De Novo Assembly and Characterization of Tall Fescue Transcriptome under Water Stress.

Water stress is a fundamental problem for tall fescue [Lolium arundinaceum (Schreb.) Darbysh.] cultivation in the south-central United States. Genetic improvement of tall fescue for water-stress tolerance is the key strategy for improving its persistence in the region. Genotypes with contrasting characteristics for relative water content and osmotic potential were identified from a tall fescue population. Transcriptome profiling between water-stress-tolerant (B400) and water-stress-susceptible (W279) genotypes was performed to unravel the genetic regulatory mechanism of water-stress responses in tall fescue. RNA samples from leaf, shoot, root, and inflorescence were pooled and sequenced through Illumina paired-end sequencing. A total of 199,399 contigs were assembled with an average length of 585 bp. Between the two genotypes, 2986 reference transcripts (RTs) were significantly differentially expressed and 1048 of them could be annotated and found to associate with metabolic pathways and enzyme coding genes. In total, 175 differentially expressed RTs were reported for various stress-related functions. Among those, 65 encoded kinase proteins, 40 each encoded transposons, and transporter proteins were previously reported to be involved with abiotic stress responses. A total of 6348 simple sequence repeats and 6658 single-nucleotide polymorphisms were identified in the contig sequences. Primers were developed from the corresponding sequences, which might be used as candidate gene markers in tall fescue. This study will lead to identification of genes or transcription factors related to water-stress tolerance and development of a comprehensive molecular marker system to facilitate marker-assisted breeding in tall fescue.

Cross-species transferability of microsatellite markers from six aphid (Hemiptera: Aphididae) species and their use for evaluating biotypic diversity in two cereal aphids.

The abundance and distribution of microsatellites, or simple sequence repeats (SSRs) were explored in the expressed sequence tag (EST) and genomic sequences of the pea aphid, Acyrthosiphon pisum (Harris), and the green peach aphid, Myzus persicae (Sulzer). A total of 108 newly developed, together with 40 published, SSR markers were investigated for their cross-species transferability among six aphid species. Genetic diversity among six greenbug, Schizaphis graminum (Rondani) and two Russian wheat aphid, Diuraphis noxia (Kurdjumov) biotypes was further examined with 67 transferable SSRs. It was found that the pea aphid genome is abundant in SSRs with a unique frequency and distribution of SSR motifs. Cross-species transferability of EST-derived SSRs is dependent on phylogenetic closeness between SSR donor and target species, but is higher than that of genomic SSRs. Neighbor-joining analysis of SSR data revealed host-adapted genetic divergence as well as regional differentiation of greenbug biotypes. The two Russian wheat aphid biotypes are genetically as diverse as the greenbug ones although it was introduced into the USA only 20 years ago. This is the first report of large-scale development of SSR markers in aphids, which are expected to have wide applications in aphid genetic, ecological and evolutionary studies.

Mapping the d1 and d2 dwarfing genes and the purple foliage color locus P in pearl millet.

The d(1) and d(2) dwarfing genes and the P purple foliage color gene were placed on the restriction fragment length polymorphism (RFLP)-based molecular marker linkage map of pearl millet [Pennisetum glaucum (L.) R. Br.] using a mapping population based on a cross of inbred lines IP 18293 (D(1)/D(1), d(2)/d(2), P/P) and Tift 238D1 (d(1)/d(1) D(2)/D(2) p/p). A skeleton genetic linkage map of 562 cM (Haldane function) was constructed using 33 RFLP markers and these three morphological markers. The D(1)/d(1) plant height locus mapped to pearl millet linkage group 1, while the D(2)/d(2) plant height locus and the P/p foliage color locus mapped to pearl millet linkage group 4. Loose genetic linkage was observed between the D(2)/d(2) and P/p loci, with 42% repulsion-phase recombination corresponding to 92 cM (Haldane). This loose linkage of morphological marker loci detected on pearl millet LG4 can likely find use in applied pearl millet breeding programs, as host plant resistances to both downy mildew and rust have previously been identified in this genomic region. Such exploitation of these morphological markers in an applied disease resistance breeding program would require development of appropriate genetic stocks, but the relatively loose genetic linkage between d(2) and P suggests that this should not be difficult.