A Transcriptional Regulatory Network of \u3cem\u3eRsv3\u3c/em\u3e-Mediated Extreme Resistance against \u3cem\u3eSoybean Mosaic Virus\u3c/em\u3e

Resistance genes are an effective means for disease control in plants. They predominantly function by inducing a hypersensitive reaction, which results in localized cell death restricting pathogen spread. Some resistance genes elicit an atypical response, termed extreme resistance, where resistance is not associated with a hypersensitive reaction and its standard defense responses. Unlike hypersensitive reaction, the molecular regulatory mechanism(s) underlying extreme resistance is largely unexplored. One of the few known, naturally occurring, instances of extreme resistance is resistance derived from the soybean Rsv3 gene, which confers resistance against the most virulent Soybean mosaic virus strains. To discern the regulatory mechanism underlying Rsv3-mediated extreme resistance, we generated a gene regulatory network using transcriptomic data from time course comparisons of Soybean mosaic virus-G7-inoculated resistant (L29, Rsv3-genotype) and susceptible (Williams82, rsv3-genotype) soybean cultivars. Our results show Rsv3 begins mounting a defense by 6 hpi via a complex phytohormone network, where abscisic acid, cytokinin, jasmonic acid, and salicylic acid pathways are suppressed. We identified putative regulatory interactions between transcription factors and genes in phytohormone regulatory pathways, which is consistent with the demonstrated involvement of these pathways in Rsv3-mediated resistance. One such transcription factor identified as a putative transcriptional regulator was MYC2 encoded by Glyma.07G051500. Known as a master regulator of abscisic acid and jasmonic acid signaling, MYC2 specifically recognizes the G-box motif (“CACGTG”), which was significantly enriched in our data among differentially expressed genes implicated in abscisic acid- and jasmonic acid-related activities. This suggests an important role for Glyma.07G051500 in abscisic acid- and jasmonic acid-derived defense signaling in Rsv3. Resultantly, the findings from our network offer insights into genes and biological pathways underlying the molecular defense mechanism of Rsv3-mediated extreme resistance against Soybean mosaic virus. The computational pipeline used to reconstruct the gene regulatory network in this study is freely available at https://github.com/LiLabAtVT/rsv3-network

Soybean mosaic virus: A successful potyvirus with a wide distribution but restricted natural host range

Taxonomy. Soybean mosaic virus (SMV) is a species within the genus Potyvirus, family Potyviridae that includes almost a quarter of all known plant RNA viruses affecting agriculturally important plants. The Potyvirus genus is the largest of all genera of plant RNA viruses with 160 species. Particle. The filamentous particles of SMV, typical of potyviruses, are about 7,500 Å long and 120 Å in diameter with a central hole of about 15 Å in diameter. Coat protein residues are arranged in helice of about 34 Å pitch having slightly less than 9 subunits per turn. Genome. The SMV genome consists of a single-stranded positive-sense polyadenylated RNA of approximately 9.6 kb with a virus-encoded protein (VPg) linked at the 5\u27 terminus. The genomic RNA contains a single large open reading frame (ORF). The polypeptide produced from the large ORF is processed proteolytically by three viral-encoded proteinases to yield about 10 functional proteins. A small ORF, partially overlapping the P3 cistron, pipo, is encoded as a fusion protein in the N-terminus of P3 (P3N+PIPO). Biological properties. SMV’s host range is restricted mostly to two plant species of a single genus; Glycine max (cultivated soybean) and G. soja (wild soybean). SMV is transmitted by aphids non-persistently and by seeds. Variability of SMV is recognized by reactions on cultivars with dominant resistance (R) genes. Recessive resistance genes are not known. Geographical distribution and economic importance. As a consequence of its seed transmissibility, SMV is present in all soybean growing areas of the world. SMV infections can reduce significantly seed quantity and quality (e.g., mottled seed coats, reduced seed size and viability, and altered chemical composition). Control. The most effective means of managing losses from SMV are planting virus-free seeds and cultivars containing single or multiple R genes. Key attractions. The interactions of SMV with soybean genotypes containing different dominant R genes and understanding functional role(s) of SMV-encoded proteins in virulence, transmission and pathogenicity have been intensively investigated. The SMV-soybean pathosystem has become an excellent model for examining the genetics and genomics of uniquely complex gene-for-gene resistance model in a crop of worldwide importance

A Method for Combining Isolates of Phytophthora sojae to Screen for Novel Sources of Resistance to Phytophthora Stem and Root Rot in Soybean

Soybean cultivars with specific single resistance genes (Rps) are grown to reduce yield loss due to Phytophthora stem and root rot caused by the oomycete pathogen Phytophthora sojae. To identify novel Rps loci, soybean lines are often screened several times, each time with an isolate of P. sojae that differs in virulence on various Rps genes. The goal of this study was to determine whether several isolates of P. sojae that differ in virulence on Rpsgenes could be combined into a single source of inoculum and used to screen soybean lines for novel Rps genes. A set of 14 soybean differential lines, each carrying a specific Rps gene, was inoculated with three isolates of P. sojae, which differed in virulence on 6 to 10 Rps genes, individually or in a 1:1:1 mixture. Inoculum containing the 1:1:1 mixture of isolates was virulent on 13 Rps genes. The mixed-inoculum method was used to screen 1,019 soybean accessions in a blind assay for novel sources of resistance. In all, 17% of Glycine max accessions and 11% of G. soja accessions were resistant (≤30% dead plants), suggesting that these accessions may carry a novel Rps gene or genes. Advantages of combining isolates into a single source of inoculum include reduced cost, ability to screen soybean germplasm with inoculum virulent on all known Rps genes, and ease of identifying novel sources of resistance. This study is a precursor to identifying novel sources of resistance to P. sojae in soybean using RXLR effectors

Chloroplast DNA and molecular phylogeny

The small, relatively constant size and conservative evolution of chloroplast DNA (cpDNA) make it an ideal molecule for tracing the evolutionary history of plant species. At lower taxonomic levels, cpDNA variation is easily and conveniently assayed by comparing restriction patterns and maps, while at higher taxonomic levels, DNA sequencing and inversion analysis are the methods of choice for comparing chloroplast genomes. The study of cpDNA variation has already yielded important new insights into the origin and evolution of many agriculturally important crop plants, and promises to significantly enhance our phylogenetic understanding of the major lines of descent among land plants and algae.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/50180/1/950020607_ftp.pd

Mapping net blotch resistance in ‘Nomini’ and CIho 2291 barley

Net blotch (Pyrenophora teres) is one of the most devastating diseases of barley (Hordeum vulgare L.) worldwide. Identification of diagnostic molecular markers associated with genes and quantitative trait loci (QTL) for net blotch resistance will facilitate pyramiding of independent genes. Linkage mapping was used to identify chromosomal locations of the independent, dominant genes conditioning net blotch resistance in the winter barley ‘Nomini’ (PI 566929) and spring barley CIho 2291. The F2 populations of 238 and 193 individuals, derived from crosses between the susceptible spring barley parent ‘Hector’ (CIho 15514) and the resistant parents Nomini and CIho 2291, respectively, were used to map the genes governing resistance in the resistant parents. The dominant gene governing resistance in Nomini, temporarily designated Rpt-Nomini, was mapped to a 9.2-cM region of barley chromosome 6H between the flanking microsatellite markers Bmag0344a (r2 = 0.7) and Bmag0103a (r2 = 0.9), which were 6.8 and 2.4 cM away from Rpt-Nomini, respectively. The dominant gene governing resistance in CIho 2291, temporarily designated Rpt-CIho2291, was mapped to a 34.3-cM interval on the distal region of barley chromosome 6H between the flanking microsatellite markers Bmag0173 (r2 = 0.65) and Bmag0500 (r2 = 0.26), which were 9.9 and 24.4 cM away from Rpt-CIho2291, respectively. Identification of the chromosomal location of Rpt-Nomini and Rpt-CIho2291 will facilitate efforts in pyramiding multiple genes for net blotch resistance

Infection and genotype remodel the entire soybean transcriptome

<p>Abstract</p> <p>Background</p> <p>High throughput methods, such as high density oligonucleotide microarray measurements of mRNA levels, are popular and critical to genome scale analysis and systems biology. However understanding the results of these analyses and in particular understanding the very wide range of levels of transcriptional changes observed is still a significant challenge. Many researchers still use an arbitrary cut off such as two-fold in order to identify changes that may be biologically significant. We have used a very large-scale microarray experiment involving 72 biological replicates to analyze the response of soybean plants to infection by the pathogen <it>Phytophthora sojae </it>and to analyze transcriptional modulation as a result of genotypic variation.</p> <p>Results</p> <p>With the unprecedented level of statistical sensitivity provided by the high degree of replication, we show unambiguously that almost the entire plant genome (97 to 99% of all detectable genes) undergoes transcriptional modulation in response to infection and genetic variation. The majority of the transcriptional differences are less than two-fold in magnitude. We show that low amplitude modulation of gene expression (less than two-fold changes) is highly statistically significant and consistent across biological replicates, even for modulations of less than 20%. Our results are consistent through two different normalization methods and two different statistical analysis procedures.</p> <p>Conclusion</p> <p>Our findings demonstrate that the entire plant genome undergoes transcriptional modulation in response to infection and genetic variation. The pervasive low-magnitude remodeling of the transcriptome may be an integral component of physiological adaptation in soybean, and in all eukaryotes.</p

Tissue-specific patterns of a maize Myb transcription factor are epigenetically regulated

The maize p1 gene encodes a Myb-homologous regulator of red pigment biosynthesis. To investigate the tissue-specific regulation of the p1 gene, maize plants were transformed with constructs combining promoter and cDNA sequences of two alleles which differ in pigmentation patterns: P1-wr (white pericarp/red cob) and P1-rr (red pericarp/red cob). Surprisingly, all promoter/cDNA combinations produced transgenic plants with red pericarp and red cob (RR pattern), indicating that the P1-wr promoter and encoded protein can function in pericarp. Some of the RR patterned transgenic plants produced progeny plants with white pericarp and red cob (WR pattern), and this switch in tissue-specificity correlated with increased transgene methylation. A similar inverse correlation between pericarp pigmentation and DNA methylation was observed for certain natural p1 alleles, which have a gene structure characteristic of standard P1-wr alleles, but which confer red pericarp pigmentation and are consistently less methylated than standard P1-wr alleles. Although we cannot rule out the possible existence of tissue-specific regulatory elements within the p1 non-coding sequences or flanking regions, the data from transgenic and natural alleles suggest that the tissue-specific pigmentation pattern characteristic of the P1-wrphenotype is epigenetically controlled