Sert Çekirdeklilerde Bakteriyel Kansere Neden Olan Pseudomonas syringae Pathovarlarının Klasik ve Moleküler Yöntemlerle Tanısı

Year 2020, Volume: 49 Issue: 3, 55 - 61, 22.12.2020

Damla Ertimurtaş , Hatice Özaktan

Abstract

Bakteriyel kanser belirtisi görülen şeftali, badem ve kiraz gibi sert çekirdekli meyve ağaçlarından izole edilmiş ve Ege Üniversitesi Ziraat Fakültesi Bitki Koruma Bölümü Bakteriyoloji laboratuar stoklarına alınmış 15 Pseudomonas syringae (Ps) pathovarlarının LOPAT (levan oluşumu, oxidase, patateste pektolitik aktivite, arginin dehidrolaz, tütünde aşırı duyarlılık reaksiyonu), GATTa (jelatinin hidrolizi, aesculinin hidrolizi, tyrosin aktivitesi, tartaric acid), karbon kaynaklarının kullanımı klasik tanı testleri ile tanısı yapılmıştır. Ayrıca patojenisite, in-vitro’da syringomycin üretimi ve buz çekirdeği oluşturma aktivitesi (INA) tanı testleri yapılmıştır. Klasik tanı testleri sonucunda kesin tanısı yapılamayan Pseudomonas syringae pathovarı izolatlarının moleküler tanısı yapılmıştır. Pseudomonas syringae pv. syringae (Pss)’nin syringomycin sentezinden sorumlu syrB geni ve Pseudomonas syringae pv. morsprunorum (Psm)’un coronatine sentezinden sorumlu cfl geni klasik PCR ile belirlenmiştir. Pss ve Psm’nin ayırt edilmesinde 4 housekeeping geni (cts, rpoD, gapA, gyrB) sekans analizleri yapılmıştır. Klasik ve moleküler tanı sonucunda 15 Ps pathovarının 9’u Pss olarak kesin tanısı yapılmıştır.

Keywords

tanılama, PCR, Sert çekirdekliler, Pseudomonas syringae, Bakteriyel hastalık

Classical and Molecular Diagnosis of Pseudomonas syringae Pathovars Causing Bacterıal Canker on Stone Fruits

Abstract

15 Pseudomonas syringae (Ps) pathovars isolated from stone fruit trees such as peach, almond and cherry with bacterial cancer symptoms and kept in the Bacteriology laboratory stocks of Ege University Faculty of Agriculture, Department of Plant Protection were identified based on LOPAT (levan, oxidase, potato rot pectolytic activity, arginin dihydrolase, hypersensitive reaction on tobacco leaves), GATTa (gelatin liquefaction, aesculin hydrolysis, tyrosinase activity, utilization of tartaric acid), utilization of carbon tests classical methods. In addition, pathogenicity, syringomycin production and ice nucleation activity (INA) were used for the identification. Pseudomonas syringae pathovars were identified by PCR that cannot be diagnosed with classical tests. Molecular diagnosis of Pseudomonas syringae pv. syringae (Pss) strains were based on syringomycin synthesis (syrB) and Pseudomonas syringae pv. morsprunorum (Psm) strains were based on coronatine synthesis (cfl) were diagnosed using PCR. Also sequence analysis of 4 housekeeping genes (cts, rpoD, gapA, gyrB) were used to distinguish Pss and Psm. As a result of classical and molecular identification 9 of 15 Ps pathovars were identified as Pss.

Keywords

identification, PCR, Stone fruits, Pseudomonas syringae, Bacterial disease

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