Multilocus Phylogenetic Characterization of Mesorhizobium-like Nodule-associated Isolates from Root Nodules of Genista acanthoclada

Öz

The root nodule symbiosis between legumes and rhizobia plays a fundamental role in biological nitrogen fixation and ecosystem sustainability. Genista acanthoclada, a perennial woody legume native to arid and rocky slopes of Western Türkiye, has not previously been investigated regarding to its symbiotic microsymbionts. In this study, five bacterial isolates were obtained from root nodules of G. acanthoclada collected from Yamanlar Mountain (İzmir, Türkiye) on Yeast Extract Mannitol Agar (YEMA) and rhizobial strains (GA.7.1 and GA.9) were rapidly selected by colony PCR targeting the nodC gene. Both isolates exhibited typical rhizobial phenotypic characteristics, including Gram-negative rod-shaped morphology, positive catalase and oxidase activities, and weak Congo red absorption on YEMA medium. Taxonomic analyses based on 16S rRNA, recA, and atpD gene sequences revealed that both isolates belong to the genus Mesorhizobium. The incongruence between the phylogenetic analysis based on nodC amino acid sequences and the core gene phylogenies suggests that horizontal gene transfer may play a role in the evolution of nodulation-related genes. In contrast, the nifH-based phylogeny was largely congruent with housekeeping gene phylogenies, indicating a more conserved evolutionary history of nitrogen fixation genes. These findings provide the first data on the rhizobia associated with the nodules of G. acanthoclada and contribute to a better understanding of the evolutionary dynamics of legume–rhizobium interactions.

Anahtar Kelimeler

• Rhizobia
• legume
• symbiotic nitrogen fixation (SNF)
• nodC
• nifH

Destekleyen Kurum

Ege University

Proje Numarası

75599

Etik Beyan

Ethics committee approval is not required for this study.

Teşekkür

This study was conducted as part of Betül Dağlı’s master’s thesis and was funded by the Ege University Scientific Research Projects Coordination Unit (BAP), Project No. 75599.

Genista acanthoclada'nın Kök Nodüllerinden Elde Edilen Mesorhizobium Benzeri Nodül Ilişkili Izolatların Çoklu Lokus Filogenetik Karakterizasyonu

Öz

Baklagiller ve Rhizobia arasındaki kök nodül simbiyozu, biyolojik azot fiksasyonunda ve ekosistem sürdürülebilirliğinde temel bir rol oynamaktadır. Batı Türkiye’de kurak ve kayalık yamaçlara özgü çok yıllık odunsu bir baklagil olan Genista acanthoclada, bugüne kadar simbiyotik mikrosimbiyontları açısından incelenmemiştir. Bu çalışmada, Yamanlar Dağı’ndan (İzmir, Türkiye) toplanan G. acanthoclada kök nodüllerinden Yeast Extract Mannitol Agar (YEMA) besiyerinde beş bakteri izolatı elde edilmiş ve nodC genine yönelik koloni PCR analizi ile rhizobiyal suşlar (GA.7.1 ve GA.9) hızlı biçimde seçilmiştir. Her iki izolat, Gram-negatif çubuk şekilli hücre morfolojisi, pozitif katalaz ve oksidaz aktiviteleri ve YEMA ortamında zayıf Kongo kırmızısı emilimi gibi tipik rhizobiyal fenotipik özellikler sergilemiştir. 16S rRNA, recA ve atpD gen dizilerine dayalı taksonomik analizler, her iki izolatın Mesorhizobium cinsine ait olduğunu ortaya koymuştur. nodC amino asit dizilerine dayalı filogenetik analiz ile çekirdek gen filogenileri arasındaki uyumsuzluk, nodülasyonla ilişkili genlerin evriminde yatay gen transferinin rol oynayabileceğini düşündürmektedir. Buna karşılık nifH genine dayalı filogeni, evrimsel kronometre olarak kabul edilen diğer genlerle büyük ölçüde uyumlu bulunmuş ve azot fiksasyon genlerinin daha korunmuş bir evrimsel geçmişe sahip olduğunu göstermiştir. Bu bulgular, G. acanthoclada'nın nodülleriyle ilişkili rhizobia hakkında ilk verileri sağlamakta ve baklagil-rizobiyum etkileşimlerinin evrimsel dinamiklerinin daha iyi anlaşılmasına katkıda bulunmaktadır.

Anahtar Kelimeler

• Rhizobia
• baklagil
• simbiyotik azot fiksasyonu (SNF)
• nodC
• nifH

Destekleyen Kurum

Ege Üniversitesi

Proje Numarası

75599

Etik Beyan

Bu çalışma için etik kurul onayı gerekmemektedir.

Teşekkür

Bu çalışma, Betül Dağlı'nın yüksek lisans tezinin bir parçası olarak yürütülmüş ve Ege Üniversitesi Bilimsel Araştırma Projeleri Koordinasyon Birimi (BAP) tarafından 75599 numaralı proje kapsamında finanse edilmiştir.

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