The microbial nitrogen cycling, bacterial community composition, and functional potential in a natural grassland are stable from breaking dormancy to being dormant again

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Authors

Das, Bikram K.
Ishii, Satoshi
Antony, Linto
Smart, Alexander J.
Scaria, Joy
Brözel, Volker Siegfried

Journal Title

Journal ISSN

Volume Title

Publisher

MDPI

Abstract

The quantity of grass-root exudates varies by season, suggesting temporal shifts in soil microbial community composition and activity across a growing season. We hypothesized that bacterial community and nitrogen cycle-associated prokaryotic gene expressions shift across three phases of the growing season. To test this hypothesis, we quantified gene and transcript copy number of nitrogen fixation (nifH), ammonia oxidation (amoA, hao, nxrB), denitrification (narG, napA, nirK, nirS, norB, nosZ), dissimilatory nitrate reduction to ammonia (nrfA), and anaerobic ammonium oxidation (hzs, hdh) using the pre-optimized Nitrogen Cycle Evaluation (NiCE) chip. Bacterial community composition was characterized using V3-V4 of the 16S rRNA gene, and PICRUSt2 was used to draw out functional inferences. Surprisingly, the nitrogen cycle genes and transcript quantities were largely stable and unresponsive to seasonal changes. We found that genes and transcripts related to ammonia oxidation and denitrification were different for only one or two time points across the seasons (p < 0.05). However, overall, the nitrogen cycling genes did not show drastic variations. Similarly, the bacterial community also did not vary across the seasons. In contrast, the predicted functional potential was slightly low for May and remained constant for other months. Moreover, soil chemical properties showed a seasonal pattern only for nitrate and ammonium concentrations, while ammonia oxidation and denitrification transcripts were strongly correlated with each other. Hence, the results refuted our assumptions, showing stability in N cycling and bacterial community across growing seasons in a natural grassland.

Description

DATA AVAILABILITY STATEMENT : The raw data is available on NCBI’s Sequence Read Archive (SRA) database under BioProject: PRJNA803487.
SUPPLEMENTARY MATERIALS : FIGURE S1: Soil textures of the sampling sites; FIGURE S2: Distribution of plant groups across the six study sites; FIGURE S3: Plant species distribution across the study sites. The legume species are represented by asterisk (*) on the legend. The abbreviations represent: BRIN, Bromus inermis; POPR, Poa pratensis; PHAR, Phalaris arundinacea; CA, Carex sp.; NA, Nassella sp.; ANGE, Andropogon gerardii; SCSC, Schizachyrium scoparium; BOCU, Bouteloua curtipendula; DIOL, Dichanthelium oligosanthes; CIFL, Cirsium flodmanii; ASSP, Asclepias speciose; SOMI, Solidago missouriensis; HEMA, Helianthus maximiliani; ANCA, Anemone canadensis; SOCA, Solidago canadensis; GLLE, Glycyrrhiza lepidota; PH, Physalis sp.; AMCA*, Amorpha canescens; DAPU*, Dalea purpurea; PE*, Pediomelum sp.; RO, Rosa sp.; FIGURE S4: Soil chemical properties across sampling sites within the sampling time points with Kruskal–Wallis test results; FIGURE S5: Alpha-diversity of the bacterial communities across 6 months. (a) Shannon diversity and (b) Pielou Evenness.; FIGURE S6: Taxa differences across at least one time point across the seasons (p < 0.01). The names of the phyla shown here are based on the taxonomic profile downloaded from the Greengenes database, however, some of the phylum names have recently been changed [38]; TABLE S1. G-Block solutions; TABLE S2. Primers used for nitrogen cycle genes. SUPPLEMENTARY DATA S3. R 2 values of the qPCR assays. SUPPLEMENTARY DATA S4. OTU table.

Keywords

Prairie, Grassland, Nitrogen cycle, Soil bacterial community, SDG-15: Life on land

Sustainable Development Goals

Citation

Das, B.K.; Ishii, S.; Antony, L.; Smart, A.J.; Scaria, J.; Brözel, V.S. The Microbial Nitrogen Cycling, Bacterial Community Composition, and Functional Potential in a Natural Grassland Are Stable from Breaking Dormancy to Being Dormant Again. Microorganisms 2022, 10, 923. https://DOI.org/10.3390/microorganisms10050923.