נגישות
menu      
Advanced Search
Syntax
Search...
Volcani treasures
About
Terms of use
Manage
Community:
אסיף מאגר המחקר החקלאי
Powered by ClearMash Solutions Ltd -
Molecular tools for isolate and community studies of Pyrenomycete fungi
Year:
2004
Source of publication :
Mycologia
Authors :
Freeman, Stanley
;
.
Minz, Dror
;
.
Volume :
96
Co-Authors:
Green, S.J., Dept. Microbiol. and Plant Pathol., Fac. Agric., Food Environ. Qual. S., Hebrew University of Jerusalem, Rehovot, Israel, Inst. Soil, Water and Environ. Sci., Agriculture Research Organization, Volcani Center, P.O. Box 6, Bet-Dagan, 50-250, Israel
Freeman, S., Department of Plant Pathology, Agriculture Research Organization, Volcani Center, P.O. Box 6, Bet-Dagan, 50-250, Israel
Hadar, Y., Dept. Microbiol. and Plant Pathol., Fac. Agric., Food Environ. Qual. S., Hebrew University of Jerusalem, Rehovot, Israel
Minz, D., Inst. Soil, Water and Environ. Sci., Agriculture Research Organization, Volcani Center, P.O. Box 6, Bet-Dagan, 50-250, Israel
Facilitators :
From page:
439
To page:
451
(
Total pages:
13
)
Abstract:
The Pyrenomycetes, defined physiologically by the formation of a flask-shaped fruiting body present in the sexual form, are a monophyletic group of fungi that consist of a wide diversity of populations including human and plant pathogens. Based on sequence analysis of 18S ribosomal DNA (rDNA), rDNA regions conserved among the Pyrenomycetes but divergent among other organisms were identified and used to develop selective PCR primers and a highly specific primer set. The primers presented here were used to amplify large portions of the 18S rDNA as well as the entire internal transcribed spacer (ITS) region (ITS 1, 5.8S rDNA, and ITS 2). In addition to database searches, the specificity of the primers was verified by PCR amplification of DNA extracted from pure culture isolates and by sequence analysis of fungal rDNA PCR-amplified from environmental samples. In addition, denaturing gradient gel electrophoresis (DGGE) analyses were performed on closely related Colletotrichum isolates serving as a model pathogenic genus of the Pyrenomycetes. Although both ITS and 18S rDNA DGGE analyses of Colletotrichum were consistent with a phylogeny established from sequence analysis of the ITS region, DGGE analysis of the ITS region was found to be more sensitive than DGGE analysis of the 18S rDNA. This study introduces molecular tools for the study of Pyrenomycete fungi by the development of two specific primers, demonstration of the enhanced sensitivity of ITS-DGGE for typing of closely related isolates and application of these tools to environmental samples.
Note:
Related Files :
18S rDNA
Colletotrichum
DGGE
fungi
fungus
ITS
molecular analysis
Nested PCR
PCR Primers
pyrenomycetes
Show More
Related Content
More details
DOI :
Article number:
Affiliations:
Database:
Scopus
Publication Type:
article
;
.
Language:
English
Editors' remarks:
ID:
31451
Last updated date:
02/03/2022 17:27
Creation date:
17/04/2018 01:02
Scientific Publication
Molecular tools for isolate and community studies of Pyrenomycete fungi
96
Green, S.J., Dept. Microbiol. and Plant Pathol., Fac. Agric., Food Environ. Qual. S., Hebrew University of Jerusalem, Rehovot, Israel, Inst. Soil, Water and Environ. Sci., Agriculture Research Organization, Volcani Center, P.O. Box 6, Bet-Dagan, 50-250, Israel
Freeman, S., Department of Plant Pathology, Agriculture Research Organization, Volcani Center, P.O. Box 6, Bet-Dagan, 50-250, Israel
Hadar, Y., Dept. Microbiol. and Plant Pathol., Fac. Agric., Food Environ. Qual. S., Hebrew University of Jerusalem, Rehovot, Israel
Minz, D., Inst. Soil, Water and Environ. Sci., Agriculture Research Organization, Volcani Center, P.O. Box 6, Bet-Dagan, 50-250, Israel
Molecular tools for isolate and community studies of Pyrenomycete fungi
The Pyrenomycetes, defined physiologically by the formation of a flask-shaped fruiting body present in the sexual form, are a monophyletic group of fungi that consist of a wide diversity of populations including human and plant pathogens. Based on sequence analysis of 18S ribosomal DNA (rDNA), rDNA regions conserved among the Pyrenomycetes but divergent among other organisms were identified and used to develop selective PCR primers and a highly specific primer set. The primers presented here were used to amplify large portions of the 18S rDNA as well as the entire internal transcribed spacer (ITS) region (ITS 1, 5.8S rDNA, and ITS 2). In addition to database searches, the specificity of the primers was verified by PCR amplification of DNA extracted from pure culture isolates and by sequence analysis of fungal rDNA PCR-amplified from environmental samples. In addition, denaturing gradient gel electrophoresis (DGGE) analyses were performed on closely related Colletotrichum isolates serving as a model pathogenic genus of the Pyrenomycetes. Although both ITS and 18S rDNA DGGE analyses of Colletotrichum were consistent with a phylogeny established from sequence analysis of the ITS region, DGGE analysis of the ITS region was found to be more sensitive than DGGE analysis of the 18S rDNA. This study introduces molecular tools for the study of Pyrenomycete fungi by the development of two specific primers, demonstration of the enhanced sensitivity of ITS-DGGE for typing of closely related isolates and application of these tools to environmental samples.
Scientific Publication
You may also be interested in