Molecular Characterization of Some Saprophyte Agrobacterium spp. Strains Isolated from Root Nodules of C. arietinum L. Cultivated in Central Anatolia Region of Turkey

Yıl 2020, , 475 - 483, 31.12.2020

Cem Tolga Gürkanlı

https://doi.org/10.35229/jaes.744909

Öz

ABSTRACT
In the present study twelve plant associated bacteria which isolated from root nodules of C. arietinum L. collected from Central-Anatolia region of Turkey were identified with current molecular techniques. None of the isolates were produced root nodules or showed pathogenic effects (gall or hairy root) on the original host as the result of authentication and pathogenicity tests, respectively. These results have suggested that all these isolates are root endophytic bacteria. Additionally, PCR amplifications for nodD and virA genes did not revealed any bands. These results showed that these isolates have not harbour symbiotic (pSym) or pathogenicity (Ti: Tumor inducing or Ri: Root inducing) plasmids which are necessary for nodulation or virulence, respectively. TP-RAPD analysis revealed three patterns indicating three genetically distinct group within the isolate collection. From each pattern one representative isolate was selected for further molecular analyses. Phylogenetic analyses depending on nucleotide sequences of 16S rDNA and recA genes assigned representative isolates of Pattern-A (n: 4) and Pattern-B (n: 6) to A. radiobacter. On the other hand, the representative isolate of Pattern-C (n: 2) appeared as related to A. nepotum. As a result, this study presents the first phylogenetically identified root-endophytic Agrobacterium radiobacter and A. nepotum isolates from root nodules of C. arietinum L. grown in Central Anatolia part of Turkey. Additionally the first molecular data of A. radiobacter for Turkey also presented.

ÖZ
Bu çalışmada Türkiye’nin İç Anadolu bölgesinden toplanan C. arietinum L. bitkisinin kök nodüllerin’den izole edilmiş on iki bitki ilişkili bakteri moleküler teknikler kullanılarak teşhis edilmiştir. Bu izolatların hiç birisi otantikasyon ve patojenite testlerinin sonucunda orijinal konak üzerinde nodül oluşturmamış veya patojenik etki (gall oluşumu veya saçak kök oluşumu) göstermemiştir. Bu sonuçlar bütün bu izolatların kök endofitik bakteriler olduğunu ortaya koymuştur. Ek olarak nodD ve virA genleri için yapılan PCR amplifikasyonları herhangi bir bant ortaya koymamıştır. Bu sonuçlar bu izolatların nodülasyon veya virülans için gerekli olan sırasıyla simbiyotik (pSym) veya patojenite (Ti: Tümör oluşturan veya Ri: Saçak kök oluşturan) pilazmitlerini taşımadıklarını göstermiştir. TP-RAPD analizi izolat kolleksiyonu içerisinde genetik olarak ayrı üç gruba işaret eden üç şablon ortaya koymuştur. İleri moleküler analizler için her şablondan bir temsilci izolat seçilmiştir. 16S rDNA ve recA genlerinin nükleotid dizilerine dayalı filogenetik analizler Şablon-A (n: 4) ve Şablon-B’nin (n: 6) temsilci izolatlarını Agrobacterium radiobacter türü ile ilişkilendirmiştir. Diğer taraftan, Şablon-C’nin (n: 2) temsilci izolatı A. nepotum olarak ortaya çıkmıştır. Sonuç olarak bu çalışma Türkiye’nin İç Anadolu Bölgesinde yetiştirilen C. arietinum L.’nin kök nodüllerin’den filogenetik olarak tanımlanmış ilk kök endofitik Agrobacterium radiobacter and A. nepotum izolatlarını sunmaktadır. Ek olarak, A. radiobacter için Türkiye’den ilk moleküler verileri de sunmaktadır.

Anahtar Kelimeler

Agrobacterium, Chickpeas, Nodulation, Phylogeny

Kaynakça

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