Molecular identification of ascochyta blight of Cicer montbretii Jaub. & Spach
Fevzi BULAT, Duygu Sarı, Hatice Sari, Tuba Eker, Hilal ÖZAY, Cengiz Toker
Abstract
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Fevzi BULAT, Duygu Sarı, Hatice Sari, Tuba Eker, Hilal ÖZAY, Cengiz Toker
Abstract
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Molecular detection of ascochyta blight caused by Ascochyta rabiei (Pass.) Labr. is important for effective control of the ascochyta blight and efficient chickpea (Cicer arietinum L.) breeding program. The present research was therefore aimed to diagnose ascochyta blight of C. montbretii Jaub. & Spach via molecular techniques. Infected plant samples were collected and placed on potato dextrose agar (PDA) medium for 1 week at 20-24℃, and colonies with typical ascochyta blight symptoms were transferred to new PDA medium and incubated for 1 week at 25℃. DNA was isolated from small parts of fungus isolates via the CTAB method. Internal transcribed spacer (ITS) regions (ITS-1, 5.8S rDNA subunit, ITS-2) were amplified with ITS 5 and ITS 4 primers for molecular characterization. Based on the BLAST analysis, the sequence had 99 and 100% nucleotide identity with the corresponding sequence of A. rabiei in GeneBank. To our knowledge, this is the first report of ascochyta blight of C. montbretii in Turkiye. The pathogen is considered to be co-evolved with C. montbretii. Molecular techniques, as in the present study, can be diagnosed with great accuracy, in a short time, and with relatively little effort and expense.
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Molecular detection of ascochyta blight caused by Ascochyta rabiei (Pass.) Labr. is important for effective control of the ascochyta blight and efficient chickpea (Cicer arietinum L.) breeding program. The present research was therefore aimed to diagnose ascochyta blight of C. montbretii Jaub. & Spach via molecular techniques. Infected plant samples were collected and placed on potato dextrose agar (PDA) medium for 1 week at 20-24℃, and colonies with typical ascochyta blight symptoms were transferred to new PDA medium and incubated for 1 week at 25℃. DNA was isolated from small parts of fungus isolates via the CTAB method. Internal transcribed spacer (ITS) regions (ITS-1, 5.8S rDNA subunit, ITS-2) were amplified with ITS 5 and ITS 4 primers for molecular characterization. Based on the BLAST analysis, the sequence had 99 and 100% nucleotide identity with the corresponding sequence of A. rabiei in GeneBank. To our knowledge, this is the first report of ascochyta blight of C. montbretii in Turkiye. The pathogen is considered to be co-evolved with C. montbretii. Molecular techniques, as in the present study, can be diagnosed with great accuracy, in a short time, and with relatively little effort and expense.
Key concepts: Ascochyta, Blight, Biology, Potato dextrose agar, Botany, Fungus, Horticulture, Agar