Kubota Kenji, Ng James C K
First author: NARO Agricultural Research Center, Kannondai, Tsukuba, Ibaraki 305-8666, Japan, and Department of Plant Pathology and Microbiology, University of California, Riverside 92521; second author: Department of Plant Pathology and Microbiology, University of California, Riverside 92521.
Phytopathology. 2016 Jun;106(6):653-62. doi: 10.1094/PHYTO-09-15-0219-R. Epub 2016 Apr 13.
RNA silencing functions as an antivirus defense strategy in plants, one that plant viruses counter by producing viral suppressors of RNA silencing (VSRs). VSRs have been identified in three members of the genus Crinivirus but they do not all share identical suppression mechanisms. Here, we used Agrobacterium co-infiltration assays to investigate the suppressor activity of proteins encoded by Lettuce chlorosis virus (LCV). Of 7 LCV proteins (1b, P23, HSP70 homolog, P60, CP, CPm, and P27) tested for the suppression of silencing of green fluorescent protein (GFP) expression in wild-type Nicotiana benthamiana plants, only P23 suppressed the onset of local silencing. Small-interfering (si)RNA accumulation was reduced in leaves co-infiltrated with P23, suggesting that P23 inhibited the accumulation or enhanced the degradation of siRNA. P23 also inhibited the cell-to-cell and systemic movement of RNA silencing in GFP-expressing transgenic N. benthamiana plants. Expression of P23 via agroinfiltration of N. benthamiana leaves induced local necrosis that increased in severity at elevated temperatures, a novelty given that a direct temperature effect on necrosis severity has not been reported for the other crinivirus VSRs. These results further affirm the sophistication of crinivirus VSRs in mediating the evasion of host's antiviral defenses and in symptom modulation.
RNA沉默在植物中作为一种抗病毒防御策略发挥作用,植物病毒通过产生RNA沉默抑制因子(VSR)来对抗这种策略。在毛形病毒属的三个成员中已鉴定出VSR,但它们并非都具有相同的抑制机制。在这里,我们使用农杆菌共浸润试验来研究莴苣黄化病毒(LCV)编码蛋白的抑制活性。在对野生型本氏烟草植株中绿色荧光蛋白(GFP)表达沉默抑制进行测试的7种LCV蛋白(1b、P23、热休克蛋白70同源物、P60、外壳蛋白、外壳蛋白移动蛋白和P27)中,只有P23抑制了局部沉默的发生。与P23共浸润的叶片中小干扰(si)RNA积累减少,这表明P23抑制了siRNA的积累或增强了其降解。P23还抑制了表达GFP的转基因本氏烟草植株中RNA沉默的细胞间和系统移动。通过农杆菌浸润本氏烟草叶片表达P23会诱导局部坏死,坏死严重程度在高温下增加,鉴于尚未报道其他毛形病毒VSR对坏死严重程度有直接温度影响,这是一个新发现。这些结果进一步证实了毛形病毒VSR在介导逃避宿主抗病毒防御和症状调节方面的复杂性。