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Soybean mosaic virus (SMV), a member of Potyvirus, is the most destructive and widespread viral disease in soybean production. Our earlier studies identified a soybean 40S ribosomal protein S8 (GmRPS8) using the 6K1 protein of SMV as the bait to screen a soybean cDNA library. The present study aims to identify the interactions between GmRPS8 and SMV and characterize the role of GmRPS8 in SMV infection in soybean. Expression analysis showed higher SMV-induced GmRPS8 expression levels in a susceptible soybean cultivar when compared with a resistant cultivar, suggesting that GmRPS8 was involved in the response to SMV in soybean. Subcellular localization showed that GmRPS8 was localized in the nucleus. Moreover, the yeast two-hybrid (Y2H) experiments showed that GmRPS8 only interacted with 6K1 among the eleven proteins encoded by SMV. The interaction between GmRPS8 and 6K1 was further verified by a bimolecular fluorescence complementation (BiFC) assay, and the interaction was localized in the nucleus. Furthermore, knockdown of GmRPS8 by a virus-induced gene silencing (VIGS) system retarded the growth and development of soybeans and inhibited the accumulation of SMV in soybeans. Together, these results showed that GmRPS8 interacts with 6K1 and contributes to soybean susceptibility to SMV. Our findings provide new insights for understanding the role of GmRPS8 in the SMV infection cycle, which could help reveal potyviral replication mechanisms. [ABSTRACT FROM AUTHOR]

Soybean mosaic virus (SMV) is one of the most devastating viral pathogens of soybean (Glycine max (L.) Merr). In total, 22 Chinese SMV strains (SC1–SC22) have been classified based on the responses of 10 soybean cultivars to these pathogens. However, although several SMV‐resistance loci in soybean have been identified, no gene conferring SMV resistance in the resistant soybean cultivar (cv.) Kefeng No.1 has been cloned and verified. Here, using F2‐derived F3 (F2:3) and recombinant inbred line (RIL) populations from a cross between Kefeng No.1 and susceptible soybean cv. Nannong 1138‐2, we localized the gene in Kefeng No.1 that mediated resistance to SMV‐SC3 strain to a 90‐kb interval on chromosome 2. To study the functions of candidate genes in this interval, we performed Bean pod mottle virus (BPMV)‐induced gene silencing (VIGS). We identified a recombinant gene (which we named RSC3K) harboring an internal deletion of a genomic DNA fragment partially flanking the LOC100526921 and LOC100812666 reference genes as the SMV‐SC3 resistance gene. By shuffling genes between infectious SMV DNA clones based on the avirulent isolate SC3 and virulent isolate 1129, we determined that the viral protein P3 is the avirulence determinant mediating SMV‐SC3 resistance on Kefeng No.1. P3 interacts with RNase proteins encoded by RSC3K, LOC100526921, and LOC100812666. The recombinant RSC3K conveys much higher anti‐SMV activity than LOC100526921 and LOC100812666, although those two genes also encode proteins that inhibit SMV accumulation, as revealed by gene silencing in a susceptible cultivar and by overexpression in Nicotiana benthamiana. These findings demonstrate that RSC3K mediates the resistance of Kefeng No.1 to SMV‐SC3 and that SMV resistance of soybean is determined by the antiviral activity of RNase proteins. [ABSTRACT FROM AUTHOR]

Key message: A putative candidate gene conferring resistance to SMV strain SC1 was identified on chromosome 2, and the linked marker was validated in soybean cultivars Soybean mosaic, caused by the soybean mosaic virus, is the most common disease in soybean and a significant impediment to soybean production in the Huanghuai and Yangtze River regions of China. Kefeng No.1, a soybean cultivar, showed high resistance to soybean mosaic virus strain (SC1) collected from Huanghuai and Yangtze River regions. Genetic analysis based on the Mendelian genic population derived from the cross Kefeng No.1 × Nannong 1138-2 revealed that Kefeng No.1 possesses a single dominant gene. Furthermore, genetic fine-mapping using an F2 population containing 281 individuals delimited resistant gene to a genomic region of 186 kb flanked by SSR markers BS020610 and BS020620 on chromosome 2. Within this region, there were 14 genes based on the Williams 82 reference genome. According to sequence analysis, six of the 14 genes have amino acid differences, and one of these genes is the Rsv4 allele designated as Rsc1-DR. The functional analysis of candidate genes using the bean pod mottle virus (BPMV)-induced gene silencing (VIGS) system revealed that Rsc1-DR was accountable for Kefeng No.1's resistance to SMV-SC1. Based on the genome sequence of Rsc1-DR, an Insertion/Deletion (InDel) molecular marker, JT0212, was developed and genotyped using 100 soybean cultivars, and the coincidence rate was 89%. The study enriched our understanding of the SMV resistance mechanism. The marker developed in this study could be directly used by the soybean breeders to select the genotypes with favorable alleles for making crosses, and also it will facilitate marker-assisted selection of SMV resistance in soybean breeding. [ABSTRACT FROM AUTHOR]

E3-ubiquitin ligases are known to confer abiotic stress responses in plants. In the present study, GmPUB21, a novel U-box E3-ubiquitin ligase-encoding gene, was isolated from soybean and functionally characterized. The expression of GmPUB21, which possesses E3-ubiquitin ligase activity, was found to be significantly up-regulated by drought, salinity, and ABA treatments. The fusion protein GmPUB21-GFP was localized in the cytoplasm, nucleus, and plasma membrane. Transgenic lines of the Nicotiana benthamiana over-expressing GmPUB21 showed more sensitive to osmotic, salinity stress and ABA in seed germination and inhibited mannitol/NaCl-mediated stomatal closure. Moreover, higher reactive oxygen species accumulation was observed in GmPUB21 overexpressing plants after drought and salinity treatment than in wild-type (WT) plants. Contrarily, silencing of GmPUB21 in soybean plants significantly enhanced the tolerance to drought and salinity stresses. Collectively, our results revealed that GmPUB21 negatively regulates the drought and salinity tolerance by increasing the stomatal density and aperture via the ABA signaling pathway. These findings improved our understanding of the role of GmPUB21 under drought and salinity stresses in soybean. [ABSTRACT FROM AUTHOR]

Soybean mosaic virus (SMV) is a worldwide disease of soybean (Glycine max (L.) Merr.) that can cause serious reduction in yield and seed quality. Soybean cv. Qihuang-1 is an important source of resistance to SMV in China, carrying a resistance gene (RSC3Q) against SMV strain SC3. In order to discover genes and networks regulated by RSC3Q -mediated resistance in Qihuang-1, we analysed transcriptome data of a pair of near-isogenic lines, R (RSC3Q) and S (rSC3Q), from the cross Qihuang-1 × Nannong 1138-2 (rSC3Q), after SC3 inoculation. Many differentially expressed genes (DEGs) were identified in the R and S lines at 6, 20 and 48 h post-inoculation. Based on pathway-enrichment analysis of DEGs, three genes encoding calmodulin-like protein (Glyma03g28650 , Glyma19g31395 and Glyma11g33790) with downregulated expression in the S line were identified in the plant–pathogen interaction pathway at 6 h post-inoculation. Analyses by quantitative real-time PCR were performed to verify that these three genes were not beneficial for SMV infection. Our results also revealed a complex plant-hormone signal network in RSC3Q -mediated resistance during the early stage of SMV infection. Expression of jasmonic acid repressor genes (TIFY/JAZ) and abscisic acid-induced genes (PP2C3a) was upregulated in the R line but not the S line. More DEGs related to indole-3-acetic acid were found in the R line than the S line, and no salicylic acid-related DEGs were identified. These results suggest that suppression of jasmonic acid or promotion of abscisic acid is important for RSC3Q -mediated resistance against SC3, and that salicylic acid may not act as a main regulator of RSC3Q -mediated resistance during early stages of SC3 infection. Growth and development were greatly affected through RSC3Q -mediated resistance responses after SC3 infection. Our understanding would be enhanced by identification of factors associated with RSC3Q that help to trigger the resistance response. SMV is a worldwide soybean disease that can cause serious reduction in yield and seed quality; however, the mechanism underlying SMV resistance is not clear. We applied transcriptome analysis of a pair of near-isogenic lines from Chinese cultivars Qihuang-1(R) and Nannong 1138-2(r). Some genes and networks in Qihuang-1 that are regulated by R -mediated resistance were discovered. [ABSTRACT FROM AUTHOR]

Soybean mosaic virus (SMV) infections can cause significant reduction in soybean yield and seed quality. Identifying SMV resistance genes in soybean and accordingly breeding resistant soybean varieties is an important approach to control soybean loss caused by SMV. Here, whole genome resequencing was performed on a pair of near‐isogenic line (NIL), one resistant (R) line and one susceptible (S) line to SC3, from Qihuang‐1 (resistant to SC3, RSC3Q) × Nannong 1138‐2 (susceptible to SC3, rSC3Q). Based on the distribution of single nucleotide polymorphisms (SNPs) and InDels in each chromosome, the RSC3Q was mapped to a region between 27.4 and 29.8 Mbp in chromosome 13. Comparing the sequence and transcriptome data between R line and susceptible lines (S line and Willams82) to SC3, three RSC3Q candidate genes (Glyma13g25950, Glyma13g25970 and Glyma13g26380) were identified. Silencing Glyma13g26380 in Qihuang‐1 increased the accumulation of SMV at 6 and 24 hpi. Subsequently, the SMV concentration in silenced plants decreased to the same level as that of the control. Thus, Glyma13g26380 is involved in the early resistance response of Qihuang No. 1 to SC3. These results provide a basis for elucidating the soybean resistance mechanism against SMV and breeding more resistant soybean varieties against SMV. [ABSTRACT FROM AUTHOR]

Abstract: Soybean mosaic virus (SMV) can cause serious yield losses in soybean. Soybean cultivar ‘RN‐9’ is resistant to 15 of 21 SMV strains. To well‐characterize this invaluable broad‐spectrum SMV‐resistance, populations (F1, F2 and F2:3) derived from resistant (R) × susceptible (S) and R × R crosses were tested for SMV‐SC18 resistance. Genetic analysis revealed that SC18 resistance in ‘RN‐9’ plus two elite SMV‐resistant genotypes (‘Qihuang No.1’ and ‘Kefeng No.1’) are controlled by independently single dominant genes. Linkage analysis showed that the resistance of ‘RN‐9’ to SMV strains SC10, SC14, SC15 and SC18 is controlled by more than one gene(s). Moreover, Rsc10‐r and Rsc18‐r were both positioned between the two simple sequence repeats markers Satt286 and Satt277, while Rsc14‐r was fine‐mapped in 136.8‐kb genomic region containing sixteen genes, flanked by BARCSOYSSR_06_0786 and BARCSOYSSR_06_0790 at genetic distances of 3.79 and 4.14 cM, respectively. Allelic sequence comparison showed that Cytochrome P450‐encoding genes (Glyma.06g176000 and Glyma.06g176100) likely confer the resistance to SC14 in ‘RN‐9’. Our results would facilitate the breeding of broad‐spectrum and durable SMV resistance in soybeans. [ABSTRACT FROM AUTHOR]

Key message: The Mendelian gene conferring resistance to Soybean mosaic virus Strain SC20 in soybean was fine-mapped onto a 79-kb segment on Chr.13 where two closely linked candidate genes were identified and qRT-PCR verified. Abstract: Soybean mosaic virus (SMV) threatens the world soybean production, particularly in China. A country-wide SMV strain system composed of 22 strains was established in China, among which SC20 is a dominant strain in five provinces in Southern China. Resistance to SC20 was evaluated in parents, F, F and the F RIL (recombinant inbred line) population derived from a cross between Qihuang-1 (resistant) and NN1138-2 (susceptible). The segregation ratio of resistant to susceptible in the populations suggested a single dominant gene involved in the resistance to SC20 in Qihuang-1. A 'partial genome mapping strategy' was used to map the resistance gene on Chromosome 13. Linkage analysis between 178 RILs and genetic markers showed that the SC20-resistance gene located at 3.9 and 3.8 cM to the flanking markers BARCSOYSSR_13_1099 and BARCSOYSSR_13_1185 on Chromosome 13. Subsequently, a residual heterozygote segregating population with 346 individuals was developed by selfing four plants heterozygous at markers adjacent to the tentative SC20-resistance gene; then, the candidate region was delimited to a genomic interval of approximately 79 kb flanked by the new markers gm-ssr_13-14 and gm-indel_13-3. Among the seven annotated candidate genes in this region, two genes, Glyma.13G194700 and Glyma.13G195100, encoding Toll Interleukin Receptor-nucleotide-binding-leucine-rich repeat resistance proteins were identified as candidate resistance genes by quantitative real-time polymerase chain reaction and sequence analysis. The two closely linked genes work together to cause the phenotypic segregation as a single Mendelian gene. These results will facilitate marker-assisted selection, gene cloning and breeding for the resistance to SC20. [ABSTRACT FROM AUTHOR]

Key message : Rsc15, a novel locus underlying soybean resistance to SMV, was fine mapped to a 95-kb region on chromosome 6. The Rsc15- mediated resistance is likely attributed to the gene GmPEX14 , the relative expression of which was highly correlated with the accumulation of H O along with the activities of POD and CAT during the early stages of SMV infection in RN-9. Abstract: Soybean mosaic virus (SMV) causes severe yield losses and seed quality deterioration in soybean [ Glycine max (L.) Merr.] worldwide. A series of single dominant SMV resistance genes have been identified on respective soybean chromosomes 2, 13 and 14, while one novel locus, Rsc15, underlying resistance to the virulent SMV strain SC15 from soybean cultivar RN-9 has been recently mapped to a 14.6-cM region on chromosome 6. However, candidate gene has not yet been identified within this region. In the present study, we aimed to fine map the Rsc15 region and identify candidate gene(s) for this invaluable locus. High-resolution fine-mapping revealed that the Rsc15 gene was located in a 95-kb genomic region which was flanked by the two simple sequence repeat (SSR) markers SSR_06_17 and BARCSOYSSR_06_0835. Allelic sequence comparison and expression profile analysis of candidate genes inferred that the gene Glyma.06g182600 (designated as GmPEX14) was the best candidate gene attributing for the resistance of Rsc15, and that genes encoding receptor-like kinase (RLK) (i.e., Glyma.06g175100 and Glyma.06g184400) and serine/threonine kinase (STK) (i.e., Glyma.06g182900 and Glyma.06g183500) were also potential candidates. High correlations were established between the relative expression level of GmPEX14 and the hydrogen peroxide (HO) concentration and activities of catalase (CAT) and peroxidase (POD) during the early stages of SMV-SC15 infection in RN-9. The results of the present study will be useful in marker-assisted breeding for SMV resistance and will lead to further understanding of the molecular mechanisms of host resistance against SMV. [ABSTRACT FROM AUTHOR]

The Ferredoxin-NADP reductase (FNR) gene plays a significant role in NADPH production, carbon assimilation, antioxidation, and cross-talking between chloroplasts and mitochondria in plants. This study aims to know the functional response of the soybean FNR gene (GmFNR) during a soybean mosaic virus (SMV) infection. For this purpose, we developed the bean pod mottle virus (BPMV)-based gene construct (BPMV-GmFNR) and used it to silence the GmFNR gene in resistant and susceptible lines. The results showed that GmFNR expression decreased to 50% in the susceptible line, compared to 40% in the resistant line. The silencing of GmFNR reduces the photosynthetic capacity and CAT activity of both lines compared to their respective controls. In addition, the H2O2 content increased significantly in the susceptible line, whereas the resistant line did not exhibit any change. Further, an SMV infection in the silencing plants of the susceptible line resulted in serious morphological changes and increased the SMV NIa-protease transcript accumulation compared to its control plants. However, the same impact was not observed in the resistant line. The yeast two-hybrid system, BIFC assay, and quantitative real-time polymerase chain reaction (qRT-PCR) analyses revealed that the GmFNR was interacting with EF1A and coincided with the increased SMV accumulation. The results obtained in this study improve the understanding of the soybean FNR gene response during SMV infection and provide a novel insight into the SMV resistance mechanism. [ABSTRACT FROM AUTHOR]