Molecular identification of ascochyta blight of Cicer montbretii Jaub. & Spach

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.

Molecular identification of ascochyta blight of Cicer montbretii Jaub. & Spach

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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  • Abbo S, Redden RJ, Yadav SS (2007) Utilization of wild relatives. In: Yadav SS, Redden R, Chen W, Sharma B (Eds), Chickpea Breeding and Management. CABI International, Wallingford, UK, pp. 338-368.
  • Ahlawat IPS, Gangaiah B, Zahid MA (2007) Nutrient management in chickpea. In: Yadav SS, Chen W (Eds), Chickpea Breeding and Management. Trowbridge, UK, pp. 213-232.
  • Akamatsu HO, Chilvers MI, Kaiser WJ, Peever TL (2012) Karyotype polymorphism and chromosomal rearrangement in populations of the phytopathogenic fungus, Ascochyta rabiei. Fungal Biology 116(11): 1119-1133.
  • Bahr L, Castelli MV, Barolo MI, Mostacero NR, Tosello ME, López SN (2016) Ascochyta blight: Isolation, characterization, and development of a rapid method to detect inhibitors of the chickpea fungal pathogen Ascochyta rabiei. Fungal Biology 120(3): 424-432.
  • Baite MS, Dubey SC, Upadhyay BK (2017) Genetic diversity of Ascochyta rabiei causing blight of chickpea in India. Research Journal of Biotechnology 12: 29-37.
  • Barve MP, Arie T, Salimath SS, Muehlbauer FJ, Peever TL (2003) Cloning and characterization of the mating type (MAT) locus from Ascochyta rabiei (teleomorph: Didymella rabiei) and a MAT phylogeny of legume-associated Ascochyta spp. Fungal Genetics and Biology 39: 151-67.
  • Bayraktar H, Dolar FS, Tör M (2007) Determination of genetic diversity within Ascochyta rabiei (Pass.) Labr., the cause of ascochyta blight of chickpea in Turkey. Journal of Plant Pathology 89: 341-347.
  • Chen W, Coyne CJ, Peever TL, J. Muehlbauer F (2004) Characterization of chickpea differentials for pathogenicity assay of ascochyta blight and identification of chickpea accessions resistant to Didymella rabiei. Plant Pathology 53(6): 759-769.
  • Cho S, Chen W, Muehlbauer FJ (2005) Constitutive expression of the Flavanone 3-hydroxylase gene related to pathotype-specific ascochyta blight resistance in Cicer arietinum L. Physiological and Molecular Plant Pathology 67(2): 100-107.
  • Donmez A (2011) Cicer uludereensis Donmez: A new species of Cicer (Chickpea) (Fabaceae) from around the Fertile Crescent, SE Turkey. Turkish Journal of Botany 35: 71-76.
  • Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12: 13-15.
  • Eker T, Sari D, Sari H, Tosun HS, Toker CA (2022) Kabuli chickpea ideotype. Scientific Reports 12(1): 1-17.
  • FAOSTAT (2023) http://faostat.fao.org/site/567/default.aspx#ancor Accessed 29 June, 2023.
  • Frenkel O, Shtienberg D, Abbo S, Sherman A (2007) Sympatric ascochyta complex of wild Cicer judaicum and domesticated chickpea. Plant Pathology 56(3): 464-471.
  • Frenkel O, Peever TL, Chilvers MI, Ozkilinc H, Can C, Abbo S, Sherman A (2010) Ecological genetic divergence of the fungal pathogen Didymella rabiei on sympatric wild and domesticated Cicer spp. (Chickpea). Applied and Environmental Microbiology 76(1): 30-39.
  • Guler U (2018) Cicer anatolicum Alef. için yanıklık hastalığının [Ascochyta rabiei (Pass.) labr.] moleküler tekniklerle tanısı. Yüksek Lisans Tezi, Akdeniz Üniversitesi Fen Bilimleri Enstitüsü, Antalya.
  • Guner N, Kaygusuz R, Ventura G (2012) Taxation and household labour supply. The Review of Economic Studies 79(3): 1113-1149.
  • Hawtin GC, Singh, KB (1984) Prospects and potential of winter sowing of chickpea in the Mediterranean region. In: Saxena MC, Singh KB (Eds), Ascochyta blight and winter sowing of chickpeas. Netherlands, pp. 7-16.
  • Imtiaz M, Abang MM, Malhotra RS, Ahmed S, Bayaa B et al. (2011) Pathotype IV, a new and highly virulent pathotype of Didymella rabiei, causing Ascochyta blight in chickpea in Syria. Plant Disease 95(9): 1192-1192.
  • Islam W, Qasim M, Noman A, Idrees A, Wang L (2017) Genetic resistance in chickpea against Ascochyta blight: historical efforts and recent accomplishments. Journal of Animal and Plant Sciences 27(6): 1941-1957.
  • Jukanti AK, Gaur PM, Gowda CLL, Chibbar RN (2012) Nutritional quality and health benefits of chickpea (Cicer arietinum L.): A review. British Journal of Nutrition 108: 11-26.
  • Kafadar FN, Özkan A, Can C, Kar Y, Mart D, Ceyhan E (2019) Genetic and biochemical properties of Cicer spp. reveal distinction between wild and cultivated chickpea genotypes. Legume Research-An International Journal 42(1): 1-9.
  • Kaiser WJ, Muehlbauer FJ (1988) An outbreak of ascochyta blight of chickpea in the Pacific Northwest USA in 1987, International Chickpea Newsletter 18: 16­17.
  • Kaiser WJ, Hannan RM, Muehlbauer FJ, Mihov M (1998) First report of Ascochyta blight of Cicer montbretii, a wild perennial chickpea in Bulgaria. Plant Disease 82(7): 830-830.
  • Kantar F, Hafeez FY, Shivakumar BG, Sundaram SP, Tejera NA, Aslam A, Bano A, Raja P (2007) Chickpea: Rhizobium management and nitrogen fixation. In: Yadav SS, Chen W (Eds), Chickpea Breeding and Management. Trowbridge, UK, pp. 179-192.
  • Khaliq I, Moore K, Sparks AH (2021) The relationship between natural rain intensity and Ascochyta blight in chickpea development. AgriRxiv.
  • Kumar G, Bajpai R, Rashid MM (2020) Management of Blight in Chickpea Caused by Ascochyta rabiei: A Review. Bioscience Biotechnology Research Communications 13: 60-64.
  • Misra JP, Rao GM, Kumar A, Yadav A, Kumar S, Yadav R, Kumar R, Srivastava S (2016) Molecular assisted breeding for Ascochyta Blight resistance in chickpea (Cicer arietinum L.)-A review. Journal of Pure and Applied Microbiology 10(2): 1469-1476.
  • Muehlbauer FJ, Kaiser WJ (1994) Using host plant resistance to manage biotic stresses in cool season food legumes. In: Muehlbauer FJ, Kaiser WJ (Eds), Expanding the production and use of cool season food legumes. Springer, Dordrecht, pp. 233-246.
  • Nalcaci N, Turan A, Basbuga S, Kafadar FN, Isler Ceyhan D, Anay A, Sarpkaya, K (2021) Virulence and mating type distribution of Didymella rabiei in chickpea growing areas of Turkey. Journal of Agricultural Science and Technology 23(1): 209-220.
  • Nene YL, Sheila VK, Sharma SB (1996) A world list of chickpea and pigeonpea pathogens. 5th ed. ICRISAT, Patancheru, India, pp. 27.
  • Ozturk M, Duran A, Hakkı EE (2011) Cicer floribundum var. amanicola Fabaceae, a new variety from south Anatolia, Turkey. Biyolojik Çeşitlilik ve Koruma 4(3): 44-51.
  • Ozturk M, Duran A, Hakki EE (2013) Cladistic and phylogenetic analyses of the genus Cicer in Turkey. Plant Systematics and Evolution 299(10): 1955-1966.
  • Pande S, Sharma M, Gaur PM, Gowda CLL (2010) Host Plant Resistance to Ascochyta Blight of Chickpea. Information Bulletin No. 82. International Crops Research Institute for the Semi-Arid Tropics.
  • Peever TL, Barve MP, Stone LJ, Kaiser WJ (2007) Evolutionary relationships among Ascochyta species infecting wild and cultivated hosts in the legume tribes Cicereae and Vicieae. Mycologia 99(1): 59-77.
  • Phan HTT, Ford R, Bretag T, Taylor PWJ (2002) A rapid and sensitive polymerase chain reaction (PCR) assay for detection of Ascochyta rabiei, the cause of ascochyta blight of chickpea. Australasian Plant Pathology 31(1): 31-39.
  • Savolainen V, Cowan RS, Vogler AP, Roderick GK, Lane R (2005) Towards writing the encyclopaedia of life: an introduction to DNA barcoding. Philosophical Transactions of the Royal Society B: Biological Sciences 360(1462): 1805-1811.
  • Shahid AA, Husnain T, Riazuddin S (2008) Ascochyta blight of chickpea: Production of phytotoxins and disease management. Biotechnology Advances 26(6): 511-515.
  • Sharma M, Babu TK, Gaur PM, Ghosh R, Rameshwar T, Chaudhary RG, Pande S (2012) Identification and multi-environment validation of resistance to Fusarium oxysporum f. sp. ciceris in chickpea. Field Crops Research 135: 82-88.
  • Singh G, Singh K, Kapoor S (1982) Screening for sources of resistance to ascochyta blight of chickpea. International Chickpea Newsletter 6: 15-17.
  • Singh KB, Saxena MC (1993) Breeding for stress tolerance in cool-season food legumes (No. 635.65 S52). Chichester, UK: Wiley.
  • Singh G, Chen W, Rubiales D, Moore K, Sharma YR, Gan Y (2007) Diseases and their management. In: Yadav SS, Redden R, Chen W, Sharma B (Eds), Chickpea Breeding and Management. CAB International, UK, pp. 497-519.
  • Tekin M, Sari D, Catal M, Ikten C, Smykal P, Penmetsa RV, Toker C (2018) Eco-geographic distribution of Cicer isauricum PH Davis and threats to the species. Genetic Resources and Crop Evolution 65(1): 67-77.
  • Toker C, Berger J, Kahraman A, Aydogan A, Can C, Bukun B, Cook DR (2014) Cicer reticulatum Ladizinsky, progenitor of the cultivated chickpea (C. arietinum L.). Legume Perspect 5: 26-27.
  • Toker C, Berger J, Eker T, Sari D, Sari H, Gokturk RS, Von Wettberg EJ (2021) Cicer turcicum: A new Cicer species and its potential to improve chickpea. Frontiers in Plant Science 12: 662891.
  • Turkkan M (2008) Türkiye’deki Ascochyta rabiei (Pass.) Patotiplerinin Ürettiği Solanapyrone Toksinlerinin Belirlenmesi. Doktora Tezi, Ankara Üniversitesi Fen Bilimleri Enstitüsü, Ankara.
  • Udupa SM, Weigand F, Saxena MC, Kahl G (1998) Genotyping with RAPD and microsatellite markers resolves pathotype diversity in the ascochyta blight pathogen of chickpea. Theoretical and Applied Genetics 97(1): 299-307.
  • van der Maesen LJG (1972) Cicer L. A Monograph of the Genus, with Special Reference to the Chickpea (Cicer arietinum L.). Its Ecology and Cultivation: Meded. Landbouwhogesch. Wageningen, pp. 342.
  • van der Maesen LJG (1987) Origin, history and taxonomy of chickpea. In: Saxena, MC, Singh KB. (Eds), The Chickpea. CABI International, Wallingford, UK, pp. 11-34.
  • van der Maesen LJG, Maxted N, Javadi F, Coles S, Davies AMR (2007) Taxonomy of the Genus Cicer Revisited. In: Yadav SS, Redden R, Chen W, Sharma B (Eds), Chickpea Breeding and Management. CABI International, Wallingford, UK, pp.14-46.
  • Verma MM, Singh G, Sandhu TS, Brar HS, Singh K and Bhullar BS (1981) Sources of resistance to gram blight and mold. International Chickpea Newsletter 4: 14-16.
  • von Wettberg EJ, Chang PL, Başdemir F, Carrasquila-Garcia N, Korbu LB, Moenga SM, ... & Cook DR (2018) Ecology and genomics of an important crop wild relative as a prelude to agricultural innovation. Nature Communications 9(1): 1-13.
  • Walter DG (2009) Studies on the disinfection of chickpea seeds (Cicer arietinum) infected with Ascochyta rabiei (Pass.) Labr. (in France) The Annals of Benaki Phytopathological Institute 5: 76-87.
  • White TJ, Bruns T, Lee SJWT, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. PCR protocols: A Guide to Methods and Applications 18(1): 315-322.