Endophytic bacterial diversity in grapevine (Vitis vinifera L.) leaves described by 16S rRNA gene sequence analysis and length heterogeneity-PCR

Daniela Bulgari, Paola Casati, Lorenzo Brusetti, Fabio Quaglino, Milena Brasca, Daniele Daffonchio, Piero Attilio Bianco

Research output: Contribution to journalArticlepeer-review

60 Scopus citations

Abstract

Diversity of bacterial endophytes associated with grapevine leaf tissues was analyzed by cultivation and cultivation-independent methods. In order to identify bacterial endophytes directly from metagenome, a protocol for bacteria enrichment and DNA extraction was optimized. Sequence analysis of 16S rRNA gene libraries underscored five diverse Operational Taxonomic Units (OTUs), showing best sequence matches with γ-Proteobacteria, family Enterobacteriaceae, with a dominance of the genus Pantoea. Bacteria isolation through cultivation revealed the presence of six OTUs, showing best sequence matches with Actinobacteria, genus Curtobacterium, and with Firmicutes genera Bacillus and Enterococcus. Length Heterogeneity-PCR (LH-PCR) electrophoretic peaks from single bacterial clones were used to setup a database representing the bacterial endophytes identified in association with grapevine tissues. Analysis of healthy and phytoplasma-infected grapevine plants showed that LH-PCR could be a useful complementary tool for examining the diversity of bacterial endophytes especially for diversity survey on a large number of samples.

Original languageEnglish (US)
Pages (from-to)393-401
Number of pages9
JournalJournal of Microbiology
Volume47
Issue number4
DOIs
StatePublished - Sep 1 2009

Keywords

  • 16S rRNA gene
  • Bacterial enrichment
  • Length heterogeneity-PCR
  • Plant endophytes
  • Vitis vinifera L.

ASJC Scopus subject areas

  • Microbiology
  • Applied Microbiology and Biotechnology

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