Development of specific primers for genus Fusarium and F. solani using rDNA sub-unit and transcription elongation factor (TEF-1α) gene

Abstract

Detection of Fusarium solani causal agent of wilt and rots in many plant species are difficult if based only on morphological characteristics. Beside this, morphological discrimination requires special skill and the expertise of taxonomists or specialists. To simplify the detection and discrimination of F. solani from other Fusarium species, end-point polymerase chain reaction (PCR) assays were developed. Consensus sequences obtained from multiple alignments of target genes, internal transcribed spacer (ITS), rDNA and transcription elongation factor (TEF-1α), were used to design the primers for rapid detection of genus Fusarium (amplified product 420 and 466) and F. solani (amplified product 658, 595 and 485). BLASTn was used for in silico specificity. No cross reactivity was observed when primers were checked against the near-neighbor plant pathogens. The described primer sets allowed accurate identification and discrimination of genus Fusarium and F. solani. All tests have multiple applications including screening of infected plants, breeding programs and disease diagnosis.

Key words: Inter transcribed spacer (ITS), rDNA, transcription elongation factor, beta tubulin, Fusarium solani.