ÇEŞİTLİ SUCUL HİPERSALİN HABİTATLARDAKİ (TÜRKİYE) PROKARYOTİK TOPLULUKLARIN VE NANOHALOARKEAL SOYLARIN ÇEŞİTLİLİĞİNİN YÜKSEK-VERİMLİ DİZİLEME VE KLONLAMA KULLANILARAK DEĞERLENDİRİLMESİ

Year 2021, , 57 - 68, 25.01.2021

Seval Çınar , Mehmet Burçin Mutlu

https://doi.org/10.18036/estubtdc.777824

Cited By: 2

Abstract

Bu çalışmada, 16S rRNA geni hedefli yaklaşımlar kullanılarak prokaryotik grupların Tuz Gölü, Ayvalık ve Tuzlagözü güneş tuzlalarındaki dağılımı araştırılmıştır. Numunelerdeki prokaryotik taksonların nispi bolluğu, yüksek-verimli dizileme kullanılarak saptanmıştır. Haloquadratum ile ilişkili operasyonel taksonomik birimler (OTU'lar) MiSeq okumalarında en bol saptananlardı. Nanohaloarchaeota-ilişkili OTU’lar Ayvalık ve Tuzlagözü güneş tuzlalarında nadirdi (<%1), ve Tuz Gölü'nde yaklaşık %5 idi. Örneklerde sıklıkla bulunan ve paylaşılan diğer OTU'lar Halorubrum, Halonotius ve Salinibacter cinsleri ile ilişkiliydi.

Filotipleri çeşitli hipersalin ortamlarda sıklıkla tespit edilen Nanohaloarchaeota soyları, gruba özgü primer kullanılarak 16S rRNA gen klonlama ile daha ayrıntılı olarak incelenmiştir. Klon kütüphanelerinde yüksek oranda temsil edilen nanohaloarkeal filotiplerden bazıları, veri tabanındaki dizilere düşük benzerlik göstererek iki farklı dal oluşturmuştur. Ayvalık örneğinden oluşturulan klon kütüphanesinde yeni soylardan birinin öne çıktığı saptanmıştır. Ca. Nanopetramus'a % 95-97 dizi benzerliği gösteren filotipler de Ayvalık’ta yüksek oranda temsil edilmiştir. Tuz Gölü ve Tuzlagözü numunelerinin klon kütüphanelerinde sıklıkla saptanan filotipler, Ca. Nanosalina ve akrabalarının yanı sıra yeni bir soy ile ilişkilendirilmiştir.

Keywords

Nanohaloarchaeota, Yüksek-Verimli Dizileme, Klonlama, Halofiller

ASSESSING THE DIVERSITY OF PROKARYOTIC COMMUNITIES AND NANOHALOARCHAEAL LINEAGES IN VARIOUS AQUATIC HYPERSALINE HABITATS (TURKEY) USING HIGH-THROUGHPUT SEQUENCING AND CLONING

Abstract

IIn this study, distribution of the prokaryotic groups in Tuz Lake, Ayvalık and Tuzlagözü solar salterns was investigated using 16S rRNA gene targeted approaches. The relative abundance of prokaryotic taxa in the samples was detected by using high-throughput sequencing. Operational taxonomic units (OTUs) associated with Haloquadratum were the most abundant in MiSeq reads. Nanohaloarchaeota-related OTUs were rare (<1%) in Ayvalık and Tuzlagözü solar salterns, and around 5% in Tuz Lake. Other OTUs frequently found and shared in the samples were associated with Halorubrum, Halonotius and Salinibacter genera.

Nanohaloarchaeota lineages, whose phylotypes have been frequently detected in diverse hypersaline environments, were examined in more detail by 16S rRNA gene cloning using group-specific primer. Some of the highly represented nanohaloarchaeal phylotypes in the clone libraries showed low similarity to any sequence in the database, generating two distinct clades. One of the novel lineages was found to be prominent in the clone library constructed from Ayvalık sample. Phylotypes showing 95-97% sequence similarity to Ca. Nanopetramus were also highly represented in Ayvalık. Phylotypes frequent in the clone libraries of Tuz Lake and Tuzlagözü samples were associated with a novel lineage, as well as Ca. Nanosalina and its relatives.

Keywords

Nanohaloarchaeota, High-Throughput Sequencing, Cloning, Halophiles

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