Virus-Induced Gene Silencing in Diverse Maize Lines Using the Brome Mosaic Virus-based silencing vector

Virus-induced gene silencing (VIGS) is a widely used tool for gene function studies in many plant species, though its use in cereals has been limited. In addition, within cereal species the varieties that best respond during VIGS screens are often not known. Using a Brome mosaic virus (BMV) vector designed to silence the maize phytoene desaturase (PDS) gene, a genetically diverse set of maize inbred lines was screened for development of gene silencing after inoculation of seeds through the novel use of a vascular puncture inoculation technique. In addition to Va35, which previously was shown to support silencing, maize lines NC300, Ki11, Oh7b, M162W and CML52 displayed significant visible photobleaching when challenged with the BMV-PDS. In these plants, targeted PDS mRNA expression was decreased 50-80% relative to levels in plants that were inoculated with BMV containing a fragment of the GUS gene or were mock-inoculated

On the genome constitution and evolution of intermediate wheatgrass (Thinopyrum intermedium: Poaceae, Triticeae)

<p>Abstract</p> <p>Background</p> <p>The wheat tribe Triticeae (Poaceae) is a diverse group of grasses representing a textbook example of reticulate evolution. Apart from globally important grain crops, there are also wild grasses which are of great practical value. Allohexaploid intermediate wheatgrass, <it>Thinopyrum intermedium </it>(2n = 6x = 42), possesses many desirable agronomic traits that make it an invaluable source of genetic material useful in wheat improvement. Although the identification of its genomic components has been the object of considerable investigation, the complete genomic constitution and its potential variability are still being unravelled. To identify the genomic constitution of this allohexaploid, four accessions of intermediate wheatgrass from its native area were analysed by sequencing of chloroplast <it>trn</it>L-F and partial nuclear GBSSI, and genomic <it>in situ </it>hybridization.</p> <p>Results</p> <p>The results confirmed the allopolyploid origin of <it>Thinopyrum intermedium </it>and revealed new aspects in its genomic composition. Genomic heterogeneity suggests a more complex origin of the species than would be expected if it originated through allohexaploidy alone. While <it>Pseudoroegneria </it>is the most probable maternal parent of the accessions analysed, nuclear GBSSI sequences suggested the contribution of distinct lineages corresponding to the following present-day genera: <it>Pseudoroegneria</it>, <it>Dasypyrum</it>, <it>Taeniatherum</it>, <it>Aegilops </it>and <it>Thinopyrum</it>. Two subgenomes of the hexaploid have most probably been contributed by <it>Pseudoroegneria </it>and <it>Dasypyrum</it>, but the identity of the third subgenome remains unresolved satisfactorily. Possibly it is of hybridogenous origin, with contributions from <it>Thinopyrum </it>and <it>Aegilops</it>. Surprising diversity of GBSSI copies corresponding to a <it>Dasypyrum</it>-like progenitor indicates either multiple contributions from different sources close to <it>Dasypyrum </it>and maintenance of divergent copies or the presence of divergent paralogs, or a combination of both. <it>Taeniatherum</it>-like GBSSI copies are most probably pseudogenic, and the mode of their acquisition by <it>Th. intermedium </it>remains unclear.</p> <p>Conclusions</p> <p>Hybridization has played a key role in the evolution of the Triticeae. Transfer of genetic material via extensive interspecific hybridization and/or introgression could have enriched the species' gene pools significantly. We have shown that the genomic heterogeneity of intermediate wheatgrass is higher than has been previously assumed, which is of particular concern to wheat breeders, who frequently use it as a source of desirable traits in wheat improvement.</p

The Genetics and Genomics of Virus Resistance in Maize

Viruses cause significant diseases on maize worldwide. Intensive agronomic practices, changes in vector distribution, and the introduction of vectors and viruses into new areas can result in emerging disease problems. Because deployment of resistant hybrids and cultivars is considered to be both economically viable and environmentally sustainable, genes and quantitative trait loci for most economically important virus diseases have been identified. Examination of multiple studies indicates the importance of regions of maize chromosomes 2, 3, 6, and 10 in virus resistance. An understanding of the molecular basis of virus resistance in maize is beginning to emerge, and two genes conferring resistance to sugarcane mosaic virus, Scmv1 and Scmv2, have been cloned and characterized. Recent studies provide hints of other pathways and genes critical to virus resistance in maize, but further work is required to determine the roles of these in virus susceptibility and resistance. This research will be facilitated by rapidly advancing technologies for functional analysis of genes in maize

Glutamine Synthetase and Ferredoxin-Dependent Glutamate Synthase Expression in the Maize (Zea mays) Root Primary Response to Nitrate (Evidence for an Organ-Specific Response).

To define further the early, or primary, events that occur in maize (Zea mays) seedlings exposed to NO3-, accumulation of chloroplast glutamine synthetase (GS2; EC 6.3.1.2) and ferredoxin-dependent glutamate synthase (Fd-GOGAT; EC 1.4.7.1), transcripts were examined in roots and leaves. In roots, NO3- treatment caused a rapid (within 30 min), transient, and cycloheximide-independent accumulation of GS2 and Fd-GOGAT transcripts. In addition, 10 [mu]M external NO3- was sufficient to cause transcript accumulation. The induction was NO3- specific, since NH4Cl treatment did not affect mRNA levels. GS2 and Fd-GOGAT mRNA accumulation in roots was similar to that observed for nitrate reductase (NR) mRNA. Therefore, the four genes involved in NO3- assimilation (NR, nitrite reductase, GS2, and Fd-GOGAT) are expressed in the root primary response to NO3-, suggesting that all four genes can respond to the same signal transduction system. In contrast, relatively high levels of GS2 and Fd-GOGAT mRNAs were present in untreated leaf tissue, and NO3- treatment had little or no influence on transcript accumulation. Rapid, transient, and cycloheximide-independent NR mRNA expression was seen in the NO3--treated leaves, demonstrating that NO3- was not limiting. The NO3--independent constitutive expression of GS2 and Fd-GOGAT is likely due to the requirement for reassimilation of photorespiratory NH4+ in these young leaves