Isolation of a species-specific mitochondrial DNA sequence for identification of Tilletia indica, the karnal bunt of wheat fungus
Loading...
Date
Authors
Ferreira, M. A. S. V.
Tooley, P. W.
Hatziloukas, E.
Castro, C.
Schaad, N. W.
Journal Title
Journal ISSN
Volume Title
Publisher
Abstract
Type
Type of the conference item
Journal type
peer reviewed
Educational material type
Conference Name
Journal name
Appl Environ Microbiol
Book name
Book series
Book edition
Alternative title / Subtitle
Description
Mitochondrial DNA (mtDNA) from five isolates of Tilletia indica was isolated and digested with several restriction enzymes. A 2.3-kb EcoRI fragment was chosen, cloned, and shown to hybridize with total DNA restricted with EcoRI from T, indica and not from a morphologically similar smut fungus, Tilletia barclayana. The clone was partially sequenced, and primers were designed and tested under high-stringency conditions in PCR assays. The primer pair Ti1/Ti4 amplified a 2,3-kb fragment from total DNA of 17 T. indica isolates from India, Pakistan, and Mexico, DNA from 25 isolates of other smut fungi (T. barclayana, Tilletia foetida, Tilletia caries, Tilletia fusca, and Tilletia controversa) did not produce any bands, as detected by ethidium bromide-stained agarose gels and Southern hybridizations. The sensitivity of the assay was determined and increased by using a single nested primer in a second round of amplification, so that 1 pg of total mycelial DNA could be detected, The results indicated that the primers which originated from a cloned mtDNA sequence can be used to differentiate T. indica from other Tilletia species and have the potential to identify teliospores contaminating wheat seeds.
Description
Keywords
polymerase chain-reaction, fragment-length-polymorphisms, gaeumannomyces-graminis, phytophthora, nuclear, disease, differentiation, hybridization, amplification, diversity
Subject classification
Citation
Link
<Go to ISI>://A1996TN22600014
Language
en
Publishing department/division
Advisor name
Examining committee
General Description / Additional Comments
Institution and School/Department of submitter
Πανεπιστήμιο Ιωαννίνων. Σχολή Επιστημών και Τεχνολογιών. Τμήμα Βιολογικών Εφαρμογών και Τεχνολογιών