SONER SOYLU, Merve KARA, Osman TOKETTİ, EMİNE MİNE SOYLU, Aysun UYSAL, Şener KURT

Konya İlinde Yetiştirilen Patates Yumrularında Lastik Çürüklük Hastalık Etmeni Geotrichum candidum’un İzolasyonu, Morfolojik ve Moleküler Karakterizasyonu

Patates (Solanum tuberosum L.), Türkiye’de en fazla tüketilensebzelerin başında gelmektedir. Konya ilinin Çumra ilçesinde 2019yılının Temmuz ayında patates yetiştirilen alanlarda gerçekleştirilensörveyler sırasında ekşi koku yayan patates yumrularında beyazmiselyum ile kaplanmış suya batırılmış yumuşak lastik çürüklüklezyonları şeklinde yeni hastalık belirtileri gözlenmiştir. Yüzeydezenfeksiyonu yapılan enfekteli yumrulardan alınan dokularPatates Dekstroz Agar (PDA) besiyeri üzerine yerleştirilmiş ve25°C’de inkübasyona bırakılmıştır. Enfekteli dokulardan hızlıbüyüyen beyaz, düz ve süetimsi-tozlu görünümlü pigmentsiz fungalkoloniler gelişmiştir. Gelişen kolonilerin miselyumları dichotomousolarak dallanmış, şeffaf ve bölmeli yapıda olup, hiflerden oldukçafarklı boyutlarda şeffaf, tek hücreli ve silindirik yapıda havaiartrokonidilerin geliştiği gözlenmiştir. Fungal etmenin kültürel vemorfolojik özellikleri Geotrichum candidum ile tutarlıdır. Yapayinokulasyon yapılan patates yumruları üzerinde doğal enfektelipatates yumrularında görülen belirtilerin benzeri gözlenmiş olupfungal etmen bu dokulardan yeniden izole edilmiştir. İzole edilenfungal etmenin morfolojik tanısı, Matrix Assisted Laser DesorptionIonization-Time of Flight Mass Spectrometry (MALDI-TOF MS)cihazı kullanarak protein profili ve Internal Transcribed Spacer (ITS)gen bölgesi (ITS1-5.8S rDNA-ITS4) çoğaltılıp dizilenmesi ilemoleküler olarak da teyit edilmiştir. Gen bankasına kaydedilentemsili izolatın sekans sonucu G. candidum izolatı ile %99 oranındabenzerlik göstermiştir. G. candidum’un sebep olduğu patates lastikçürüklük hastalığının varlığı Türkiye’de önceden bildirilmemiştir.Türkiye’de yetiştirilen patates yumrularında lastik çürüklüğühastalığı etmeni G. candidum’un morfolojik ve molekülerkarakterizasyonu ilk kez bu çalışma ile ortaya konulmuştur.

Isolation, Morphological and Molecular Characterization of Rubbery Rot Disease Disease Agent Geotrichum candidum on Potato Tubers Grown in Konya Province

Potato is the one of the most consumed vegetable in Turkey. During field surveys in June of 2019, new disease symptoms on potato tubers, displaying a water-soaked soft rubbery rot with a sour, fermented smell, and white mycelial growth, were observed in commercial fields in Çumra district of Konya, Turkey. Diseased tubers were collected and small tissue pieces from surface disinfected symptomatic tuber were placed on potato dextrose agar (PDA) and incubated at 25°C. Fast growing, flat, white to cream, dry and finely suede-like with no reverse pigmented fungal colonies were subsequently developed from infected tissues. Hyphae were hyaline, dichotomously branched and septate, producing aerial chains of hyaline, smooth, one-celled, cylindrical arthroconidia. Cultural and morphological characters were consistent with Geotrichum candidum. All artificially inoculated tubers displayed the same symptoms as seen for naturally infected tubers and the fungus having same morphological characters was reisolated from inoculated tubers. Morphological identification of disease agent was confirmed molecularly based on protein profiling by using MALDI-TOF MS and also by sequencing amplified Internal Transcribed Spacer (ITS) region ((ITS1-5.8S rDNA-ITS4). The sequence was 99% identical and a representative sequence of isolate was submitted to GenBank. The presence of potato rubbery rot disease caused by G. candidum has not been previously reported in Turkey. This is the first report of morphological and molecular characterization of G. candidum causal disease agent of rubber rot on potato tubers grown in Turkey.

PDF

___

Aktan ZC, Soylu S 2020. Diyarbakır İlinde Yetişen Badem Ağaçlarından Endofit ve Epifit Bakteri Türlerinin İzolasyonu ve Bitki Gelişimini Teşvik Eden Mekanizmalarının Karakterizasyonu. KSU Tarım ve Doğa Dergisi 23 (3): 641-654.
Alam MW, Rehman A, Malik AU, Iqbal Z, Amin M, Ali S, Hameed A, Sarfraz S 2017. First Report of Geotrichum candidum Causing Postharvest Sour Rot of Peach in Punjab, Pakistan. Plant Disease 101(8): 1543-1544.
Anonim 2017. TUİK Bitkisel Üretim İstatistikleri. https://biruni.tuik.gov.tr/medas/?kn=92&locale=tr (Erişim Tarihi: 21.02.2020)
Anonymous, 2017. FAOSTAT, World Production Quantities of Crops http://www.fao.org (Erişim tarihi: 21.02.2020).
Aydın M.H, Pala F, Kaplan C 2016. Potato Tuber Sprout Rot Caused by Fusarium sambucinum in Turkey. Scientific Papers - Series A Agronomy 59: 189-193.
Boratyn GM, Camacho C, Cooper PS, Coulouris G, Fong A, Ma N, Madden TL, Matten WT, McGinnis SD, Merezhuk Y, Raytselis Y, Sayers EW, Tao T, Ye J, Zaretskaya I 2013. BLAST: A More Efficient Report with Usability Improvements. Nucleic Acids Research 41: 29–33.
Bozkurt İA, Soylu S, Kara M, Soylu EM 2020. Chemical Composition and Antibacterial Activity of Essential Oils Isolated from Medicinal Plants against Gall Forming Plant Pathogenic Bacterial Disease Agents. KSU Tarım ve Doğa Dergisi 23 (6): 1474-1482.
Çakır E 2005. First Report of Potato Wart Disease in Turkey. Plant Pathology 54: 584.
Çakır E, Demirci F 2012. First Report of Phytophthora cryptogea on Potato Tubers in Turkey. Plant Disease 96: 1224-1225.
Çakır E, Karahan A, Kurbetli İ 2019. Involvement of Colletotrichum coccodes Causing Atypical Symptoms of Potato Tubers in Turkey. Journal of Plant Diseases and Protection 126: 173–176.
Carolis ED, Posteraro B, Lass-Flo C, Vella A, Florio AR, Torelli R, Girmenia C, Colozza C, Tortorano AM, Sanguinetti M, Fadda G 2012. Species Identification of Aspergillus, Fusarium and Mucorales with Direct Surface Analysis by MatrixAssisted Laser Desorption Ionization Time-ofFlight Mass Spectrometry. Clinical Microbiology and Infection 18: 475–484.
Chalupová J, Raus M, Sedlářová M, Šebela M 2014. Identification of Fungal Microorganisms by MALDI-TOF Mass Spectrometry. Biotechnology Advances 32: 230–241.
De Hoog GS, Smith MT, Gueho E 1986. A Revision of the Genus Geotrichum and Its Teleomorphs. Studies in Mycology 29: 1-131.
Eken C, Demirci E, Șahin F 2000. Pathogenicity of the Fungi Determined on Tubers from Potato Storages in Erzurum. Journal of Turkish Phytopathology 29: 61-69.
Ezekiel R, Singh N, Sharma S, Kaur A 2013. Beneficial Phytochemicals in Potato – A Review. Food Research International 50: 487–496.
Farr DF, Rossman AY 2020. Fungal Databases. Systematic Mycology and Microbiology Laboratory, USDA ARS. Retrieved from http://nt.arsgrin.gov/fungaldatabases/ (Erişim Tarihi: 21.02.2020)
Gore ME, Senkal BC, Berk SK, Onaran H, Altin N, Ay E, Tuna S, Zencirci N 2015. Recovery of Verticillium dahliae from Commercially Available Potato Seed Lots Planted in Turkey and Characterization of Isolates by Vegetative Compatibility and Aggressiveness. Phytoparasitica 43: 241-251.
Hafeez R, Akhtar N, Shoaib A, Bashir U, Haidar MS, Awan ZA 2015. First Report of Geotrichum candidum from Pakistan Causing Postharvest Sour Rot in Loquat (Eriobotrya japonica). The Journal of Animal & Plant Sciences 25(6): 1737-1740.
Hameed A, Alam MW, Rehman A, Naveed K, Atiq M, Rajput NA, Sarfraz S, Liaqat N, Tahir FA 2019. First Report of Geotrichum candidum Causing Postharvest Sour Rot of Carrot in Punjab, Pakistan. Journal of Plant Pathology 101(3): 763.
Horita H, Hatta Y 2016. Sour Rot of Carrot Caused by Geotrichum candidum in Japan. Journal of General Plant Pathology 82: 65-68.
Hussain M, Hamid MI, Ghazanfar MU, Akhtar N, Raza M 2016. First Report of Fruit Rot of Strawberry Caused by Geotrichum candidum in Pakistan. Plant Disease 100(9): 1948-1949.
Kahn MFR, Haque ME, Brueggeman R, Zhong S, Bhuiyan MZR, Poudel RS, Gross T, Hakk P, Liu Y, 2019. First Report of Geotrichum candidum Causing Postharvest Rot of Sugar Beet (Beta vulgaris) Roots in Minnesota and North Dakota. Plant Disease 103(12): 3278.
Kara M, Uysal A, Soylu S, Kurt Ş, Soylu, EM 2017. Identification of Plant-Associated Microorganisms Employing MALDI TOF Mass Spectrometry as a Rapid Detection Technique. International Conference on Agriculture, Forest, Food Sciences and Technologies (ICAFOF), 15-17 May, 2017. Nevşehir, p. 1111.
Kara M, Soylu EM 2020. Assessment of Glucosinolate‐ Derived Isothiocyanates as Potential Natural Antifungal Compounds Against Citrus Sour Rot Disease Agent Geotrichum citri‐aurantii. Journal of Phytopathology 168(5): 279-289.
Kim YK, Kim TS, Shim HS, Park KS, Yeh WH, Hong SJ, Shim CK, Kim JS, Park JH, Han EJ, Lee MH, Jee HJ 2011. First Report of Sour Rot on PostHarvest Oriental Melon, Tomato, Cucumber, Potato, Pumpkin and Carrot Caused by Geotrichum candidum. Research in Plant Disease 17(2): 232- 234.
Koike ST, Gladders P, Paulus, AO 2007. Vegetable Diseases: A Colour Handbook. Manson Publishing, London, pp. 95-115.
Kumar S, Stecher G, Tamura K 2016. MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. Molecular Evolutionary Genetics Analysis 33(7): 1870–1874.
Kurt Ş, Uysal A, Kara M, Soylu S, Soylu EM 2017. Natural Infection of Potato by Sclerotinia sclerotiorum Causing Stem Rot Disease in Turkey. Australasian Plant Disease Notes 12: 39.
Ozer G, Bayraktar H, Tsror L, Yaman T, Kabakci H, Gore ME 2018. Vegetative Compatibility Groups in Colletotrichum coccodes from Turkey and Their Aggressiveness to Potato. Plant Pathology 67: 1735- 1739.
Pulcrano G, Roscetto E, Iula VD, Panellis D, Rossano F, Catania MR 2012. MALDI-TOF Mass Spectrometry and Microsatellite Markers to Evaluate Candida parapsilosis Transmission In Neonatal Intensive Care Units. European Journal of Clinical Microbiology & Infectious Diseases 31: 2919–2928.
Punja ZK, Rodriguez G, Tirajoh A, Formby S 2016. Role of Fruit Surface Mycoflora, Wounding and Storage Conditions on Post-Harvest Disease Development on Greenhouse Tomatoes. Canadian Journal of Plant Pathology 38(4): 448-459.
Shaw DE 1984. Microorganisms in Papua New Guinea. Research Bulletin of the Department of Primary Industries Port Moresby 33: 1-344. (6277)
Soylu EM, Soylu S, Kara M, Kurt Ş 2020. Sebzelerde Sorun Olan Önemli Bitki Fungal Hastalık Etmenlerine Karşı Vermikomposttan Izole Edilen Mikrobiyomların in vitro Antagonistik Etkilerinin Belirlenmesi. KSU Tarım ve Doğa Dergisi 23(1): 7- 18.