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为检测福建省马铃薯纺锤块茎类病毒及了解其分子变异情况,以感染马铃薯纺锤块茎类病毒的植株为试验材料,提取RNA,反转录cDNA,利用设计合成的特异性引物进行RT-PCR检测,扩增产物经回收纯化后克隆和测序。结果表明,PCR扩增出359bp的特异片段,将扩增产物经回收和纯化并进行克隆与测序,测序结果经BLAST比对后,该片段为PSTVd基因序列,PSTVd分离物命名为Fujian PTSTd(KM588065.1),与国内外报道的PSTVd分离物同源性达98%以上。该分离物与弱毒株系(M14814.1)、中间株系(AY492084.1)和强毒株系(U23058.1)比对,有11个核苷酸具有多态性,其中5个发生在致病区。不同国家和地区序列进化树分析表明,该分离物与我国北方及欧美国家分离物亲缘关系最近,与新西兰分离物亲缘关系最远。
In order to detect potato spindle tuber-like virus in Fujian Province and understand its molecular variation, the plants infected with potato spindle tuber-like virus were used as experimental material to extract RNA and reverse transcribed cDNA. The specific primers designed and synthesized were used for RT- The amplified product was recovered and cloned and sequenced. The results showed that PCR amplified 359bp specific fragment, the amplified product was recovered and purified and cloned and sequenced. After BLAST alignment of the sequencing results, the fragment was PSTVd gene sequence, PSTVd isolate named Fujian PTSTd (KM588065 .1), which shared 98% homology with PSTVd isolates reported at home and abroad. The isolate was compared with the attenuated strain (M14814.1), the intermediate strain (AY492084.1) and the virulent strain (U23058.1), and 11 nucleotides were polymorphic, of which 5 occurred in the Pathogenic zone. The phylogenetic tree analysis of different countries and regions showed that this isolate has the closest genetic relationship with isolates from northern China, Europe and the United States, and the farthest genetic relationship with New Zealand isolates.