Pathotypes in the Entomophaga grylli species complex of grasshopper pathogens differentiated with random amplification of polymorphic DNA and cloned-DNA probes

The zygomycetous fungus Entomophaga grylli is a pathogen that shows host-specific variance to grasshopper subfamilies. Three pathotypes of the E. grylli species complex were differentiated by three molecular techniques. In the first method, the three pathotypes showed different fragment patterns gen...

Full description

Saved in:

Bibliographic Details
Published in:	Applied and Environmental Microbiology Vol. 61; no. 2; pp. 556 - 560
Main Authors:	BIDOCKHA, M. J, WALSH, S. R. A, RAMOS, M. E, ST LEGER, R. J, SILVER, J. C, ROBERTS, D. W
Format:	Journal Article
Language:	English
Published:	Washington, DC American Society for Microbiology 01-02-1995
Subjects:	Acrididae adn analytical methods Animals Bacteriology Base Sequence Biological and medical sciences champignon entomopathogene Cloning, Molecular Deoxyribonucleic acid DNA DNA Primers - genetics DNA Probes DNA, Fungal - genetics DNA, Fungal - isolation & purification entomogenous fungi entomophaga Entomophaga grylli Environment esporas Fundamental and applied biological sciences. Psychology Fungi Fungi - classification Fungi - genetics Fungi - pathogenicity genetic markers genetic polymorphism Grasshoppers - microbiology hongos entomogenos identificacion identification insecticidas insecticide insecticides isolation techniques marcadores geneticos marqueur genetique Melanoplinae melanoplus melanoplus sanguinipes Microbiology Molecular Sequence Data Molecules Mycology Orthoptera Pathogenicity, host-agent relations, miscellaneous strains, epidemiology pathotype pathotypes patotipos Pesticides polimorfismo genetico Polymerase Chain Reaction Polymorphism, Restriction Fragment Length polymorphisme genetique spore spores Spores, Fungal - chemistry Spores, Fungal - genetics technique analytique technique de l' isolement tecnicas analiticas tecnicas de aislamiento Fungi Entomopathogen DNA Pathotype Molecular probe Method Differentiation Thallophyta
Online Access:	Get full text
Tags:	Add Tag No Tags, Be the first to tag this record!

Description
Summary:	The zygomycetous fungus Entomophaga grylli is a pathogen that shows host-specific variance to grasshopper subfamilies. Three pathotypes of the E. grylli species complex were differentiated by three molecular techniques. In the first method, the three pathotypes showed different fragment patterns generated by random amplification of polymorphic DNA (RAPD). There was little or no interisolate variability in RAPD fragment patterns within each pathotype. Passage of an isolate of pathotype 3, originally from an Australian grasshopper (Praxibulas sp.), through a North American grasshopper resulted in no differences in the resultant RAPD fragment patterns. In the second method, polymorphic RAPD fragments were used to probe the genomic DNA from the three pathotypes, and pathotype-specific fragments were found. In the third method, restriction fragments from genomic DNA of the three pathotypes were cloned and screened for pathotype specificity. A genomic probe specific for each pathotype was isolated. These probes did not hybridize to DNA from Entomophaga aulicae or from grasshoppers. To facilitate the use of RAPD analysis and other molecular tools to identify pathotypes, a method for extracting DNA from resting spores from infected grasshoppers was developed. The DNA from the fractured resting spores was of sufficient integrity to be blotted and probed with the pathotype-specific DNA probes, thus validating the use of these probes for pathotype identification in field-collected grasshoppers.
Bibliography:	H H10 ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 ObjectType-Article-1 ObjectType-Feature-2
ISSN:	0099-2240 1098-5336
DOI:	10.1128/aem.61.2.556-560.1995