Transcriptome Profiling of Buffalograss Challenged with the Leaf Spot Pathogen Curvularia inaequalis

Buffalograss (Bouteloua dactyloides) is a low maintenance United States native turfgrass species with exceptional drought, heat and cold tolerance. Leaf spot caused by Curvularia inaequalis negatively impacts buffalograss visual quality. Two leaf spot susceptible and two resistant buffalograss lines were challenged with C. inaequalis. Samples were collected from treated and untreated leaves when susceptible lines showed symptoms. Transcriptome sequencing was done and differentially expressed genes were identified. Approximately 27 million raw sequencing reads were produced per sample. More than 86% of the sequencing reads mapped to an existing buffalograss reference transcriptome. De novo assembly of unmapped reads was merged with the existing reference to produce a more complete transcriptome. There were 461 differentially expressed transcripts between the resistant and susceptible lines when challenged with the pathogen and 1552 in its absence. Previously characterized defense-related genes were identified among the differentially expressed transcripts. Twenty one resistant line transcripts were similar to genes regulating pattern triggered immunity and 20 transcripts were similar to genes regulating effector triggered immunity. There were also nine up-regulated transcripts in resistance lines which showed potential to initiate systemic acquired resistance (SAR) and three transcripts encoding pathogenesis-related proteins which are downstream products of SAR. This is the first study characterizing changes in the buffalograss transcriptome when challenged with C. inaequalis

Transcriptome Profiling of Buffalograss Challenged with the Leaf Spot Pathogen \u3ci\u3eCurvularia inaequalis\u3c/i\u3e

Buffalograss (Bouteloua dactyloides) is a low maintenance U. S. native turfgrass species with exceptional drought, heat, and cold tolerance. Leaf spot caused by Curvularia inaequalis negatively impacts buffalograss visual quality. Two leaf spot susceptible and two resistant buffalograss lines were challenged with C. inaequalis. Samples were collected from treated and untreated leaves when susceptible lines showed symptoms. Transcriptome sequencing was done and differentially expressed genes were identified. Approximately 27 million raw sequencing reads were produced per sample. More than 86% of the sequencing reads mapped to an existing buffalograss reference transcriptome. De novo assembly of unmapped reads was merged with the existing reference to produce a more complete transcriptome. There were 461 differentially expressed transcripts between the resistant and susceptible lines when challenged with the pathogen and 1552 in its absence. Previously characterized defense-related genes were identified among the differentially expressed transcripts. Twenty one resistant line transcripts were similar to genes regulating pattern triggered immunity and 20 transcripts were similar to genes regulating effector triggered immunity. There were also nine upregulated transcripts in resistance lines which showed potential to initiate systemic acquired resistance (SAR) and three transcripts encoding pathogenesis-related proteins which are downstream products of SAR. This is the first study characterizing changes in the buffalograss transcriptome when challenged with C. inaequalis

Editorial: Genomic Approaches for Improvement of Understudied Grasses

Grasses are diverse, spanning native prairies to high-yielding grain cropping systems. They are valued for their beauty and useful for soil stabilization, pollution mitigation, biofuel production, nutritional value, and forage quality; grasses encompass the most important grain crops in the world. There are thousands of distinct grass species and many have promiscuous hybridization patterns, blurring species boundaries. Resources for advancing the science and knowledgebase of individual grass species or their unique characteristics varies, often proportional to their perceived value to society. For many grasses, limited genetic information hinders research progress. Presented in this research topic is a brief snapshot of creative efforts to apply modern genomics research methodologies to the study of several minor grass species

Genomic Approaches for Improvement of Understudied Grasses

Grasses are diverse, spanning native prairies to high-yielding grain cropping systems. They are valued for their beauty and useful for soil stabilization, pollution mitigation, biofuel production, nutritional value, and forage quality; grasses encompass the most important grain crops in the world. There are thousands of distinct grass species and many have promiscuous hybridization patterns, blurring species boundaries. Resources for advancing the science and knowledgebase of individual grass species or their unique characteristics varies, often proportional to their perceived value to society. For many grasses, limited genetic information hinders research progress. Presented in this research topic is a brief snapshot of creative efforts to apply modern genomics research methodologies to the study of several minor grass species

Gene Expression Profiling of Iron Deficiency Chlorosis Sensitive and Tolerant Soybean Indicates Key Roles for Phenylpropanoids under Alkalinity Stress

Alkaline soils comprise 30% of the earth and have low plant-available iron (Fe) concentration, and can cause iron deficiency chlorosis (IDC). IDC causes soybean yield losses of $260 million annually. However, it is not known whether molecular responses to IDC are equivalent to responses to low iron supply. IDC tolerant and sensitive soybean lines provide a contrast to identify specific factors associated with IDC.We used RNA-seq to compare gene expression under combinations of normal pH (5.7) or alkaline pH (7.7, imposed by 2.5mM bicarbonate, or pH 8.2 imposed by 5mM bicarbonate) and normal (25μM) or low (1μM) iron conditions from roots of these lines. Thus, we were able to treat pH and Fe supply as separate variables. We also noted differential gene expression between IDC sensitive and tolerant genotypes in each condition. Classical iron uptake genes, including ferric-chelate reductase (FCR) and ferrous transporters, were upregulated by both Fe deficiency and alkaline stress, however, their gene products did not function well at alkaline pH. In addition, genes in the phenylpropanoid synthesis pathway were upregulated in both alkaline and low Fe conditions. These genes lead to the production of fluorescent root exudate (FluRE) compounds, such as coumarins. Fluorescence of nutrient solution increased with alkaline treatment, and was higher in the IDC tolerant line. Some of these genes also localized to previously identified QTL regions associated with IDC. We hypothesize that FluRE become essential at alkaline pH where the classical iron uptake system does not function well. This work could result in new strategies to screen for IDC tolerance, and provide breeding targets to improve crop alkaline stress tolerance

Transcriptomic and physiological characterization of the \u3ci\u3efefe\u3c/i\u3e mutant of melon (\u3ci\u3eCucumis melo\u3c/i\u3e) reveals new aspects of iron-copper crosstalk

Iron (Fe) and copper (Cu) homeostasis are tightly linked across biology. In previous work, Fe deficiency interacted with Cu-regulated genes and stimulated Cu accumulation. The C940-fe (fefe) Fe-uptake mutant of melon (Cucumis melo) was characterized, and the fefe mutant was used to test whether Cu deficiency could stimulate Fe uptake. Wild-type and fefe mutant transcriptomes were determined by RNA-seq under Fe and Cu deficiency. FeFe-regulated genes included core Fe uptake, metal homeostasis, and transcription factor genes. Numerous genes were regulated by both Fe and Cu. The fefe mutant was rescued by high Fe or by Cu deficiency, which stimulated ferric-chelate reductase activity, FRO2 expression, and Fe accumulation. Accumulation of Fe in Cu-deficient plants was independent of the normal Fe-uptake system. One of the four FRO genes in the melon and cucumber (Cucumis sativus) genomes was Fe-regulated, and one was Cu-regulated. Simultaneous Fe and Cu deficiency synergistically up-regulated Fe-uptake gene expression. Overlap in Fe and Cu deficiency transcriptomes highlights the importance of Fe-Cu crosstalk in metal homeostasis. The fefe gene is not orthologous to FIT, and thus identification of this gene will provide clues to help understand regulation of Fe uptake in plants

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

Genetic diversity of \u3ci\u3eDanthonia spicata\u3c/i\u3e (L.) Beauv. based on genomic simple sequence repeat markers

Danthonia spicata (L.) Beauv., commonly known as poverty oatgrass, is a perennial bunch-type grass native to North America. D. spicata is often found in low input turfgrass areas on the East Coast of the United States and has potential for development as a new native low input turfgrass species. Roche 454 sequenced randomly sheared genomic DNA reads of D. spicata were mined for SSR markers using the MIcroSAtellite identification tool. A total of 66,553 singlet sequences (approximately 37.5 Mbp) were examined, and 3454 SSR markers were identified. Trinucleotide motifs with greater than six repeats and possessing unique PCR priming sites within the genome, as determined by Primer-BLAST, were evaluated visually for heterozygosity and mutation consistent with stepwise evolution using CLC Genomics software. Sixty-three candidate markers were selected for testing from the trinucleotide SSR marker sites meeting these in silico criteria. Ten primer pairs that amplified polymorphic loci in preliminary experiments were used to screen 91 individual plants composed of at least 3–5 plants from each of 23 different locations. The primer pairs amplified 54 alleles ranging in size from 71 to 246 bp. Minimum and maximum numbers of alleles per locus were two and 12, respectively, with an average of 5.4. A dendrogram generated by unweighted pair group method with arithmetic mean cluster analysis using the Jaccard’s similarity coefficient was in agreement with the grouping obtained by Structure v2.3. The analyses were dominated by clonal groupings and lack evidence for gene flow with some alleles present in a single plant from a single location. Fourteen multilocus genotype groups were observed providing strong evidence for asexual reproduction in the studied D. spicata populations

Feasibility Study of the Health Empowerment Intervention to Evaluate the Effect on Self-Management, Functional Health, and Well-Being in Older Adults with Heart Failure

abstract: ABSTRACT The population of older adults in the United States is growing disproportionately, with corresponding medical, social and economic implications. The number of Americans 65 years and older constitutes 13.7% of the U.S. population, and is expected to grow to 21% by 2040. As the adults age, they are at risk for developing chronic illness and disability. According to the Centers for Disease Control and Prevention, 5.7 million Americans have heart failure, and almost 80% of these are 65 years and older. The prevalence of heart failure will increase with the increase in aging population, thus increasing the costs associated with heart failure from 34.7 billion dollars in 2010 to 77.7 billion dollars by 2020. Of all cardiovascular hospitalizations, 28.9% are due to heart failure, and almost 60,000 deaths are accounted for heart failure. Marked disparities in heart failure persist within and between population subgroups. Living with heart failure is challenging for older adults, because being a chronic condition, the responsibility of day to day management of heart failure principally rests with patient. Approaches to improve self-management are targeted at adherence, compliance, and physiologic variables, little attention has been paid to personal and social contextual resources of older adults, crucial for decision making, and purposeful participation in goal attainment, representing a critical area for intervention. Several strategies based on empowerment perspective are focused on outcomes; paying less attention to the process. To address these gaps between research and practice, this feasibility study was guided by a tested theory, the Theory of Health Empowerment, to optimize self-management, functional health and well-being in older adults with heart failure. The study sample included older adults with heart failure attending senior centers. Specific aims of this feasibility study were to: (a) examine the feasibility of the Health Empowerment Intervention in older adults with heart failure, (b) evaluate the effect of the health empowerment intervention on self-management, functional health, and well-being among older adults with heart failure. The Health Empowerment Intervention was delivered focusing on strategies to identify and building upon self-capacity, and supportive social network, informed decision making and goal setting, and purposefully participating in the attainment of personal health goals for well-being. Study was feasible and significantly increased personal growth, and purposeful participation in the attainment of personal health goals.Dissertation/ThesisDoctoral Dissertation Nursing and Healthcare Innovation 201

Response of \u3ci\u3eAmaranthus\u3c/i\u3e spp. following exposure to sublethal herbicide rates via spray particle drift

The adverse consequences of herbicide drift towards sensitive crops have been extensively reported in the literature. However, little to no information is available on the consequences of herbicide drift onto weed species inhabiting boundaries of agricultural fields. Exposure to herbicide drift could be detrimental to long-term weed management as several weed species have evolved herbicide-resistance after recurrent selection with sublethal herbicide rates This study investigated the deposition of glyphosate, 2,4-D, and dicamba spray particle drift from applications with two different nozzles in a low speed wind tunnel, and their impact on growth and development of Amaranthus spp. Herbicide drift resulted in biomass reduction or complete plant mortality. Inflection points (distance to 50% biomass reduction) for Amaranthus tuberculatus were 7.7, 4.0, and 4.1 m downwind distance for glyphosate, 2,4-D, and dicamba applications with the flat-fan nozzle, respectively, whereas these values corresponded to 2.8, 2.5, and 1.9 m for applications with the air-inclusion nozzle. Inflection points for Amaranthus palmeri biomass reduction were 16.3, 10.9, and 11.5 m for glyphosate, 2,4-D, and dicamba applications with the flat-fan nozzle, respectively, whereas these values corresponded to 7.6, 5.4, and 5.4 m for applications with the air-inclusion nozzle. Plants were more sensitive to glyphosate at higher exposure rates than other herbicides, whereas plants were more sensitive to 2,4-D and dicamba at lower exposure rates compared to glyphosate. Applications with the flat-fan nozzle resulted in 32.3 and 11.5% drift of the applied rate at 1.0 and 3.0 m downwind, respectively, whereas the air-inclusion nozzle decreased the dose exposure in the same distances (11.4 and 2.7%, respectively). Herbicide drift towards field boundaries was influenced by nozzle design and exposed weeds to herbicide rates previously reported to select for herbicide-resistant biotypes