title: Real-time PCR assay based on topoisomerase-II gene for detection of Fusarium udum.
abstract: Fusarium wilt is an important soilborne disease of pigeonpea, caused by Fusarium udum. In this study, we have designed a real-time PCR assay for the detection of Fusarium udum from infected pigeonpea plants. Based on Topoisomerase-II gene sequence data from Fusarium udum and other related Fusarium species, a pair of primer was designed. The species-specific primers were tested in real-time PCR SYBR green assay. No increasing fluorescence signals exceeding the baseline threshold was observed with tested microbes, except Fusarium udum DNA. A single dissociation peak of increased fluorescence was obtained for the specific primers at melting temperature of 81.25°C. The real-time PCR showed a lowest detection of 0.1 pg genomic DNA. The assay was more sensitive, accurate and less time consuming for detection of Fusarium udum in infected plants root.
authors.affiliation: National Bureau of Agriculturally Important Microorganisms, Mau, UP, 27510, India. muki_yadav@rediffmail.com
authors.forename: Mukesh Kumar
authors.initials: MK
authors.lastname: Yadav
authors.forename: Bandavari Kishore
authors.initials: BK
authors.lastname: Babu
authors.forename: Anil Kumar
authors.initials: AK
authors.lastname: Saxena
authors.forename: Bhim Pratap
authors.initials: BP
authors.lastname: Singh
authors.forename: Kiran
authors.initials: K
authors.lastname: Singh
authors.forename: Dilip Kumar
authors.initials: DK
authors.lastname: Arora
mesh_terms: D036561:Cajanus / Q000382:microbiology*; D017931:DNA Primers / Q000235:genetics; D004250:DNA Topoisomerases, Type II / Q000235:genetics*; D005656:Fungal Proteins / Q000235:genetics*; D005670:Fusarium / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification*; D010935:Plant Diseases / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D017931:DNA Primers; D005656:Fungal Proteins; D004250:DNA Topoisomerases, Type II