Archaeal community structure and diversity in Urumqi No.10 cold sulfur Spring analyzed by culture-independent approach

Abstract:

Abstract:［Objective］We explored the composition and diversity of archaea in a cold sulfur spring water in Xinjiang earthquake fault zone．［Methods］Environmental total DNA was extracted directly with enzymatic lysis method from a cold sulfur spring water． We constructed clone library of 16S rRNA gene amplified with archaeal-specific primers． A total of 115 positive clones were selected randomly from the library and identified by restriction length polymorphism(RFLP) with enzyme Alu I and Afa I．The unique RFLP patterns corresponded clones were selected for sequencing，BLAS alignment and constructing 16S rRNA gene phylogenetic tree．［Results］ In total，44 operational taxonomic units (OTUs) were determined from the library． BLAST and phylogenetic analysis indicated that these OTUs were affiliated with Euryarchaeota (94.78%) and Thaumarchaeota (4.35%)．Only one Thaumarchaeotal clone was detected and most related to the genus Nitrosopumilus with 93% similarity． Euryarchaeotal clones were abundant and diverse．Of them，42.61% of clones belonged to RC-V cluster; 13.91% of clones，20.87% of clones were classified into LDS cluster and Methanomicrobiales respectively; 4.35% of clones had high similarity with ANME-1a-FW，which were involved in Anaerobic oxidation of methane (AOM)．In addition，we also detected some (13.05%) unknown Euryarchaeotal clones．Conclusion］ Euryarchaeota in the environment were diverse，and possibly with a large fraction of potential novel species．