Biodiversity Data Journal 9: e60245 CO) doi: 10.3897/BDJ.9.e60245 open access Data Paper Revealing the active microbiome connected with the rhizosphere soil of maize plants in Ventersdorp, South Africa Olubukola O. Babalola*, Rebaona R. Molefe?, Adenike E. Amoo*t $ Food Security and Safety Niche, Faculty of Natural and Agricultural Sciences, North-West University, Private Bag X2046, Mafikeng, South Africa Corresponding author: Olubukola O. Babalola (olubukola.babalola@nwu.ac.za) Academic editor: Anna Sandionigi Received: 01 Nov 2020 | Accepted: 05 Feb 2021 | Published: 25 Feb 2021 Citation: Babalola OO, Molefe RR, Amoo AE (2021) Revealing the active microbiome connected with the rhizosphere soil of maize plants in Ventersdorp, South Africa. Biodiversity Data Journal 9: e60245. https://doi.org/10.3897/BDJ.9.e60245 Abstract We conducted shotgun metagenomics sequencing of the maize rhizosphere and bulk soils in Ventersdorp, South Africa. Information on the structural composition and functional capabilities of microbial communities in the maize rhizosphere are provided by the data. Characterising the functional potentials of rhizosphere microbiomes gives an opportunity to link the microbiome to plant growth and health and provides the possibility of discovering new plant-beneficial genes that could enhance agricultural sustainability. Keywords Illumina sequencing, metagenomics, rhizosphere, maize plants Introduction Maize is one of South Africa's most economically-valuable crops. Globally, it fills the diets of billions of people with basic carbohydrates. Poor management practices, such as over- © Babalola O et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 2 Babalola O et al fertilisation, have gone up significantly due to the quest to feed the ever-increasing human population. Therefore, it is imperative to identify eco-friendly fertilisers that do not have adverse effects on soil and maize development. Plants establish associations with soil microorganisms for various functions including nutrient cycling, stress tolerance and pathogen immunity (Liu et al. 2019). Increased knowledge of these mechanisms is a productive and positive way for the improvement of sustainable agriculture (Babalola et al. 2021). The rhizosphere, which is the medium between plants and soil, has been labelled a ‘hotspot’ for new genes and biomolecules (Babalola et al. 2020). Plant-root exudates generate nourishing conditions for microbial growth and easily attract a selection of soil microorganisms (Adedeji and Babalola 2020; Canarini et al. 2019; Chukwuneme et al. 2021). Microbial communities in the rhizosphere are recruited from the large and diverse pool of microbes in bulk soils through root exudate chemical signalling (Adedeji and Babalola 2020;Hartman and Tringe 2019). This has contributed to an increase in microbial activity and quantity in rhizosphere soils compared to bulk soils. In contrast, microbial diversity significantly reduces in the rhizosphere soil relative to bulk soil (Praeg et al. 2019; Hartman and Tringe 2019). Rhizosphere microbes exist to protect against pathogens and improve growth by developing phytohormones. These organisms enable plants to handle environmental disruptions, such as irregular climate-related changes in temperature, drought and salinity (Lu et al. 2018). It has been shown that nitrogen-fixing rhizobia and the mycorrhizal fungi in the rhizosphere have significant impacts on plant nutrient status (Mendes et al. 2013; Lu et al. 2018). For example, symbiotics, such as mycorrhizal fungi, are important for the absorption of nutrients and minerals from the soil to plants. Therefore, studies on the rhizospheric microbes and their functions could open several appealing features, from alleviating several of the consequences of climate change and environmental stress on plants by modifying plant features using microbial inocula to enhancing crop production. Therefore, the discovery of new genes in the maize rhizosphere could be an incentive to fix food insecurity and promote agricultural sustainability. Value of the dataset The dataset contains raw sequences (FASTQ format files) obtained using shotgun metagenomic sequencing of the maize rhizosphere and bulk soils. Samples were collected from the maize rhizosphere (F3R1) and bulk (F3B1) soils to understand the microbial community structure, function and plant-beneficial genes in maize plantations. These data can be used alone or along with other datasets to achieve a larger scale view with more power for maize-associated microbiome research. Revealing the active microbiome connected with the rhizosphere soil of ... 3 Methods Sampling Soil samples were collected from the rhizosphere soil (F3R1) and the bulk soil (F3B1) of maize plants on 16 June 2019 from a farm situated at Ventersdorp, South Africa. The rhizosphere soil samples were collected at 8 cm diameter, 15 cm depth of maize plants. The bulk soils were also collected within the maize farms. Environmental profile The maize field being investigated in this study is a private farm in Ventersdorp in the North West Province of South Africa. The farm was intentionally selected, based on the geographic location and the availability of maize plants. Ventersdorp has summer temperatures ranging from 17°C to 31°C and winter temperatures ranging from 3°C to 21°C. The annual rainfall ranges between 300 mm and 600 mm with more rain falling in summer than in winter. Geographic range Ventersdorp, North West Province (approximately 26°19'36.9"S, 26°53'19.1"E). Coordinates: -26°18'60.00"S; 26°48'59.99"E. Sample processing The soil samples were transported to the laboratory on ice and stored until further use. Genomic DNA extraction was conducted using the DNeasy PowerSoil® DNA isolation kit (MoBio Laboratories, Carlsbad, CA) in accordance with the manufacturer's directions. The extracted DNA was sent for shotgun metagenome sequencing to the Molecular Research Laboratory (www.mrdnalab.com) in Texas, USA. The initial concentration of DNA was evaluated using the Qubit® dsDNA HS Assay Kit (Life Technologies). The libraries were prepared using Nextera DNA Flex library preparation kit (Illumina), following the manufacturer's user guide. Using 50 ng of DNA from each sample, libraries were prepared according to the Illumina NovaSeq DNA library preparation protocol. The determination of library average insert size was determined using the Agilent 2100 Bioanalyzer (Agilent Technologies). The library insert size ranged from 617 bp to 873 bp. The libraries were pooled, diluted (to 0.6 nM) and sequenced paired-end for 300 cycles using the NovaSeq system (Illumina). Data processing The raw metagenome sequences were subjected to quality control using Metagenomic Rapid Annotations using Subsystems Technology (MG-RAST) online server (Meyer et al. 2008). This resulted in evacuation of artificial sequences generated by sequencing errors, exclusion of sequences of host-specific organisms, unclear base filtering (abolition of 4 Babalola O et al sequences of > 5 questionable base pairs with a cut-off score of 15 Q) and filtering of length (abolition of sequences of > 2 standard deviations from mean length). Following the quality control (QC), the sequences were annotated using the BLAT (the BLAST-like alignment tool) algorithm (Kent 2002) against the M5NR database (Wilke et al. 2012), which provides a non-redundant integration of many databases. For taxonomic profiling of microbial communities, the SEED subsystem was used and evaluation of their functional profiles was performed using SEED subsystem level 1. The subsystem database revealed bacteria (98.76%) had the highest taxonomical representation compared with eukaryote (0.72%) and archaea (0.73%). Annotation revealed that F3R1 had 15,713,893 sequences totalling 2,338,704,495 bp size and 64.11% G+C content. F3B1 had 12,463,113 sequences totalling 1,850,061,852 bp size and G+C 66.11%. Technologies used MG-RAST (https://mg-rast.org). Source: The National Human Genome Research Institute (NHGRI) Biodiversity scope The maize rhizosphere soil sample had more microorganisms than the bulk soil sample. Target The rhizosphere microbiome and their functional potentials. Taxonomic range All soil microbiomes were identified to genus or species level. The study revealed that the most abundant phyla were Proteobacteria and Actinobacteria in the rhizosphere and bulk soils. Ascomycota and Basidiomycota were distributed fungal reads, while Thauarcheota and Euyarchaeota were distributed as archaeal reads, respectively, but with an abundance of < 1%.Table 1 Table 1. Taxonomic classification of microorganisms in the maize rhizosphere and bulk soils Domain Phyla F3R1 F3B1 Bacteria Acidobacteria 368735 226709 Bacteria Actinobacteria 2511153 2794515 Bacteria Aquificae 15800 10882 Bacteria Bacteroidetes 347509 261611 Bacteria Candidatus Poribacteria 3480 1944 Revealing the active microbiome connected with the rhizosphere soil of ... Domain Phyla F3R1 F3B1 Bacteria Chlamydiae 6644 4512 Bacteria Chlorobi 35848 24915 Bacteria Chloroflexi 203916 170908 Bacteria Chrysiogenetes 2318 1452 Bacteria Cyanobacteria 190459 138395 Bacteria Deferribacteres 6452 4206 Bacteria Deinococcus-Thermus 66466 56965 Bacteria Dictyoglomi 4479 3216 Bacteria Elusimicrobia 1759 1224 Bacteria Fibrobacteres 1452 1021 Bacteria Firmicutes 393062 304682 Bacteria Fusobacteria 5707 4073 Bacteria Gemmatimonadetes 153957 140020 Bacteria Lentisphaerae 4635 3029 Bacteria Nitrospirae 27120 15636 Bacteria Planctomycetes 185528 121559 Bacteria Proteobacteria 3443718 2362316 Bacteria Spirochaetes 17790 12214 Bacteria Synergistetes 9010 6373 Bacteria Tenericutes 1687 1121 Bacteria Thermotogae 15919 11866 Bacteria Verrucomicrobia 179156 100394 Bacteria unclassified (derived from Bacteria) 22169 18869 Fungi Ascomycota 30221 31523 Fungi Basidiomycota 3615 2657 Fungi Blastocladiomycota 14 20 Fungi Chytridiomycota 44 32 Fungi Glomeromycota 38 6 Fungi Microsporidia 121 57 Fungi unclassified (derived from Fungi) 43 20 Archaea Crenarchaeota 10423 7705 Archaea Euryarchaeota 59001 45925 Archaea Korarchaeota 728 451 Archaea Nanoarchaeota 60 44 Archaea Thaumarchaeota 6596 5229 Viruses unclassified (derived from Viruses) 1244 1109 6 Babalola O et al Functional range The functional annotation using SEED subsystems revealed that reads were more ascribed to carbohydrates metabolism (15.76 to 15.90%), amino acids and derivatives (11.53 to 11.61%) and clustering-based systems (13.63 to 13.78%) in the maize rhizosphere and bulk soils samples. Data Resources Maize associated microbiome studies (Suppl. material 1). Resource 1 Download URL https://trace.ncbi.nim.nih.gov/Traces/sra/?run=SRR12288319 Resource identifier SRR12288319 Data format FASTQ Resource 2 Download URL https://trace.ncbi.nim.nih.gov/Traces/sra/?run=SRR12288317 Resource identifier SRR12288317 Data format FASTQ Usage Rights Creative Commons Public Domain Waiver (CC-Zero) Revealing the active microbiome connected with the rhizosphere soil of ... if Acknowledgements OOB would like to thank the National Research Foundation of South Africa for a grant (Grant Ref: UID123634; OOB) that has supported work in our laboratory. RRM would like to thank the North-West University for the postgraduate bursary that was granted during her MSc programme. AEA is grateful to the North-West University for postdoctoral bursary and research support. Hosting institution North-West University Author contributions All the mentioned authors contributed substantially and intellectually to the work. OOB designed the research, revised the work critically for important intellectual content, performed quality assurance, provided funding acquisition, project administration and resources. RRM was involved in data curation, formal analysis, investigation, visualisation of data and writing of the original draft of the manuscript. AEA was involved in data curation, visualisation of data, reviewing and thoroughly editing of the original draft, validation and formal analysis. Conflicts of interest The authors declare that they have no conflict of interest, either financial or commercial wise. References ° Adedeji AA, Babalola OO (2020) Secondary metabolites as plant defensive strategy: a large role for small molecules in the near root region. Planta 61: 252. https://doi.org/ 10.1007/s00425-020-03468-1 ° Babalola OO, Alawiye TT, Rodriguez Lopez CM, Ayangbenro AS (2020) Shotgun metagenomic sequencing data of sunflower rhizosphere microbial community in South Africa. Data in Brief 31: 105831. https://doi.org/10.1016/j.dib.2020.105831 ° Babalola OO, Omotayo OP, Igiehon NO (2021) Survey of Maize Rhizosphere Microbiome Using Shotgun Metagenomics. Microbiology Resource Announcements 10 (1): €01309-20. https://doi.org/10.1128/MRA.01309-20 ° Canarini A, Kaiser C, Merchant A, Richter A, Wanek W (2019) Root Exudation of Primary Metabolites: Mechanisms and Their Roles in Plant Responses to Environmental Stimuli. Frontiers in Plant Science (10)157. https://doi.org/10.3389/fpls. 2019.00157 8 Babalola O et al ° Chukwuneme FC, Ayangbenro AS, Babalola OO, Kutu FR (2021) Functional diversity of microbial communities in two contrasting maize rhizosphere soils. 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Frontiers in Microbiology (10)1429. https://doi.org/10.3389/fmicb.2019.01429 ° Wilke A, Harrison T, Wilkening J, Field D, Glass EM, Kyrpides N, Mavrommatis K, Meyer F (2012) The M5nr: a novel non-redundant database containing protein sequences and annotations from multiple sources and associated tools. BMC Bioinformatics 13 (1): 141. https://doi.org/10.1186/1471-2105-13-141 Supplementary material Suppl. material 1: BioSample metadata file EE} Authors: Olubukola O. Babalola; Rebaona R. Molefe; Adenike E. Amoo Data type: Metagenomic data Download file (2.25 kb)