Nerva Luca, Silvestri Alessandro, Ciuffo Marina, Palmano Sabrina, Varese Givanna Cristina, Turina Massimo
Istituto per la Protezione Sostenibile delle Piante, CNR, Strada delle Cacce 73, Torino 10135, Italy.
Department of Life Sciences and Systems Biology, University of Turin, Mycotheca Universitatis Taurinensis (MUT), Viale Mattioli 25, Torino 10125, Italy.
Environ Microbiol. 2017 Nov;19(11):4480-4492. doi: 10.1111/1462-2920.13894. Epub 2017 Oct 13.
We attempted to transfect six recently characterized virus species to protoplasts of Penicillium janczewskii and Chryphonectria parasitica. None of the recovered P. janczewskii colonies was positive for the transfected viruses, but Penicillium aurantiogriseum partiti-like virus 1 (PaPLV1) was detected in three distinct regenerated C. parasitica colonies. We screened the phenotype of the infected strains in up to 45 different conditions combining different media, salinity and temperatures: our results show that the infected strains grow slower than the virus- free in most of the tested conditions with the exception of halophilic stress in a specific nutrient combination media. We proceeded to characterize molecularly the population of distinct isolates of PaPLV1 infected C. parasitica through RNAseq: comparison to the viral population present in the original host - P. auratiogriseum - showed that two isolates accumulated non-synonymous mutations suggesting adaptation to the new host. RNAseq analyses identified a second genomic RNA segment and northern blot of RNA extracted from purified virus suspensions allowed establishing that PaPLV1 is at least bipartite in nature and that it forms isometric virions of circa 36-38 nm in diameter. In light of these new acquisitions, we discuss the taxonomic placement of PaPLV1 inside the Partitiviridae.
我们尝试将六种最近鉴定出的病毒种类转染到简氏青霉(Penicillium janczewskii)和寄生隐孢壳菌(Chryphonectria parasitica)的原生质体中。回收的简氏青霉菌落中没有一个对转染病毒呈阳性,但在三个不同的再生寄生隐孢壳菌菌落中检测到了橙灰青霉类双分病毒1(PaPLV1)。我们在多达45种不同条件下(结合不同培养基、盐度和温度)筛选了感染菌株的表型:我们的结果表明,在大多数测试条件下,感染菌株的生长速度比未感染病毒的菌株慢,但在特定营养组合培养基中的嗜盐胁迫条件下除外。我们通过RNA测序对感染寄生隐孢壳菌的不同PaPLV1分离株群体进行了分子特征分析:与原始宿主——橙灰青霉中存在的病毒群体相比,表明有两个分离株积累了非同义突变,这表明其对新宿主有适应性。RNA测序分析鉴定出了第二个基因组RNA片段,从纯化病毒悬液中提取的RNA的Northern印迹分析确定PaPLV1本质上至少是双分体的,并且它形成直径约为36 - 38纳米的等轴病毒粒子。鉴于这些新发现,我们讨论了PaPLV1在双分病毒科中的分类地位。