Plant Virology Group, ICGEB Biosafety Outstation, Ca' Tron di Roncade, I-31056, Italy.
Mol Plant Microbe Interact. 2013 Jul;26(7):801-11. doi: 10.1094/MPMI-02-13-0057-R.
Recombination is a major source of virus variability, and the question of whether novel recombinant viruses would emerge in transgenic plants expressing viral sequences has been a biosafety issue. We describe the results of pyrosequencing the recombinant viral RNAs appearing in transgenic plants expressing the coat protein (CP) gene and 3' noncoding region of Cucumber mosaic virus RNA3, as well as in nontransgenic controls. The populations of recombinants in both transgenic and nontransgenic plants were similar to those previously described from Sanger sequencing but many more recombinant types were observed, including a novel class of large deletions removing all or nearly the entire CP gene. These results show that populations of recombinant viral genomes arising de novo can be characterized in detail by pyrosequencing, and confirm that the transgenic plants did not harbor novel recombinants of biosafety concern.
重组是病毒变异的主要来源，在表达病毒序列的转基因植物中是否会出现新型重组病毒一直是一个生物安全问题。我们描述了对表达黄瓜花叶病毒 RNA3 外壳蛋白（CP）基因和 3'非编码区的转基因植物以及非转基因对照中出现的重组病毒 RNA 进行焦磷酸测序的结果。重组体在转基因和非转基因植物中的群体与之前通过 Sanger 测序描述的群体相似，但观察到更多的重组类型，包括一种新型的大片段缺失，几乎完全去除了整个 CP 基因。这些结果表明，通过焦磷酸测序可以详细描述从头出现的重组病毒基因组群体，并证实转基因植物不含有新的引起生物安全关注的重组体。
