Symbiotic association of Photobacterium fischeri with the marine luminous fish Monocentris japonica; a model of symbiosis based on bacterial studies.

Isolation of bacteria from the luminous organ of the fish Monocentris japonica has revealed that the organ contains a pure culture of luminous bacteria. For the four fish examined, all contained Photobacterium fischeri as their luminous bacterial symbiont. This is the first time that P. fischeri has been identified in a symbiotic association. A representative isolate (MJl) of the light organ population was selected for in vivo studies of its luminous system. Several physiological features suggest adaptation for symbiotic existence. First, MJl has been shown to produce and respond to an inducer of luciferase that could accumulate in the light organ. Secondly, the specific activity of light production was seen to be maximal under low, growth-limiting concentrations of oxygen. Thirdly, unlike another luminous species (Beneckea harveyi), synthesis of the light production system of these bacteria is not catabolite repressed by glucose--a possible source of nutrition in the light organ. Fourthly, when grown aerobically on glucose these bacteria excrete pyruvic acid into the medium. This production of pyruvate is a major process, accounting for 30-40% of the glucose utilized and may serve as a form of regulatory and nutritional communication with the host.     

Morphology of the Luminous Organ of the Squid Loligo duvaucelid'Orbigny, 1839

The present paper deals with the morphology of the luminous organ of the squid Loligo duvauceli, caught in the sea off Indonesia and Thailand. Two luminous organs are situated on the ventral surface of the ink sac, near the anus. Each organ consists of a luminous sac divided into numerous discrete chambers containing bacteria. On the ventral side the organ is bordered by a lens-like structure, consisting of muscle cells. On the remaining sides the bacterial chambers are limited by a cup-shaped reflector layer with numerous parallel lamellae. The reflector separates the bacterial chamber from the ink sac. A ciliated channel connects the interior of the bacterial chamber with the mantle cavity.     

Dual bioluminescent systems in the anglerfish genus Linophryne (Pisces: Ceratioidea)

Females of the anglerfish genus Linophryne bear barbels containing luminous organs, in addition to an escal light organ. Luminescence has been observed from the barbels of four species of Linophryne , and the morphology of the luminous organs investigated. The barbel light organs do not contain bacteria but complex paracrystalline photogenic granules. The esca contains luminous bacteria. The esca is ectodermal in origin whereas the barbel organs may be derived from the mesoderm.
The possible significance of this unique dual system of luminous organs is discussed.     

The Luminescent Systems of Pony Fishes

The anatomy of bioluminescent organs and mode of light production in 18 species of pony fish have been investigated using fresh and preserved material. The luminescent systems are similarly arranged in all. Basically, the system consists of a light organ located at the distal end of the esophagus, and a series of abdominal accessory structures positioned in tandem for controlling light intensity and for directing and dispersing the light. Light is produced by numerous symbiotic luminous bacteria in the light organ. A simple classification of the luminescent systems is proposed. The light organs of Leiognathus elongatus and L. rivulatus show marked sexual dimorphism. The bacteria present in the light organs of many pony fishes are easily culturable, but not those from L. elongatus. Electron micrographs of the light organs of L. elongatus and L. rivulatus show the presence of numerous rod-shaped bacteria measuring approximately 0.8 µ x 2.4 µ and 0.8 µ x 7.3 µ, respectively. It is concluded that the light organ of L. elongatus contains another example of a type of non-culturable luminous bacteria that have been found elsewhere. Such bacteria appear to require from the host some special factor for growth and luminescence.     

A New Species of Luminous Percoids of the Acropoma Genus (Acropomatidae)

Journal of Ichthyology - A new species, Acropoma leobergi sp. n. from the Arafura Sea, belonging to a group of species with a short U-shaped luminous organ slightly receding behind the anus, is...     

Competition between Vibrio fischeri strains during initiation and maintenance of a light organ symbiosis.

Colonization of the light-emitting organ of the Hawaiian squid Euprymna scolopes is initiated when the nascent organ of a newly hatched squid becomes inoculated with Vibrio fischeri cells present in the ambient seawater. Although they are induced for luminescence in the light organ, these symbiotic strains are characteristically non-visibly luminous (NVL) when grown in laboratory culture. The more typical visibly luminous (VL) type of V. fischeri co-occurs in Hawaiian seawater with these NVL strains; thus, two phenotypically distinct groups of this species potentially have access to the symbiotic niche, yet only the NVL ones are found there. In laboratory inoculation experiments, VL strains, when presented in pure culture, showed the same capability for colonizing the light organ as NVL strains. However, in experiments with mixed cultures composed of both VL and NVL strains, the VL ones were unable to compete with the NVL ones and did not persist within the light organ as the symbiosis became established. In addition, NVL strains entered light organs that had already been colonized by VL strains and displaced them. The mechanism underlying the symbiotic competitiveness exhibited by NVL strains remains unknown; however, it does not appear to be due to a higher potential for siderophore activity. While a difference in luminescence phenotype between VL and NVL strains in culture is not likely to be significant in the symbiosis, it has helped identify two distinct groups of V. fischeri that express different colonization capabilities in the squid light organ. This competitive difference provides a useful indication of important traits in light organ colonization.     

Genomic and phylogenetic characterization of luminous bacteria symbiotic with the deep-sea fish Chlorophthalmus albatrossis (Aulopiformes: Chlorophthalmidae)

Bacteria forming light-organ symbiosis with deep-sea chlorophthalmid fishes (Aulopiformes: Chlorophthalmidae) are considered to belong to the species Photobacterium phosphoreum. The identification of these bacteria as P. phosphoreum, however, was based exclusively on phenotypic traits, which may not discriminate between phenetically similar but evolutionarily distinct luminous bacteria. Therefore, to test the species identification of chlorophthalmid symbionts, we carried out a genomotypic (repetitive element palindromic PCR genomic profiling) and phylogenetic analysis on strains isolated from the perirectal light organ of Chlorophthalmus albatrossis. Sequence analysis of the 16S rRNA gene of 10 strains from 5 fish specimens placed these bacteria in a cluster related to but phylogenetically distinct from the type strain of P. phosphoreum, ATCC 11040(T), and the type strain of Photobacterium iliopiscarium, ATCC 51760(T). Analysis of gyrB resolved the C. albatrossis strains as a strongly supported clade distinct from P. phosphoreum and P. iliopiscarium. Genomic profiling of 109 strains from the 5 C. albatrossis specimens revealed a high level of similarity among strains but allowed identification of genomotypically different types from each fish. Representatives of each type were then analyzed phylogenetically, using sequence of the luxABFE genes. As with gyrB, analysis of luxABFE resolved the C. albatrossis strains as a robustly supported clade distinct from P. phosphoreum. Furthermore, other strains of luminous bacteria reported as P. phosphoreum, i.e., NCIMB 844, from the skin of Merluccius capensis (Merlucciidae), NZ-11D, from the light organ of Nezumia aequalis (Macrouridae), and pjapo.1.1, from the light organ of Physiculus japonicus (Moridae), grouped phylogenetically by gyrB and luxABFE with the C. albatrossis strains, not with ATCC 11040(T). These results demonstrate that luminous bacteria symbiotic with C. albatrossis, together with certain other strains of luminous bacteria, form a clade, designated the kishitanii clade, that is related to but evolutionarily distinct from P. phosphoreum. Members of the kishitanii clade may constitute the major or sole bioluminescent symbiont of several families of deep-sea luminous fishes.     

STUDIES ON BIOLUMINESCENCE : XVII. FLUORESCENCE AND INHIBITION OF LUMINESCENCE IN CTENOPHORES BY ULTRA-VIOLET LIGHT.

1. Small dumps of the luminous cells of Mnemiopsis cannot readily be stimulated mechanically but will luminesce on treatment with saponin solution. Larger groups of luminous cells (such as are connected with two paddle plates) luminesce on mechanical stimulation. This suggests that mechanical stimulation to luminesce occurs chiefly through a nerve mechanism which has been broken up in the small dumps of luminous tissue. 2. The smallest bits of luminous tissue, even cells freed from the animal by agitation, that will pass through filter paper, lose their power to luminesce in daylight and regain it (at least partially) in the dark. 3. Luminescence of the whole animal and of individual cells is suppressed by near ultra-violet light (without visible light). 4. Inhibition in ultra-violet light is not due to stimulation (by the ultra-violet light) of the animal to luminesce, thereby using up the store of photogenic material. 5. Animals stimulated mechanically several times and placed in ultra-violet light show a luminescence along the meridians in the same positions as the luminescence that appears on stimulation. This luminescence in the ultra-violet or "tonic luminescence," is not obtained with light adapted ctenophores and is interpreted to be a fluorescence of the product of oxidation of the photogenic material. 6. Marked fluorescence of the luminous organ of the glowworm (Photuris) and of the luminous slime of Chatopterus may be observed in ultra-violet but no marked fluorescence of the luminous substances of Cypridina is apparent. 7. Evidence is accumulating to show a close relation between fluorescent and chemiluminescent substances in animals, similar to that described for unsaturated silicon compounds and the Grignard reagents.     

Animal-Bacterial Interactions in the Early Life History of Marine Invertebrates: The Euprymna scolopes/Vibrio fischeri Symbiosis

SYNOPSIS. The symbiotic association between the Hawaiian sepiolidsquid Euprymna scolopes and the marine luminous bacterium Vibriofischeri is being developed as a model system for the studyof animal-bacterial interactions during development. Changesin light organ morphology during embryogenesis foster successfulinfection of the light organ with the proper bacterial partner.These embryonic events of light organ morphogenesis includethe elaboration of an epithelial surface with a complex ciliated,microvillous field. The squid host hatches without the bacterialsymbionts, but acquires them within hours from the free-livingpopulation of the bacteria in the water column. Upon exposureto the proper symbionts, the host organ undergoes a series ofmorphogenetic changes, including loss of the ciliated, microvillousfield. The light organ then goes on to mature into a morphologicalconfiguration that serves to promotethe maintenance of a stableassociation with the bacteria and that correlates with the useof the bacterial bioluminescence in behavior of the host. Thissymbiosis is discussed in light of other cyclically transmittedanimal-bacterial associations.     

Mechanisms of light organ occlusion in flashlight fishes, family Anomalopidae (Teleostei:Beryciformes), and the evolution of the group

The circumtropical, nocturnal, shore-fish family Anomalopidae is characterized by a subocular luminous organ containing symbiotic luminous bacteria. The five known species are placed in four genera, one of which is new. Phthanophaneron is restricted to the eastern Pacific, Kryptophanaron to the western Atlantic, Photoblepharon is Indo-West Pacific in distribution and Anomalops is west Pacific. The symbiotic bacteria emit light continuously, and two superficially different mechanisms of occluding the glowing face of the organ are found. In Photoblepharon a black shutter of elastic skin is drawn up over the face of the organ, whereas in Anomalops the organ is rotated downward, so that only the heavily pigmented back of the organ is exposed. In Phthanophaneron and Kryptophanaron, both rotational and shutter mechanisms are present. Elucidation of the structures and linkages involved in light-organ occlusion reveals that the superficially different mechanisms are based on a common functional complex. In all four genera, the light organ is supported by a cartilaginous cup that articulates anteriorly with a cartilaginous stalk. Motive power for both the shutter and rotational mechanisms is supplied by the adductor mandibulae through a complex biomechanical linkage involving the ethmomaxillary ligament and a ligament unique to anomalopids, the Ligament of Diogenes. The structures involved in shutter erection and organ rotation are illustrated and described in detail for Photoblepharon and Anomalops and are compared with those in the other two forms; a functional hypothesis is advanced. Extrafamilial relationships of the Anomalopidae are discussed, and a hypothesis of the phylogenetic relationships of the four genera is derived from a cladistic analysis involving 19 non-light-organ characters and corroborated by some light-organ characters. Most characters associated with the light-organ complex cannot be polarized by conventional outgroup comparison, and the evolution of the light organ occlusion mechanisms is interpreted in light of the hypothesized phylogeny and a hypothesized ancestral mechanism. We propose that the common ancestor of anomalopids possessed a forced rotational mechanism like that of Phthanophaneron and Kryptophanaron. This was refined to a more efficient flipping rotational mechanism in Anomalops, the sister group of the lineage comprising the other three genera, within which the shutter mechanism was progressively refined. The ostensibly unnecessary complexity of the shutter mechanism is apparently a result of functional-morphological constraints imposed on the system by the pre-existence of a rotational mechanism. A brief zoogeographic scenario is proposed.     

Luminous bacteria and light emitting fish: ultrastructure of the symbiosis

The luminescent fish Monocentris japonicus uses symbiotic luminous bacteria as a source of light. These bacteria live in light organs, complex tissue compartments, consisting of richly vascularized tubules or canals (in which the bacteria are cultured) lined with mitochondria-rich epithelial cells. The structure is consistent with a proposed model of symbiosis in which nutrients and oxygen are supplied by the vertebrate blood (vascular system). The nutrients, oxidized by the bacteria for growth and light production, are returned in part to the fish as pyruvate, which by reacting with mitochondrial oxygen regulates the light organ oxygen tensions. The luminous bacteria provide steady light that is modulated by passage through the melanocyte-containing dermis of the fish. Both the fish and the bacteria are highly adapted for their symbiotic coexistence.     

Squid-derived chitin oligosaccharides are a chemotactic signal during colonization by Vibrio fischeri

Chitin, a polymer of N-acetylglucosamine (GlcNAc), is noted as the second most abundant biopolymer in nature. Chitin serves many functions for marine bacteria in the family Vibrionaceae ("vibrios"), in some instances providing a physical attachment site, inducing natural genetic competence, and serving as an attractant for chemotaxis. The marine luminous bacterium Vibrio fischeri is the specific symbiont in the light-emitting organ of the Hawaiian bobtail squid, Euprymna scolopes. The bacterium provides the squid with luminescence that the animal uses in an antipredatory defense, while the squid supports the symbiont's nutritional requirements. V. fischeri cells are harvested from seawater during each host generation, and V. fischeri is the only species that can complete this process in nature. Furthermore, chitin is located in squid hemocytes and plays a nutritional role in the symbiosis. We demonstrate here that chitin oligosaccharides produced by the squid host serve as a chemotactic signal for colonizing bacteria. V. fischeri uses the gradient of host chitin to enter the squid light organ duct and colonize the animal. We provide evidence that chitin serves a novel function in an animal-bacterial mutualism, as an animal-produced bacterium-attracting synomone.     

Enlightenment of old ideas from new investigations: more questions regarding the evolution of bacteriogenic light organs in squids

Bioluminescence is widespread among many different types of marine organisms. Metazoans contain two types of luminescence production, bacteriogenic (symbiotic with bacteria) or autogenic, via the production of a luminous secretion or the intrinsic properties of luminous cells. Several species in two families of squids, the Loliginidae and the Sepiolidae (Mollusca: Cephalopoda) harbor bacteriogenic light organs that are found central in the mantle cavity. These light organs are exceptional in function, that is, the morphology and the complexity suggests that the organ has evolved to enhance and direct light emission from bacteria that are harbored inside. Although light organs are widespread among taxa within the Sepiolidae, the origin and development of this important feature is not well studied. We compared light organ morphology from several closely related taxa within the Sepiolidae and combined molecular phylogenetic data using four loci (nuclear ribosomal 28S rRNA and the mitochondrial cytochrome c oxidase subunit I and 12S and 16S rRNA) to determine whether this character was an ancestral trait repeatedly lost among both families or whether it evolved independently as an adaptation to the pelagic and benthic lifestyles. By comparing other closely related extant taxa that do not contain symbiotic light organs, we hypothesized that the ancestral state of sepiolid light organs most likely evolved from part of a separate accessory gland open to the environment that allowed colonization of bacteria to occur and further specialize in the eventual development of the modern light organ.     

发光甲虫与生物荧光

生物荧光是活体生物自身可以发光的有趣生命现象。具有这一现象的生物存在于生物四界中,但目前关于这一现象的研究报道主要来自于昆虫,尤其是以萤火虫为代表的发光甲虫的研究。文章对发光甲虫的分类地位、生物荧光发生的原理、发光器官的类型、闪光的“开关”机制、生物荧光的生物学意义及其相关行为学研究进展等进行了详细介绍。此外,还简要提及了荧光生物及其荧光酶的应用。这对了解及探讨生物荧光现象、加强对中国的发光甲虫及其它发光生物的研究及保护利用具有一定的借鉴作用。     

Detection of the Light Organ Symbiont, Vibrio fischeri, in Hawaiian Seawater by Using lux Gene Probes

Symbiotic bacteria that inhabit the light-emitting organ of the Hawaiian squid Euprymna scolopes are distinctive from typical Vibrio fischeri organisms in that they are not visibly luminous when grown in laboratory culture. Therefore, the abundance of these bacteria in seawater samples cannot be estimated simply by identifying them among luminous colonies that arise on nutrient agar plates. Instead, we have used luxR and polymerase chain reaction generated luxA gene probes to identify both luminous and non-visibly luminous V. fischeri colonies by DNA-DNA hybridization. The probes were specific, hybridizing at least 50 to 100 times more strongly to immobilized DNAs from V. fischeri strains than to those of pure cultures of other related species. Thus, even non-visibly luminous V. fischeri colonies could be identified among colonies obtained from natural seawater samples by their probe-positive reaction. Bacteria in seawater samples, obtained either within or distant from squid habitats, were collected on membrane filters and incubated until colonies appeared. The filters were then observed for visibly luminous V. fischeri colonies and hybridized with the lux gene probes to determine the number of total V. fischeri colonies (both luminous and non-visibly luminous). We detected no significant differences in the abundance of luminous V. fischeri CFU in any of the water samples observed (≤1 to 3 CFU/100 ml). However, probe-positive colonies of V. fischeri (up to 900 CFU/100 ml) were found only in seawater collected from within the natural habitats of the squids. A number of criteria were used to confirm that these probe-positive strains were indistinguishable from symbiotic V. fischeri. Therefore, the luxA and luxR gene probes were species specific and gave a reliable estimate of the number of culturable V. fischeri colonies in natural water samples.     

The specificity of symbiosis: Pony fish and luminescent bacteria

Luminescent bacteria isolated from light organs of seven different species (3 genera) of fishes of the family Leiognathidae were subjected to taxonomic analysis. Of the 733 isolated all but seven were identified as Photobacterium leiognathi; the others are considered to be either chance contaminants of the sampling procedure or transients within the organ. In most fish, the luminous organ appeared to contain a single predominating strain of P. leiognathi with small numbers of one to three other strains of the same species, differing by only one or two characters.     

Light production by the arm tips of the deep-sea cephalopod Vampyroteuthis infernalis

The archaic, deep-sea cephalopod Vampyroteuthis infernalis occurs in dark, oxygen-poor waters below 600 m off Monterey Bay, California. Living specimens, collected gently with a remotely operated vehicle (ROV) and quickly transported to a laboratory ashore, have revealed two hitherto undescribed means of bioluminescent expression for the species. In the first, light is produced by a new type of organ located at the tips of all eight arms. In the second, a viscous fluid containing microscopic luminous particles is released from the arm tips to form a glowing cloud around the animal. Both modes of light production are apparently linked to anti-predation strategies. Use of the tip-lights is readily educed by contact stimuli, while fluid expulsion has a much higher triggering threshold. Coelenterazine and luciferase are the chemical precursors of light production. This paper presents observations on the structure and operation of the arm-tip light organs, the character of the luminous cloud, and how the light they produce is incorporated into behavioral patterns.