Fuhrman J A, McCallum K, Davis A A
Department of Biological Sciences, University of Southern California, Los Angeles 90089-0371.
Appl Environ Microbiol. 1993 May;59(5):1294-302. doi: 10.1128/aem.59.5.1294-1302.1993.
The extent of the diversity of marine prokaryotes is not well known, primarily because of poor cultivability. However, new techniques permit the characterization of such organisms without culturing, via 16S rRNA sequences obtained directly from biomass. We performed such an analysis by polymerase chain reaction amplification with universal primers on five oligotrophic open-ocean samples: from 100-m (three samples) and 500-m depths in the western California Current (Pacific Ocean) and from a 10-m depth in the Atlantic Ocean near Bermuda. Of 61 clones, 90% were in clusters of two or more related marine clones obtained by ourselves or others. We report 15 clones related to clone SAR 11 found earlier near Bermuda (S. J. Giovannoni, T. B. Britschgi, C. L. Moyer, and K. G. Field, Nature [London] 345:60-63, 1990), 11 related to marine cyanobacteria, 9 clustered in a group affiliated with gram-positive bacteria, 9 in an archaeal cluster we recently described (mostly from the 500-m sample), 4 in a novel gamma-proteobacterial cluster, and 6 in three two-membered clusters (including other archaea). One clone was related to flavobacteria. Only the cyanobacteria plus one other clone, related to Roseobacter denitrificans (formerly Erythrobacter longus Och114), were within 10% sequence identity to any previously sequenced cultured organism in a major data base. We never found more than two occurrences of the same sequence in a sample, although four times we found identical sequences between samples, two of which were between oceans; one of these sequences was also identical to SAR 11.(ABSTRACT TRUNCATED AT 250 WORDS)
海洋原核生物的多样性程度尚不为人所知,主要是因为其可培养性较差。然而,新技术使人们能够在不进行培养的情况下,通过直接从生物量中获得的16S rRNA序列对这类生物进行特征描述。我们通过聚合酶链反应扩增,使用通用引物对五个贫营养开阔海洋样本进行了此类分析:来自加利福尼亚洋流(太平洋)西部100米深度(三个样本)和500米深度,以及来自百慕大附近大西洋10米深度的样本。在61个克隆中,90%属于我们自己或他人获得的两个或更多相关海洋克隆的簇。我们报告了15个与之前在百慕大附近发现的克隆SAR 11相关的克隆(S. J. 乔瓦诺尼、T. B. 布里奇吉、C. L. 莫耶尔、K. G. 菲尔德,《自然》[伦敦]345:60 - 63,1990),11个与海洋蓝细菌相关,9个聚集在一个与革兰氏阳性菌相关的组中,9个在我们最近描述的古菌簇中(大多来自500米深度的样本),4个在一个新的γ-变形菌簇中,6个在三个二元簇中(包括其他古菌)。一个克隆与黄杆菌相关。只有蓝细菌加上另一个与反硝化玫瑰杆菌(以前的长红赤杆菌Och114)相关的克隆,与主要数据库中任何先前测序的培养生物的序列同一性在10%以内。我们在一个样本中从未发现同一序列出现超过两次,尽管有四次我们在样本之间发现了相同序列,其中两次是在不同海洋之间;这些序列中有一个也与SAR 11相同。(摘要截短至250字)