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Phylogenetic and Functional Analysis of Gut Microbiota of a Fungus-Growing Higher Termite: Bacteroidetes from Higher Termites Are a Rich Source of β-Glucosidase Genes

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Abstract

Fungus-growing termites, their symbiotic fungi, and microbiota inhibiting their intestinal tract comprise a highly efficient cellulose-hydrolyzing system; however, little is known about the role of gut microbiota in this system. Twelve fosmid clones with β-glucosidase activity were previously obtained by functionally screening a metagenomic library of a fungus-growing termite, Macrotermes annandalei. Ten contigs containing putative β-glucosidase genes (bgl1–10) were assembled by sequencing data of these fosmid clones. All these contigs were binned to Bacteroidetes, and all these β-glucosidase genes were phylogenetically closed to those from Bacteroides or Dysgonomonas. Six out of 10 β-glucosidase genes had predicted signal peptides, indicating a transmembrane capability of these enzymes to mediate cellulose hydrolysis within the gut of the termites. To confirm the activities of these β-glucosidase genes, three genes (bgl5, bgl7, and bgl9) were successfully expressed and purified. The optimal temperature and pH of these enzymes largely resembled the environment of the host’s gut. The gut microbiota composition of the fungus-growing termite was also determined by 454 pyrosequencing, showing that Bacteroidetes was the most dominant phylum. The diversity and the enzyme properties of β-glucosidases revealed in this study suggested that Bacteroidetes as the major member in fungus-growing termites contributed to cello-oligomer degradation in cellulose-hydrolyzing process and represented a rich source for β-glucosidase genes.

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Acknowledgments

This work was supported by grants of the National Natural Science Foundation of China (31070098). We appreciate Professor Shubiao Wu from the University of New England for his critical reading and kind suggestions. We also give our thanks to Prashanth Singanallur and Lauren Christine Radlinski from the University of Illinois at Urbana-Champaign for proofreading. We appreciated Dr. Haokui Zhou from Department of Microbiology, The Chinese University of Hongkong and Lei zhang from Logic Informatics Co.,Ltd., for their kind help in data analysis.

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Authors and Affiliations

  1. School of Life Sciences, East China Normal University, Shanghai, China

    Meiling Zhang

  2. Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China

    Changli Qian, Qianfu Wang, Zhihua Zhou & Xing Yan

  3. Key Laboratory of Insect Development and Evolutionary Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China

    Ning Liu, Qian Wang & Yongping Huang

  4. Zhejiang Academy of Agricultural Sciences, Hangzhou, China

    Ning Liu

  5. School of Life Science, Anhui Agricultural University, Hefei, Anhui, China

    Yanhua Long

  6. Key Laboratory of Synthetic Biology, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, 300 Fenglin Road, 200032, Shanghai, China

    Xing Yan

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  1. Meiling Zhang
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Correspondence to Xing Yan.

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Zhang, M., Liu, N., Qian, C. et al. Phylogenetic and Functional Analysis of Gut Microbiota of a Fungus-Growing Higher Termite: Bacteroidetes from Higher Termites Are a Rich Source of β-Glucosidase Genes. Microb Ecol 68, 416–425 (2014). https://doi.org/10.1007/s00248-014-0388-3

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