Endophytic Bacterial Diversity in Grapevine (Vitis vinifera L.) Leaves Described by 16S rRNA Gene Sequence Analysis and Length Heterogeneity-PCR

被引：71

作者：

Bulgari, Daniela ^{[1
]}

Casati, Paola ^{[1
]}

Brusetti, Lorenzo ^{[2
]}

Quaglino, Fabio ^{[1
]}

Brasca, Milena ^{[3
]}

Daffonchio, Daniele ^{[4
]}

Bianco, Piero Attilio ^{[1
]}

机构：

[1] Univ Milan, Dipartimento Prod Vegetale, Sez Patol Vegetale, I-20133 Milan, Italy

[2] Libera Univ Bolzano, Fac Sci & Technol, I-39100 Bolzano, Italy

[3] CNR, Ist Sci Prod Alimentari, I-20133 Milan, Italy

[4] Univ Milan, DISTAM, I-20133 Milan, Italy

来源：

JOURNAL OF MICROBIOLOGY | 2009年 / 47卷 / 04期

关键词：

Vitis vinifera L; plant endophytes; 16S rRNA gene; length heterogeneity-PCR; bacterial enrichment; POLYMERASE-CHAIN-REACTION; GROWTH-PROMOTING RHIZOBACTERIA; PANTOEA-AGGLOMERANS; BIOLOGICAL-CONTROL; MICROBIAL COMMUNITIES; FLAVESCENCE DOREE; UNIVERSAL PRIMERS; ERWINIA-AMYLOVORA; SUGAR-BEET; ROOTS;

D O I：

10.1007/s12275-009-0082-1

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

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 gamma-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.

引用

页码：393 / 401

页数：9

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