Windows .NET Network Distributed Basic Local Alignment Search Toolkit (W.ND-BLAST)

BACKGROUND: BLAST is one of the most common and useful tools for Genetic Research. This paper describes a software application we have termed Windows .NET Distributed Basic Local Alignment Search Toolkit (W.ND-BLAST), which enhances the BLAST utility by improving usability, fault recovery, and scalability in a Windows desktop environment. Our goal was to develop an easy to use, fault tolerant, high-throughput BLAST solution that incorporates a comprehensive BLAST result viewer with curation and annotation functionality. RESULTS: W.ND-BLAST is a comprehensive Windows-based software toolkit that targets researchers, including those with minimal computer skills, and provides the ability increase the performance of BLAST by distributing BLAST queries to any number of Windows based machines across local area networks (LAN). W.ND-BLAST provides intuitive Graphic User Interfaces (GUI) for BLAST database creation, BLAST execution, BLAST output evaluation and BLAST result exportation. This software also provides several layers of fault tolerance and fault recovery to prevent loss of data if nodes or master machines fail. This paper lays out the functionality of W.ND-BLAST. W.ND-BLAST displays close to 100% performance efficiency when distributing tasks to 12 remote computers of the same performance class. A high throughput BLAST job which took 662.68 minutes (11 hours) on one average machine was completed in 44.97 minutes when distributed to 17 nodes, which included lower performance class machines. Finally, there is a comprehensive high-throughput BLAST Output Viewer (BOV) and Annotation Engine components, which provides comprehensive exportation of BLAST hits to text files, annotated fasta files, tables, or association files. CONCLUSION: W.ND-BLAST provides an interactive tool that allows scientists to easily utilizing their available computing resources for high throughput and comprehensive sequence analyses. The install package for W.ND-BLAST is freely downloadable from . With registration the software is free, installation, networking, and usage instructions are provided as well as a support forum

The rehydration transcriptome of the desiccation-tolerant bryophyte Tortula ruralis: transcript classification and analysis

BACKGROUND: The cellular response of plants to water-deficits has both economic and evolutionary importance directly affecting plant productivity in agriculture and plant survival in the natural environment. Genes induced by water-deficit stress have been successfully enumerated in plants that are relatively sensitive to cellular dehydration, however we have little knowledge as to the adaptive role of these genes in establishing tolerance to water loss at the cellular level. Our approach to address this problem has been to investigate the genetic responses of plants that are capable of tolerating extremes of dehydration, in particular the desiccation-tolerant bryophyte, Tortula ruralis. To establish a sound basis for characterizing the Tortula genome in regards to desiccation tolerance, we analyzed 10,368 expressed sequence tags (ESTs) from rehydrated rapid-dried Tortula gametophytes, a stage previously determined to exhibit the maximum stress induced change in gene expression. RESULTS: The 10, 368 ESTs formed 5,563 EST clusters (contig groups representing individual genes) of which 3,321 (59.7%) exhibited similarity to genes present in the public databases and 2,242 were categorized as unknowns based on protein homology scores. The 3,321 clusters were classified by function using the Gene Ontology (GO) hierarchy and the KEGG database. The results indicate that the transcriptome contains a diverse population of transcripts that reflects, as expected, a period of metabolic upheaval in the gametophyte cells. Much of the emphasis within the transcriptome is centered on the protein synthetic machinery, ion and metabolite transport, and membrane biosynthesis and repair. Rehydrating gametophytes also have an abundance of transcripts that code for enzymes involved in oxidative stress metabolism and phosphorylating activities. The functional classifications reflect a remarkable consistency with what we have previously established with regards to the metabolic activities that are important in the recovery of the gametophytes from desiccation. A comparison of the GO distribution of Tortula clusters with an identical analysis of 9,981 clusters from the desiccation sensitive bryophyte species Physcomitrella patens, revealed, and accentuated, the differences between stressed and unstressed transcriptomes. Cross species sequence comparisons indicated that on the whole the Tortula clusters were more closely related to those from Physcomitrella than Arabidopsis (complete genome BLASTx comparison) although because of the differences in the databases there were more high scoring matches to the Arabidopsis sequences. The most abundant transcripts contained within the Tortula ESTs encode Late Embryogenesis Abundant (LEA) proteins that are normally associated with drying plant tissues. This suggests that LEAs may also play a role in recovery from desiccation when water is reintroduced into a dried tissue. CONCLUSION: The establishment of a rehydration EST collection for Tortula ruralis, an important plant model for plant stress responses and vegetative desiccation tolerance, is an important step in understanding the genome level response to cellular dehydration. The type of transcript analysis performed here has laid the foundation for more detailed functional and genome level analyses of the genes involved in desiccation tolerance in plants

An integrated approach to use genetic resources for resurrection plants to enhance drought tolerance in breeding-extension programs [abstract]

Only abstract of poster available.Track V: BiomassThe ultimate goals of this project are to gain a basic understanding of the unique gene and gene regulatory networks that are necessary and sufficient for vegetative tissues to withstand dehydration and then rapidly recover upon rehydration and to use the knowledge gained to develop crops, maize and forage grasses that maintain biomass production under drought condition. Our approach is to combine comparative genomics and phylogenetics to identify genes and gene networks that are adaptive and central to the tolerance of cellular dehydration. This involves the use of resurrection species as models for dehydration tolerance coupled with a suite of comparative bioinformatic tools that allows for the phylogenetic assessment of gene expression patterns in response to dehydration and rehydration. Once the key genetic elements have been identified and assessed we will use a transgenic functional assessment of their involvement in the phenotype, both at a molecular and physiological level, of drought tolerance. One of our key resurrection species is the South African grass Sporobolus stapfianus, which is capable of surviving -240 MPa of water deficit (a hundred times lower than most crop plants). This plant not only serves as a model for monocot crops such as maize and switchgrass, our major targets for crop improvement, but also serves as a direct possibility for an alternate forage grass and biomass source. The improvement of biomass production under drought conditions is not only important for sustainable biofuel production but also for food and energy security. Funded by a CSREES-NRI Grant of $450,000 over three years to PI Mel Oliver USDA-ARS-PGRU Columbia, CoPIs Robert Sharp, University of Missouri; John Cushman, University of Nevada, Reno; Paxton Payton, USDA-ARS-PSRU Lubbock

Expression of an Arabidopsis Sodium/Proton Antiporter Gene

Abstract Salinity is a major environmental stress that affects agricultural productivity worldwide. One approach to improving salt tolerance in crops is through high expression of the Arabidopsis gene AtNHX1, which encodes a vacuolar sodium/proton antiporter that sequesters excess sodium ion into the large intracellular vacuole. Sequestering cytosolic sodium into the vacuoles of plant cells leads to a low level of sodium in cytosol, which minimizes the sodium toxicity and injury to important enzymes in cytosol. In the meantime, the accumulation of sodium in vacuoles restores the correct osmolarity to the intracellular milieu, which favors water uptake by plant root cells and improves water retention in tissues under soils that are high in salt. To improve the yield and quality of peanut under high salt conditions, AtNHX1 was introduced into peanut plants through Agrobacterium-mediated transformation. The AtNHX1-expressing peanut plants displayed increased tolerance of salt at levels up to 150 mM NaCl. When compared to wild-type plants, AtNHX1-expressing peanut plants suffered less damage, produced more biomass, contained more chlorophyll, and maintained higher photosynthetic rates under salt conditions. These data indicate that AtNHX1 can be used to enhance salt tolerance in peanut

Gene expression profiling in peanut using high density oligonucleotide microarrays

<p>Abstract</p> <p>Background</p> <p>Transcriptome expression analysis in peanut to date has been limited to a relatively small set of genes and only recently has a significant number of ESTs been released into the public domain. Utilization of these ESTs for oligonucleotide microarrays provides a means to investigate large-scale transcript responses to a variety of developmental and environmental signals, ultimately improving our understanding of plant biology.</p> <p>Results</p> <p>We have developed a high-density oligonucleotide microarray for peanut using 49,205 publicly available ESTs and tested the utility of this array for expression profiling in a variety of peanut tissues. To identify putatively tissue-specific genes and demonstrate the utility of this array for expression profiling in a variety of peanut tissues, we compared transcript levels in pod, peg, leaf, stem, and root tissues. Results from this experiment showed 108 putatively pod-specific/abundant genes, as well as transcripts whose expression was low or undetected in pod compared to peg, leaf, stem, or root. The transcripts significantly over-represented in pod include genes responsible for seed storage proteins and desiccation (e.g., late-embryogenesis abundant proteins, aquaporins, legumin B), oil production, and cellular defense. Additionally, almost half of the pod-abundant genes represent unknown genes allowing for the possibility of associating putative function to these previously uncharacterized genes.</p> <p>Conclusion</p> <p>The peanut oligonucleotide array represents the majority of publicly available peanut ESTs and can be used as a tool for expression profiling studies in diverse tissues.</p

Physiology and transcriptomics of water-deficit stress responses in wheat cultivars TAM 111 and TAM 112

Citation: Reddy, S. K., Liu, S., Rudd, J. C., Xue, Q., Payton, P., Finlayson, S. A., … Lu, N. (2014). Physiology and transcriptomics of water-deficit stress responses in wheat cultivars TAM 111 and TAM 112. Retrieved from http://krex.ksu.eduHard red winter wheat crops on the U.S. Southern Great Plains often experience moderate to severe drought stress, especially during the grain filling stage, resulting in significant yield losses. Cultivars TAM 111 and TAM 112 are widely cultivated in the region, share parentage and showed superior but distinct adaption mechanisms under water-deficit (WD) conditions. Nevertheless, the physiological and molecular basis of their adaptation remains unknown. A greenhouse study was conducted to understand the differences in the physiological and transcriptomic responses of TAM 111 and TAM 112 to WD stress. Whole-plant data indicated that TAM 112 used more water, produced more biomass and grain yield under WD compared to TAM 111. Leaf-level data at the grain filling stage indicated that TAM 112 had elevated abscisic acid (ABA) content and reduced stomatal conductance and photosynthesis as compared to TAM 111. Sustained WD during the grain filling stage also resulted in greater flag leaf transcriptome changes in TAM 112 than TAM 111. Transcripts associated with photosynthesis, carbohydrate metabolism, phytohormone metabolism, and other dehydration responses were uniquely regulated between cultivars. These results suggested a differential role for ABA in regulating physiological and transcriptomic changes associated with WD stress and potential involvement in the superior adaptation and yield of TAM 112

Genome-wide transcriptome and physiological analyses provide new insights into peanut drought response mechanisms

Drought is one of the main constraints in peanut production in West Texas and eastern New Mexico regions due to the depletion of groundwater. A multi-seasonal phenotypic analysis of 10 peanut genotypes revealed C76-16 (C-76) and Valencia-C (Val-C) as the best and poor performers under deficit irrigation (DI) in West Texas, respectively. In order to decipher transcriptome changes under DI, RNAseq was performed in C-76 and Val-C. Approximately 369 million raw reads were generated from 12 different libraries of two genotypes subjected to fully irrigated (FI) and DI conditions, of which ~329 million (90.2%) filtered reads were mapped to the diploid ancestors of peanut. The transcriptome analysis detected 4,508 differentially expressed genes (DEGs), 1554 genes encoding transcription factors (TFs) and a total of 514 single nucleotide polymorphisms (SNPs) among the identified DEGs. The comparative analysis between the two genotypes revealed higher and integral tolerance in C-76 through activation of key genes involved in ABA and sucrose metabolic pathways. Interestingly, one SNP from the gene coding F-box protein (Araip.3WN1Q) and another SNP from gene coding for the lipid transfer protein (Aradu.03ENG) showed polymorphism in selected contrasting genotypes. These SNPs after further validation may be useful for performing early generation selection for selecting drought-responsive genotypes

Effect of elevated CO2 on peanut performance in a semi-arid production region

With the intensification and frequency of heat waves and periods of water deficit stress, along with rising atmospheric carbon dioxide [CO2], understanding the seasonal leaf-gas-exchange responses to combined abiotic factors will be important in predicting crop performance in semi-arid production systems. In peanut (Arachis hypogaea L.), the availability of developmental stage physiological data on the response to repeated water deficit stress periods in an elevated [CO2] (EC) environment is limited and necessary to improve crop model predictions. Here, we investigated the effects of season-long EC (650 µmol CO2 m−2 s−1) on the physiology and productivity of peanut in a semi-arid environment. This study was conducted over two-growing seasons using field-based growth chambers to maintain EC conditions, and impose water-stress at three critical developmental stages. Our results showed that relative to ambient [CO2] (AC), long-term EC during water-stress episodes, increased leaf-level light-saturated CO2 assimilation (Asat), transpiration efficiency (TE), vegetative biomass, and pod yield by 58%, 73%, 58%, and 39%, respectively. Although leaf nitrogen content was reduced by 16%, there was 41% increase in maximum Rubisco carboxylation efficiency in EC, indicating that there was minimal photosynthetic down-regulation. Furthermore, long-term EC modified the short-term physiological response (Asat) to rapid changes in [CO2] during the water-stress episodes, generating a much greater change in EC (54%) compared to AC (10%). Additionally, long-term EC generated a 23% greater Asat compared to the short-term EC during the water-stress episodes. These findings indicate high levels of physiological adjustment in EC, which may increase drought resilience. We concluded that EC may reduce the negative impacts of repeated water-stress events at critical developmental stages on rain-fed peanut in semi-arid regions. These results can inform current models to improve the projections of peanut response to future climates

ESTs, cDNA microarrays, and gene expression profiling : tools for dissecting plant physiology and development

Gene expression profiling holds tremendous promise for dissecting the regulatory mechanisms and transcriptional networks that underlie biological processes. Here we provide details of approaches used by others and ourselves for gene expression profiling in plants with emphasis on cDNA microarrays and discussion of both experimental design and downstream analysis. We focus on methods and techniques emphasizing fabrication of cDNA microarrays, fluorescent labeling, cDNA hybridization, experimental design, and data processing. We include specific examples that demonstrate how this technology can be used to further our understanding of plant physiology and development (specifically fruit development and ripening) and for comparative genomics by comparing transcriptome activity in tomato and pepper fruit

Primed acclimation of cultivated peanut (Arachis hypogaea L.) through the use of deficit irrigation timed to crop developmental periods

Water-deficits and high temperatures are the predominant factors limiting peanut production across the U.S., either because of regional aridity or untimely rainfall events during crucial crop developmental periods. In the southern High Plains of west Texas and eastern New Mexico, low average annual rainfall (450. mm) and high evaporative demand necessitates the use of significant irrigation in production systems. In this west Texas study, the primary objective was to develop irrigation schemes that maximized peanut yield and grade while reducing overall water consumption. Therefore, a large-scale field experiment was established in 2005 and 2006 that utilized 15 treatment combinations of differing rates of irrigation (50, 75, and 100% of grower applied irrigation) applied at different periods of peanut development (early, middle, and late season). Precipitation patterns and ambient temperatures showed greater stress levels in 2006 which likely reduced yields across all treatments in comparison to 2005. Yields were reduced 26 (2005) and 10% (2006) in the lowest irrigation treatment (50% full season) compared with full irrigation (100% full season); but early-season water deficit (50 and 75% in the first 45. days after planting) followed by 100% irrigation in the mid- and late-seasons were successful at sustaining yield and/or crop value. Root growth was significantly enhanced at 50% irrigation compared with 100% irrigation, through greater root length, diameter, surface area, and depth, suggesting greater access to water during mid- and late-season periods. These results suggest that early to mid-season deficit irrigation has the potential to maintain peanut yield without altering quality, and to substantially reduce water use in this semi-arid environment