Isolation of a species-specific mitochondrial DNA sequence for identification of Tilletia indica, the karnal bunt of wheat fungus
Ferreira, M.A.S.V.
Isolation of a species-specific mitochondrial DNA sequence for identification of Tilletia indica, the karnal bunt of wheat fungus - 1996 - Printed
references US (DNAL 448.3 Ap5)
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
English
0099-2240
Acids
Basidiomycotina
Cell structure
Cytoplasm
Cytoplasmic organelles
Developmental stages
Fungi
Genetic engineering
Genomes
Gramineae
Molecular hybridization
Nucleic acids
Nucleic compounds
Organic acids
Pathogenesis
Plant developmental stages
Plant diseases
Plant genetics and breeding
Protoplasm
Tilletia
Ustilaginales
Triticum
96-097098
Isolation of a species-specific mitochondrial DNA sequence for identification of Tilletia indica, the karnal bunt of wheat fungus - 1996 - Printed
references US (DNAL 448.3 Ap5)
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
English
0099-2240
Acids
Basidiomycotina
Cell structure
Cytoplasm
Cytoplasmic organelles
Developmental stages
Fungi
Genetic engineering
Genomes
Gramineae
Molecular hybridization
Nucleic acids
Nucleic compounds
Organic acids
Pathogenesis
Plant developmental stages
Plant diseases
Plant genetics and breeding
Protoplasm
Tilletia
Ustilaginales
Triticum
96-097098