Van Gölü Havzası Micromonospora Cinsi Bakterilerinin Moleküler Karakterizasyonu ve Teşhisi

Yıl 2025, Cilt: 30 Sayı: 3, 932 - 967, 24.12.2025

Metin Ertaş , Kerem Özdemir

https://doi.org/10.53433/yyufbed.1739734

Öz

Bu çalışmada, Van Gölü Havzası’ndan toprak ve sediment örneklerinden izole edilen Micromonospora türleri, filogenetik ilişkilerini ve çeşitliliklerini ortaya koymak amacıyla fenotipik, biyokimyasal ve moleküler (16S rRNA gen bölgesi) analizleri bir arada içeren polifazik taksonomik bir yaklaşımla karakterize edilmiştir. İzolasyon için dilüsyon plak yöntemi kullanılmış ve toplam 141 saf Micromonospora izolatı elde edilmiştir. Farklı morfolojiye sahip koloniler seçilerek numaralandırılmış ve kriyojenik tüplerde muhafaza edilmiştir. Toprak ve sediment örneklerinin nem ve pH değerleri ölçülmüş, izolatlar koloni pigmentasyonuna göre 11 renk grubuna ayrılmıştır. Nümerik taksonomi için 83 morfolojik ve biyokimyasal karakter değerlendirilmiş ve MVSP 3.1 yazılımında Simple Matching Coefficient (SSM) yöntemiyle dendrogram oluşturulmuştur. %78 benzerlik oranına göre 7’si majör, 15’i minör olmak üzere toplam 22 küme tespit edilmiştir. Aynı kümede yer alan izolatların genellikle benzer rakım ve lokalitelerden geldiği gözlemlenmiştir. Spor zinciri morfolojisi Taramalı Elektron Mikroskobu (SEM) ile incelenerek teşhis sürecine katkı sağlanmıştır.
Moleküler karakterizasyon için seçilen 13 izolatın genomik DNA’sı izole edilmiş ve 16S rRNA gen bölgesi, 27F ve 1492R evrensel primerleri kullanılarak çoğaltılmıştır. Maximum Likelihood ve Bayesian çıkarım algoritmaları ile filogenetik ağaçlar oluşturulmuş, elde edilen ağaç topolojilerinin genellikle benzerlik gösterdiği belirlenmiştir. İzolatların NCBI’dan alınan referans türlerle güçlü bir homoloji gösterdiği ve genetik pozisyonlarının ortaya konduğu tespit edilmiştir.

Anahtar Kelimeler

Micromonospora , Moleküler karakterizasyon , Van Gölü Havzası , 16S rRNA

Destekleyen Kurum

Van Yüzüncü Yıl Üniversitesi, Bilimsel Araştırma Projeleri Koordinasyon Birimi

Proje Numarası

2014-FBE-D092

Teşekkür

Van Yüzüncü Yıl Üniversitesi Bilimsel Araştırma Projeleri Koordinasyon Birimine teşekkürlerimi sunarım.

Molecular Characterization and Identification of Micromonospora Genus Bacteria from the Van Lake Basin

Öz

In this study, Micromonospora species were isolated from soil and sediment samples collected from the Van Lake Basin and characterized through a polyphasic taxonomic approach combining phenotypic, biochemical, and molecular (16S rRNA gene) analyses to elucidate their phylogenetic relationships and diversity. The dilution plate method was employed for isolation, yielding 141 purified Micromonospora isolates. Colonies with distinct morphologies were selected, numbered, and preserved in cryogenic tubes. The moisture and pH levels of the soil and sediment samples were also measured. Based on colony pigmentation, the isolates were classified into 11 color groups. For numerical taxonomy, 83 morphological and biochemical characteristics were assessed, and a dendrogram was constructed using the Simple Matching Coefficient (SSM) in MVSP 3.1 software. As a result, 22 clusters (7 major and 15 minor) were formed based on 78% similarity. Cluster members generally showed similarity in altitude and geographic origin. Scanning Electron Microscopy (SEM) was used to analyze the spore chain morphology, contributing to more accurate identification. For molecular characterization, genomic DNA was extracted from 13 selected isolates, and the 16S rRNA gene was amplified using universal primers 27F and 1492R. Phylogenetic trees were constructed using Maximum Likelihood and Bayesian inference algorithms. The resulting topologies were largely congruent, and Micromonospora isolates clustered with reference strains from NCBI with high sequence homology. These findings revealed the genetic positions of the isolates and provided insight into their evolutionary relationships, contributing to the microbial diversity knowledge of alkaline environments such as the Van Lake Basin.

Anahtar Kelimeler

Micromonospora , Molecular characterization , Van Lake Basin , 16S rRNA

Etik Beyan

The authors declare that the materials and methods used in this study did not require ethics committee approval and/or specific legal permission.

Destekleyen Kurum

Van Yüzüncü Yıl University, Scientific Research Projects Coordination Unit

Proje Numarası

2014-FBE-D092

Teşekkür

I would like to express my gratitude to Van Yüzüncü Yıl University, Scientific Research Projects Coordination Unit, for supporting this study with project number 2014-FBE-D092. This study is derived from Metin ERTAŞ's Doctoral dissertation.

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