Does Stress Causes Changes In Bacterial Community Associated With Soft Coral Plexaura homomalla?
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Does Stress Causes Changes In Bacterial Community Associated With Soft Coral Plexaura homomalla?
- 1. Does Stress Causes Changes In Bacterial Community Associated With Soft Coral Plexaura homomalla? Zeenat Tinwala University of Houston-Clear Lake Houston- Texas zeenat16@gmail.com
- 2. CORAL REEFS AND SOFT CORAL • Most diverse ecosystems - Economically valuable – Food, fishing, tourism and costal protection • Corals: marine invertebrates (animals), Phylum Cnidaria, Class Anthozoa • Soft corals: lack calcium carbonate skeleton • Producers of marine natural products (MNPs) with biological activity Eunicea fusca: fuscosides Plexaura homomalla: Prostaglandins 2
- 3. CORAL STRESS: BLEACHING • Corals - Stress • Bleaching – Loss of symbiotic dinoflagellate Symbiodinium sp. • Possible disruption of beneficial bacteria & pathogen susceptibility • Prolonged bleaching - coral death Photo: Great Barrier Reef Marine Parks Authority Bleaching Disease 3
- 4. PROJECT AIM • Aim: To identify bacterial species associated in the soft coral Plexaura homomalla during normal and stress conditions • Culture-independent technique (CIT) - 16S rDNA clone library analysis 4
- 5. • Non-stressed samples: Population of mostly Proteobacteria, especially beneficial gamma- Proteobacteria. • Atypical bacteria such as other Proteobacteria (especially pathogenic Vibrios), Firmicutes and Actinobacteria expected in stressed organisms. 5 HYPOTHESIS
- 6. Samples collected Summerland Key Reef, FL, June 2010 Three stress treatments (12 hours): • Heat: 31 C • LPS (Lipopolysaccharides): 10µg/ml • Control (27 C) Flash Frozen and stored at -80oC DNA Isolation NucleoSpin Soil – Macherey-Nagel PCR: 16S rDNA bacterial primers Clone Library (TOPO TA vector) RFLP (Restriction Fragment Length Polymorphism) HhaI and TaqI DNA Sequencing & Analysis 6
- 7. RFLP RESULTS CONTROL A A AA BB 500 100 bp HEAT A A A A AB B B B 100 bp 500 C D LPS A A A A AB 100 500 bp B AA 7
- 8. RESULTS RFLP Pattern # Control Clones # Heat Clones # LPS clones Pattern A 7 6 10 Pattern B 4 6 1 Pattern C - 1 3 Pattern D 2 - 1 Pattern E - 1 - Undefinable 6 5 4 8
- 9. FUTURE WORK 9 • Screen more clones • Sequence samples with different RFLP patterns • Construct a phylogenetic tree with sequence data • Compare results from other culture dependent (classic plate culture), independent techniques (DGGE) and to production of
- 10. CONCLUSION • Culture Independent techniques like RFLP and DGGE analysis are helpful preliminary basis in determining changes of bacteria in soft corals • Studying the changes in microbial flora of the corals might allow the design of biomarkers for prediction of early stress 10
- 11. ACKNOWLEDGEMENTS • Dr. Lory Santiago-Vazquez for guiding and providing with the materials and lab space • Research students Semhar Kesete & Catherine Sampson • University of Houston-Clear Lake 11
Editor's Notes
- Stress to corals-Rise of ocean temperature, ocean acidification, UV radiations Healthy tissue of most stony corals ranges from yellow to brownish in color, a function of the photosynthetic pigments of their symbiotic zooxanthellae. When corals are inordinately stressed, they often expel their zooxanthellae, or the concentration of photosynthetic pigments declines. This response is known as bleaching
- Should I keep this slide hidden or go through it quickly?
- 2% agarose gels stained withethidiumbrominde-? Does this come on the side?
- What to write for conclusion?Future goals: To correlate these changes to 16s rDNA changes of the same specimens analyzed by denaturing gradient gel electrophoresis (DGGE), to production of prostaglandins, and to changes in the soft coral transcriptome.