Two flavodoxin genes in Trichodesmium (Oscillatoriales, Cyanophyceae): Remarkable sequence divergence and possible functional diversification

Abstract

We analyzed flavodoxin gene sequences and transcription patterns in the marine nitrogen-fixing cyanobacterium Trichodesmium Ehrenberg ex Gomont 1892. While a typical cyanobacterial flavodoxin ortholog ( fld1) was identified from cultured (strain IMS101) and environmental Trichodesmium by PCR and Southern blot hybridization, a second flavodoxin ( fld2) was identified in the genome sequence. BLAST and phylogenetic analyses indicated that the two fld genes were highly divergent: fld1 was most closely related to fld common in cyanobacteria, wheras fld2 formed a distinct cluster with a fld so far only found in the unicellular diazotrophic cyanobacteria Cyanothece and Crocosphaera (in which fld2 was the only fld). The fld2 cluster was more closely allied with a fld in non-cyanobacterial diazotrophs such as Azotobacter and Azoarcus than fld1 in other cyanobacteria. This result suggests different evolutionary history of these two genes. Real-Time RT–PCR analysis on iron-replete and iron-deplete Trichodesmium cultures revealed remarkable diel dynamics in transcription of both flds but surprisingly failed to show induction under iron stress. Addition of nitrate to these cultures depressed nifH and fld2 transcription while elevating the transcription of fld1 in iron-stressed cultures. A positive correlation between fld and nifH transcript abundances was found, which was stronger in the case of fld2 than fld1. Our data suggest that these two flavodoxins in Trichodesmium may be functionally diversified and that use of flavodoxin as an iron stress indicator needs to be validated for each organism with consideration of diel dynamics of gene expression.