Transcriptome analysis of two buffalograss cultivars

Background: Buffalograss [Buchloë dactyloides (Nutt.) Engel. syn. Bouteloua dactyloides (Nutt.) Columbus] is a United States native turfgrass species that requires less irrigation, fungicides and pesticides compared to more commonly used turfgrass species. In areas where water is limited, interest in this grass species for lawns is increasing. While several buffalograss cultivars have been developed through buffalograss breeding, the timeframe for new cultivar development is long and is limited by a lack of useful genetic resources. Two high throughput next-generation sequencing techniques were used to increase the genomic resources available for buffalograss. Results: Total RNA was extracted and purified from leaf samples of two buffalograss cultivars. ‘378’ and ‘Prestige’ cDNA libraries were subjected to high throughput sequencing on the Illumina GA and Roche 454 Titanium FLX sequencing platforms. The 454 platform (3 samples) produced 1,300,885 reads and the Illumina platform (12 samples) generated approximately 332 million reads. The multiple k-mer technique for de novo assembly using Velvet and Oases was applied. A total of 121,288 contigs were assembled that were similar to previously reported Ensembl commelinid sequences. Original Illumina reads were also mapped to the high quality assembly to estimate expression levels of buffalograss transcripts. There were a total of 325 differentially expressed genes between the two buffalograss cultivars. A glycosyl transferase, serine threonine kinase, and nb-arc domain containing transcripts were among those differentially expressed between the two cultivars. These genes have been previously implicated in defense response pathways and may in part explain some of the performance differences between ‘Prestige’ and ‘378’. Conclusions: To date, this is the first high throughput sequencing experiment conducted on buffalograss. In total, 121,288 high quality transcripts were assembled, significantly expanding the limited genetic resources available for buffalograss genetic studies. Additionally, 325 differentially expressed sequences were identified which may contribute to performance or morphological differences between ‘Prestige’ and ‘378’ buffalograss cultivars

Transcriptomic comparison of invasive bigheaded carps (\u3ci\u3eHypophthalmichthys nobilis\u3c/i\u3e and \u3ci\u3eHypophthalmichthys molitrix\u3c/i\u3e) and their hybrids

Bighead carp (Hypophthalmichthys nobilis) and silver carp (Hypophthalmichthys molitrix), collectively called bigheaded carps, are invasive species in the Mississippi River Basin (MRB). Interspecific hybridization between bigheaded carps has been considered rare within their native rivers in China; however, it is prevalent in the MRB. We conducted de novo transcriptome analysis of pure and hybrid bigheaded carps and obtained 40,759 to 51,706 transcripts for pure, F1 hybrid, and backcross bigheaded carps. The search against protein databases resulted in 20,336–28,133 annotated transcripts (over 50% of the transcriptome) with over 13,000 transcripts mapped to 23 Gene Ontology biological processes and 127 KEGG metabolic pathways. More transcripts were detected in silver carp than in bighead carp; however, comparable numbers of transcripts were annotated. Transcriptomic variation detected between two F1 hybrids may indicate a potential loss of fitness in hybrids. The neighbor-joining distance tree constructed using over 2,500 one-to-one orthologous sequences suggests transcriptomes could be used to infer the history of introgression and hybridization. Moreover, we detected 24,792 candidate SNPs that can be used to identify different species. The transcriptomes, orthologous sequences, and candidate SNPs obtained in this study should provide further knowledge of interspecific hybridization and introgression

The complete mitochondrial genome of the shoal chub, Macrhybopsis hyostoma

The complete mitochondrial genome of the shoal chub (Macrhybopsis hyostoma) was determined to be 16,899 bp and contained 22 tRNA genes, 2 rRNA genes and 1 control region. The whole genome base composition was 30.5% A, 28.5% T, 24.9% C and 16.1 G. This complete mitochondrial genome provides essential molecular markers for resolving phylogeny and future conservation efforts

The complete mitochondrial genome of the silver chub, Macrhybopsis storeriana

The complete mitochondrial genome of the silver chub M. storeriana was determined to be 16,709 bp and contained 22 tRNA genes, 2 rRNA genes and one control region. The whole genome base composition was 30.3% A, 28% T, 25.5% C and 16.2 G. This complete mitochondrial genome provides essential molecular markers for resolving phylogeny and aiding future conservation efforts

Resolving the genetic paradox of invasions: Preadapted genomes and postintroduction hybridization of bigheaded carps in the Mississippi River Basin

The genetic paradox of biological invasions is complex and multifaceted. In particular, the relative role of disparate propagule sources and genetic adaptation through postintroduction hybridization has remained largely unexplored. To add resolution to this paradox, we investigate the genetic architecture responsible for the invasion of two invasive Asian carp species, bighead carp (Hypophthalmichthys nobilis) and silver carp (H. molitrix) (bigheaded carps) that experience extensive hybridization in the Mississippi River Basin (MRB). We sequenced the genomes of bighead and silver carps (~1.08G bp and ~1.15G bp, respectively) and their hybrids collected from the MRB. We found moderate‐to‐high heterozygosity in bighead (0.0021) and silver (0.0036) carps, detected significantly higher dN/dS ratios of single‐copy orthologous genes in bigheaded carps versus 10 other species of fish, and identified genes in both species potentially associated with environmental adaptation and other invasion‐related traits. Additionally, we observed a high genomic similarity (96.3% in all syntenic blocks) between bighead and silver carps and over 90% embryonic viability in their experimentally induced hybrids. Our results suggest intrinsic genomic features of bigheaded carps, likely associated with life history traits that presumably evolved within their native ranges, might have facilitated their initial establishment of invasion, whereas ex-situ interspecific hybridization between the carps might have promoted their range expansion. This study reveals an alternative mechanism that could resolve one of the genetic paradoxes in biological invasions and provides invaluable genomic resources for applied research involving bigheaded carps

Transcriptome analysis of two buffalograss cultivars

Background: Buffalograss [Buchloë dactyloides (Nutt.) Engel. syn. Bouteloua dactyloides (Nutt.) Columbus] is a United States native turfgrass species that requires less irrigation, fungicides and pesticides compared to more commonly used turfgrass species. In areas where water is limited, interest in this grass species for lawns is increasing. While several buffalograss cultivars have been developed through buffalograss breeding, the timeframe for new cultivar development is long and is limited by a lack of useful genetic resources. Two high throughput next-generation sequencing techniques were used to increase the genomic resources available for buffalograss. Results: Total RNA was extracted and purified from leaf samples of two buffalograss cultivars. ‘378’ and ‘Prestige’ cDNA libraries were subjected to high throughput sequencing on the Illumina GA and Roche 454 Titanium FLX sequencing platforms. The 454 platform (3 samples) produced 1,300,885 reads and the Illumina platform (12 samples) generated approximately 332 million reads. The multiple k-mer technique for de novo assembly using Velvet and Oases was applied. A total of 121,288 contigs were assembled that were similar to previously reported Ensembl commelinid sequences. Original Illumina reads were also mapped to the high quality assembly to estimate expression levels of buffalograss transcripts. There were a total of 325 differentially expressed genes between the two buffalograss cultivars. A glycosyl transferase, serine threonine kinase, and nb-arc domain containing transcripts were among those differentially expressed between the two cultivars. These genes have been previously implicated in defense response pathways and may in part explain some of the performance differences between ‘Prestige’ and ‘378’. Conclusions: To date, this is the first high throughput sequencing experiment conducted on buffalograss. In total, 121,288 high quality transcripts were assembled, significantly expanding the limited genetic resources available for buffalograss genetic studies. Additionally, 325 differentially expressed sequences were identified which may contribute to performance or morphological differences between ‘Prestige’ and ‘378’ buffalograss cultivars

Transcriptional Profiling of Resistant and Susceptible Buffalograsses in Response to \u3ci\u3eBlissus occiduus\u3c/i\u3e (Hemiptera: Blissidae) Feeding

ABSTRACT Understanding plant resistance mechanisms at a molecular level would provide valuable insights into the biological pathways impacted by insect feeding, and help explain specific plant tolerance mechanisms. As a first step in this process, we conducted next-generation sequencing using RNA extracted from chinch bug-tolerant and -susceptible buffalograss genotypes at 7 and 14 d after chinch bug feeding. Sequence descriptions and gene ontology terms were assigned to 1,701 differentially expressed genes. Defense-related transcripts were differentially expressed within the chinch bug-tolerant buffalograss, Prestige, and susceptible buffalograss, 378. Interestingly, four peroxidase transcripts had higher basal expression in tolerant control plants compared with susceptible control plants. Defense-related transcripts, including two peroxidase genes, two catalase genes, several cytochrome P450 transcripts, a glutathione s-transferase, and a WRKY gene were upregulated within the Prestige transcriptome in response to chinch bug feeding. The majority of observed transcripts with oxidoreductase activity, including nine peroxidase genes and a catalase gene, were downregulated in 378 in response to initial chinch bug feeding. The observed difference in transcript expression between these two buffalograss genotypes provides insight into the mechanism(s) of resistance, specifically buffalograss tolerance to chinch bug feeding

Analysis of the Skin Transcriptome in Two Oujiang Color Varieties of Common Carp

<div><p>Background</p><p>Body color and coloration patterns are important phenotypic traits to maintain survival and reproduction activities. The Oujiang color varieties of common carp (<i>Cyprinus carpio</i> var. <i>color</i>), with a narrow distribution in Zhejiang Province of China and a history of aquaculture for over 1,200 years, consistently exhibit a variety of body color patterns. The molecular mechanism underlying diverse color patterns in these variants is unknown. To the practical end, it is essential to develop molecular markers that can distinguish different phenotypes and assist selective breeding.</p><p>Methodology/Principal Findings</p><p>In this exploratory study, we conducted Roche 454 transcriptome sequencing of two pooled skin tissue samples of Oujiang common carp, which correspond to distinct color patterns, red with big black spots (RB) and whole white (WW), and a total of 737,525 sequence reads were generated. The reads obtained in this study were co-assembled jointly with common carp Roche 454 sequencing reads downloaded from NCBI SRA database, resulting in 43,923 isotigs and 546,676 singletons. Over 31 thousand (31,445; 71.6%) isotigs were found with significant BLAST matches (E<1e-10) to the nr protein database, which corresponds to 12,597 annotated zebrafish genes. A total of 70,947 isotigs and singletons (transcripts) were annotated with Gene Ontology, and 60,221 transcripts were found with corresponding EC numbers. Out of 145 zebrafish pigmentation genes, orthologs for 117 were recovered in Oujiang color carp transcriptome, including 18 found only among singletons. Our transcriptome analysis revealed over 52,902 SNPs in Oujiang common carp, and identified 63 SNP markers that are putatively unique either for RB or WW.</p><p>Conclusions</p><p>The transcriptome of Oujiang color varieties of common carp obtained through this study, along with the pigmentation genes recovered and the color pattern-specific molecular markers developed, will facilitate future research on the molecular mechanism of color patterns and promote aquaculture of Oujiang color varieties of common carp through molecular marker assisted-selective breeding.</p></div

Analysis pipeline of Oujiang color varieties of common carp (RB - red with big black spots and WW – whole white) and gynogenic common carp (CC) transcriptomes, with the number of raw 454 reads, singletons and isotigs denoted.

<p>The pictures of RB, WW and CC are shown at the top panel. CC raw sequencing reads were downloaded from NCBI SRA database (SRA 050545).</p

Putative color pattern-specific SNP markers found in Oujiang color varieties of common carp isotig sequences generated from the skin transcriptome^*.

<p>* See <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0090074#pone.0090074.s003" target="_blank">Table S3</a> for the complete list of candidate SNP markers.</p><p>** RB - red with big black spots, WW - whole white.</p