Specific PCR-based detection of Alternaria helianthi: the cause of blight and leaf spot in sunflower

Udayashankar, A. C. and Chandra Nayaka, S. and Archana, B. and Anjana, G. and Niranjana, S. R. and Mortensen, C. N. and Lund, Ole S. and Prakash, H. S. (2012) Specific PCR-based detection of Alternaria helianthi: the cause of blight and leaf spot in sunflower. Archives of Microbiology, 194 (11). pp. 923-932. ISSN 1432-072X

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Abstract

Alternaria helianthi is an important seed-borne pathogenic fungus responsible for blight disease in sunflower. The current detection methods, which are based on culture and morphological identification, are time-consuming, laborious and are not always reliable. A PCR-based diagnostic method was developed with species-specific primers designed based on the sequence data of a region consisting of the 5.8S RNA gene and internal transcribed spacers---ITS 1 and ITS 2 of nuclear ribosomal RNA gene (rDNA) repeats of A. helianthi. The specificity of the primer pairs AhN1F and AhN1R designed was verified by PCR analysis of DNA from 18 Alternaria helianthi strains isolated from India, 14 non-target Alternaria spp. and 11 fungal isolates of other genera. A single amplification product of 357-bp was detected from DNA of A. helianthi isolates. No cross-reaction was observed with any of the other isolates tested. The detection limit of the PCR method was of 10 pg from template DNA. The primers could also detect the pathogen in infected sunflower seed. This species-specific PCR method provides a quick, simple, powerful and reliable alternative to conventional methods in the detection and identification of A. helianthi. This is the first report of an A. helianthi-specific primer set.

Item Type:	Article
Uncontrolled Keywords:	Alternaria blight, Seed-borne fungi, Seed health, Molecular diagnosis, PCR, Specific primers, ITS sequences
Subjects:	B Life Science > Biotechnology
Divisions:	Department of > Biotechnology
Depositing User:	C Swapna Library Assistant
Date Deposited:	15 Jul 2019 10:11
Last Modified:	15 Jul 2019 10:11
URI:	http://eprints.uni-mysore.ac.in/id/eprint/5202

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