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. 2019 Sep 27;10:4033. doi: 10.1038/s41467-019-12052-5

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — Rsv4 encodes an RNase H family protein. a Symptoms of SMV-inoculated soybean plants. 211c-2 is a transgenic line derived from susceptible cv. Jack, overexpressing Rsv4 from Peking. Bar = 5 cm. b SMV coat protein (CP) accumulation in control and transgenic soybean plants. 211c expresses Rsv4 cDNA driven by the cauliflower mosaic virus 35 S promoter, while 219 g carries a genomic DNA fragment of Peking containing the putative native promoter of Rsv4. SMV CP accumulation in non-inoculated upper leaves was detected by western blotting at 10 days post-inoculation (dpi) (upper panel). Rsv4 mRNA was detected by RT-PCR (lower panel). Lanes represent individual plants. c SMV strains SMV-A, -D, -E, and -L(Q1033K) were used to inoculate 211c, and CP accumulation was analyzed as in panel b. d Co-segregation of PeM-1211 and PeM-0484 (identical nonsense mutations in Peking Rsv4 gene) and SMV-susceptible phenotype. W wild-type, M mutant-type, H heterozygous. e Rsv4, but not the Rsv4(D99N) mutant protein, inhibits SMV multiplication in N. benthamiana leaves. SMV CP accumulation was detected by western blotting at 5 dpi. f SMV-L(Q1033K) is less sensitive to Rsv4 than SMV-C. Agrobacterium strains containing constructs to express Rsv4 or Rsv4(D99N) were infiltrated at the indicated concentrations. 1 day after infiltration, Agrobacterium harboring SMV cDNA was infiltrated at OD₆₀₀ = 0.005. CP accumulation was detected at 7 dpi. The source data of Fig. 2b-f are provided as a Source Data file