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The leaves of soybean cultivar ZheA8901 show various symptoms (necrosis, mosaic, and symptomless) when infected with different strains of soybean mosaic virus (SMV). Based on a proteomic analysis performed with tandem mass tags (TMT), 736 proteins were differentially expressed from soybean samples that showed asymptomatic, mosaic, and necrosis symptoms induced by SMV strains SC3, SC7, and SC15, respectively. Among these, GmGSTU13 and ascorbate peroxidase (APX) were only upregulated in mosaic and symptomless leaves, respectively. The protein level of GmGSTU13 determined by western blot analysis was consistent with TMT analysis, and quantitative reverse transcriptase PCR analysis showed that GmGSTU13 mRNA levels in mosaic plants were 5.26- and 3.75-fold higher than those in necrotic and symptomless plants, respectively. Additionally, the expression of the viral coat protein (CP) gene was increased, and serious mosaic symptoms were observed in GmGSTU13-overexpressing plants inoculated with all three SMV strains. These results showed that GmGSTU13 is associated with the development of SMV-induced mosaic symptoms in soybean and that APX is upregulated in symptomless leaves at both the transcriptional and protein levels. In APX gene-silenced soybean plants, the relative expression of the viral CP gene was 1.50, 7.59, and 1.30 times higher than in positive control plants inoculated with the three SMV strains, suggesting that the upregulation of APX may be associated with lack of symptoms in soybean infected with SMV. This work provides a useful dataset for identifying key proteins responsible for symptom development in soybean infected with different SMV strains.

Host proteins are essential during virus infection, and viral factors must target numerous host factors to complete their infectious cycle. The mature 6K1 protein of potyviruses is required for viral replication in plants. However, the interaction between 6K1 and host factors is poorly understood. The present study aims to identify the host interacting proteins of 6K1. Here, the 6K1 of Soybean mosaic virus (SMV) was used as the bait to screen a soybean cDNA library to gain insights about the interaction between 6K1 and host proteins. One hundred and twenty-seven 6K1 interactors were preliminarily identified, and they were classified into six groups, including defense-related, transport-related, metabolism-related, DNA binding, unknown, and membrane-related proteins. Then, thirty-nine proteins were cloned and merged into a prey vector to verify the interaction with 6K1, and thirty-three of these proteins were confirmed to interact with 6K1 by yeast two-hybrid (Y2H) assay. Of the thirty-three proteins, soybean pathogenesis-related protein 4 (GmPR4) and Bax inhibitor 1 (GmBI1) were chosen for further study. Their interactions with 6K1 were also confirmed by bimolecular fluorescence complementation (BiFC) assay. Subcellular localization showed that GmPR4 was localized to the cytoplasm and endoplasmic reticulum (ER), and GmBI1 was located in the ER. Moreover, both GmPR4 and GmBI1 were induced by SMV infection, ethylene and ER stress. The transient overexpression of GmPR4 and GmBI1 reduced SMV accumulation in tobacco, suggesting their involvement in the resistance to SMV. These results would contribute to exploring the mode of action of 6K1 in viral replication and improve our knowledge of the role of PR4 and BI1 in SMV response.

Additional file 1: Fig. S1. Schematic diagram of T-DNA region of recombinant plasmid pB7GWIWG2(II)-GmVma12i,Fig. S2. The morphology of Fig. S3. The transcript levels of two isforms of GmVma12 by qRT-PCR in T1 RNAi silenced transgenic plants qRT-PCR. X-axes indicate T1 transgenic plants and non-transformed (Mock) plants. Data are expressed as the means of three biological replicates with error bars indicating the SD (n = 3). Asterisks denote significant difference from mock, as determined by the t- test, p

Blue light (BL) plays an important role in regulation of the growth and development of aquatic plants and land plants. Aureochrome (AUREO), the recent BL photoreceptor identified in photosynthetic stramenopile algae, is involved in the photomorphogenesis and early development of Saccharina japonica porophytes (kelp). However the factors that interact with the SjAUREO under BL conditions specifically are not clear. Here in our study, three high quality cDNA libraries with CFU over 5 × 106 and a recombination rate of 100% were constructed respectively through white light (WL), BL and darkness (DK) treatments to the juvenile sporophytes. Based on the constructed cDNA libraries, the interactors of SjAUREO were screened and analyzed. There are eighty-four genes encoding the sixteen predicted proteins from the BL cDNA library, sixty-eight genes encoding eighteen predicted proteins from the DK cDNA library, and seventy-four genes encoding nineteen proteins from the WL cDNA library. All the predicted proteins are presumed to interact with SjAUREO when co-expressed with SjAUREO seperately. The 40S ribosomal protein S6 (RPS6), which only exists in the BL treated cDNA library except for two other libraries, and which is essential for cell proliferation and is involved in cell cycle progression, was selected for detailed analysis. We showed that its transcription was up-regulated by BL, and was highly transcribed in the basal blade (meristem region) of juvenile sporophytes but less in the distal part. Taken together, our results indicated that RPS6 was highly involved in BL-mediated kelp cellular division and photomorphogenesis by interacting with SjAUREO.

Soybean mosaic virus (SMV) is one of the most destructive pathogens of soybean (Glycine max (L.) Merr.) worldwide. In this study, 184 F7:11 recombinant inbred line (RIL) populations derived from Kefeng No. 1 × Nannong 1138-2 were used to study the inheritance and linkage mapping of resistance genes against SMV strains SC7 and SC13 in Kefeng No. 1. Two independent dominant genes (designated Rsc7 and Rsc13) that control resistance to SC7 and SC13 were located on a molecular linkage group (MLG) of chromosome 2 (D1b). A mixed segregating population was developed by self-pollination of three heterozygous plants of residual heterozygous lines (RHL3-27, RHL3-30, RHL3-53) with five markers linked to the loci, and was used in fine-mapping of Rsc7 and Rsc13. In addition, Rsc7 was fine-mapped between BARCSOYSSR_02_0667 and BARCSOYSSR_02_0670 on MLG D1b. The genetic distance between the two closest markers was 0.7 cM and the physical distance of the interval was ~77 kb, which included one LRR gene and another gene containing an F-box region. Two SSR markers (BARCSOYSSR_02_0610 and BARCSOYSSR_02_0621) were closely linked to the SC13 resistance gene. The physical distance where Rsc13 was located was ~191 kb. Sequence analysis showed that there were two K-box region types of transcription factor genes; GmHSP40 and two serine/threonine protein kinase (STK) genes were the most likely candidate genes. These results will facilitate map-based cloning of the Rsc7 and Rsc13 genes and development of transgenic disease-resistant varieties, and will provide SMV-resistance breeding systems with excellent resistance germplasm.

The biochemical function of the potyviral P3 protein is not known, although it is known to regulate virus replication, movement, and pathogenesis. We show that P3, the putative virulence determinant of soybean mosaic virus (SMV), targets a component of the translation elongation complex in soybean. Eukaryotic elongation factor 1A (eEF1A), a well-known host factor in viral pathogenesis, is essential for SMV virulence and the associated unfolded protein response (UPR). Silencing GmEF1A inhibits accumulation of SMV and another ER-associated virus in soybean. Conversely, endoplasmic reticulum (ER) stress-inducing chemicals promote SMV accumulation in wild-type, but not GmEF1A-knockdown, plants. Knockdown of genes encoding the eEF1B isoform, which is important for eEF1A function in translation elongation, has similar effects on UPR and SMV resistance, suggesting a link to translation elongation. P3 and GmEF1A promote each other’s nuclear localization, similar to the nuclear-cytoplasmic transport of eEF1A by the Human immunodeficiency virus 1 Nef protein. Our results suggest that P3 targets host elongation factors resulting in UPR, which in turn facilitates SMV replication and place eEF1A upstream of BiP in the ER stress response during pathogen infection.

Soybean mosaic virus (SMV), a member of the Potyvirus genus, is one of the most prevalent and devastating viral pathogens in soybean-growing regions worldwide. It is generally accepted that symptom development of a viral plant disease results from molecular interactions between the virus and its host plant. P3 protein is the most variable polyprotein in potyviruses, which potentially plays an important role in the process of the evolution of virus type specialization. However, P3 not only plays a major role in virus replication and movement, but it is also responsible for symptom development in SMV-infected plants. This study provides evidence that actin-depolymerizing factor 2 (designated as ADF2) of soybean interacts with SMV P3 via a two-hybrid yeast system by screening a soybean cDNA library. Bimolecular fluorescence complementation assay further confirmed the interaction, which occurred in both the cytomembrane and cytoskeleton of Nicotiana benthamiana cells. The results support the hypothesis that SMV P3 might have a role in virus movement within cells.