Michael P. Schellenberg

Diversity and functionality of arbuscular mycorrhizal fungi in three plant communities in semiarid Grasslands National Park, Canada.

Septate endophytes proliferating in the roots of grasslands’ plants shed doubts on the importance of arbuscular mycorrhizal (AM) symbioses in dry soils. The functionality and diversity of the AM symbioses formed in four replicates of three adjacent plant communities (agricultural, native, and restored) in Grasslands National Park, Canada were assessed in periods of moisture sufficiency and deficiency typical of early and late summer in the region. The community structure of AM fungi, as determined by polymerase chain reaction-denaturing gradient gel electrophoresis, varied with sampling time and plant community. Soil properties other than soil moisture did not change significantly with sampling time. The DNA sequences dominating AM extraradical networks in dry soil apparently belonged to rare taxa unreported in GenBank. DNA sequences of Glomus viscosum, Glomus mosseae, and Glomus hoi were dominant under conditions of moisture sufficiency. In total, nine different AM fungal sequences were found suggesting a role for the AM symbioses in semiarid areas. Significant positive linear relationships between plant P and N concentrations and active extraradical AM fungal biomass, estimated by the abundance of the phospholipid fatty acid marker 16:1ω5, existed under conditions of moisture sufficiency, but not under dry conditions. Active extraradical AM fungal biomass had significantly positive linear relationship with the abundance of two early season grasses, Agropyron cristatum (L.) Gaertn. and Koeleria gracilis Pers., but no relationship was found under dry conditions. The AM symbioses formed under conditions of moisture sufficiency typical of early summer at this location appear to be important for the nutrition of grassland plant communities, but no evidence of mutualism was found under the dry conditions of late summer.

The potential of legume-shrub mixtures for optimum forage production in southwestern Saskatchewan: A greenhouse study

Grazing in fall and early winter decreases the cost of beef production in southwe stern Saskatchewan. This grazing system can be improved by utilizing legume and native shrub species, which exhibit high nutritive value in the fall. To realize the systems full potential, a better understanding of optimum mixtures of legumes and shrubs is required. A greenhouse study was conducted to optimize mixtures of legumes and shrubs for economical pasture production. The first goal was to obtain better understanding of synergy from mixtures of legumes and native shrubs. The second goal was to estimate the changes in soil quality caused by growing legumes and shrubs in monocultures or mixtures. Legume species studied were: alfalfa (Medicago sativa L.) (Alf), purple prairie clover [Petalostemon purpureum (Vert.) Rydb] (Pr Cl) and American vetch (Vicia americana Muhl.) (Vetch); shrubs were: winterfat [Krascheninnkovia lanata (Pursh) Guldenstaedt] (Wf) and Gardners saltbush [Atriplex gardneri (Moq.) D. Dietr.] (Sb). Tr...

Soil Respiration in Semiarid Temperate Grasslands under Various Land Management

Soil respiration, a major component of the global carbon cycle, is significantly influenced by land management practices. Grasslands are potentially a major sink for carbon, but can also be a source. Here, we investigated the potential effect of land management (grazing, clipping, and ungrazed enclosures) on soil respiration in the semiarid grassland of northern China. Our results showed the mean soil respiration was significantly higher under enclosures (2.17μmol.m−2.s−1) and clipping (2.06μmol.m−2.s−1) than under grazing (1.65μmol.m−2.s−1) over the three growing seasons. The high rates of soil respiration under enclosure and clipping were associated with the higher belowground net primary productivity (BNPP). Our analyses indicated that soil respiration was primarily related to BNPP under grazing, to soil water content under clipping. Using structural equation models, we found that soil water content, aboveground net primary productivity (ANPP) and BNPP regulated soil respiration, with soil water content as the predominant factor. Our findings highlight that management-induced changes in abiotic (soil temperature and soil water content) and biotic (ANPP and BNPP) factors regulate soil respiration in the semiarid temperate grassland of northern China.

Early productivity and crude protein content of establishing forage swards composed of combinations of native grass and legume species in mixed-grassland ecoregions

Mischkolz, J. M., Schellenberg, M. P. and Lamb, E. G. 2013. Early productivity and crude protein content of establishing forage swards composed of combinations of native grass and legume species in mixed-grassland ecoregions. Can. J. Plant Sci. 93: 445-454. We evaluated the early establishment productivity of forage swards of native, perennial, cool and warm season grasses, and legumes as they have the potential to provide non-invasive, productive, and drought resistant rangelands. Seven species with agronomic potential and a broad native geographic distribution were selected for testing including: nodding brome [Bromus anomalus (Coult.)], blue bunch wheatgrass [Pseudoregneria spicata (Pursh)], western wheatgrass [Pascopyrum smithii (Rydb.)], side oats grama [Bouteloua curtipendula (Michx.)], little blue stem [Schizachyrium scoparium (Michx.)], purple prairie clover [Dalea purpurea (Vent.)], and white prairie clover [Dalea candida (Willd.)]. Forage swards, including all seven monocultures, 21 two-species mixtures and a mixture with all species, were planted in two sites, Saskatoon and Swift Current, Saskatchewan. Western wheatgrass (WWG) had the highest overall plant density and the strongest effect on the forage yield of the forage swards; however, productivity and crude protein content were not reduced when other species were also included in the forage sward. Dalea spp. did not establish as well as the other species, but had the highest crude protein concentrations. This work provides insight into forage sward development at the establishment stage; additional work is required to determine long-term species impacts for well established forage swards.

Different responses of plant species to deferment of sheep grazing in a desert steppe of Inner Mongolia, China

This paper reports the effects of different deferments of sheep grazing on plant variables of desert steppe vegetation in Inner Mongolia during 2010–2012. The study was initiated in May 2010 and comprised five treatments: no grazing (UG), grazing throughout the growing season (G) and grazing deferment for 40, 50 and 60 days (40UG, 50UG and 60UG) from the start of the growing season in a completely randomised block design replicated three times. The plant species were classified into two functional groups (C3 and C4 species) and the relationships among species composition, biomass and annual precipitation were examined in different deferments of sheep grazing. Treatment G significantly decreased the biomass of several species compared with some of the deferred treatments and treatment UG, because of differences in grazing preferences, and, consequently, decreased the aboveground net primary productivity (ANPP). Grazing deferment for 40, 50 and 60 days significantly increased C3-species richness and biomass compared with treatment UG, whereas grazing sheep’s preferences for C4 plants at some growth stages compared with C3 plants, resulted in lower species richness and biomass of C4 plants in treatments G, 40UG and 50UG in a dry year. Similarly, different species responses to treatment 60UG resulted in an increase in ANPP in a year with more precipitation during the growing season. The annual precipitation patterns strongly affected the temporal changes in biomass as well as the responses to grazing, indicating that the plant responses in the desert steppe were co-limited by grazing and precipitation. These findings provide important insights into the management and conservation of desert steppe vegetation in Inner Mongolia.

Responses of Plant Community and Soil Properties to Inter-Annual Precipitation Variability and Grazing Durations in a Desert Steppe in Inner Mongolia

Grazing can dramatically affect arid grassland communities that are very vulnerable to environmental changes due to its relatively short and sparse ground coverage, low biomass, sandy soil and inter-annual precipitation found in the desert steppe. The study investigates the effects of different grazing durations on vegetation and soil properties of a desert steppe community. The experiment was conducted in Xisu Banner in Inner Mongolia with five treatments: CG (continuous grazing), 40UG (40 d ungrazed), 50UG (50 d ungrazed), 60UG (60 d ungrazed) and UG (ungrazed). The biomass of both shrub and annual-biennial plant communities were significantly decreased by CG. Continuous grazing and 40UG significantly reduced the ANPP (aboveground net primary productivity) by the end of the three year study. 60UG treatment increased soil organic carbon (OC), total nitrogen concentration (TN) and total phosphorus concentration (TP) concentrations and 50UG increased the TN and total phosphorus concentration (TK) concentrations, whereas CG, 40UG and 50UG decreased soil OC, TP and available phosphorus concentration (AP) concentrations. The perennial plant species of the desert steppe were generally tolerant for grazing. The annual-biennial plant species had large variability in ANPP because of the inter-annual precipitation. Our results highlight that inter-annual precipitation variations could strongly modify the community responses to grazing in arid ecosystems.

RNA-Seq analysis of gene expression for floral development in crested wheatgrass (Agropyron cristatum L.).

Crested wheatgrass [Agropyron cristatum L. (Gaertn.)] is widely used for early spring grazing in western Canada and the development of late maturing cultivars which maintain forage quality for a longer period is desired. However, it is difficult to manipulate the timing of floral transition, as little is known about molecular mechanism of plant maturity in this species. In this study, RNA-Seq and differential gene expression analysis were performed to investigate gene expression for floral initiation and development in crested wheatgrass. Three cDNA libraries were generated and sequenced to represent three successive growth stages by sampling leaves at the stem elongation stage, spikes at boot and anthesis stages. The sequencing generated 25,568,846; 25,144,688 and 25,714,194 qualified Illumina reads for the three successive stages, respectively. De novo assembly of all the reads generated 311,671 transcripts with a mean length of 487 bp, and 152,849 genes with an average sequence length of 669 bp. A total of 48,574 (31.8%) and 105,222 (68.8%) genes were annotated in the Swiss-Prot and NCBI non-redundant (nr) protein databases, respectively. Based on the Kyoto Encyclopedia of Genes and Genome (KEGG) pathway database, 9,723 annotated sequences were mapped onto 298 pathways, including plant circadian clock pathway. Specifically, 113 flowering time-associated genes, 123 MADS-box genes and 22 CONSTANS-LIKE (COL) genes were identified. A COL homolog DN52048-c0-g4 which was clustered with the flowering time genes AtCO and OsHd1 in Arabidopsis (Arabidopsis thaliana L.) and rice (Oryza sativa L.), respectively, showed specific expression in leaves and could be a CONSTANS (CO) candidate gene. Taken together, this study has generated a new set of genomic resources for identifying and characterizing genes and pathways involved in floral transition and development in crested wheatgrass. These findings are significant for further understanding of the molecular basis for late maturity in this grass species.

Long-Term Land Use Affects Phosphorus Speciation and the Composition of Phosphorus Cycling Genes in Agricultural Soils

Agriculturally-driven land transformation is increasing globally. Improving phosphorus (P) use efficiency to sustain optimum productivity in diverse ecosystems, based on knowledge of soil P dynamics, is also globally important in light of potential shortages of rock phosphate to manufacture P fertilizer. We investigated P chemical speciation and P cycling with solution 31P nuclear magnetic resonance, P K-edge X-ray absorption near-edge structure spectroscopy, phosphatase activity assays, and shotgun metagenomics in soil samples from long-term agricultural fields containing four different land-use types (native and tame grasslands, annual croplands, and roadside ditches). Across these land use types, native and tame grasslands showed high accumulation of organic P, principally orthophosphate monoesters, and high acid phosphomonoesterase activity but the lowest abundance of P cycling genes. The proportion of inositol hexaphosphates (IHP), especially the neo-IHP stereoisomer that likely originates from microbes rather than plants, was significantly increased in native grasslands than croplands. Annual croplands had the largest variances of soil P composition, and the highest potential capacity for P cycling processes based on the abundance of genes coding for P cycling processes. In contrast, roadside soils had the highest soil Olsen-P concentrations, lowest organic P, and highest tricalcium phosphate concentrations, which were likely facilitated by the neutral pH and high exchangeable Ca of these soils. Redundancy analysis demonstrated that IHP by NMR, potential phosphatase activity, Olsen-P, and pH were important P chemistry predictors of the P cycling bacterial community and functional gene composition. Combining chemical and metagenomics results provides important insights into soil P processes and dynamics in different land-use ecosystems.

RNA-Seq Analysis of Plant Maturity in Crested Wheatgrass (Agropyron cristatum L.)

Crested wheatgrass (Agropyron cristatum L.) breeding programs aim to develop later maturing cultivars for extending early spring grazing in Western Canada. Plant maturity is a complex genetic trait, and little is known about genes associated with late maturity in this species. An attempt was made using RNA-Seq to profile the transcriptome of crested wheatgrass maturity and to analyze differentially expressed genes (DEGs) between early and late maturing lines. Three cDNA libraries for each line were generated by sampling leaves at the stem elongation stage, spikes at the boot and anthesis stages. A total of 75,218,230 and 74,015,092 clean sequence reads were obtained for early and late maturing lines, respectively. De novo assembly of all sequence reads generated 401,587 transcripts with a mean length of 546 bp and N50 length of 691 bp. Out of 13,133 DEGs detected, 22, 17, and eight flowering related DEGs were identified for the three stages, respectively. Twelve DEGs, including nine flowering related DEGs at the stem elongation stage were further confirmed by qRT-PCR. The analysis of homologous genes of the photoperiod pathway revealed their lower expression in the late maturing line at the stem elongation stage, suggesting that their differential expression contributed to late maturity in crested wheatgrass.

Genotyping-by-sequencing data of 272 crested wheatgrass (Agropyron cristatum) genotypes

Crested wheatgrass [Agropyron cristatum L. (Gaertn.)] is an important cool-season forage grass widely used for early spring grazing. However, the genomic resources for this non-model plant are still lacking. Our goal was to generate the first set of next generation sequencing data using the genotyping-by-sequencing technique. A total of 272 crested wheatgrass plants representing seven breeding lines, five cultivars and five geographically diverse accessions were sequenced with an Illumina MiSeq instrument. These sequence datasets were processed using different bioinformatics tools to generate contigs for diploid and tetraploid plants and SNPs for diploid plants. Together, these genomic resources form a fundamental basis for genomic studies of crested wheatgrass and other wheatgrass species. The raw reads were deposited into Sequence Read Archive (SRA) database under NCBI accession SRP115373 (https://www.ncbi.nlm.nih.gov/sra?term=SRP115373) and the supplementary datasets are accessible in Figshare (10.6084/m9.figshare.5345092).