scholarly journals IsoSeq transcriptome assembly of C3 panicoid grasses provides tools to study evolutionary change in the Panicoideae

2019 ◽  
Author(s):  
Daniel S. Carvalho ◽  
James C. Schnable
Keyword(s):  
Wild Species ◽  
Sequence Data ◽  
Transcriptome Assembly ◽  
Crop Improvement ◽  
Grass Species ◽  
Crop Species ◽  
Long Read ◽  
Syntenic Gene ◽  
Panicoid Grass ◽  
Reference Genomes

AbstractThe number of plant species with genomic and transcriptomic data has been increasing rapidly. The grasses – Poaceae – have been well represented among species with published reference genomes. However, as a result the genomes of wild grasses are less frequently targeted by sequencing efforts. Sequence data from wild relatives of crop species in the grasses can aid the study of domestication, gene discovery for breeding and crop improvement, and improve our understanding of the evolution of C4 photosynthesis. Here we used long read sequencing technology to characterize the transcriptomes of three C3 panicoid grass species: Dichanthelium oligosanthes, Chasmanthium laxum, and Hymenachne amplexicaulis. Based on alignments to the sorghum genome we estimate that assembled consensus transcripts from each species capture between 54.2 and 65.7% of the conserved syntenic gene space in grasses. Genes co-opted into C4 were also well represented in this dataset, despite concerns that, because these genes might play roles unrelated to photosynthesis in the target species, they would be expressed at low levels and missed by transcript-based sequencing. A combined analysis using syntenic orthologous genes from grasses with published reference genomes and consensus long read sequences from these wild species was consistent with previously published phylogenies. It is hoped that this data, targeting under represented classes of species within the PACMAD grasses – wild species and species utilizing C3 photosynthesis – will aid in futurue studies of domestication and C4 evolution by decreasing the evolutionary distance between C4 and C3 species within this clade, enabling more accurate comparisons associated with evolution of the C4 pathway.

scholarly journals Re-examination of two diatom reference genomes using long-read sequencing

BMC Genomics ◽  
2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Gina V. Filloramo ◽  
Bruce A. Curtis ◽  
Emma Blanche ◽  
John M. Archibald
Keyword(s):  
Repetitive Dna ◽  
Dna Sequences ◽  
Optical Mapping ◽  
Sequence Data ◽  
Thalassiosira Pseudonana ◽  
Rapid Expansion ◽  
Model Organisms ◽  
Algal Group ◽  
Long Read ◽  
Reference Genomes

Abstract Background The marine diatoms Thalassiosira pseudonana and Phaeodactylum tricornutum are valuable model organisms for exploring the evolution, diversity and ecology of this important algal group. Their reference genomes, published in 2004 and 2008, respectively, were the product of traditional Sanger sequencing. In the case of T. pseudonana, optical restriction site mapping was employed to further clarify and contextualize chromosome-level scaffolds. While both genomes are considered highly accurate and reasonably contiguous, they still contain many unresolved regions and unordered/unlinked scaffolds. Results We have used Oxford Nanopore Technologies long-read sequencing to update and validate the quality and contiguity of the T. pseudonana and P. tricornutum genomes. Fine-scale assessment of our long-read derived genome assemblies allowed us to resolve previously uncertain genomic regions, further characterize complex structural variation, and re-evaluate the repetitive DNA content of both genomes. We also identified 1862 previously undescribed genes in T. pseudonana. In P. tricornutum, we used transposable element detection software to identify 33 novel copia-type LTR-RT insertions, indicating ongoing activity and rapid expansion of this superfamily as the organism continues to be maintained in culture. Finally, Bionano optical mapping of P. tricornutum chromosomes was combined with long-read sequence data to explore the potential of long-read sequencing and optical mapping for resolving haplotypes. Conclusion Despite its potential to yield highly contiguous scaffolds, long-read sequencing is not a panacea. Even for relatively small nuclear genomes such as those investigated herein, repetitive DNA sequences cause problems for current genome assembly algorithms. Determining whether a long-read derived genomic assembly is ‘better’ than one produced using traditional sequence data is not straightforward. Our revised reference genomes for P. tricornutum and T. pseudonana nevertheless provide additional insight into the structure and evolution of both genomes, thereby providing a more robust foundation for future diatom research.

scholarly journals Transcript-targeted analysis reveals isoform alterations and double-hop fusions in breast cancer

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Shinichi Namba ◽  
Toshihide Ueno ◽  
Shinya Kojima ◽  
Kenya Kobayashi ◽  
Katsushige Kawase ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cancer Biology ◽  
Sequence Data ◽  
Short Form ◽  
Transcriptome Assembly ◽  
Full Length ◽  
Clinical Samples ◽  
Aberrant Expression ◽  
Fusion Transcripts ◽  
Long Read

AbstractAlthough transcriptome alteration is an essential driver of carcinogenesis, the effects of chromosomal structural alterations on the cancer transcriptome are not yet fully understood. Short-read transcript sequencing has prevented researchers from directly exploring full-length transcripts, forcing them to focus on individual splice sites. Here, we develop a pipeline for Multi-Sample long-read Transcriptome Assembly (MuSTA), which enables construction of a transcriptome from long-read sequence data. Using the constructed transcriptome as a reference, we analyze RNA extracted from 22 clinical breast cancer specimens. We identify a comprehensive set of subtype-specific and differentially used isoforms, which extended our knowledge of isoform regulation to unannotated isoforms including a short form TNS3. We also find that the exon–intron structure of fusion transcripts depends on their genomic context, and we identify double-hop fusion transcripts that are transcribed from complex structural rearrangements. For example, a double-hop fusion results in aberrant expression of an endogenous retroviral gene, ERVFRD-1, which is normally expressed exclusively in placenta and is thought to protect fetus from maternal rejection; expression is elevated in several TCGA samples with ERVFRD-1 fusions. Our analyses provide direct evidence that full-length transcript sequencing of clinical samples can add to our understanding of cancer biology and genomics in general.

scholarly journals Development of Genomic Tools for Cucumber, Cucumis sativus L.

HortScience ◽  
2004 ◽  
Vol 39 (4) ◽  
pp. 774C-774
Author(s):  
Rebecca Grumet* ◽  
Xiaofeng Wang ◽  
Mohamed Tawfik ◽  
Mitch McGrath
Keyword(s):  
Cucumis Sativus ◽  
Genomic Library ◽  
Sequence Data ◽  
Crop Improvement ◽  
Limited Attention ◽  
Yeast Two Hybrid ◽  
High Quality ◽  
Crop Species ◽  
Genomic Tools ◽  
Two Hybrid

Genomics tools have become increasingly varied and valuable for crop improvement. While several species have been targeted for concerted genomic efforts, the majority of horticultural species have received limited attention. Despite the wide variety of important cucurbit crop species, the Cucurbitaceae family has had minimal effort. We have initiated projects to develop genomic tools for cucumber, Cucumis sativus L. Efforts include production of cDNA, yeast two-hybrid, and genomic libraries, and development of an EST database and website for cucumber genomics. Sequences of cucumber leaf ESTs so far indicate that the cDNA library is of high quality and has modest redundancy. Distribution of sequences, as nominally predicted from GeneBank BLAST analysis, indicates that expressed genes fall in the following general categories: photosynthesis (21%), DNA/RNA/protein synthesis (20%), metabolism (15%), signaling (5%), other (16%), and unknown proteins (23%). Cucumber sequence data have been deposited into GenBank and are available on the Michigan State Univ. website (http://genomics.msu.edu/cucumberdb). The yeast two-hybrid library has been successfully used to identify and characterize several genes based on interaction with key proteins of interest, including genes interacting with viral replicases and poly(A) binding protein. The genomic library has been verified to be of high quality and has been used to identify clones of interest.

scholarly journals Comparative DNA Sequence Analysis of Wheat and Rice Genomes

2003 ◽  
Vol 13 (8) ◽  
pp. 1818-1827 ◽  
Author(s):  
Mark E. Sorrells ◽  
Mauricio La Rota ◽  
Catherine E. Bermudez-Kandianis ◽  
Robert A. Greene ◽  
Ramesh Kantety ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Dna Sequence ◽  
Sequence Data ◽  
Chromosomal Rearrangements ◽  
Crop Improvement ◽  
Rice Genome ◽  
Wheat Chromosome ◽  
Comparative Sequence Analysis ◽  
Crop Species ◽  
Transfer Of Information

The use of DNA sequence-based comparative genomics for evolutionary studies and for transferring information from model species to crop species has revolutionized molecular genetics and crop improvement strategies. This study compared 4485 expressed sequence tags (ESTs) that were physically mapped in wheat chromosome bins, to the public rice genome sequence data from 2251 ordered BAC/PAC clones using BLAST. A rice genome view of homologous wheat genome locations based on comparative sequence analysis revealed numerous chromosomal rearrangements that will significantly complicate the use of rice as a model for cross-species transfer of information in nonconserved regions.

scholarly journals Long-read sequencing reveals widespread intragenic structural variants in a recent allopolyploid crop plant

2020 ◽  
Author(s):  
Harmeet Singh Chawla ◽  
HueyTyng Lee ◽  
Iulian Gabur ◽  
Suriya Tamilselvan-Nattar-Amutha ◽  
Christian Obermeier ◽  
...  
Keyword(s):  
Agronomic Traits ◽  
Crop Improvement ◽  
Screening Methods ◽  
Crop Species ◽  
Gene Variation ◽  
Plant Genomes ◽  
Sequencing Technologies ◽  
Long Read ◽  
Genome Screening ◽  
High Level

SummaryGenome structural variation (SV) contributes strongly to trait variation in eukaryotic species and may have an even higher functional significance than single nucleotide polymorphism (SNP). In recent years there have been a number of studies associating large, chromosomal scale SV ranging from hundreds of kilobases all the way up to a few megabases to key agronomic traits in plant genomes. However, there have been little or no efforts towards cataloging small (30 to 10,000 bp) to mid-scale (10,000 bp to 30,000 bp) SV and their impact on evolution and adaptation related traits in plants. This might be attributed to complex and highly-duplicated nature of plant genomes, which makes them difficult to assess using high-throughput genome screening methods. Here we describe how long-read sequencing technologies can overcome this problem, revealing a surprisingly high level of widespread, small to mid-scale SV in a major allopolyploid crop species, Brassica napus. We found that up to 10% of all genes were affected by small to mid-scale SV events. Nearly half of these SV events ranged between 100 bp to 1000 bp, which makes them challenging to detect using short read Illumina sequencing. Examples demonstrating the contribution of such SV towards eco-geographical adaptation and disease resistance in oilseed rape suggest that revisiting complex plant genomes using medium-coverage, long-read sequencing might reveal unexpected levels of functional gene variation, with major implications for trait regulation and crop improvement.

scholarly journals Genome sequences of human cytomegalovirus strain TB40/E variants propagated in fibroblasts and epithelial cells

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Ahmed Al Qaffas ◽  
Salvatore Camiolo ◽  
Mai Vo ◽  
Alexis Aguiar ◽  
Amine Ourahmane ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Human Cytomegalovirus ◽  
Viral Entry ◽  
Sequence Data ◽  
Laboratory Strain ◽  
Serial Passage ◽  
Wild Type Virus ◽  
Protein Coding ◽  
Genetic Changes ◽  
Long Read

AbstractThe advent of whole genome sequencing has revealed that common laboratory strains of human cytomegalovirus (HCMV) have major genetic deficiencies resulting from serial passage in fibroblasts. In particular, tropism for epithelial and endothelial cells is lost due to mutations disrupting genes UL128, UL130, or UL131A, which encode subunits of a virion-associated pentameric complex (PC) important for viral entry into these cells but not for entry into fibroblasts. The endothelial cell-adapted strain TB40/E has a relatively intact genome and has emerged as a laboratory strain that closely resembles wild-type virus. However, several heterogeneous TB40/E stocks and cloned variants exist that display a range of sequence and tropism properties. Here, we report the use of PacBio sequencing to elucidate the genetic changes that occurred, both at the consensus level and within subpopulations, upon passaging a TB40/E stock on ARPE-19 epithelial cells. The long-read data also facilitated examination of the linkage between mutations. Consistent with inefficient ARPE-19 cell entry, at least 83% of viral genomes present before adaptation contained changes impacting PC subunits. In contrast, and consistent with the importance of the PC for entry into endothelial and epithelial cells, genomes after adaptation lacked these or additional mutations impacting PC subunits. The sequence data also revealed six single noncoding substitutions in the inverted repeat regions, single nonsynonymous substitutions in genes UL26, UL69, US28, and UL122, and a frameshift truncating gene UL141. Among the changes affecting protein-coding regions, only the one in UL122 was strongly selected. This change, resulting in a D390H substitution in the encoded protein IE2, has been previously implicated in rendering another viral protein, UL84, essential for viral replication in fibroblasts. This finding suggests that IE2, and perhaps its interactions with UL84, have important functions unique to HCMV replication in epithelial cells.

scholarly journals ValSten: a new wild species derived allotetraploid for increasing genetic diversity of the peanut crop (Arachis hypogaea L.)

2021 ◽  
Author(s):  
Dongying Gao ◽  
Ana C. G. Araujo ◽  
Eliza F. M. B. Nascimento ◽  
M. Carolina Chavarro ◽  
Han Xia ◽  
...  
Keyword(s):  
High Resistance ◽  
Wild Species ◽  
Chromosome Doubling ◽  
Crop Improvement ◽  
Snp Array ◽  
Great Majority ◽  
Morphological Variations ◽  
Peanut Crop ◽  
Cultivated Peanut ◽  
Wild Peanut

AbstractIntrogression of desirable traits from wild relatives plays an important role in crop improvement, as wild species have important characters such as high resistance to pests and pathogens. However, use of wild peanut relatives is challenging because almost all wild species are diploid and sexually incompatible with cultivated peanut, which is tetraploid (AABB genome type; 2n = 4x = 40). To overcome the ploidy barrier, we used 2 wild species to make a tetraploid with the same allotetraploid genome composition as cultivated peanut. Crosses were made between 2 diploid wild species, Arachis valida Krapov. and W.C. Greg. (BB genome; 2n = 2x = 20) and Arachis stenosperma Krapov. and W.C. Greg. (AA genome; 2n = 2x = 20). Cuttings from the diploid F1 AB hybrid were treated with colchicine to induce chromosome doubling thus generating an induced allotetraploid. Chromosome counts confirmed polyploidy (AABB genome; 2n = 4x = 40). We named the new allotetraploid ValSten. Plants had well-developed fertile pollen, produced abundant seed and were sexually compatible with cultivated peanut. ValSten exhibits the same high resistance to early and late leaf spot and rust as its diploid parents. Notably, we observed morphological variations, including flower width and branch angles in the earliest generation (S0) of allotetraploids. A SNP array was used to genotype 47 S0 allotetraploids. The great majority of markers showed the additive allelic state from both parents (AABB). However, some loci were AAAA or BBBB, indicating homeologous recombination. ValSten provides a new, vigorous, highly fertile, disease resistant germplasm for peanut research and improvement.

Cytogenetic analyses of intergeneric hybrids between barley and nine species of Elymus

Genome ◽  
2008 ◽  
Vol 51 (11) ◽  
pp. 897-904 ◽  
Author(s):  
N.-S. Kim ◽  
G. Fedak ◽  
F. Han ◽  
W. Cao
Keyword(s):  
Wild Species ◽  
Meiotic Chromosome ◽  
Abiotic Stress Tolerance ◽  
Chromosome Analysis ◽  
Intergeneric Hybrids ◽  
Biotic And Abiotic Stress ◽  
Crop Species ◽  
Barley Cultivars ◽  
Bivalent Formation ◽  
Gish Analysis

Wild species in the Triticeae tribe are very valuable resources for agronomic improvement in cereal crop species. Intergeneric hybrids were produced between several barley cultivars and perennial species in the genera Elymus , Thinopyrum , and Pseudoroegneria . Caryopsis formation and subsequent plantlet regeneration from embryo culture were variable depending on the hybrid combinations. Chromosome numbers and hybrid identity were confirmed by GISH analysis on the somatic cells of the hybrids. While the hybrids showed very robust vegetative growth and exceeded the parental spikes in size, their floral morphologies resembled that of the wild species. Meiotic chromosome analysis revealed that the bivalent formation frequency per cell ranged from 0.06 in Hordeum vulgare ‘Betzes’ × Elymus curvatus to 3.0 in Elymus humidus  × H. vulgare ‘Manley’. By GISH analysis on the meiocytes of the hybrid E. humidus × ‘Manley’, the frequency of autosyndetic bivalents exceeded the allosyndetic bivalent formation, which gave an insight into the genome constitution of E. humidus as an autoallohexploid species. Regardless of the low allosyndetic chromosome pairing between barley and E. humidus, this combination may be useful for further input, since E. humidus is known to carry many valuable genes for biotic and abiotic stress tolerance.

scholarly journals Reference transcriptome assembly and annotation for perennial ryegrass

Genome ◽  
2017 ◽  
Vol 60 (12) ◽  
pp. 1086-1088 ◽  
Author(s):  
Hiroshi Shinozuka ◽  
Noel O.I. Cogan ◽  
German C. Spangenberg ◽  
John W. Forster
Keyword(s):  
Perennial Ryegrass ◽  
Transcriptome Assembly ◽  
Genotyping By Sequencing ◽  
Whole Genome Sequence ◽  
Grass Species ◽  
Rna Seq ◽  
Pasture Grass ◽  
Tissue Samples ◽  
Genome Sequence Assembly ◽  
Transcriptome Resource

RNA-Seq methodology has been used to generate a comprehensive transcriptome sequence resource for perennial ryegrass, an important temperate pasture grass species. A total of 931 547 255 reads were obtained from libraries corresponding to 19 distinct tissue samples, including both vegetative and reproductive stages of development. Assembly of data generated a final filtered reference set of 48 713 contigs and scaffolds. The transcriptome resource will support whole genome sequence assembly, comparative genomics, implementation of genotyping-by-sequencing (GBS) methods based on transcript sampling, and identification of candidate genes for multiple biological functions.

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