Detection of Fusarium oxysporum F. sp. ciceris from chickpea with rt-pcr

Chickpea has an important place among food legumes and because of that Fusarium oxysporum f. sp. ciceris, causal agent of Fusarium wilt, is also possesses importance. Due to this and many other similar reasons, large losses occur annually and studies to prevent these losses are being continued. Aim of this study was quantitative determination of causal agent F. oxysporum f. sp. ciceris, in infected chickpeas via Real Time-Polymerase Chain Reaction (RT-PCR) using specific primers. At the same time, determination of the disease that is not observed in infected tissues on the first days, this method would elucidate the host-pathogen relationship. For the study carried out with quantitative polymerase chain reaction (qPCR) method, ILC482 chickpea cultivar susceptible to the disease, was infected with both disease forms, wilting and yellowing and then fresh infected plant samples were collected for further utilization in total RNA isolation. Total mRNA were converted to cDNAs and were assayed in RT-PCR studies. Assays were performed in SYBR Green Master Mix using specially designed primer sets. cDNA obtained from mycelial culture was used as a positive control in RT-PCR assay to create standard curves. The disease was observed with this method from the first day and the results were statistically significant for both sets of primers (p<0.05). The J5 primer set provided a better identification than H3. Taken together, these data suggest that ribosomal (J5) and histone (H3) gene regions have correlation with the disease and both regions were determined as a useful markers for identification of chickpea Fusarium wilt through by qRT-PCR studies.