J. Terr. Mar. Res. Vol.

1(4) 2017:71-76
Journal of Terrestrial and Marine Research
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Short Communication

Construction of hypothetical lux operon of ancestral luminous bacteria

CH. RAMESH* and R. MOHANRAJU

1
Department of Ocean Studies and Marine Biology, Pondicherry Central University,
Port Blair-744112, Andaman & Nicobar Islands, India.
2
Present address: Andaman Nicobar Center for Ocean Science and Technology (ANCOST),
National Institute of Ocean Technology (NIOT), Dolygunj, Port Blair-744103,
Andaman & Nicobar Islands
Abstract
Notions are always essential to inspect the past, present and future events. The present study constructed the
hypothetical lux operon of luminous bacterial ancestor i.e. “luxZYLOPUMNQRSTICDABFEGH-ribEBHA” by
assembling the regulatory lux genes, structural lux genes and other flanking genes that exists at upstream and
downstream of the lux operons of different luminous bacterial species. All the genes on lux operon were arranged
according to base pairs. This kind of lux operon carrying ancestor might not been isolated yet or extincted, and/or we
expect its possible existence. This hypothetical lux operon forms a sort of connecting link between luminous bacterial
ancestor (LBA) and modern luminescent bacteria (MLB). However, functional variation of lux genes of LBA and MLB
are not addressed here. The processes of “Horizontal gene transfer, insertion, deletion, and interchromosomal
rearrangements, geological processes are appear to support the origin of MLB. Some possible hypothetical theories on
propagation of MLB to different environments are also detailed.

Keywords: Lux operon, lux genes, ancestral luminous bacteria, horizontal gene transfer.

Introduction
Bacterial bioluminescence is an enzymatic production of
light which is regulated by the lux genes. The presence
of these lux genes on chromosome are depend on the
species since some species possess one chromosome and
some with two (Okada et al., 2005). Lux genes act as
accessory genes which are related to environmental
factors. Accessory genomes are able to transfer from one
bacterium to the other for better adaptations to specific
niche (Laing et al., 2010). Genetic exchange among the
species of Vibrionaceae appears to be benefit in adapting
new environmental conditions (Lilburn et al., 2010).
The presence and absence of some lux genes in
different luminous bacterial species (Dunlap, 2009)
appear to be a concern that these luminescent bacteria
how actually originated and why some bacteria does not
carry some lux genes. Different assumptions have related
the evolution of bacterial bioluminescence on basis of
lux genes and other protein paralogous (O' Kane and
Prasher, 1992) and oxygen based mechanism (Rees et
al., 1998; Timmins, 2001). In a different view we
propose a new concept on the existence of ancestral
luminous bacterial lux operon that may be the origin for
all MLB.
1. Horizontal gene transfer (HGT)
Luminescent bacteria are the members of 3 families of
Vibrionaceae, Shewanellaceae and Enterobacteriaceae.
Principally, inheritance of lux genes among these
luminescent strains is due to conjugation. However, the
transfers of lux genes (accessory genes) among
luminescent bacteria are also apparent due to “HGT or
chromosomal exchange mechanism or gene acquisition”.
In this phenomenon the mobility of unstable DNA
fragments transfers within the species or even to other
non-related bacteria (Ziebuhr et al., 1999). Without the
requirement of cell division, the unstable or stable
mobile genetic elements are found to transfer as entire
operon or as individual gene into recipients (Rapa and
Labbate, 2013; Kirkup et al., 2010; Gray and Garey,
2001). These mobile genetic elements are a wide source

Manuscript Information
Received: 14th July 2017 / Revised: 08th August 2017 / Accepted: 12th August 2017 / Published: 15 th August 2017
/ Corresponding Author: chrameshpu@gmail.com

Journal of Terrestrial and Marine Research 71

 Construction of hypothetical lux operon of ancestral luminous bacteria
for evolution (Frost et al., 2005). According to Kasai et
al. (2007), the lux gene cluster was distributed from
Vibrio cholerae to the other luminous bacteria. They also
deduced that it is difficult to find the pathway through
which the origin of original lux operon was actually
transferred. Recent reports on luminescent strains of P.
aquimaris has revealed natural vertical inheritance of
lux-rib operon and also horizontally acquired lux-rib
operon (Urbanczyk et al., 2012). Whereas, in certain
strains HGT process (recruitment time of lux genes)
under certain unknown environmental factors is not
understood (Urbanczyk et al., 2008). HGT is not only
within the luminescent bacteria (HGT of lux genes) but
also found in other non-luminescent bacteria (HGT of
16s rRNA) (Passel et al., 2006). It indicates that HGT
possibly be an evolutionary mechanism that is why the
lux gene sequences of some luminescent and non-
luminescent bacteria are appear to be homologous.
2. Other Genetical factors
Several genetical factors like inversion of genes
(as in A. fisheri) (Kirkup et al., 2010), gene duplications
(from luxA to luxB and luxB to luxF), gene loss (luxF in
P. leiognathi, and ribE in P. phosphoreum), mutations in
gene (luxF of P. mandapamensis), gene recruitment (1.
ribB in V. harveyi and A. fischeri, 2. luxR and luxI in A.
fischeri and A. salmonicida, 3. Lux genes in V. chagasii,
P. damselae, and V. vulnificus), gene shift (luxI in A.
salmonicida), unlinked genes (luxR of V. harveyi and
ribB in A. fischeri), HGT (1. as in A. fischeri and S.
hanedai, 2. Lux gene transfer into V. chagasii, P.
damselae, and V. vulnificus,), transposon-mediated
transfer (rib operon placement in P. leiognathi starins)
(Dunlap, 2009), and interchromosomal rearrangements
in low frequency (observed in certain Vibrio species)
(Makino et al., 2003) have arose MLB.
Natural environment strains are appear to show
good intensity of luminescence, while those strains
isolated in laboratory are found to become dim or dark
(Dunlap and Urbanczyk, 2013). Some nonluminous
bacterial members of Vibrio cholerae are known to
possess lux genes and emit luminescence (Grim et al.,
2008), and some were found to remain nonluminescent
(Ramaiah et al., 2000; Zo et al., 2009). A non-
luminescent strain of V. campbellii found to bear luxA
gene (Hedreyda and Orata, 2011), and some
nonluminescent Vibrio strains with incomplete lux
operon (O’Grady and Wimpee, 2008). All these
incidents give an idea that luminous bacterial strains
have adapted themselves to the surrounding milieus by
acquiring physical and genetical modifications.
3. Geological processes
The extinction of some species and rise of new
species during the biological evolution is due to changes
in environmental conditions. Overtime as Pangaea began
splitting, changes and adaptations had been acquired by
microorganisms and macroorganisms. The changes in
global and local environmental climatic conditions are
potential sources for origin of genetically adopted
Journal of Terrestrial and Marine Research
microorganisms (Franks and Hoffmann, 2012). It was
inferred that nucleotide composition of microbial
communities are actively influenced by the environment
for adaptation (Foerstner et al., 2005). Bacterial
bioluminescence is appear to sensitive to environmental
osmolarity (Peat and Adams, 2008), indicating that
environmental factors might have influenced the genetic
content of LBA to give raise MLB.
4. Hypothetical theories on the origin and propagation
of MLB
Wind -water theory
This theory supports the belief of many researchers
that the ancestor of luminescent bacteria has originated
from marine (Dunlap and Kita-Tsukamoto, 2006). In
order to survive under different environmental
conditions LBA got changes in its lux operon and gave
origin to many new species. These new species started
spreading from marine to estuarine, fresh water and
terrestrial environments by sea breeze phenomenon or by
waves and currents, or by marine snow (Herren et al.,
2004). This theory also stresses that sea breezing
phenomenon might helped the propagation of surface
water luminescent bacterial strains into atmosphere or
from atmosphere to ocean. However no atmospheric
luminescent bacterial strains were reported so far, and
the authors believes the existence of these bacteria.
Migration theory
This theory is observed with salmon fish
Oncorhynchus kisutch that show anadromous migration.
According to Budsberg et al. (2003), during anadromous
migration, epizoic luminescent bacteria were protected
from environmental factors by salmon fish slime. We
assume that during anadromous migration marine
luminous bacteria might have adopted to estuarine and
freshwater environmental conditions by gradual changes
in physiological, metobolical, lux gene lose or
duplications and other factors as mentioned above.
Food web theory
According to this theory marine LBA that associated
with dissolved organic particles, dead or decaying tissue
pieces found on the beach and fish intestinal faeces
might have propagated to estuarine, freshwater and
terrestrial environments through food web or dietary
links. Luring nature of LBA also might have assisted its
distribution through anadromous and catadromous
organisms. This theory is also supported by the research
observations of Zarubin et al. (2012). Propagation of
LBA to terrestrial environment may be due to
amphibians, or cadavers (Kakizak et al., 2009), or other
anthropogenic activities- like in olden days many people
eat shrimps without cooking.
Bacteriophage theory
This theory defines the transduction of lux gens into
different environments by bacteriophages. It is known
that marine bacteriophages are one of the source for
HGT process (Weinbauer and Rassoulzadegan, 2004;
72

Arber, 2000). Luminous bacteria like V. harveyi are
known to host marine bacteriophages (Chrisolite et al.,
2008; Sekar and Kandasamy, 2013), and some luminous
Vibrio strains are appear to be sensitive to infection
caused by phages (Yetinson and Shilo, 1979). These
reports give a clue that LBA specific bacteriophages or
its associated phages in natural environment (not yet
known about luminescent bacterial species specific
bacteriophages) might have transferred LBA lux genes
into other bacterial species [e.g. luxS presence in more
than 40 bacterial species (Santiago-Rodriguez et al.,
2013)]. Another concept is that bacteriophages might
have uptake lux genes for developing photosynthesis
mechanism to be autotrophic.
Omnipotent Alpha-Omega cell (protocell) theory
According to this theory the protocell is the source
for spreading of “bags of genes” to single cell organisms
to multicellular organisms. We postulate that the
protocell might have possessed lux genes and in
evolutionary adaptation process these genes given origin
for different types of light emitting mechanisms. The
gene loss by protocell, and acquisition and gene
modifications by new born cells might have caused
slowly evolutionary variations in luminous bacteria.
Otherwise, according to central dogma- replication
process might have given origin to diverse hierarchical
lux operons, where as in transcription process different
proteins that are involved in luminescence emission
might have originated systematically. Therefore
ancestral marks are seen in anatomical structures, cell
organs, enzymes and genes of different micro and macro
organisms with identical or modified identities.
Figure 1. Lux genes of luminescent ancestor and the origin of MLB .
Journal of Terrestrial and Marine Research
J. Terr. Mar. Res. 1(3) 2017:1-76
Discussion
Based on the above phenomenons and using lux
genes (Meighen, 1994; Lin et al., 2001; Freeman and
Bassler, 1999; Herring, 2002), we have constructed
approximate lux operon order of LBA based on base
pairs of respective lux genes i.e. as follows
luxZYLOPUMNQRSTICDABFEGH-ribEBHA (Figure
1). There is a perfect view for this kind of LBA, when
we consider that the LBA had not undergone through
any of the processes like HGT or CEM, PDE, ICR, gene
duplication, gene loss, gene recruitment, gene mutations,
transposon mediated transfer and merodiploidy of gene.
While, when we include the above processes, there
appears the MLB with changes in their lux operon. We
believe that MLB might have arisen from this
constructed ancestor lux operon and adapted to
respective environments. This lux operon order may be
seen in other bacterial species as discordance
arrangement (Kasai et al., 2007). The presence of dual
luxR genes in A. salmonicida and A. logei give some
possible postulation that the existence of multiple lux
genes other than luxR can also be found (it is not yet
found in other lux genes). It is unknown that in which
conditions luminous bacteria use these processes like
HGT, PDE, gene loss, gene recruitment, duplication etc.
Apart from the mentioned processes, integrons cassette
that code for proteins related to adaptations might have
also involved in evolution of speciation of luminescent
bacteria and to adapt to different environments (Cambray
et al., 2010). Further, environmental factors are also
known to influence the nucleotide content of bacteria
(Thompson et al., 2009).
73
 Construction of hypothetical lux operon of ancestral luminous bacteria
However, the logical reasons behind
these genes ever were all together and evidence on
divergence of these gene contents over time are yet to be
solved. Our idea is that lux genes of LBA might be
regulating many functions, and overtime due to gene loss
or gene acquisition, the lux operon functions of MLB
might have restricted to few functions for betterment
(Figure 2). Therefore we assume a theory “One gene
Figure 2. Functions of lux genes of MLB (Red marked genes are non-lux genes which are linked to the lux operon).
Acknowledgements
The author grateful to the Department of Science and
Technology for providing the Inspire fellowship
DST/IF120230. Ramesh thanks the Science and
Engineering Research Board (SERB) for granting the
travel award SB/ITS-Y/01418/14-15 to present this work
in “The 18th International Symposium on
Bioluminescence and Chemiluminescence” on June 23 rd
to 28th, 2014 held at Uppsala Biomedical Centre – BMC,
Husargatan 3, Sweden.
Journal of Terrestrial and Marine Research
with multifunction to one gene single function- for better
cellular metabolism and gene regulation”. Further we
infer that whole genome maps of luminescent bacterial
strains will certainly be more reliable for understanding
accurate evolutionary studies and to inspect this type
hypothetical LBA.
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