Genetic Diversity and Molecular Characterization of Worldwide Prairie Grass (Bromus catharticus Vahl) Accessions Using SRAP Markers

Prairie grass (Bromus catharticus Vahl) is an important grass species that could be used in the production systems of certified seed and high-quality forage for grazing ruminants. In the present research, a sequence-related amplified polymorphism (SRAP) marker was employed to detect the genetic variability and structure of 80 prairie grass accessions from all over the world. Altogether, 460 reliable bands were amplified from 47 SRAP primer pairs with 345 (75%) polymorphic bands. The average values of discrimination power (DP) and polymorphic information content (PIC) were 0.753 and 0.317, respectively. Both the UPGMA clustering and PCoA analyses grouped the 80 accessions into five clusters, whereas the STRUCTURE analysis showed that 80 prairie grass accessions possessed three genetic memberships (K = 3). The results of the Mantel test showed that the distance matrix has a moderately positive correlation between the morphological and molecular data sets (r = 0.524). A poor genetic differentiation (Fst = 0.045) was discovered among the six geo-groups of accessions. Besides, the highest intragroup genetic diversity was found in the North America group (He = 0.335). This study provides a genetic structure and diversity case for prairie grass, and supplies new clues for the study and utilization of prairie grass.

Transcriptomic resources for prairie grass (Bromus catharticus): expressed transcripts, tissue-specific genes, and identification and validation of EST-SSR markers

Background Prairie grass (Bromus catharticus) is a typical cool-season forage crop with high biomass production and fast growth rate during winter and spring. However, its genetic research and breeding has remained stagnant due to limited available genomic resources. The aim of this study was to generate large-scale genomic data using high-throughput transcriptome sequencing, and perform a preliminary validation of EST-SSR markers of B. catharticus. Results Eleven tissue samples including seeds, leaves, and stems were collected from a new high-yield strain of prairie grass BCS1103. A total of 257,773 unigenes were obtained, of which 193,082 (74.90%) were annotated. Comparison analysis between tissues identified 1803, 3030, and 1570 genes specifically and highly expressed in seed, leaf, and stem, respectively. A total of 37,288 EST-SSRs were identified from unigene sequences, and more than 80,000 primer pairs were designed. We synthesized 420 primer pairs and selected 52 ones with high polymorphisms to estimate genetic diversity and population structure in 24 B. catharticus accessions worldwide. Despite low diversity indicated by an average genetic distance of 0.364, the accessions from South America and Asia and wild accessions showed higher genetic diversity. Moreover, South American accessions showed a pure ancestry, while Asian accessions demonstrated mixed internal relationships, which indicated a different probability of gene flow. Phylogenetic analysis clustered the studied accessions into four clades, being consistent with phenotypic clustering results. Finally, Mantel analysis suggested the total phenotypic variation was mostly contributed by genetic component. Stem diameter, plant height, leaf width, and biomass yield were significantly correlated with genetic data (r > 0.6, P < 0.001), and might be used in the future selection and breeding. Conclusion A genomic resource was generated that could benefit genetic and taxonomic studies, as well as molecular breeding for B. catharticus and its relatives in the future.

Transcriptomic resources for prairie grass (Bromus catharticus): expressed transcripts, tissue-specific genes, and identification and validation of EST-SSR markers

Background: Prairie grass (Bromus catharticus) is a typical cool-season forage crop with high biomass production and fast growth rate during winter and spring. However, its genetic research and breeding has remained stagnant due to limited available genomic resources. The aim of this study was to generate large-scale genomic data using high-throughput transcriptome sequencing, and perform a preliminary validation of EST-SSR markers of B. catharticus.Results: Eleven tissue samples including seeds, leaves, and stems were collected from a new high-yield strain of prairie grass BCS1103. A total of 257,773 unigenes were obtained, of which 193,082 (74.90%) were annotated. Comparison analysis between tissues identified 1803, 3030, and 1570 genes specifically expressed in the seed, leaf, and stem, respectively. A total of 37,288 EST-SSRs were identified from unigene sequences, and more than 80,000 primer pairs were designed. We synthesized 420 primer pairs and selected 52 ones with high polymorphisms to estimate genetic diversity and population structure in 24 B. catharticus accessions worldwide. Despite low diversity indicated by an average genetic distance of 0.358, the accessions from South America and Asia and wild accessions showed higher genetic diversity. Moreover, South American accessions showed a pure ancestry, but Asian accessions demonstrated mixed relationships, which indicated a different probability of gene flow among the two regions. Phylogenetic analysis clustered the studied accessions into four clades. Finally, phenotypic clustering and Mantel analysis suggested the total phenotypic variation was mostly contributed by the genetic component. Stem diameter, plant height, leaf width, and biomass yield significantly correlated with genetic data (r > 0.6, P < 0.001), and might be genetically stable in the future selection and breeding.Conclusion: A genomic resource was generated that could benefit genetic and taxonomic studies, as well as molecular breeding for B. catharticus and it relatives in the future.

A Statistical Evaluation of WRF-LES Trace Gas Dispersion Using Project Prairie Grass Measurements

In recent years, new measurement systems have been deployed to monitor and quantify methane emissions from the natural gas sector. Large-eddy simulation (LES) has complemented measurement campaigns by serving as a controlled environment in which to study plume dynamics and sampling strategies. However, with few comparisons to controlled-release experiments, the accuracy of LES for modeling natural gas emissions is poorly characterized. In this paper, we evaluate LES from the Weather Research and Forecasting (WRF) model against Project Prairie Grass campaign measurements and surface layer similarity theory. Using WRF-LES, we simulate continuous emissions from 30 near-surface trace gas sources in two stability regimes: strong and weak convection. We examine the impact of grid resolutions ranging from 6.25 m to 52 m in the horizontal dimension on model results. We evaluate performance in a statistical framework, calculating fractional bias and conducting Welch’s t-tests. WRF-LES accurately simulates observed surface concentrations at 100 m and beyond under strong convection; simulated concentrations pass t-tests in this region irrespective of grid resolution. However, in weakly convective conditions with strong winds, WRF-LES substantially overpredicts concentrations – the magnitude of fractional bias often exceeds 30%, and all but one C-test fails. The good performance of WRF-LES under strong convection correlates with agreement with local free convection theory and a minimal amount of parameterized turbulent kinetic energy. The poor performance under weak convection corresponds to misalignment with Monin-Obukhov similarity theory and a significant amount of parameterized turbulent kinetic energy.

Effect of pH on xylitol production by Candida species from a prairie cordgrass hydrolysate

Using hydrolysates of the North American prairie grass prairie cordgrass buffered at pH 4.5, 5.0, 5.5 or 6.0, xylitol production, xylitol yield, cell biomass production and productivity were investigated for three strains of yeast Candida. Of the three strains, the highest xylitol concentration of 20.19 g xylitol (g xylose consumed)−1 and yield of 0.89 g xylitol (g xylose consumed)−1 were produced by Candida mogi ATCC 18364 when grown for 120 h at 30° C on the pH 5.5-buffered hydrolysate-containing medium. The highest biomass level being 7.7 g cells (kg biomass)−1 was observed to be synthesized by Candida guilliermondii ATCC 201935 after 120 h of growth at 30° C on a pH 5.5-buffered hydrolysate-containing medium. The highest xylitol specific productivity of 0.73 g xylitol (g cells h)−1 was determined for C. guilliermondii ATCC 20216 after 120 h of growth at 30°C on a pH 5.0-buffered hydrolysate-containing medium. Xylitol production and yield by the three Candida strains was higher on prairie cordgrass than what was previously observed for the same strains after 120 h at 30° C when another North American prairie grass big bluestem served as the plant biomass hydrolysate indicating that prairie cordgrass may be a superior plant biomass substrate.

Effect of yeast extract addition to a mineral salts medium containing hydrolyzed plant xylan on fungal pullulan production

The ability of the fungus Aureobasidium pullulans ATCC 42023 to produce pullulan from yeast extract-supplemented xylan hydrolysates of the prairie grass prairie cordgrass was examined relative to polysaccharide and cell biomass production, yield, and pullulan content of the polysaccharide. A pullulan concentration of 11.2 g L−1 and yield of 0.79 g g−1 was produced by ATCC 42023 when grown for 168 h at 30°C on the phosphate-buffered hydrolysate supplemented with yeast extract. The highest biomass level being 8.8 g L−1 was produced by ATCC 42023 after 168 h on a yeast extract-supplemented, hydrolysate-containing complete medium lacking sodium chloride. The highest pullulan content of the polysaccharide produced by ATCC 42023 after 168 h on the hydrolysate medium supplemented with yeast extract and ammonium sulfate was 70%. The findings indicate that a polysaccharide with a high pullulan content can be produced at a relatively high yield by the fungus grown on a yeast extract-supplemented xylan hydrolysate, suggesting that pullulan could be produced using a biomass-based process.