DNA-Dependent Detection of the Grapevine Fungal Endophytes Aureobasidium pullulans and Epicoccum nigrum

被引:86
作者
Martini, M. [1 ]
Musetti, R. [1 ]
Grisan, S. [1 ]
Polizzotto, R. [1 ]
Borselli, S. [1 ]
Pavan, F. [1 ]
Osler, R. [1 ]
机构
[1] Univ Udine, Dept Biol & Protez Piante, I-33100 Udine, Italy
关键词
BOTRYTIS-CINEREA; PCR; DIVERSITY; STRAINS; POTATO; AGENT;
D O I
10.1094/PDIS-93-10-0993
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
Aureobasidium pullulons and Epicoccum nigrum are frequently reported as endophytes of various crops, including grapevine (Vitis vinifera). Because of their potential role as biological control agents against grapevine pathogens, we examined the occurrence of A. pullulans and E. nigrum in two grapevine varieties (Merlot and Prosecco) in Italian vineyards where spontaneous recovery from phytoplasma disease is recurrent. Species-specific primers for A. pullulans and two genetically distinct strains of E. nigrum were designed in variable regions of ITS1 and ITS2. Primer specificity was confirmed by polymerase chain reaction using purified DNA from other fungal endophytes that are usually encountered during isolation attempts from grapevine tissues and from several other strains of A. pullulans and E. nigrum isolated from other sources. In order to determine the occurrence of the two endophytes in grapevine plants, DNA was extracted from shoots of 44 grapevines collected in six vineyards from different localities of northeast Italy. Both endophytes were detected and their identity was confirmed by restriction fragment length polymorphism (RFLP) patterns obtained from reference strains. RFLP analyses confirmed the presence of two E. nigrum strains belonging to different RFLP groups in grapevine. The molecular methods described allowed a sensitive, specific, and reliable identification of the two endophytes in grapevine.
引用
收藏
页码:993 / 998
页数:6
相关论文
共 39 条
[1]   Population biology of Aureobasidium pullulans on apple leaf surfaces [J].
Andrews, JH ;
Spear, RN ;
Nordheim, EV .
CANADIAN JOURNAL OF MICROBIOLOGY, 2002, 48 (06) :500-513
[2]  
[Anonymous], 1990, PCR Protocols. A Guide to Methods and Applications
[3]  
Arenal F, 2004, MYCOTAXON, V89, P465
[4]   Evaluation of different PCR-based DNA fingerprinting techniques for assessing the genetic variability of isolates of the fungus Epicoccum nigrum [J].
Arenal, F ;
Platas, G ;
Martín, J ;
Salazar, O ;
Peláez, F .
JOURNAL OF APPLIED MICROBIOLOGY, 1999, 87 (06) :898-906
[5]  
Arnold A. Elizabeth, 2007, Fungal Biology Reviews, V21, P51, DOI 10.1016/j.fbr.2007.05.003
[6]  
CAMPANILE G, 2006, FORESTA, V3, P78
[7]   Strains of Aureobasidium pullulans Can Lower Ochratoxin A Contamination in Wine Grapes [J].
de Felice, D. V. ;
Solfrizzo, M. ;
De Curtis, F. ;
Lima, G. ;
Visconti, A. ;
Castoria, R. .
PHYTOPATHOLOGY, 2008, 98 (12) :1261-1270
[8]   A PCR-based technique for detection of Neotyphodium endophytes in diverse accessions of tall fescue [J].
Doss, RP ;
Clement, SL ;
Kuy, SR ;
Welty, RE .
PLANT DISEASE, 1998, 82 (07) :738-740
[9]  
Doyle J.J., 1987, Focus, V19, P11, DOI DOI 10.2307/2419362
[10]   Incidence, aggressiveness and In planta interactions of Botrytis cinerea and other filamentous fungi quiescent in grape berries and dormant buds in Central Washington State [J].
Dugan, FM ;
Lupien, SL ;
Grove, GG .
JOURNAL OF PHYTOPATHOLOGY-PHYTOPATHOLOGISCHE ZEITSCHRIFT, 2002, 150 (07) :375-381