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Sensitivity of Alternaria spp. Isolates to Some Fungicides Used on Sultani Çekirdeksiz Grapes in Aegean Region Vineyards

Year 2022, Volume: 4 Issue: 2, 23 - 35, 30.05.2022

Abstract

This study aimed to identify Alternaria species morphologically and molecularly, that is one of the important pathogens causing cluster rot, and determine their reactions to some fungicides in in vitro. The fungal isolates obtained from leaves, flowers, stems, and shoots in almost every development period of the Sultani Çekirdeksiz vineyards in 2009 and 2010 from Manisa province in the Aegean region were identified as Alternaria alternata (Fr.) Keissler according to morphological and molecular methods. In molecular studies, the ITS (Internal Transcribed Spacer) gene region was amplified by real-time PCR and the nucleotide sequences of 570 bp long isolates were stored in the NCBI GenBank database. ITS sequences were aligned using the Mega X software, and a phylogenetic tree was constructed. Sensitivity levels of isolates to cyprodinil+fludioxanil, mancozeb, pyrimethanil, kresoxim-methyl+boscalid, azoxystrobin, and basic copper sulfate were tested under laboratory conditions. In vitro antifungal effect of different concentrations (0.01-100 µg ml-1 and 3-3000 µg ml-1) of six fungicides against 24 A. alternata isolates were evaluated. Cyprodinil+fludioxanil was recorded as the most effective fungicide that, according to ED50 value, 23 isolates were in the range of 0.01-0.1 µg ml-1. BAC 1/2 and GMK 2/1 isolates were found to have the highest virulence with values of 100% and 86.67%, respectively.

Thanks

This study is the Ege University Graduate School of Natural and Applied Sciences PhD Thesis Project. The authors would like to thank Ege University, Faculty of Agriculture, Department of Plant Protection, and TOPAR laboratory.

References

  • Abbott, W.S. 1925. A method of computing the effectiveness of an insecticide. J. Econ. Entomol. 18: 265-267.
  • Basım, E., Basım, H., Abdulai, M., Baki, D., Oztürk, N., 2017. Identification and characterization of Alternaria alternata causing leaf spot of olive tree (Olea europaea) in Turkey. Crop Protection 92: 79-88.
  • Baroffio, C.A., Siegfried, W., Hilber, U.W., 2003. Long-term monitoring for resistance of Botryotinia fuckeliana to anilinopyrimidine, phenylpyrrole and hydroxyanilide fungicides in Switzerland. Plant Disease 87: 662-666.
  • Barnett, H.L., Hunter, B.B., 1998. Illustrated Genera of Imperfect Fungi. American Phytopathological Society, 4ed., p. 240.
  • Cenis, J.L., 1992. Rapid extraction of fungal DNA for PCR amplification. Nucleic Acid Research 20(9): 2380.
  • Delen, N., Yıldız, M., Maraite, H., 1984. Benzimidazole and Dithiocarbamate resistance of Botrytis cinerea on greenhouse crops in Turkey. Mededelingen in Viticulture Ed Enologia Universita Torino 9: 278-279.
  • Delen, N., Tosun, N., Yılmaz, O., Yıldız, Z., 2000. Variation in the sensitivities of Botrytis cinerea isolates to some fungicides with non-specific mode of action. XIIth International Botrytis Symposium. July 3-7, p: 64, France.
  • Delen, N., 2001. Studies on bunch rots of fungal in the vineyards. Turkey IX. Phytopathology Congress, pp: 347-353, Tekirdağ.
  • Delen, N., 2008. Fungicides. Nobel press. Ankara, Turkey. pp. 318.
  • Ellis, M.B., 1993. Dematiaceous Hyphomycetes. Cab Intern. p. 608.
  • Erdoğan, O., Çelik, A., Yıldız, Ş., Kökten, K., 2014. Antifungal Effect of Some Plant Extracts and Essential Oils Against Root Rot Factors of Seedlings in Cotton. Turkish Journal of Agricultural and Natural Sciences 1(3): 398-404.
  • Erdoğan, O., Celik, A., Zeybek, A., 2016. In Vitro Antifungal Activity of Mint, Thyme, Lavender Extracts and Essential Oils on Verticillium dahliae Kleb. Fresenius Environmental Bulletin. 25(11):4856-4862.
  • FAO, 2020. Food and Agriculture Organization of the United Nations. FAST. Available from: http://faostat.fao.org/site/291/default.aspx. (Accessed: 28 June 2020).
  • Farra, J. J., Pryor, B.M., Davis, R.M., 2004. Alternaria diseases of carrot. Plant disease 88(8): 776-784.
  • Forster, B., Staub, T., 1996. Basis of use strategies of anilinopyrimidine and phenylpyrrole fungicides against Botrytis cinerea. Crop. Prot., 15: 529-537.
  • Henry, T., Iwen, P.C., Hinrichs, S.H., 2000. Identification of Aspergillus species using internal transcribed spacer regions 1 and 2. Journal of Clin. Microbiol. 38(4):1510-1515.
  • Izumi, Y., Kamei, E., Miyamoto, Y., Ohtani, K., Masunaka, A., Fukumoto, T., 2012. Role of the pathotype-specific ACRTS1 gene encoding a hydroxylase involved in the biosynthesis of host-selective ACR-toxin in the rough lemon pathotype of Alternaria alternata. Phytopathology 102: 741–748.
  • Kakalíková, L., Jankura, E., Šrobárová, A., 2009. First report of Alternaria bunch rot of grapevines in Slovakia. Australasian Plant Disease Notes 4: 68–69.
  • Koç, İ., Yardım, E.N., Çelik, A., Mendeş, M., Mirtagioğlu, H., Namlı, A., 2018. Determination of Antifungal Effect of Wood Vinegar Obtained from Hazelnut Shells Against Mold Factors in In-Vitro Conditions. The Science Journal of Bitlis Eren University 7(2): 296-300.
  • Koç, İ., Çelik, A., Durak, E. D., Demir, İ., Bayman, S., Mirtagioğlu, H., & Demir, S. 2020. Morphological and Molecular Characterization of Walnut Anthracnose (Ophiognomonia leptostyla) Disease in Walnut Growing Areas of Bitlis Province. Journal of the Institute of Science and Technology 10(1): 28-38.
  • Koplay, C., 2004. Studies on determination of fungal pathogens causing rots on Sultanina table grapes and their control with fungicides in vitro conditions. İzmir. Turkey. University of Ege. MS thesis. pp. 105.
  • Leroux, P., Fritz, R., Debieu, D., Albertini, C., Lanen, C., Bach, J., Gredit, M., Chapeland, F., 2002. Mechanisms of resistance to fungicides in field strains of Botrytis cinerea. Pest. Man. Sci. 58:876-888.
  • Lorbeer, J.W. and Vincelli, P.C., 1990. Efficacy of dicarboximide fungicides and fungicide combinations for control of Botrytis leaf blight of onion in New York. Plant Diseases 74:235-237.
  • Lorenz, H.D., Eichhorn, K.W., 1978. Untersuchungen zur möglichen Resistenzbildung von Botrytis cinerea an Reben gegen die Wirkstoffe Vinclozolin und Iprodion. Die Wein-Wissenschatt. 33:2-10.
  • Lu, Y., Ma, J., Sutton, T.B., Ypema, H., 2004. Comparision of assay methods for evaluating the sensitivity of Alternaria mali to boscalid. Phytopathology p. 94.
  • Mostert, L., Crous, P.W., Petrini, O., 2000. Endophytic fungi associated with shots and 19 leaves of Vitis vinifera, with specific reference to the Phomopsis viticola complex. Sydowia 54: 46-58.
  • Pary, K.E., Wood, R.K.S., 1958. The adaptation of fungi to fungicides: adaptation to captan Ann. Appl. Biol. 46: 446-456.
  • Pashley, C.H., Fairs, A., Free, R.C., Wardlaw, A.J., 2012. DNA analysis of outdoor air reveals a high degree of fungal diversity, temporal variability, and genera not seen by spore morphology. Fungal Biol. 116: 214-224.
  • Peever, T.L., Ibañez, A., Akimitsu, K., Timmer, L.W., 2002. Worldwide phylogeography of the citrus brown spot pathogen. Alternaria alternata. Phytopathology 92: 794-802. doi:10.1094/PHYTO.2002.92.7.794
  • Pitt, J.I., Hockıng, A.D., 1997. Fungi and Food Spoilage 2nd ed. London: Blackie Academic & Professional. p. 593 ISBN 0-8342-1306-0.
  • Polizzotto, R., Andresen, B., Martini, M., Grisan, S., Assante, G., Musetti, R., 2012. A polyphasic approach for the characterization of endophytic Alternaria strains isolated from grapevines. Journal of Microbiological Methods 88:162-171. http://dx.doi.org/10.1016/j.mimet.2011.11.009.
  • Prusky, D., Fuchs, Y., Yanko, U., 1983. Assessment of latent infections as a basis for control of post-harvest disease of mango. Plant Diseases 67: 816-818.
  • Ranganath, C., Yonelinas, A. P., Cohen, M. X., Dy, C. J., Tom, S.M., D’Esposito, M.D., 2003. Dissociable correlates of recollection and familiarity with the medial temporal lobes. Neuropsychologia 42: 2-13.
  • Rotem, J., 1994. The genus Alternaria: biology, epidemiology and pathogenicity. St. Paul, MN: The American Phytopathological Society p. 326.
  • Rotondo, F., Collina, M., Brunelli, A., and Pryor, B.M, 2012. Compari-son of Alternaria spp. collected in Italy from apple with A. mali and other AM-toxin producing strains. Phytopathology 102:1130-1142.
  • Saitou, N., Nei, M., 1987. The neighbor-joining method: A new method for reconstructing phylogenetic trees. Molecular Biology and Evolution 4: 406-425.
  • Serra, R., Braga, A., Venâncio, A., 2005. Mycotoxin producing and other fungi isolated from grapes for wine production, with particular emphasis on ochratoxin A. Research in Microbiology 156(4): 515-521.
  • Sharma, M., Ghosh, R., Pande, S., 2013. Occurrence of Alternaria alternata causing Alternaria blight in pigeon pea in India. Advances in Bioscience and Biotechnology 4: 702-705.
  • Simmons, E.G., 1992. Alternaria themes and variations. Mycotaxon 48(73): 109-140.
  • Simmons, E.G., 2007. Alternaria: an identification manual, CBS Biodiversity series no. 6. Utrecht, The Netherlands, p: 775.
  • Swart, A.E., Lennox, C.L., Holz, G., 1995. Infection of table grape bunches by Alternaria alternata. South African Journal for Enology and Viticulture 16: 3–6.
  • White, T.J., Bruns, T., S-Lee, S., Taylor, J.W., 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In PCR Protocols: A Guide to Methods and Applications. Innis MA, Gel-fand DH, Sninsky JJ, White TJ, eds. Academic Press, Inc., New York, pp: 315–322.
  • Zahavi, T., Cohen, L., Weiss, B., Schena, L., Daus, A., Kaplunov, T., Jutkhi, J., Ben-Arie, R., Droby, S. 2000. Biological control of Botrytis. Aspergillus and Rhizopus rots on table and wine grapes in Israel. Postharvest Biology and Technology. 20: 115-124.

Ege Bölgesi Bağlarında Sultaniye Çekirdeksiz Üzüm Çeşidinde Kullanılan Bazı Fungisitlere Alternaria spp'i İzolatlarının duyarlılığı

Year 2022, Volume: 4 Issue: 2, 23 - 35, 30.05.2022

Abstract

Bu çalışmada, salkım çürüklüğüne neden olan patojenlerden biri olan Alternaria türlerinin morfolojik ve moleküler olarak tanımlanması ve in vitro olarak bazı fungisitlere karşı etkilerinin belirlenmesi amaçlanmıştır. Ege bölgesinde Manisa ilindeki, Sultani Çekirdeksiz bağlarının 2009 ve 2010 yıllarında hemen her gelişme döneminde yaprak, çiçek, gövde ve sürgünlerinden elde edilen izolatlar morfolojik ve moleküler yöntemlere göre Alternaria alternata (Fr.) Keissler olarak tanımlanmıştır. Moleküler tanıda ITS (Internal Transcribed Spacer) gen bölgesi real-time PCR ile amplifiye edilmiş ve 570 bp uzunluğunda nükleotid dizisi elde edilerek, NCBI Gen bankasında kayıt altına alınmıştır. ITS dizileri Mega X yazılımı kullanılarak hizalanmış ve bir filogenetik ağaç çizilmiştir. İzolatların cyprodinil+fludioxanil, mancozeb, pyrimethanil, kresoxim-methyl+boscalid, azoxystrobin ve temel bakır sülfata karşı duyarlılık düzeyleri laboratuvar koşullarında test edilmiştir. Altı fungisitin farklı konsantrasyonlarda (0.01-100 µg ml-1 ve 3-3000 µg ml-1) 24 A. alternata izolatına karşı in vitro antifungal etkisi değerlendirilmiştir. EC50 değerine göre 23 izolatın 0.01-0.1 µg ml-1 aralığında olması ile cyprodinil+fludioxanil en etkili fungisit olarak kaydedilmiştir. BAC 1/2 ve GMK 2/1 izolatları sırasıyla %100 ve %86,67 değerleri ile en yüksek virülans değerine sahip bulunmuştur.

References

  • Abbott, W.S. 1925. A method of computing the effectiveness of an insecticide. J. Econ. Entomol. 18: 265-267.
  • Basım, E., Basım, H., Abdulai, M., Baki, D., Oztürk, N., 2017. Identification and characterization of Alternaria alternata causing leaf spot of olive tree (Olea europaea) in Turkey. Crop Protection 92: 79-88.
  • Baroffio, C.A., Siegfried, W., Hilber, U.W., 2003. Long-term monitoring for resistance of Botryotinia fuckeliana to anilinopyrimidine, phenylpyrrole and hydroxyanilide fungicides in Switzerland. Plant Disease 87: 662-666.
  • Barnett, H.L., Hunter, B.B., 1998. Illustrated Genera of Imperfect Fungi. American Phytopathological Society, 4ed., p. 240.
  • Cenis, J.L., 1992. Rapid extraction of fungal DNA for PCR amplification. Nucleic Acid Research 20(9): 2380.
  • Delen, N., Yıldız, M., Maraite, H., 1984. Benzimidazole and Dithiocarbamate resistance of Botrytis cinerea on greenhouse crops in Turkey. Mededelingen in Viticulture Ed Enologia Universita Torino 9: 278-279.
  • Delen, N., Tosun, N., Yılmaz, O., Yıldız, Z., 2000. Variation in the sensitivities of Botrytis cinerea isolates to some fungicides with non-specific mode of action. XIIth International Botrytis Symposium. July 3-7, p: 64, France.
  • Delen, N., 2001. Studies on bunch rots of fungal in the vineyards. Turkey IX. Phytopathology Congress, pp: 347-353, Tekirdağ.
  • Delen, N., 2008. Fungicides. Nobel press. Ankara, Turkey. pp. 318.
  • Ellis, M.B., 1993. Dematiaceous Hyphomycetes. Cab Intern. p. 608.
  • Erdoğan, O., Çelik, A., Yıldız, Ş., Kökten, K., 2014. Antifungal Effect of Some Plant Extracts and Essential Oils Against Root Rot Factors of Seedlings in Cotton. Turkish Journal of Agricultural and Natural Sciences 1(3): 398-404.
  • Erdoğan, O., Celik, A., Zeybek, A., 2016. In Vitro Antifungal Activity of Mint, Thyme, Lavender Extracts and Essential Oils on Verticillium dahliae Kleb. Fresenius Environmental Bulletin. 25(11):4856-4862.
  • FAO, 2020. Food and Agriculture Organization of the United Nations. FAST. Available from: http://faostat.fao.org/site/291/default.aspx. (Accessed: 28 June 2020).
  • Farra, J. J., Pryor, B.M., Davis, R.M., 2004. Alternaria diseases of carrot. Plant disease 88(8): 776-784.
  • Forster, B., Staub, T., 1996. Basis of use strategies of anilinopyrimidine and phenylpyrrole fungicides against Botrytis cinerea. Crop. Prot., 15: 529-537.
  • Henry, T., Iwen, P.C., Hinrichs, S.H., 2000. Identification of Aspergillus species using internal transcribed spacer regions 1 and 2. Journal of Clin. Microbiol. 38(4):1510-1515.
  • Izumi, Y., Kamei, E., Miyamoto, Y., Ohtani, K., Masunaka, A., Fukumoto, T., 2012. Role of the pathotype-specific ACRTS1 gene encoding a hydroxylase involved in the biosynthesis of host-selective ACR-toxin in the rough lemon pathotype of Alternaria alternata. Phytopathology 102: 741–748.
  • Kakalíková, L., Jankura, E., Šrobárová, A., 2009. First report of Alternaria bunch rot of grapevines in Slovakia. Australasian Plant Disease Notes 4: 68–69.
  • Koç, İ., Yardım, E.N., Çelik, A., Mendeş, M., Mirtagioğlu, H., Namlı, A., 2018. Determination of Antifungal Effect of Wood Vinegar Obtained from Hazelnut Shells Against Mold Factors in In-Vitro Conditions. The Science Journal of Bitlis Eren University 7(2): 296-300.
  • Koç, İ., Çelik, A., Durak, E. D., Demir, İ., Bayman, S., Mirtagioğlu, H., & Demir, S. 2020. Morphological and Molecular Characterization of Walnut Anthracnose (Ophiognomonia leptostyla) Disease in Walnut Growing Areas of Bitlis Province. Journal of the Institute of Science and Technology 10(1): 28-38.
  • Koplay, C., 2004. Studies on determination of fungal pathogens causing rots on Sultanina table grapes and their control with fungicides in vitro conditions. İzmir. Turkey. University of Ege. MS thesis. pp. 105.
  • Leroux, P., Fritz, R., Debieu, D., Albertini, C., Lanen, C., Bach, J., Gredit, M., Chapeland, F., 2002. Mechanisms of resistance to fungicides in field strains of Botrytis cinerea. Pest. Man. Sci. 58:876-888.
  • Lorbeer, J.W. and Vincelli, P.C., 1990. Efficacy of dicarboximide fungicides and fungicide combinations for control of Botrytis leaf blight of onion in New York. Plant Diseases 74:235-237.
  • Lorenz, H.D., Eichhorn, K.W., 1978. Untersuchungen zur möglichen Resistenzbildung von Botrytis cinerea an Reben gegen die Wirkstoffe Vinclozolin und Iprodion. Die Wein-Wissenschatt. 33:2-10.
  • Lu, Y., Ma, J., Sutton, T.B., Ypema, H., 2004. Comparision of assay methods for evaluating the sensitivity of Alternaria mali to boscalid. Phytopathology p. 94.
  • Mostert, L., Crous, P.W., Petrini, O., 2000. Endophytic fungi associated with shots and 19 leaves of Vitis vinifera, with specific reference to the Phomopsis viticola complex. Sydowia 54: 46-58.
  • Pary, K.E., Wood, R.K.S., 1958. The adaptation of fungi to fungicides: adaptation to captan Ann. Appl. Biol. 46: 446-456.
  • Pashley, C.H., Fairs, A., Free, R.C., Wardlaw, A.J., 2012. DNA analysis of outdoor air reveals a high degree of fungal diversity, temporal variability, and genera not seen by spore morphology. Fungal Biol. 116: 214-224.
  • Peever, T.L., Ibañez, A., Akimitsu, K., Timmer, L.W., 2002. Worldwide phylogeography of the citrus brown spot pathogen. Alternaria alternata. Phytopathology 92: 794-802. doi:10.1094/PHYTO.2002.92.7.794
  • Pitt, J.I., Hockıng, A.D., 1997. Fungi and Food Spoilage 2nd ed. London: Blackie Academic & Professional. p. 593 ISBN 0-8342-1306-0.
  • Polizzotto, R., Andresen, B., Martini, M., Grisan, S., Assante, G., Musetti, R., 2012. A polyphasic approach for the characterization of endophytic Alternaria strains isolated from grapevines. Journal of Microbiological Methods 88:162-171. http://dx.doi.org/10.1016/j.mimet.2011.11.009.
  • Prusky, D., Fuchs, Y., Yanko, U., 1983. Assessment of latent infections as a basis for control of post-harvest disease of mango. Plant Diseases 67: 816-818.
  • Ranganath, C., Yonelinas, A. P., Cohen, M. X., Dy, C. J., Tom, S.M., D’Esposito, M.D., 2003. Dissociable correlates of recollection and familiarity with the medial temporal lobes. Neuropsychologia 42: 2-13.
  • Rotem, J., 1994. The genus Alternaria: biology, epidemiology and pathogenicity. St. Paul, MN: The American Phytopathological Society p. 326.
  • Rotondo, F., Collina, M., Brunelli, A., and Pryor, B.M, 2012. Compari-son of Alternaria spp. collected in Italy from apple with A. mali and other AM-toxin producing strains. Phytopathology 102:1130-1142.
  • Saitou, N., Nei, M., 1987. The neighbor-joining method: A new method for reconstructing phylogenetic trees. Molecular Biology and Evolution 4: 406-425.
  • Serra, R., Braga, A., Venâncio, A., 2005. Mycotoxin producing and other fungi isolated from grapes for wine production, with particular emphasis on ochratoxin A. Research in Microbiology 156(4): 515-521.
  • Sharma, M., Ghosh, R., Pande, S., 2013. Occurrence of Alternaria alternata causing Alternaria blight in pigeon pea in India. Advances in Bioscience and Biotechnology 4: 702-705.
  • Simmons, E.G., 1992. Alternaria themes and variations. Mycotaxon 48(73): 109-140.
  • Simmons, E.G., 2007. Alternaria: an identification manual, CBS Biodiversity series no. 6. Utrecht, The Netherlands, p: 775.
  • Swart, A.E., Lennox, C.L., Holz, G., 1995. Infection of table grape bunches by Alternaria alternata. South African Journal for Enology and Viticulture 16: 3–6.
  • White, T.J., Bruns, T., S-Lee, S., Taylor, J.W., 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In PCR Protocols: A Guide to Methods and Applications. Innis MA, Gel-fand DH, Sninsky JJ, White TJ, eds. Academic Press, Inc., New York, pp: 315–322.
  • Zahavi, T., Cohen, L., Weiss, B., Schena, L., Daus, A., Kaplunov, T., Jutkhi, J., Ben-Arie, R., Droby, S. 2000. Biological control of Botrytis. Aspergillus and Rhizopus rots on table and wine grapes in Israel. Postharvest Biology and Technology. 20: 115-124.
There are 43 citations in total.

Details

Primary Language English
Subjects Agricultural Engineering, Horticultural Production
Journal Section Articles
Authors

Nurdan Güngör Savaş 0000-0002-3450-4747

Figen Yıldız 0000-0002-9562-5657

Nafiz Delen 0000-0002-1634-0013

Publication Date May 30, 2022
Submission Date January 10, 2022
Published in Issue Year 2022 Volume: 4 Issue: 2

Cite

APA Güngör Savaş, N., Yıldız, F., & Delen, N. (2022). Sensitivity of Alternaria spp. Isolates to Some Fungicides Used on Sultani Çekirdeksiz Grapes in Aegean Region Vineyards. Uluslararası Anadolu Ziraat Mühendisliği Bilimleri Dergisi, 4(2), 23-35.