The Associations Between Fishes and Luminescent Bacteria

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The Associations Between Fishes and Luminescent Bacteria ONE The Associations between Fishes and Luminescent Bacteria Luminescent Bacteria ­photooxidation, hydrostatic pressure and the ability to grow in nutrient poor conditions (Yetinson and Luminescent bacteria are the most common and Shilo, 1979; Shilo and Yetinson, 1979; Herring, 1982; widely distributed of all luminous organisms, typi- Al Ali et al., 2010). Luminous Vibrionacea are found cally emitting a continuous blue-green light, peak- both free living, as saprophytes, and as gut symbionts ing around 490 nm. They occur most frequently in and symbionts contained in special light organs of the marine environment but are also found in the fishes and cephalopods. Five species of luminous freshwater and terrestrial environments (Nealson ­bacteria, Aliivibrio fisheri, Photobacterium leiognathi, and Hastings, 1990; Meighen, 1991). Currently, P. kishitanii, P. mandapamensis and Photodesmus twenty-five species of luminescent bacteria, belong- katoptron, and two groups of not yet identified bacte- ing to six genera and three families, have been iden- ria associated with deep sea anglerfishes and flashlight tified. Bacteria of the genus Schewanella (fam. fishes have been so far described from fish light organs Schewanellaceae) are commonly found free living (Table 1.1). The associations of these bacteria with in the freshwater environment. Members of the fishes is discussed throughout this chapter. A shift genus Photorhabdus (fam. Enterobacteriaceae) are from one niche to another (e.g., Photobacterium terrestrial, gut endosymbionts of nematodes of the ­leiognathi shifting from the nutrient-rich esophageal genus Heterorhabditis. The genera Photobacterium, light organ of ponyfish into the fish’s gut and, subse- Vibrio, Aliivibrio and Photodesmus are all marine quently, into the water column adapting a free living members of the Vibrionaceae, an ecologically mode in a starvation/survival habitat) is typical for diverse group of Gram-negative bacteria often asso- many of these symbiotic bacteria (Nealson and ciated with marine animals (Peat and Adams, 2008; Hastings, 1990; Urbanczyk et al., 2011). Hendry and Dunlap, 2011;COPYRIGHTED Urbanczyk et al., 2011). The MATERIAL bacterial light producing reaction is Luminescent bacteria of the Vibrionaceae are ­catalyzed by the heterodimeric enzyme luciferase, found in all marine environments from the cold polar which ­consists of two similarly structured α and β seas to the warm tropics and from surface waters to subunits with molecular masses of 40 and 37 kDa, great depths. Both geographic, seasonal and depth- respectively. This enzyme oxidizes with atmos- related differences in luminescent bacterial abundance­ pheric oxygen (O2), a reduced riboflavin phos­ and species composition have been documented. phate (FMNH2) and a long chain fatty aldehyde The ­distribution patterns were ­suggested to be con- (RCHO) into an electronically excited flavin. With trolled by temperature, salt ­tolerance, ­resistance to the release of a blue-green light (490 nm), flavin Symbiosis in Fishes: The Biology of Interspecific Partnerships, First Edition. Ilan Karplus. © 2014 Ilan Karplus. Published 2014 by John Wiley & Sons, Ltd. 0002073813.INDD 6 2/19/2014 3:27:15 PM The Associations between Fishes and Luminescent Bacteria 7 Table 1.1 Fish hosts and symbiotic luminescent bacteria. Host Fish Depth and Temperature Luminescent Order and Family Bacteria Species Anguilliformes Shallow? Cool? Not identified Congridae Argentiformes Deep, cold Photobacterium kishitanii Opisthoproctidae Aulopiformes Deep, cold Photobacterium kishitanii Chlorophthalmidae Gadiformes Moderate to deep, cold Photobacterium kishitanii Macrouridae Aliivibrio fisheri Gadiformes Moderate to deep, cold Photobacterium mandapamensis Merculidae Gadiformes Moderate to deep, cold Photobacterium kishitanii Moridae Photobacterium mandapamensis Lophiformes Deep, pelagic, cold Not identified 9 families Berciformes Shallow to moderate, Not identified* Anomalopidae warm to cool Berciformes Shallow to moderate, Aliivibrio fisheri Monocentridae warm to cool Berciformes Deep, cold Photobacterium kishitanii Trachichthyidae Perciformes Shallow to moderate, Photobacterium kishitanii Acropomatidae cool to cold Photobacterium leiognathi Photobacterium mandapamensis Perciformes Shallow, warm Photobacterium mandapamensis Apogonidae Perciformes Shallow to moderate, Photobacterium leiognathi Leiognathidae warm to cool Photobacteriun mandapamensis Vibrio harveyi (?) *Except for Photodesmus katoptron (Hendry and Dunlap 2011). Urbanczyk et al. 2011. Reproduced with permission of John Wiley & Sons. mononucleotide is produced together with water which probably resulted from gene duplication, since and a fatty acid (RCOOH): there is about 30% identity in the amino acid sequence between the α and β subunits of all bacte- FMNH2 + RCHO + O2 → FMN + H2O + RCOOH rial luciferases. The order of the lux CDE genes cod- + hv (490 nm) ing for the fatty acid reductase complex is the same in all operons. The lux C and lux D genes, which code A fatty acid reductase complex containing three for fatty acid reductase and acyl-transferase polypep- enzymes, a reductase, a transferase and a synthetase, tides, respectively, flank the luciferase genes upstream catalyzes the acid with water back into the fatty alde- and lux E, which code for acyl-protein synthetase hyde substrate to facilitate the continuous produc- being downstream (Figure 1.1). Downstream of lux tion of light (Nealson and Hastings, 1990; Meighen, E is located lux G which specifies flavin reductase. 1991). Structural genes coding for enzymes involved Upstream of the lux operon of Aliivibrio fisheri are in the bioluminescent reaction of bacteria are located found the genes lux I and lux R, which are involved in the lux operon (Figure 1.1). The genes lux A and in quorum sensing, as discussed later. Similar content lux B code for the α and β subunits of luciferase, of the bacterial lux genes, their organization, and 0002073813.INDD 7 2/19/2014 3:27:15 PM 8 Chapter 1 Photobacterium C D A B F E G Symbiotic Luminescent Bacteria phosphoreum in Fish Light Organs Photobacterium C D A B F G leiognathi The anatomical, physiological and behavioral Vibrio C D A B E G H harveyi ­expression of luminescence reaches its zenith in Vibrio fishes, being more diverse and complex than in any C D A B E G cholerae other group of organisms. Overall, the majority of Aliivibrio luminescent fishes produce their own light in R I C D A B E G fisheri intrinsic numerous photophores, whereas only a Shewanella minority forms symbiotic associations with lumi- R I C D A B E G hanedai nescent ­bacteria which they harbor in few light Photorhabdus C D A B E organs. Partnerships with luminous bacteria, the luminescens subject of this chapter are only formed by approxi- mately 500 species, constituting of less than 2% of all recognized fish species. However, these species Figure 1.1 Organization of the lux operons of are members of 21 families and 7 orders (Table 1.1). a variety of luminescent marine, freshwater and In contrast to the diversity of host fishes terrestrial bacteria including two light organ (Figure 1.2D, 1.2E, 1.2F), the symbiotic bacteria are symbionts. (Peat and Adams 2008. Reproduced few, relatively closely related, and found in three with permission of Springer Science+Business monophyletic groups (Figure 1.3). These bacteria Media B.V.) consist of one group of facultative symbionts of the genera Photobacterium (Figure 1.2A, 1.2B, 1.2C) homology, including those of the unculturable and Aliivibrio and two groups of not yet identified Photodesmus katoptron, support the view that bacteria apparently occurring obligatorily with ­bacterial luminescence arose apparently once flashlight fishes and deep sea anglerfishes. These (Meighen, 1991; Dunlap and Ast, 2005; Peat and symbionts were suggested to be unable to repro- Adams, 2008; Hendry and Dunlap, 2011). duce outside the fish light organs due to extreme Bacteria emit light in an energy conserving manner specialization and metabolic integration with their depending on population density, termed quorum hosts (Herring and Morin, 1978; Haygood, 1993; sensing. The bacteria release into the growth media a Urbanczyk et al., 2011). signal molecule, termed an inducer, which accumulates Light produced by bioluminescent organisms at and triggers luminescence only after a critical threshold the water surface is too dim to be functional in full concentration is crossed. Below a critical density the sunlight or moonlight. In shallow water ­lumi­nous generated light will probably not be seen by higher species of fishes are, therefore, crepuscular or organisms and is, therefore, nonbeneficial. The regula- ­nocturnally active (e.g., flashlight fishes) whereas tory mechanism of auto induction was first studied in deep water fishes (e.g., ceratioid anglerfishes) may Aliivibrio fischeri. This bacteria secrets a highly specific use light irrespective of the circadian light cycle auto inducer purified as β-ketocaproyle ­homoserine (Morin, 1981; Haygood, 1993). Luminescence in lactone. A. fischeri are usually nonluminous in sea water coastal water fishes is more often of a bacterial while free living when their concentration is usually than of an endogenous origin. This phenomenon below 102/ml. However, when a density of 107/ml is was suggested to be in some way related to the reached the inducer activates the lux R regulatory relative abundance of luminescent bacteria in gene, which in a simplistic manner acts as a specific
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