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ITS-RFLP fingerprinting and molecular marker for detection of Fusarium oxysporum f.sp. ciceris
S.C. Dubey, A. Tripathi, S.R. Singh
Language English Country Czech Republic
- MeSH
- Cicer microbiology MeSH
- DNA Fingerprinting methods MeSH
- DNA, Fungal genetics chemistry MeSH
- DNA Primers genetics MeSH
- Financing, Organized MeSH
- Fusarium genetics isolation & purification classification MeSH
- Genetic Variation MeSH
- DNA, Ribosomal Spacer genetics chemistry MeSH
- Molecular Sequence Data MeSH
- Mycology methods MeSH
- Plant Diseases microbiology MeSH
- Polymorphism, Restriction Fragment Length MeSH
- DNA Restriction Enzymes metabolism MeSH
- Sequence Analysis, DNA MeSH
- Sensitivity and Specificity MeSH
- Geographicals
- India MeSH
Genetic diversity of 11 representative isolates of Fusarium oxysporum f.sp. ciceris causing chickpea wilt was determined through internal transcribed spacer (ITS) region of the ribosomal DNA-restriction fragment length polymorphism (ITS-RFLP). ITS1+5.8s+ITS2 regions of the isolates were amplified with a set of primers ITS1 and ITS4 and amplified products were digested with 4 restriction enzymes (AluI, MboI, RsaI, MseI). Six different kinds of ITS-RFLP patterns were obtained. The ITS region of these isolates was sequenced and deposited to NCBI GeneBank. The nucleotide sequence homology of ITS region grouped the isolates into 5 categories. Primers were designed with sequence information using Primer 3 software. F. oxysporum f.sp. ciceris specific markers (FOC F2 and FOC R2) based on ITS region were developed for the first time for detection of the pathogen. The markers produced an amplicon of 292 bp; they were validated against the isolates of the pathogen collected from different locations of India.
Literatura
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- $a Genetic diversity of 11 representative isolates of Fusarium oxysporum f.sp. ciceris causing chickpea wilt was determined through internal transcribed spacer (ITS) region of the ribosomal DNA-restriction fragment length polymorphism (ITS-RFLP). ITS1+5.8s+ITS2 regions of the isolates were amplified with a set of primers ITS1 and ITS4 and amplified products were digested with 4 restriction enzymes (AluI, MboI, RsaI, MseI). Six different kinds of ITS-RFLP patterns were obtained. The ITS region of these isolates was sequenced and deposited to NCBI GeneBank. The nucleotide sequence homology of ITS region grouped the isolates into 5 categories. Primers were designed with sequence information using Primer 3 software. F. oxysporum f.sp. ciceris specific markers (FOC F2 and FOC R2) based on ITS region were developed for the first time for detection of the pathogen. The markers produced an amplicon of 292 bp; they were validated against the isolates of the pathogen collected from different locations of India.
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