Genetic diversity and pathogenic variability among Cercospora beticola Sacc. isolates causing leaf spot of sugar beet in Iran

Authors
Seyed Bagher Mahmoudi 1
Maryam Saffarian Abbaszadeh 2
Saeed Abbasi 3
Reza Farrokhinejad 4
1 Sugar Beet Seed Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.
2 Clemson University, South Carolina, USA.
3 Department of Plant Protection, College of Agriculture, Razi University, Kermanshah, Iran.
4 Department of Plant Protection, College of Agriculture, Shahid Chamran Universty of Ahvaz, Ahvaz, Iran.
10.48311/jcp.2018.1386
Abstract
Cercospora leaf spot caused by Cercospora beticola has a great negative impact on yield and quality of sugar beet. In the present study, pathogenic and genotypic variation of 24 C. beticola isolates collected from different regions of Iran were studied using RFLP of the Internal Transcribed Spacer (ITS-RFLP), and Random Amplified Polymorphic DNA (RAPD-PCR). Pathogenic variability and genotype × isolate interaction were evaluated in greenhouse experiments on five sugar beet cultivars (FD0018, HM1836, Puma, Eudora and Monatuna). All of the 24 isolates tested were found to be pathogenic on the cultivars with significant variation in disease severity. Results of RAPD analysis showed wide DNA polymorphism among the Iranian C. beticola isolates. Restriction pattern of the internal transcribed spacer of rDNA (ITS1-5.8-ITS4) was studied using three restriction endonucleases: EcoR1, Taq1, and Busr1. The length of undigested DNA fragment of all isolates was estimated to be 500bp without rDNA polymorphism after digestion with EcoR1 (280, 270 bp), Taq1 (330 bp) and Busr1 (240, 220, 90 bp). RAPD and ITS-RFLP markers showed the highest level of genetic diversity which confirms the variation in C. beticola detection.
Keywords
Cercospora beticola
genotype × isolate interaction
Pathogenicity
RAPD fingerprinting

Subjects

Abbasi, S. and Mahmoudi, S. B. 2010. Reaction of sugar beet cultivars to Cercospora beticola isolates. Plant Protection, 33 (1): 69-77. Abbasi, S., Mesbah, M. and Mahmoudi, S. B. 2002. Optimization of field evaluation of resistance of sugar beet cultivars to cercospora leaf spot. Journal of Sugar Beet, 18: 181-193. Ali, A. A. M. M. 2012. Rapid detection and quantification of Cercospora beticola in soil using PCR and ELISA assays. Doctoral dissertation, Christian-Albrechts Universität Kiel. Almeida, A. M. R., Piuga, F. F., Marin, S. R. R., Binneck E., Sartori, F., Costamilan, L. M., Teixeir, A. and Lopes, M. 2005. Pathogenicity, molecular characterization, and cercosporin content of Brazilian isolates of Cercospora kikuchii. Fitopatologia Brasileira, 30: 594-602. Barres, B., Halkett, F., Dutech, C., Andrieux, A., Pinon, J. and Frey, P. 2008. Genetic structure of the poplar rust fungus Melampsora larici-populina: evidence for isolation by distance in Europe and recent founder effects overseas. Infection, Genetics and Evolution, 8: 577-587. Chiusa, G., Forgher, C., Giosue, S. and Rossi, V. 1996. Preliminary studies on genetic variability of Cercospora beticola Sacc. throughout the Mediterranean area. Proceedings of the 10th Congress of the Mediterranean Phytopathological ::union::, Montpellier. Cubeta, M. A., Vilgaiys, R. and Gonzalez, D. 1996. Molecular analysis of ribosomal RNA genes in Rhizoctonia fungi. In: Sneh, B., Jabaji-Hare, S., Neate, S. and Dijst, G. (Eds.), Rhizoctonia species: Taxonomy, Molecular Biology, Ecology, Pathology and Disease Control. Dordrecht: Kluwer Academic Publishers. pp. 81-86. Cubeta, M. A. and Vilgalys, R. 1997. Population biology of the Rhizoctonia solani complex. Phytopathology, 87: 480-484. Ershad, J. 1995. Fungi of Iran. Plant Pests and Diseases Research Institute, Tehran. Gouda, M. I. M. and El-Naggar, A. A. A. 2014. Efficacy of some fungicides on controlling cercospora leaf spot and their impact on sugar beet yield components. Journal of Plant Protection and Pathology-Mansoura University, 5 (1): 79-87. Groenewald, M., Linde, C. C., Groenewald, J. Z. and Crous, P.W. 2008. Indirect evidence for sexual reproduction in Cercospora beticola populations from sugar beet. Plant Pathology, 57: 25-32. Hemayati, S. S., Akbar, M. R. J. E., Ghaemi, A. R. and Fasahat, P. 2017. Efficiency of white mustard and oilseed radish trap plants against sugar beet cyst nematode. Applied Soil Ecology, 119: 192-196. Holtschulte, B. 2000. Cercospora beticola-worldwide distribution and incidence. In: Asher, M. I. C., Holstchulte, B., Richard Molard, M., Rosso, F., Steinrucken, G. and Beckers, R. (Eds.), Cercospora beticola Sacc. biology, agronomic influence & control measures in sugar beet. Brussels: International Institute for Beet Research. pp. 5-16.Karaoglanidis, G. S., Ioannidis, P. M. and Thanassoulopoulos, C. C. 2000. Reduced sensitivity of Cercospora beticola to sterol-demethylation-inhibiting fungicides. Plant Pathology, 49: 567-572. Karaoglanidis, G. S. and Ioannidis, P. M. 2010. Fungicide resistance of Cercospora beticola in Europe. In: Lartey, R. T., Weiland, J. J., Panella, L., Crous, P. W. and Windels C. E. (Eds.), Cercospora leaf spot of sugar Beet and related species. American Phytopathological Society: St. Paul. Khan, J., del Río, L. E., Nelson, R. and Khan, M. F. R. 2007. Improving the Cercospora leaf spot management model for sugar beet in Minnesota and North Dakota. Plant Disease, 91: 1105-1108. Khare, K. B., Utlwang, R., Khonga, E. B. and Segwagwe, A. T. 2017. Assessing the effectiveness of locally available fungicides in managing Cercospora leaf spot of Swiss chard under field conditions in Botswana. International Journal of Food, Agriculture and Veterinary Sciences, 1-8. Kilicoglu, M. C. and Ozkoc, I. 2010. Molecular characterization of Rhizoctonia solani AG4 using PCR-RFLP of the rDNA-ITS region. Turkish journal of Biology, 34 (3): 261-269. Koch, G. and Jung, C. 2000. Genetic localization of Cercospora resistance genes. In: Asher, M. I. C., Holstchulte, B., Richard Molard, M., Rosso, F., Steinrucken, G. and Beckers, R. (Eds.), Cercospora beticola Sacc. biology, agronomic influence & control measures in sugar beet. Brussels: International Institute for Beet Research. pp. 5-16. Madanian- Mohammadi, R., Minassian, V., Safaie, N., Mahmoudi, B. and Sharifi, H. 2004. Modeling of disease progress in Cercospora leaf spot of sugar beet. Iranian Journal of Plant Pathology, 40: 327-343. Mahmoudi, S. B., Mesbah, M., Rahimian, H., Alizadeh A. and Noruzi, P. 2005. Genetic diversity of sugar beet isolates of Rhizoctonia solani revealed by RAPD-PCR and ITS-rDNA analysis. Iranian Journal of Plant Pathology, 41: 523-542. Matsumoto, M., Furuya, N., Takanami, Y. and Matsuyama, N. 1996. RFLP analysis of the PCR amplified 28S rDNA in Rhizoctonia solani. Mycoscience, 37: 351-356. Meinhardt, L. W., Wulff, N. A., Bellato, C. M. and Tsai, S. M. 2002. Genetic analyses of Rhizoctonia solani isolates from Phaseolus vulgaris grown in the Atlantic Rainforest Region of São Paulo, Brazil. Fitopatologia brasileira, 27 (3): 259-267. Momeni, H., Mozafari, J., Falahati-Rastegar, M. and Jafarpour, B. 2005. Analysis of DNA polymorphism in anastomosis groups of Rhizoctonia solani in sugar beet fields of Khorasan province using RAPD markers. Iranian Journal of Plant Pathology, 41: 215-219. Moretti, M., Saracchi, M. and Farina, G. 2004. Morphological, physiological and genetic diversity within a small population of Cercospora beticola. Annals of Microbiology, 54: 129-150. Panizza, G. 1998. Indaginesulla diffusione mondiale di Cercospora beticola Sacc. Dissertation, Universita Cattolicadel Sacro Cuore. Piszczek, J., Pieczul, K. and Kiniec, A. 2017. First report of G143A strobilurin resistance in Cercospora beticola in sugar beet (Beta vulgaris) in Poland. Journal of Plant Diseases and Protection, 1-3. Ruppel, E. G. 1972. Variation among isolate of Cercospora beticola from sugar beet. Phytopathology, 62: 134-136. Sambroek, J., Fritsch, E. F. and T. Maniatis. 1989. Molecular cloning: A laboratory manual. New York: Cold spring harbor laboratory. Schneider, J. H. M., Salazar, O., Rubio, V. and Keijer, J. 1997. Identification of Rhizoctonia solani associated with field grown tulips using ITS rDNA polymorphism and pectic zymograms. European Journal of Plant Pathology, 103: 607-622. Secor, G. A., Rivera, V. V., Khan, M. F. R., and Gudmestad, N. C. 2010. Monitoring fungicide sensitivity of Cercospora beticola of sugar beet for disease management decisions. Plant Disease, 94: 1272-1282. Shane, W. W. and Teng, P. S. 1992. Impact of Cercospora leaf spot on root weight, sugar yield and purity of Beta vulgaris. Plant Disease, 76: 812-820. Shaykholeslami, M., Okhovvat, M., Hejaroude, Gh., Sharifi-Tehrani, A., Seidel, M. and Nikkhah. M. 2005. Study on genetic diversity of Iranian populations of Erysiphe betae causal agent of sugar beet powdery mildew using rDNA-RFLP method. Journal of Sugar Beet, 20: 149-161. Skaracis, G. N., Pavili, O. I. and Biancardi, E. 2010. Cercospora leaf spot disease of sugar beet. Sugar Tech, 12 (3-4): 220-228. Smith, G. A. and Martin, S. S. 1978. Differential response of sugar beet cultivars to Cercospora leaf spot disease. Crop Science, 18: 38-42. Smith, G. A. 1985. Response of sugar beet in Europe and the USA to Cercospora beticola infection. Agronomy Journal, 77: 126-129. Solel, Z. and Wahl, I. 1971. Pathogenic specialization of Cercospora beticola. Phytopathology, 61: 1081-1083. Trkulja, N. R., Milosavljević, A. G., Mitrović, M. S., Jović, J. B., Toševski, I. T., Khan, M. F. and Secor, G. A. 2017. Molecular and experimental evidence of multi-resistance of Cercospora beticola field populations to MBC, DMI and QoI fungicides. European Journal of Plant Pathology, 149 (4): 895-910. Vaghefi, N., Hay, F. S., Kikkert, J. R. and Pethybridge, S. J. 2016. Genotypic diversity and resistance to azoxystrobin of Cercospora beticola on processing table beet in New York. Plant Disease. 100 (7): 1466-1473. Vaghefi, N., Nelson, S. C., Kikkert, J. R. and Pethybridge, S. J. 2017. Genetic structure of Cercospora beticola populations on Beta vulgaris in New York and Hawaii. Scientific reports, 7 (1): 1726. Weiland, J. J. 2002. Rapid procedure for the extraction of DNA from fungal spores and mycelia. Fungal Genetics Newsletter, 44: 60-63. Whitney, E. D. and Lewellen, R. T. 1976. Identification and distribution of races C1 and C2 of Cercospora beticola from sugar beet. Phytopathology, 66: 1158-1160. 
Journal of Crop Protection
Volume 7, Issue 2
June 2018
Pages 207-217