Abstract
RNA-dependent RNA polymerases (RDRs) play key roles in gene silencing. The rice RDR6 gene was analyzed in response to viral, bacterial and fungal pathogens, after inoculation of a rice mutant line of OsRDR6, shl2-rol, with Cucumber mosaic virus, Rice necrosis mosaic virus, Xanthomonas oryzae pv. oryzae or Magnaporthe oryzae. Compared with the wild type, the mutant line accumulated more viral RNA after inoculation with the viruses and developed more severe symptoms after inoculation with the bacterium or fungus. Thus, the OsRDR6-mediated RNA silencing pathway seems to participate in defense against not only viruses, but also bacterial and fungal pathogens.
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Acknowledgments
We are grateful to D. Murphy for checking the English in the manuscript and Md. Panna Ali for the RT-qPCR analysis. We also thank Drs. M. Mori and H. Ochiai for seeds of Sensyou and Asominori, respectively. This work was supported partly by the Program for Promotion of Basic and Applied Researches in Bio-oriented Industry and Science, the Technology Research Promotion Program for Agriculture, Forestry, Fisheries and Food Industry, the Ministry of Education, Culture, Sports, Science and Technology of Japan (Grant-in-Aid for Scientific Research for Scientific Research (C), no. 24580065 to MN and (B), no. 26292026 to KK) and by grants from the Ministry of Agriculture, Forestry and Fisheries of Japan (Rice Genome Project to MN).
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10327_2015_630_MOESM1_ESM.pptx
Fig. S1 Expression of OsRDR6 in response to mock inoculation with various pathogens as determined by RT-PCR. (a) Cucmber mosaic virus/Rice necrosis mosaic virus. Lane numbers represent days post inoculation. (b) Xanthomonas oryzae pv. oryzae/Magnaporthe oryzae. Lane numbers represent hours after inoculation. Total RNA was extracted from the leaves of treated plants at theh indicated times. Primers were OsRDR6-F and OsRDR6-R. The actin gene was used as a standard to show normalization of the amount of PCR templates. Fig. S2 Expression of OsRDR6 after inoculation with Xanthomonas oryzae pv. oryzae and Magnaporthe oryzae as determined by RT-PCR. Total RNA was extracted from inoculated leaves at 36 h after inoculation. RT-PCR primers were OsRDR6-F and OsRDR6-R. The actin gene was used as a standard to show normalization of the amount of PCR templates. M, mock; I, inoculated. Fig. S3 Accumulation of Cucmber mosaic virus (CMV) and Rice necrosis mosaic virus (RNMV) in WT (+/+), homozygous (-/-)/heterozygous (±) shl2-rol. Young leaves were inoculated with CMV and total RNA extracted at 7 dpi. Total RNA was extracted from the leaves of plants grown in RNMV-infested soil for 30 days. RT-PCR primers were RNMV-R1-F-5′ and RNMV-R1-R-3′ for RNMV, CMV-R3-cDNA-F-5′ and CMV-R3-cDNA-F-3′ for CMV. The actin gene was used as standard control to show the normalization of the amount of PCR templates. (PPTX 2845 kb)
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Wagh, S.G., Alam, M.M., Kobayashi, K. et al. Analysis of rice RNA-dependent RNA polymerase 6 (OsRDR6) gene in response to viral, bacterial and fungal pathogens. J Gen Plant Pathol 82, 12–17 (2016). https://doi.org/10.1007/s10327-015-0630-y
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DOI: https://doi.org/10.1007/s10327-015-0630-y