RESUMO
Biological nitrogen fixation has been suggested as an important source of nitrogen for the ultra-oligotrophic waters of the Levantine Basin of the Mediterranean Sea. In this study, we identify and characterize the spatial and temporal distribution of the N-fixing (diazotrophic) cyanobacterium Richelia intracellularis. R. intracellularis is usually found as an endosymbiont within diatoms such as Rhizosolenia spp and Hemiaulus spp. and is an important diazotroph in marine tropical oceans. In this study, two stations off the Mediterranean coast of Israel were sampled monthly during 2005-2007. R. intracellularis was identified by microscopy and by reverse transcribed-PCR which confirmed a 98.8% identity with known nifH sequences of R. intracellularis from around the world. The diatom-diazotroph associations were found throughout the year peaking during autumn (October-November) at both stations. Abundance of R. intracellularis ranged from 10 to 55 heterocysts l(-1) and correlated positively with the dissolved Si(OH)(4)/(NO(3)+NO(2)) ratio in surface waters. Although the rates of nitrogen fixation were very low, averaging approximately 1.1 nmol N l(-1) day(-1) for the R. intracellularis size fraction (>10 microm) from surface waters, they correlated positively with heterocyst counts during thermal stratification. The lack of large-scale diatom-diazotroph blooms and the low rates of nitrogen fixation by these diazotrophs may result from the P-starved conditions affecting the Levantine basin.
Assuntos
Cianobactérias/crescimento & desenvolvimento , Cianobactérias/isolamento & purificação , Água do Mar/microbiologia , Cianobactérias/fisiologia , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Diatomáceas/microbiologia , Genes de RNAr , Mar Mediterrâneo , Dados de Sequência Molecular , Nitratos/análise , Nitritos/análise , Fixação de Nitrogênio , Filogenia , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Estações do Ano , Água do Mar/química , Análise de Sequência de DNA , Homologia de Sequência do Ácido Nucleico , Ácido Silícico/análise , SimbioseRESUMO
The eastern Mediterranean Sea is one of the most extreme oligotrophic oceanic regions on earth in terms of nutrient concentrations and primary productivity. Nitrogen fixation has been suggested to contribute to the high N : P molar ratios of approximately 28:1 found in this region. Surprisingly, no molecular biological work has been performed in situ to assess whether N(2) fixation genes actually occur in the eastern Mediterranean Sea, or to determine which organisms are responsible for this process. In this study, we examined the presence and expression of nitrogenase genes (nifH) in the upper water layer of the eastern Mediterranean. Clone libraries constructed from both DNA and reverse-transcribed PCR-amplified mRNA were examined and compared. We observed different nifH genes from diverse microbial groups, such as Cyanobacteria, Proteobacteria and methanogenic Archaea. Interestingly, numerous phylotypes were observed in coastal stations at the DNA level but none were active. However, in far offshore stations, the phylotypes observed at the DNA level were the ones that were actually active. Our preliminary study revealed diverse diazotrophs that possess and express nifH genes, which may support N(2) fixation in the eastern Mediterranean Sea.
