Abstract

BackgroundWhen the filamentous cyanobacterium Anabaena variabilis grows aerobically without combined nitrogen, some vegetative cells differentiate into N2-fixing heterocysts, while the other vegetative cells perform photosynthesis. Microarrays of sequences within protein-encoding genes were probed with RNA purified from extracts of vegetative cells, from isolated heterocysts, and from whole filaments to investigate transcript levels, and carbon and energy metabolism, in vegetative cells and heterocysts in phototrophic, mixotrophic, and heterotrophic cultures.ResultsHeterocysts represent only 5% to 10% of cells in the filaments. Accordingly, levels of specific transcripts in vegetative cells were with few exceptions very close to those in whole filaments and, also with few exceptions (e.g., nif1 transcripts), levels of specific transcripts in heterocysts had little effect on the overall level of those transcripts in filaments. In phototrophic, mixotrophic, and heterotrophic growth conditions, respectively, 845, 649, and 846 genes showed more than 2-fold difference (p < 0.01) in transcript levels between vegetative cells and heterocysts. Principal component analysis showed that the culture conditions tested affected transcript patterns strongly in vegetative cells but much less in heterocysts. Transcript levels of the genes involved in phycobilisome assembly, photosynthesis, and CO2 assimilation were high in vegetative cells in phototrophic conditions, and decreased when fructose was provided. Our results suggest that Gln, Glu, Ser, Gly, Cys, Thr, and Pro can be actively produced in heterocysts. Whether other protein amino acids are synthesized in heterocysts is unclear. Two possible components of a sucrose transporter were identified that were upregulated in heterocysts in two growth conditions. We consider it likely that genes with unknown function represent a larger fraction of total transcripts in heterocysts than in vegetative cells across growth conditions.ConclusionsThis study provides the first comparison of transcript levels in heterocysts and vegetative cells from heterocyst-bearing filaments of Anabaena. Although the data presented do not give a complete picture of metabolism in either type of cell, they provide a metabolic scaffold on which to build future analyses of cell-specific processes and of the interactions of the two types of cells.Electronic supplementary materialThe online version of this article (doi:10.1186/1471-2164-14-759) contains supplementary material, which is available to authorized users.

Highlights

  • When the filamentous cyanobacterium Anabaena variabilis grows aerobically without combined nitrogen, some vegetative cells differentiate into N2-fixing heterocysts, while the other vegetative cells perform photosynthesis

  • We report the first comparison of gene expression patterns in the vegetative cells and heterocysts of a filamentous cyanobacterium, A. variabilis ATCC 29413

  • Substantial transcript levels of rbcL in heterocysts are inconsistent with imaging of transcriptional fusions of rbcL to luciferase in PCC 7120, which showed at most very slight expression in heterocysts compared to vegetative cells [39], and with diverse other data [4], A. variabilis heterocysts might use RuBisCO to reassimilate CO2 released by the oxidative pentose phosphate pathway [87,102]

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Summary

Introduction

When the filamentous cyanobacterium Anabaena variabilis grows aerobically without combined nitrogen, some vegetative cells differentiate into N2-fixing heterocysts, while the other vegetative cells perform photosynthesis. Heterocysts are thought to maintain a microoxic interior by three mechanisms: they (i) form a thick envelope of glycolipid and polysaccharide that reduces the rate of entry of O2, (ii) respire actively, and (iii) stop producing O2 [4,5]. Their microoxic interior permits N2 fixation by nitrogenase, a highly O2-sensitive enzyme. Ferredoxin reduced by photosystem I (PS I) is the likely source of electrons for N2 fixation [10], but the metabolic pathway or pathways that transfer electrons to PS I in heterocysts are not known

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