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Insights into transcriptomes of big and low sagebrush.

ABSTRACT: We report the sequencing and assembly of three transcriptomes from Big (Artemisia tridentata ssp. wyomingensis and A. tridentata ssp. tridentata) and Low (A. arbuscula ssp. arbuscula) sagebrush. The sequence reads are available in the Sequence Read Archive of NCBI. We demonstrate the utilities of these transcriptomes for gene discovery and phylogenomic analysis. An assembly of 61,883 transcripts followed by transcript identification by the program TRAPID revealed 16 transcripts directly related to terpene synthases, proteins critical to the production of multiple secondary metabolites in sagebrush. A putative terpene synthase was identified in two of our sagebrush samples. Using paralogs with synonymous mutations we reconstructed an evolutionary time line of ancient genome duplications. By applying a constant mutation rate to the data we estimate that these three ancient duplications occurred about 18, 34 and 60 million years ago. These transcriptomes offer a foundation for future studies of sagebrush, including inferences in chemical defense and the identification of species and subspecies of sagebrush for restoration and preservation of the threatened sage-grouse.

SUBMITTER: Huynh MD

PROVIDER: S-EPMC4447352 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

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Insights into transcriptomes of big and low sagebrush.

Huynh Mark D MD Page Justin T JT Richardson Bryce A BA Udall Joshua A JA

PloS one 20150528 5

We report the sequencing and assembly of three transcriptomes from Big (Artemisia tridentata ssp. wyomingensis and A. tridentata ssp. tridentata) and Low (A. arbuscula ssp. arbuscula) sagebrush. The sequence reads are available in the Sequence Read Archive of NCBI. We demonstrate the utilities of these transcriptomes for gene discovery and phylogenomic analysis. An assembly of 61,883 transcripts followed by transcript identification by the program TRAPID revealed 16 transcripts directly related ...[more]

PMID: 26020526

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Project description:BackgroundBig sagebrush (Artemisia tridentata) is one of the most widely distributed and ecologically important shrub species in western North America. This species serves as a critical habitat and food resource for many animals and invertebrates. Habitat loss due to a combination of disturbances followed by establishment of invasive plant species is a serious threat to big sagebrush ecosystem sustainability. Lack of genomic data has limited our understanding of the evolutionary history and ecological adaptation in this species. Here, we report on the sequencing of expressed sequence tags (ESTs) and detection of single nucleotide polymorphism (SNP) and simple sequence repeat (SSR) markers in subspecies of big sagebrush.ResultscDNA of A. tridentata sspp. tridentata and vaseyana were normalized and sequenced using the 454 GS FLX Titanium pyrosequencing technology. Assembly of the reads resulted in 20,357 contig consensus sequences in ssp. tridentata and 20,250 contigs in ssp. vaseyana. A BLASTx search against the non-redundant (NR) protein database using 29,541 consensus sequences obtained from a combined assembly resulted in 21,436 sequences with significant blast alignments (≤ 1e⁻¹⁵). A total of 20,952 SNPs and 119 polymorphic SSRs were detected between the two subspecies. SNPs were validated through various methods including sequence capture. Validation of SNPs in different individuals uncovered a high level of nucleotide variation in EST sequences. EST sequences of a third, tetraploid subspecies (ssp. wyomingensis) obtained by Illumina sequencing were mapped to the consensus sequences of the combined 454 EST assembly. Approximately one-third of the SNPs between sspp. tridentata and vaseyana identified in the combined assembly were also polymorphic within the two geographically distant ssp. wyomingensis samples.ConclusionWe have produced a large EST dataset for Artemisia tridentata, which contains a large sample of the big sagebrush leaf transcriptome. SNP mapping among the three subspecies suggest the origin of ssp. wyomingensis via mixed ancestry. A large number of SNP and SSR markers provide the foundation for future research to address questions in big sagebrush evolution, ecological genetics, and conservation using genomic approaches.

Project description:Big defensins are antimicrobial polypeptides believed to be the ancestors of ?-defensins, the most evolutionary conserved family of host defense peptides (HDPs) in vertebrates. Nevertheless, big defensins underwent several independent gene loss events during animal evolution, being only retained in a limited number of phylogenetically distant invertebrates. Here, we explore the evolutionary history of this fascinating HDP family and investigate its patchy distribution in extant metazoans. We highlight the presence of big defensins in various classes of lophotrochozoans, as well as in a few arthropods and basal chordates (amphioxus), mostly adapted to life in marine environments. Bivalve mollusks often display an expanded repertoire of big defensin sequences, which appear to be the product of independent lineage-specific gene tandem duplications, followed by a rapid molecular diversification of newly acquired gene copies. This ongoing evolutionary process could underpin the simultaneous presence of canonical big defensins and non-canonical (?-defensin-like) sequences in some species. The big defensin genes of mussels and oysters, two species target of in-depth studies, are subjected to gene presence/absence variation (PAV), i.e., they can be present or absent in the genomes of different individuals. Moreover, big defensins follow different patterns of gene expression within a given species and respond differently to microbial challenges, suggesting functional divergence. Consistently, current structural data show that big defensin sequence diversity affects the 3D structure and biophysical properties of these polypeptides. We discuss here the role of the N-terminal hydrophobic domain, lost during evolution toward ?-defensins, in the big defensin stability to high salt concentrations and its mechanism of action. Finally, we discuss the potential of big defensins as markers for animal health and for the nature-based design of novel therapeutics active at high salt concentrations.

Project description:Study questionWhat variations underlie the menstrual cycle length and ovulation day of women trying to conceive?Summary answerBig data from a connected ovulation test revealed the extent of variation in menstrual cycle length and ovulation day in women trying to conceive.What is known alreadyTiming intercourse to coincide with the fertile period of a woman maximises the chances of conception. The day of ovulation varies on an inter- and intra-individual level.Study design size durationA total of 32 595 women who had purchased a connected ovulation test system contributed 75 981 cycles for analysis. Day of ovulation was determined from the fertility test results. The connected home ovulation test system enables users to identify their fertile phase. The app benefits users by enabling them to understand their personal fertility information. During each menstrual cycle, users input their perceived cycle length into an accessory application, and data on hormone levels from the tests are uploaded to the application and stored in an anonymised cloud database. This study compared users' perceived cycle characteristics with actual cycle characteristics. The perceived and actual cycle length information was analysed to provide population ranges.Participants/materials setting methodsThis study analysed data from the at-home use of a commercially available connected home ovulation test by women across the USA and UK.Main results and the role of chanceOverall, 25.3% of users selected a 28-day cycle as their perceived cycle length; however, only 12.4% of users actually had a 28-day cycle. Most women (87%) had actual menstrual cycle lengths between 23 and 35 days, with a normal distribution centred on day 28, and over half of the users (52%) had cycles that varied by 5 days or more. There was a 10-day spread of observed ovulation days for a 28-day cycle, with the most common day of ovulation being Day 15. Similar variation was observed for all cycle lengths examined. For users who conducted a test on every day requested by the app, a luteinising hormone (LH) surge was detected in 97.9% of cycles.Limitations reasons for cautionData were from a self-selected population of women who were prepared to purchase a commercially available product to aid conception and so may not fully represent the wider population. No corresponding demographic data were collected with the cycle information.Wider implications of the findingsUsing big data has provided more personalised insights into women's fertility; this could enable women trying to conceive to better time intercourse, increasing the likelihood of conception.Study funding/competing interestsThe study was funded by SPD Development Company Ltd (Bedford, UK), a fully owned subsidiary of SPD Swiss Precision Diagnostics GmbH (Geneva, Switzerland). I.S., B.G. and S.J. are employees of the SPD Development Company Ltd.

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Insights into transcriptomes of big and low sagebrush.

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Insights into transcriptomes of big and low sagebrush.

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