The Journal of Microbiology

, Volume 47, Issue 4, pp 393–401 | Cite as

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
Articles

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.

Keywords

Vitis vinifera L. plant endophytes 16S rRNA gene length heterogeneity-PCR bacterial enrichment 

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Copyright information

© The Microbiological Society of Korea and Springer Berlin Heidelberg 2009

Authors and Affiliations

  • Daniela Bulgari
    • 1
  • Paola Casati
    • 1
  • Lorenzo Brusetti
    • 2
  • Fabio Quaglino
    • 1
  • Milena Brasca
    • 3
  • Daniele Daffonchio
    • 4
  • Piero Attilio Bianco
    • 1
  1. 1.Dipartimento di Produzione Vegetale (Di.Pro.Ve.) — sez. Patologia VegetaleUniversità degli StudiMilanItaly
  2. 2.Facoltà di Scienze e TecnologieLibera Università di BolzanoBozenItaly
  3. 3.CNR — Istituto di Scienze delle Produzioni AlimentariMilanItaly
  4. 4.Dipartimento di Scienze e Tecnologie Alimentari e Microbiologiche (DISTAM)Università degli StudiMilanItaly

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