates. Analysis of their secondary structure revealed that compared with the CSVd reference sequence (NC002015), the obtained CSVd isolates from China Although 21 bases were mutated in the sequence (mostly located in the secondary structure of the pathogenic zone), but did not affect its folding into a stable quasi-rod-shaped structure.