Molecular analysis of an attenuated strain of Cucumber green mottle mosaic virus using in vitro infectious cDNA clone: pathogenicity and suppression of RNA silencing

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• 作者：Md Emran Ali ; Sumyya Waliullah…
• 关键词：Cucumber green mottle mosaic virus ; Attenuated strain ; cDNA clone ; In vitro infectious cDNA ; CGMMV ; Suppression of RNA silencing
• 刊名：Journal of Plant Biochemistry and Biotechnology
• 出版年：2016
• 出版时间：January 2016
• 年：2016
• 卷：25
• 期：1
• 页码：79-86
• 全文大小：3,110 KB
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• 作者单位：Md Emran Ali (1) (2)
  Sumyya Waliullah (1)
  Masamichi Nishiguchi (1)
  
  1. Faculty of Agriculture, Ehime University, 3-5-7 Tarumi, Matsuyama, 790-8566, Japan
  2. Department of Plant Pathology, The Ohio State University, 026 Selby Hall, 1680 Madison Ave, Wooster, OH, 44691, USA
  
• 刊物主题：Life Sciences, general; Plant Biochemistry; Protein Science; Receptors; Cell Biology;
• 出版者：Springer India
• ISSN：0974-1275

文摘

Cucumber green mottle mosaic virus (CGMMV), a tobamovirus, infects cucurbitaceous plants including watermelon. An attenuated strain of CGMMV (CGMMV-SH33b) has been isolated from CGMMV-SH and used for protecting muskmelon against wild CGMMV. There are five of nine base changes that lead to amino acid substitutions in the genome between SH and SH33b: four in replicase and one in coat protein. Using cDNA clones of both strains, two recombinant clones were constructed: first clone (pCG-KK) has two base changes (one silent at nt 5089 and the other P1397S amino acid substitution in the read-through part of SH 186K replicase) and the second clone (pCG-SP) has two silent base changes and three base changes in SH 129K/186K replicase (A1124V, N1157D and P1397S). The symptom severity on Nicotiana benthamiana induced by each recombinant virus were intermediate between those by SH and by SH33b. The accumulation levels of viruses using ELISA in infected plants were coinciding with symptom severity. Northern blot analysis of Nicotiana benthamiana at 10 dpi indicated that the virus accumulations of these recombinants was in the order of SH>pCG-KK>pCG-SP>SH33b. These results suggest that base changes located in the read-through part of 186K replicase as well as other base changes are responsible for the virus attenuation although we cannot excluded the possibility of silent mutations involved in pathogenicity. Using green fluorescent protein gene (GFP) silenced transgenic tobacco; infection with CGMMV-SH significantly suppressed GFP silencing with visible symptoms, whereas CGMMV-SH33b and their chimera viruses resulted in partial suppression. The suppression was further supported by lower level of GFP mRNA and higher siRNA detected in SH33b infected leaves, than those in CGMMV-SH infections, suggesting that the levels of virus accumulation and symptom severity on N. benthamiana were reasonably parallel with the suppression of RNA silencing of GFP mRNA. Keywords Cucumber green mottle mosaic virus Attenuated strain cDNA clone In vitro infectious cDNA CGMMV Suppression of RNA silencing