[Show abstract][Hide abstract] ABSTRACT: Pineapple (Ananas comosus (L.) Merr.) is the most economically valuable crop possessing crassulacean acid metabolism (CAM), a photosynthetic carbon assimilation pathway with high water-use efficiency, and the second most important tropical fruit. We sequenced the genomes of pineapple varieties F153 and MD2 and a wild pineapple relative, Ananas bracteatus accession CB5. The pineapple genome has one fewer ancient whole-genome duplication event than sequenced grass genomes and a conserved karyotype with seven chromosomes from before the ρ duplication event. The pineapple lineage has transitioned from C3 photosynthesis to CAM, with CAM-related genes exhibiting a diel expression pattern in photosynthetic tissues. CAM pathway genes were enriched with cis-regulatory elements associated with the regulation of circadian clock genes, providing the first cis-regulatory link between CAM and circadian clock regulation. Pineapple CAM photosynthesis evolved by the reconfiguration of pathways in C3 plants, through the regulatory neofunctionalization of preexisting genes and not through the acquisition of neofunctionalized genes via whole-genome or tandem gene duplication.
[Show abstract][Hide abstract] ABSTRACT: Brycon amazonicus is an important freshwater migratory fish in the Amazon Basin. 29 Studies involving populations of B. amazonicus are of great importance for the conservation and 30 management of this species. We developed eight microsatellite loci and applied them to 31 investigate the genetic variation of 32 wild individuals from Catalan lake of the Black river. The 32 number of alleles per locus ranged from 6 to 17, with an average of 11.1. The observed and 33 expected heterozygosity values ranged from 0.654 to 0.906 (average 0.676) and from 0.527 to 34 0.887 (average 0.792), respectively. The value of f ranged from -0.018 to 0.239 (average 35 0.038). No significant linkage disequilibrium was detected. These microsatellite loci will 36 contribute towards studies of genetic diversity and conservation of B. amazonicus.
[Show abstract][Hide abstract] ABSTRACT: The family of grasses encompasses the world's most important food, feed, and bioenergy crops, yet we are only now beginning to develop the genetic resources to explore the diversity of form and function that underlies economically important traits. Two emerging model systems, Brachypodium distachyon and Setaria viridis, promise to greatly accelerate the process of gene discovery in the grasses and to serve as bridges in the exploration of panicoid and pooid grasses, arguably two of the most important clades of plants from a food security perspective. We provide both a historical view of the development of plant model systems and highlight several recent reports that are providing these developing communities with the tools for gene discovery and pathway engineering. Expected final online publication date for the Annual Review of Plant Biology Volume 66 is April 29, 2015. Please see http://www.annualreviews.org/catalog/pubdates.aspx for revised estimates.
[Show abstract][Hide abstract] ABSTRACT: Nicotiana, a member of the Solanaceae family, is one of the most important research model plants, and of high agricultural and economic value worldwide. To better understand the substantial and rapid research progress with Nicotiana in recent years, its genomics, genetics, and nicotine gene studies are summarized, with useful web links. Several important genetic maps, including a high-density map of N. tabacum consisting of ~2,000 markers published in 2012, provide tools for genetics research. Four whole genome sequences are from allotetraploid species, including N. benthamiana in 2012, and three N. tabacum cultivars (TN90, K326, and BX) in 2014. Three whole genome sequences are from diploids, including progenitors N. sylvestris and N.
tomentosiformis in 2013 and N. otophora in 2014. These and additional studies provide numerous insights into genome evolution after polyploidization, including changes in gene composition and transcriptome expression in N. tabacum. The major genes involved in the nicotine biosynthetic pathway have been identified and the genetic basis of the differences in nicotine levels among Nicotiana species has been revealed. In addition, other progress on chloroplast, mitochondrial, and NCBI-registered projects on Nicotiana are discussed. The challenges and prospects for genomic, genetic and application research are addressed. Hence, this review provides important resources and guidance for current and future research and application in Nicotiana.
Molecular Genetics and Genomics 01/2015; DOI:10.1007/s00438-015-0989-7 · 2.73 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Tef (Eragrostis tef) is the mainstay of Ethiopian agriculture, with more acres planted than in any other crop. Tef has high resilience to both drought and waterlogged soils, but yield is limited by the tiny seed size and severe susceptibility to lodging. We have used mutational and other genetic approaches to help solve these two problems. Among tef’s many benefits is the exceptional nutritional quality of its grain. Tef grain is very high in protein and mineral content, especially calcium and iron. Recently, we have generated a recombinational map of tef, with 486 SNP markers in a RIL population, using genotype-by-sequencing technology. We are using this map to investigate QTL associated with various aspects of nutritional quality, particularly those related to mineral content. When combined with two full genome sequence analyses that have recently been completed, these studies should uncover candidate genes associated with these traits, thus suggesting routes towards further nutritional improvement of tef and other cereals.
Plant and Animal Genome XXIII, San Diego, CA; 01/2015
[Show abstract][Hide abstract] ABSTRACT: Numerous instances of presence/absence variations for introns have been documented in eukaryotes, and some cases of recurrent loss of the same intron have been suggested. However, there has been no comprehensive or phylogenetically deep analysis of recurrent intron loss. Of 883 cases of intron presence/absence variation that we detected in five sequenced grass genomes, 93 were confirmed as recurrent losses and the rest could be explained by single losses (652) or single gains (118). No case of recurrent intron gain was observed. Deep phylogenetic analysis often indicated that apparent intron gains were actually numerous independent losses of the same intron. Recurrent loss exhibited extreme non-randomness, in that some introns were removed independently in many lineages. The two larger genomes, maize and sorghum, were found to have a higher rate of both recurrent loss and overall loss and/or gain than foxtail millet, rice or Brachypodium. Adjacent introns and small introns were found to be preferentially lost. Intron loss genes exhibited a high frequency of germ line or early embryogenesis expression. In addition, flanking exon A+T-richness and intron TG/CG ratios were higher in retained introns. This last result suggests that epigenetic status, as evidenced by a loss of methylated CG dinucleotides, may play a role in the process of intron loss. This study provides the first comprehensive analysis of recurrent intron loss, makes a series of novel findings on the patterns of recurrent intron loss during the evolution of the grass family, and provides insight into the molecular mechanism(s) underlying intron loss.
[Show abstract][Hide abstract] ABSTRACT: Comparative genomic analyses among closely related species can greatly enhance our understanding of plant gene and genome evolution. We report de novo-assembled AA-genome sequences for Oryza nivara, Oryza glaberrima, Oryza barthii, Oryza glumaepatula, and Oryza meridionalis. Our analyses reveal massive levels of genomic structural variation, including segmental duplication and rapid gene family turnover, with particularly high instability in defense-related genes. We show, on a genomic scale, how lineage-specific expansion or contraction of gene families has led to their morphological and reproductive diversification, thus enlightening the evolutionary process of speciation and adaptation. Despite strong purifying selective pressures on most Oryza genes, we documented a large number of positively selected genes, especially those genes involved in flower development, reproduction, and resistance-related processes. These diversifying genes are expected to have played key roles in adaptations to their ecological niches in Asia, South America, Africa and Australia. Extensive variation in noncoding RNA gene numbers, function enrichment, and rates of sequence divergence might also help account for the different genetic adaptations of these rice species. Collectively, these resources provide new opportunities for evolutionary genomics, numerous insights into recent speciation, a valuable database of functional variation for crop improvement, and tools for efficient conservation of wild rice germplasm.
Proceedings of the National Academy of Sciences 11/2014; 111:E4954-E4962. DOI:10.1073/pnas.1418307111 · 9.67 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Plants from the Zingiberaceae family are a key source of spices and herbal medicines. Species identification within this group is critical in the search for known and possibly novel bioactive compounds. To facilitate precise characterization of this group, we have sequenced chloroplast genomes from species representing five major groups within Zingiberaceae. Generally, the structure of these genomes is similar to the basal angiosperm excepting an expansion of 3 kb associated with the inverted repeat A region. Portions of this expansion appear to be shared across the entire Zingiberales order, which includes gingers and bananas. We used whole plastome alignment information to develop DNA barcodes that would maximize the ability to differentiate species within the Zingiberaceae. Our computation pipeline identified regions of high variability that were flanked by highly conserved regions used for primer design. This approach yielded hitherto unexploited regions of variability. These theoretically optimal barcodes were tested on a range of species throughout the family and were found to amplify and differentiate genera and, in some cases, species. Still, though these barcodes were specifically optimized for the Zingiberaceae, our data support the emerging consensus that whole plastome sequences are needed for robust species identification and phylogenetics within this family.
PLoS ONE 10/2014; 9(10). DOI:10.1371/journal.pone.0108581 · 3.23 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The insertion of DNA into a genome can result in the duplication and dispersal of functional sequences through the genome. In addition, a deeper understanding of insertion mechanisms will inform methods of genetic engineering and plant transformation. Exploiting structural variations in numerous rice accessions, we have inferred and analyzed intermediate length (10-1,000 bp) insertions in plants. Insertions in this size class were found to be approximately equal in frequency to deletions, and compound insertion-deletions comprised only 0.1% of all events. Our findings indicate that, as observed in humans, tandem or partially tandem duplications are the dominant form of insertion (48%), although short duplications from ectopic donors account for a sizable fraction of insertions in rice (38%). Many nontandem duplications contain insertions from nearby DNA (within 200 bp) and can contain multiple donor sources-some distant-in single events. Although replication slippage is a plausible explanation for tandem duplications, the end homology required in such a model is most often absent and rarely is >5 bp. However, end homology is commonly longer than expected by chance. Such findings lead us to favor a model of patch-mediated double-strand-break creation followed by nonhomologous end-joining. Additionally, a striking bias toward 31-bp partially tandem duplications suggests that errors in nucleotide excision repair may be resolved via a similar, but distinct, pathway. In summary, the analysis of recent insertions in rice suggests multiple underappreciated causes of structural variation in eukaryotes.
Proceedings of the National Academy of Sciences 04/2014; 111(18). DOI:10.1073/pnas.1321854111 · 9.67 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Gene expression is a complex process, requiring precise spatial and temporal regulation of transcription factor activity; however, modifications of individual cis- and trans-acting modules can be molded by natural selection to create a sizeable number of novel phenotypes. Results from decades of research indicate that developmental and phenotypic divergence among eukaryotic organisms is driven primarily by variation in levels of gene expression that are dictated by mutations either in structural or regulatory regions of genes. The relative contributions and interplay of cis- and trans-acting regulatory factors to this evolutionary process, however, remain poorly understood. Analysis of 8 genes in the Bz1-Sh1 interval of maize indicates significant allele-specific expression biases in at least one tissue for all genes, ranging from 1.3-fold to 36-fold. All detected effects were cis-regulatory in nature, although genetic background may also influence the level of expression bias and tissue specificity for some allelic combinations. Most allelic pairs exhibited the same direction and approximate intensity of bias across all four tissues; however, a subset of allelic pairs show alternating dominance across different tissue types or variation in the degree of bias in different tissues. In addition, the genes showing the most striking levels of allelic bias co-localize with a previously described recombination hotspot in this region, suggesting a naturally occurring genetic mechanism for creating regulatory variability for a subset of plant genes that may ultimately lead to evolutionary diversification.This article is protected by copyright. All rights reserved.
The Plant Journal 04/2014; 79(3). DOI:10.1111/tpj.12537 · 5.97 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Date palm (Phoenix dactylifera) has been cultivated since ancient times, but little is known about its genetic diversity and population structure. Examination of 80 date palm accessions grown in the United Arab Emirates, including a collection of varieties from around the world, using 21 microsatellite markers, indicated extensive genetic diversity, with many accessions heterozygous for most markers. The average number of alleles per locus (19), expected heterozygosity (0.7), observed heterozygosity (0.25) and fixation indices (Fst = 0.6, Rst = 0.72) demonstrated significant population structure. Analysis with a model-based Baysian method, STRUCTURE 2.4.1, indicated that the 80 accessions could be broadly divided into nine groups. Independent samples of genotypes with the same name, collected from different experimental stations, usually clustered together. The study was enriched for germplasm from the United Arab Emirates (UAE), and one STRUCTURE-derived grouping consisted mainly of UAE accessions. In a few other clusters, several genotypes from the UAE, Iraq and Oman grouped together. Two clusters included accessions from both North Africa and the Middle East. Many accessions in the STRUCTURE-derived populations appeared to be genetic admixtures. The results indicated a broad dissemination of related germplasms across date-palm growing regions of the world, with very few alleles that still correlate with particular regional germplasms.
[Show abstract][Hide abstract] ABSTRACT: Transposable elements (TEs) are the key players in generating genomic novelty by a combination of the chromosome rearrangements they cause and the genes that come under their regulatory sway. Genome size, gene content, gene order, centromere function, and numerous other aspects of nuclear biology are driven by TE activity. Although the origins and attitudes of TEs have the hallmarks of selfish DNA, there are numerous cases where TE components have been co-opted by the host to create new genes or modify gene regulation. In particular, epigenetic regulation has been transformed from a process to silence invading TEs and viruses into a key strategy for regulating plant genes. Most, perhaps all, of this epigenetic regulation is derived from TE insertions near genes or TE-encoded factors that act in trans. Enormous pools of genome data and new technologies for reverse genetics will lead to a powerful new era of TE analysis in plants. Expected final online publication date for the Annual Review of Plant Biology Volume 65 is April 29, 2014. Please see http://www.annualreviews.org/catalog/pubdates.aspx for revised estimates.
[Show abstract][Hide abstract] ABSTRACT: Carnivorous pitcher plants have modified tubular leaves that accumulate captured insects, plant secretions (e.g., digestive enzymes) and water from rain or flooding of the habitat. Pitcher fluids can serve as a model to study food web dynamics, community genetics, trophic interactions, succession, and population structure. We investigated differences in the microbial community richness and composition in the pitcher fluids of two Sarracenia species using 454 sequencing of rRNA gene amplicons. Pitcher plants were sampled during spring, summer and fall at Splinter Hill Bog in Alabama and Ponce De Leon Bog in Florida. Eubacterial phylotypes from pitchers sampled during spring and summer showed dramatic season-dependent differences, but no location-specific differences. The summer samples from S. psittacina and S. purpurea pitchers formed separate clusters in principal co-ordinates analysis, indicating a strong effect of the plant species. Much greater abundances of Rhodopseudomonas and Bacillus were found in S. psittacina compared to S. purpurea pitchers, while Pseudomonads were much more abundant in S. purpurea pitchers. The dominant archaebacterial phylotypes were related to halobacteria and methanobacteria, but most belong to a previously undiscovered archaeabacterial class. Ants (Family:Formicidae), the major food source for these carnivorous plants, were found to represent primarily one phylotype in S. psittacina, but numerous phylotypes in S. purpurea, indicating that these two plant species have adopted respective specialist and generalist predatory niches. Numerous and dynamic rotifer, mite and fungal phylotypes were also detected.
Donald Danforth Center Annual Retreat, St. Louis; 01/2014
[Show abstract][Hide abstract] ABSTRACT: Caldicellulosiruptor bescii is an anaerobic thermophilic bacterium of special interest for use in the consolidated bioprocessing of plant biomass to biofuels. In the course of experiments to engineer pyruvate metabolism in C. bescii, we isolated a mutant of C. bescii that contained an insertion in the L-lactate dehydrogenase gene (ldh). PCR amplification and sequencing of the ldh gene from this mutant revealed a 1,609-bp insertion that contained a single open reading frame of 479 amino acids (1,440 bp) annotated as a hypothetical protein with unknown function. The ORF is flanked by an 8-base direct repeat sequence. Bioinformatic analysis indicated that this ORF is part of a novel transposable element, ISCbe4, which is only intact in the genus Caldicellulosiruptor, but has ancient relatives that are present in degraded (and previously unrecognized) forms across many bacterial and archaeal clades.
[Show abstract][Hide abstract] ABSTRACT: Maize is one of the most important food crops and a key model for genetics and developmental biology. A genetically anchored and high-quality draft genome sequence of maize inbred B73 has been obtained to serve as a reference sequence. To facilitate evolutionary studies in maize and its close relatives, much like the OMAP (www.OMAP.org) BAC resource did for the rice community, we constructed BAC libraries for maize inbred lines Zheng58, Chang7-2 and Mo17 and maize wild relatives Zea mays ssp. parviglumis and Tripsacum dactyloides. Furthermore, to extend functional genomic studies to maize and sorghum, we also constructed BIBAC libraries for the maize inbred B73 and the sorghum land race Nengsi-1. The BAC/BIBAC vectors facilitate transfer of large intact DNA inserts from BAC clones to the BIBAC vector and functional complementation of large DNA fragments. These seven ZMAP BAC/BIBAC libraries have average insert sizes ranging from 92kb to 148kb, organellar DNA from 0.17% to 2.3%, empty vector rates between 0.35% and 5.56%, and genome equivalents of 4.7- to 8.4-fold. The usefulness of the Parviglumis and Tripsacum BAC libraries was demonstrated by mapping clones to the reference genome. Novel genes and alleles present in these ZMAP libraries can now be used for functional complementation studies and positional or homology-based cloning of genes for translational genomics.
[Show abstract][Hide abstract] ABSTRACT: Background and AimsAlthough monocotyledonous plants comprise one of the two major groups of angiosperms and include >65 000 species, comprehensive genome analysis has been focused mainly on the Poaceae (grass) family. Due to this bias, most of the conclusions that have been drawn for monocot genome evolution are based on grasses. It is not known whether these conclusions apply to many other monocots.Methods
To extend our understanding of genome evolution in the monocots, Asparagales genomic sequence data were acquired and the structural properties of asparagus and onion genomes were analysed. Specifically, several available onion and asparagus bacterial artificial chromosomes (BACs) with contig sizes >35 kb were annotated and analysed, with a particular focus on the characterization of long terminal repeat (LTR) retrotransposons.Key ResultsThe results reveal that LTR retrotransposons are the major components of the onion and garden asparagus genomes. These elements are mostly intact (i.e. with two LTRs), have mainly inserted within the past 6 million years and are piled up into nested structures. Analysis of shotgun genomic sequence data and the observation of two copies for some transposable elements (TEs) in annotated BACs indicates that some families have become particularly abundant, as high as 4-5 % (asparagus) or 3-4 % (onion) of the genome for the most abundant families, as also seen in large grass genomes such as wheat and maize.Conclusions
Although previous annotations of contiguous genomic sequences have suggested that LTR retrotransposons were highly fragmented in these two Asparagales genomes, the results presented here show that this was largely due to the methodology used. In contrast, this current work indicates an ensemble of genomic features similar to those observed in the Poaceae.
Annals of Botany 07/2013; 112(5). DOI:10.1093/aob/mct155 · 3.65 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Resistance Gene Homologues (RGHs) were isolated from the switchgrass variety Alamo by a combination of PCR and expressed sequence tag (EST) database mining. Fifty-eight RGHs were isolated by PCR and 295 RGHs were identified in 424,545 switchgrass ESTs. Four NBS-LRR RGHs were selected to investigate RGH haplotypic diversity in seven switchgrass varieties chosen for their representation of a broad range of the switchgrass germplasm. Lowland and upland ecotypes were found to be less similar, even from nearby populations, than were more distant populations with similar growth environments. A majority (83.5%) of the variability in these four RGHs was found to be attributable to the within-population component. The difference in nucleotide diversity between and within populations was observed to be small, while this diversity is maintained to similar degrees at both population and ecotype levels. The results also revealed that the analyzed RGHs were under positive selection in the studied switchgrass accessions. Intragenic recombination was detected in switchgrass RGHs, thereby demonstrating an active genetic process that has the potential to generate new resistance genes with new specificities that might act against newly-arising pathogen races.