Biodiversity Science ›› 2018, Vol. 26 ›› Issue (12): 1318-1324.doi: 10.17520/biods.2018184

• Original Papers • Previous Article     Next Article

Community composition of bacteria associated with ascocarps of Tuber indicum using traditional culture method and Roche 454 high-throughput sequencing

Xiaojuan Deng1, Jianli Liu1, Xingfu Yan1, Peigui Liu2, *()   

  1. 1 College of Biological Science and Engineering, Beifang University of Nationalities, Yinchuan 750021
    2 Key Laboratory of Biodiversity and Biogeography, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming 650201
  • Received:2018-07-03 Accepted:2018-12-21 Online:2019-02-11
  • Liu Peigui
  • About author:# 同等贡献作者 Contributed equally to this work

Truffles are important economic fungi, and bacteria play an important role in their growth and development. We investigated the bacterial community composition inside ascocarps of Tuber indicum. Using the traditional culture method, 532 isolates were obtained from ascocarps. Using the rarefaction curve, the 16S rRNA gene was collected and sequenced from 112 purified isolates to identify 4 genera and 40 species. Isolates of the genera Pseudomonas, Acinetobacter, Streptomyces and Variovorax accounted for 80%, 12.5%, 5% and 2.5% of the number of these sequenced isolates, respectively. On the other hand, 9,862 sequences of the bacterial V1-V3 region of 16S rRNA gene, which represented 220 species, were analyzed via Roche 454 high-throughput sequencing. Species of the phyla Proteobacteria, Bacteroidetes, and Actinobacteria were dominant, accounting for 99.7% of all identified species. Genera Flavobacterium, Agromyces, Microbacterium, Ensifer, and Stenotrophomonas were dominant among the bacteria identified with this alternative method, accounting for 86.3% of the number of total species. It was found that relatively few bacterial species were isolated from ascocarps of T. indicum when analyzed via traditional culture method. The bacterial population associated with ascocarps of T. indicum was augmented when analyzed by Roche 454 high-throughput sequencing, which indicates that this latter method provides more comprehensive results.

Key words: Tuber indicum, acocarps, bacteria, culture, high-throughput sequencing

Fig. 1

Culturable bacteria from Tuber indicum ascocarps"

Fig. 2

One of most parsimonious trees based on the analysis of 16S rRNA gene sequences of culturable bacteria from Tuber indicum ascocarps. Numbers above branches indicate bootstrap support above 50%."

Fig. 3

Rarefaction curve between species and sequences of bacteria from Tuber indicum ascocarps"

Fig. 4

Distribution of bacterial species in phylum (a) and at genus level (b)"

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