This action might not be possible to undo. Are you sure you want to continue?
These interactions are typically assumed harmful as with pathogenic infections, but are often beneficial. One of the most important relationships is the mutualistic symbiosis whereby both the host and symbiont benefit. For example, Bacteroides in the intestines of mammals acquires nutrients from their hosts while breaking down complex polysaccharides into short chained fatty acids for the host (1). Ruminococcus in the cow’s rumen digests starch while acquiring nutrients from the cow (2). Long-term associations are typically initiated early in life and the host must be able to recognize and select for its symbiont. The Hawaiian bobtail squid Euprymna scolopes and the bioluminescent bacteria Vibrio fischeri make up a good system for symbiosis studies. V. fischeri is an excellent subject for study because it is the only species that can colonize in the E. scolopes light organ. This reduces the chance for contamination and interference when studying the symbiosis. Also, the symbiotic model is tractable because both V. fischeri and the squid are easily manipulated in the lab. The basis of the symbiosis is the ability of V. fischeri to provide protective luminescence camouflage to the squid E. scolopes which is a target for predators in the ocean; the camouflage eliminates the shadow that the squid may cast from moonlight. At the same time, E. scolopes provides growth nutrients including carbon and nitrogen for V. fischeri. Hatchling squid do not initially have symbionts in their nascent light organs. The V. fischeri symbionts are acquired after exposure to sea water, which contains a plethora of bacteria. While E. scolopes does not have adaptive immunity, it can discriminate between the millions of bacteria it contacts to select V. fischeri as the light organ colonizing species (3).
Using substrates oxygen and aldehyde.1 Even though there are aspects of the symbiosis that aren’t completely understood. Iron acquisition and utilization is essential for bacterial growth and likely contributes to symbiosis (7). H2O2 may be produced which creates oxidative stress on the bacteria (5). Motility and chemotaxis are important properties that allow the bacteria to travel to the light organ (9). sugar and nitrogen metabolism.which creates oxidative stress (Figure 2). Squid hemocytes Formatted: Font: Italic . phenotypes such as resistance to host generated oxidation (6). it can only partially reduce oxygen. which continues to stimulate luminescence in the colony. Also. when luciferase is present without aldehyde. LuxAB encodes for luciferase. lLux R codes for the lux gene regulator that senses AI. fischeri. which yields necessary substrate for luminescence. andwhich turns on the rest of the lux operon. luminescence is well characterized and known to be the main factor that is necessary in the squid-Vibrio relationship. luxCDE encodes for proteins which are necessary for aldehyde production. and hemocyte binding also contribute to V. production of siderophore. LuxI encodes for the AI molecule (3-oxohexanoyl l-homoserine lactone). Siderophore is a soluble iron chelator that helps the bacteria gather iron in low iron level environments. fischeri colonization of squid. Luminescence is directed by quorum sensing. (Figure 1). Nutrient utilization is important for growth in the light organ (8) by V. producing O2. autoinducer (AI) molecules accumulate to a high enough concentration to activate the lux operon which encodes for factors necessary for luminescence. which is responsible for catalyzing luminescence. luciferase emits light. In the presence of high amounts of oxygen and without luciferase. In addition to luminescence. which is a cell-to cell signaling mechanism typical of interacting bacterial species (4). When the bacterial population reaches high density. motility.
oxidative response. siderophore production. it is possible that luminescence is linked to these other traits. and linking light to other necessary trait would mechanistically prevent cheaters from arising. other traits could also directly affect the symbiosis relationship. fischeri (10). evaluate luminescence of various Vibrio fischeri mutants. or hemocyte binding. fischeri mutants and other bacterial species than they do to wild-type V. the linkage could be the factor that makes both luminescence and the associated trait essential to V. I think your prior description of the defect of lux mutants is necessary here. Because of this. necessary for survival linked to the lux gene expression or bioluminescence that to promotes symbiosis. fischeri colonization. scolopes. So you need to talk about cheaters. Hypothesis: While luminescence is essential for colonization. there are other traits. such as metabolism. And then you can say the defect is either due directly to bioluminescence because it enhances fitness/survival directly. While luminescence is necessary for V. To verify this idea. fischeri colonization of E. Y then need to go into the idea of how selection ma lead to this evolution (recall we discussed the difference between population pressure and squi pressure.2 bind better to V. various V. discriminating between symbionts and other bacteria. Objectives: To identify mutations that influence luminescence To identify other traits associated with those mutation To develop and test specific hypotheses about how traits linked to light production influence decreased survival of LuxA mutants in squid. fischeri mutants could be studied using luminescence and molecular techniques to identify linkage. or some other attribute i linked to light production that promotes fitness. Furthermore. . Methods: Location: Rudman Hall room 222 Comment [CW1]: There is really no explanati here for why other traits would be linked to luminescence.
Figure 2. luxI. .3 9. bacteria secrete autoinducers which instigate activation of lux gene in V. fischeri population in wild-type colonization after 48 hours. Black bars represent the population after 24 hours and striped bars represent the population after 48 hours. luxA. a) In quorum sensing. b) Larger population would have stronger luminescence (13). and luxR . fischeri. dropped in population significantly after 48 hours (2). all of which are dark mutants. There is no significant drop in V. Appendices Figure 1.
the expelled bacteria are used to colonize new bacteria. This process is done for 15 squid (4) . luciferase only partially reduces oxygen to O2. Procedure for evolution of mutants. Figure 4.4 Figure 3. Hatchling squid are colonized with bacteria and then each day they expel 95% of the bacteria and re-grow the population. Without aldehyde. With aldehyde.and does not generate light (12). luciferase enzyme reduces oxygen and produces light. After 4 days.
5. 2009. Microbiol. K. Photobiol. 2: 632-642 4. Tu. J. 2008. Journal Of Applied Microbiology 103: 2065-2073. Photochem.S. McFall-Ngai. L. 1998. 187:3603-3606. The periplamic group III catalase of Vibrio fischeri is required for normal symbiotic competence and is induced both by oxidative stress and by approach to stationary phase. 3. The Winnowing: establishing the squid–vibrio symbiosis. Bacteriol. Nyholm. identification and isolation as a dominant community member in the rumen of cattle fed a barley diet. J. J. S. . Sci. and E. and E. 1998. L. 95:1818-1822. 2004. 5:522-526 2. Proc. Ouwerkerk. Ruminococcus bromii. Activity coupling and complex formation between bacterial luciferase and flavin reduction. Bacteriol. Layers of Signaling in a Bacterium-Host Association. Nat.. K. 2000. Microbe. Cell Host &. Ruby. Visick. G. Acad. Klieve A. and M. L. M. Importance of Glycans to the Host-Bacteroides Mutualism in the Mammalian Intestine. Graf. McMillen. 2007. O’Leary . 37:168-179. Host-derived amino acids support the proliferation of symbiotic bacteria.5 Bibliography Vibrio fischeri lux Genes Play an Important Role in Colonization and Development of the Host Light Organ 1. Sci. U. 8. L. 2005. Comstock. 180:2087-2092. G. Novel effects of a transposon insertion in the Vibrio fischeri glnD gene: defects in iron uptake and symbiotic persistence in addition to nitrogen utilization. Mol. G. S. Graf. 6. J. and D. Visick. Ruby. 7. Ruby. and E.E.A.
J Bacteriol. J. 10:414-20.D. 12. Nature Reviews Microbiology. Bacteriol. McFall-Ngai. 1999. 11:483-493. 2004. Heterogeneous Response to a Quorum-Sensing Signal in the Luminescence of Individual Vibrio fischeri.. B. 11. J. Ruby. Perry. D. E. M.. Vibrio fischeri Flagellin A Is Essential for Normal Motility and for Symbiotic Competence during Initial Squid Light Organ Colonization. and E. 2010. Doino. Nyholm. 2000. Hagen.A. K. McFall-Ngai. Ruby. G.J. P. Schuster. M. 186:4315-4325. L. J.L. Whistler. Visick. and E. Millikan. . Ruby. Perez. McFall-Ngai. Environmental Microbiology. Oxygen-utilizing reactions and symbiotic colonization of the squid light organ by Vibrio fischeri. 15. 2:632-642. 10. 14. S. Cooper. G. 13. and M. Vibrio fischeri lux Genes Play an Important Role in Colonization and Development of the Host Light Organ. Ruby.S. E.G. 2009. and S. 182: 4578-4586. Breaking the language barrier: and experimental evolution of non-native Vibrio fischeri in squid tailors luminescence to the host. and C. Recognition between symbiotic Vibrio fischeri and the haemocytes of Euprymna scolopes. Microbiol.6 9. 2004. J. Symbiosis. Stabb E. S.. Trends. V. Personal Communication. J.A. Stewart. Foster.
This action might not be possible to undo. Are you sure you want to continue?
We've moved you to where you read on your other device.
Get the full title to continue reading from where you left off, or restart the preview.