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 nuclearcytoplasmic 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.

Soybean mosaic virus (SMV) is one of the most prevalent and important pathogens of soybean, which produces 11 proteins, and the third protein, P3, was suggested to be involved in virus movement and replication, as well as host infection. During the virus infection, host proteins are essential in the virus cycle. However, there is no comprehensive report on the network of host proteins that interact with P3. Fifty-one interactors were identified by using the P3 protein as the bait against the SMV SC15 strain-challenged soybean cDNA library. These proteins were classified into five groups, including transport and protein transport-related proteins, defense and disease-related proteins, photosynthesis proteins, cellular metabolic proteins, and unknown proteins. Among these proteins, the protein defined as hypersensitive response-like lesion-inducing (HRLI) appeared multiple times and showed strong affinity with P3, which indicated its important role in SMV infection. Thus, it was chosen for further investigation. Phylogenetic classification showed that paralog proteins GmHRLI-1 and GmHRLI-2 clustered together and shared 90% homologous identity. Bimolecular fluorescence complementation (BiFC) assay was carried out to confirm the interaction, and fluorescence was detected at the cell periplasmic as well as at the nucleus. Subcellular localization showed that GmHRLI was localized to the cell periplasmic, while the co-localization of GmHRLI and P3 signals was also observed in the nucleus, suggesting that GmHRLI could interact with P3 and promoted the translation of P3 to the nucleus. Moreover, the gene expression of GmHRLI was abundant in the roots, leaves, and flowers, and could be induced by SMV infection, suggesting its involvement in SMV infection. Our results together lay the foundation to explore the mechanisms of P3 in the HR process and the HRLI protein function in SMV response.

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.