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DUNKER, B. P., KOOPS, D. M., WALKER, V. K. and DAVIES. P. L. (1995). Low temperature persistence of type I antifreeze protein is mediated by cold-specific mRNA stability. FEBS Letters. 377,185-188. EDWARDS, A. R., VAN DEN BUSSCHE, R. A. and WICHMAN. H. A. (1994). Unusual pattern of bacterial ice nucleation gene evolution.. lioleculur Biology and Evolution. 11,6,911-920. EKONG, R. and WOLFE, J. (1998). Advances in fluorescent in situ hybridisation. Current Opinion in Biotechnology. 9,19-24. ELLIS-EVANS, J. C. (1985a). Interactions of bacterio- and phyto-plankton in nutrient cycling within eutrophic Heywood Lake, Signy Island. In: Antarctic Nutrient Cycles and Food Webs. (Eds. W. R. Siegfried, P. R. Condy. and R. M. Laws). Springer-Verlag, Berlin Heidelberg. ELLIS-EVANS, J. C. (1985b). Fungi from maritime Antarctic freshwater environnments. British Antarctic Survey Bulletin. 68,37-45. ELLIS-EVANS, J. C., LAYBOURN-PARRY, J., BAYLISS, P. R. and PERRISS, S. J. (1998). Physical, chemical and microbial community characteristics of the lakes of the Larsemann Hills, Continental Antarctica. Archives of Hydrobiology. 141,2, 209-230. EKONG, R. and WOLFE, J. (1998). Advances in fluorescent in situ hybridisation. Current Opinion in Biotechnology. 9,19-24. ERCOLINI, D., MOSCHETTI, G., BLAIOTTA, G. and COPPOLA, S. (2001a). Behaviour of variable V3 region from 16S rDNA of lactic acid bacteria in denaturing gradient gel electrophoresis. Current Microbiology, 42.3.199-202. ERCOLINI, D., MOSCHETTI, G., BLAIOTTA. G. and COPPOLA. S. (2001b). The potential of a polyphasic PCR-DGGE approach in evaluating microbial diversity of natural whey cultures for water-buffalo Mozzarella cheese production: bias of culture-dependent and culture-independent analyses. Systematic and Applied Microbiology, 24,4,610-617. ETZLER, F. M. (1983). A statistical thermodynamic model for water near solid interfaces. Journal of Colloid Interface Science. 92.43-56. EVANS, R. P. and FLETCHER, G. L. (2001). Isolation and characterisation of type I 204
antifreeze proteins from Atlantic snailfish (Liparis atlanticus) and dusky snailfish (Liparis gibbus). Biochimica et Biophysica Acta. 1547,235-244. EWART, K. V. and FLETCHER, G. L. (1990). Isolation and characterisation of antifreeze proteins from smelt (Osmerus mordax) and Atlantic herring (Clupea harengus). The Canadian Journal of Microbiology. 41.109-117. FARRELL, J. and ROSE, A. H. (1967). Temperature effects on microorganisms. In: Thermobiology. (Ed. A. H. Rose). Academic Press. London and New York. Pp. 147-218. FARRELLY, V., RAINEY, F. A. and STACKERBRANDT, E. (1995). Effect of genome size and rrn gene copy number on PCR amplification of 16S rRNA genes from a mixture of bacterial species. Applied and Environmental Microbiology. 61.7, 2798-2801. FEENEY, R. E. (1988). Inhibition and promotion of freezing: Fish anti-freeze proteins and ice-nucleating proteins. Comments on Agricultural Food Chemnistry. 1,147- 181. FEENEY, R. E. and YEH, Y. (1993). Antifreeze Proteins: Properties, mechanism of action, and possible applications. Food Technology. 47,1,82-90. FEENEY, R. E. and YEH, Y. (1998). Antifreeze proteins: Current status and possible food uses. Trends in Food Science and Technology. 9,102-106. FEGATELLA, F. I. LIM, J., KJELLEBERG, S. and CAVICCHIOLI, R. (1998). Implications of rRNA operon copy number and ribosome content in the marine oligotrophic Ultramicrobacterium Sphingomonas sp. strain RB2256. Applied und Environmental Microbiology. 64,11,4433-4438. FELLER, G., NARINX, E., ARPIGNY, J. L., ALTTALEB, M., BAISE. E., SABINE. G. and GERDAY, C. (1996). Enzymes from psychrophilic organisms. FEMS Microbiology Reviews. 18,189-202. FELLER, G. & GERDAY, C. (1997). Psychrophilic enzymes: molecular basis of cold adaptation. Cellular and Molecular Life Sciences. 53,830-841. FERRIS, M. J. and BURTON, H. R. (1988). The annual cycle of heat content and mechanical stability of hypersaline Deep Lake. Vestfold Hills, Antarctica. Hydrobiologia. 165,115-128. 205
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COLD ADAPTATION STRATEGIES AND DIVE
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, 71 "If you march your Winter Jour
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1.6.3. Molecular typing techniques
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3.2.1.2.3. DOC and chlorophyll a ..
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5.3.2.5. The Sphingomonas group, is
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LIST OF FIGURES Figure I. I. Diagra
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Figure 3.17a Mean bacterial abundan
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LIST OF TABLES Table 1.1. Types of
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List of Abbreviations AFP AFGP AFLP
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ACKNOWLEDGEMENTS This study was fun
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Chapter 1: INTRODUCTION 1.1 - Tempe
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1999 ; Hand & Burton, 1981) or iden
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Cold adaptation is dependant on the
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not at 25°C is a function of the a
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The most prolific literature on col
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process known as thermal hysteresis
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dominant, role in the ice-growth in
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explanation for this diversity is t
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1.5.3.5 - Membrane bound solute tra
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een well established (Russell & Nic
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& Bowman, 2001; Labrenz & Hirsch, 2
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The reaction mix contains both deox
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gene mixtures and hence can be used
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displaying a high number of common
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AACGTCGAAA AACCTCGAAA (Organism A)
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particular set of environmental par
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Lake Maximum Depth (m) Approximate
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Larsemann Hills __ rp3rtrnent of th
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Sampling trips took two days during
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Sigma, Sigma-Aldrich Company Ltd.,
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oxidized and the sample then conver
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sample. An appropriate volume was l
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min to ensure complete extension of
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Figure 2.3 - Lane delineation image
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Figure 2.6b - Molecular weights are
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RNA 41F (5'-GCT CAG ATT GAA CGC TGG
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I min. The spin column was removed
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compiled using Seqman and any erron
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ecorded and related to the level of
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information regarding the cold adap
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50ppt to 186ppt. Thirteen were sali
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M N N 't M M \p \p 00 kf) M kf) bA
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as would normally be expected, as b
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1444 6ý L CC vý O Q O O NO u 'c z
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5 - 250 3 245 240 G (, -3 235 a 0-
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3.2.2.2.5 - Dissolved organic carbo
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d 14 12 10 14 12 7 10 8 6 4 2 0 00/
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SRP ('g L- 05 10 15 20 2 3 Depth4 5
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3.2.2.3 - Biological characteristic
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a 0 Bacterial Abundance (x 106 ml-1
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3.3 - Discussion 3.3.1 - Summer sam
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to high surface water temperatures
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chemocline (Rankin et al., 1999). M
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a 250 200 150 100 50 70 60 50 40 ý
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The temperature (Fig 3.2a) through
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et al., 1999). No melt out was obse
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increase in ammonium is probably du
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10 7 9 6 8 E ö 7 5 Co u c m c 40 w
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main pigments were chlorophylls a a
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Inorganic nitrogen (NO2, NO3 and NH
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744 -- 7 644 6 19 ü SE 400-- 4 2s
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salinity decreased from July to Sep
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a Bacterial abundance (x 106 1.1) 0
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8 ö a N Ö oý L _> Q O O 4 0 rn O
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undoubtedly true of Club Lake. The
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suggested that input of bacteria an
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Pendant Lake is an unstratified, sa
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non-AFP active peptides from inhibi
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. ter rý :. 'ý Figure 4.2 -Fryka
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-ý- ---- ,. r. __-_-. ___. r. ýý
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4.2 - Results 4.2.1 - Sampling and
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SPLAT score. It has been shown that
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3'E t ý _d M! .PJ r rn 15 3.5 4 3
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4.3 - Discussion Of the 866 bacteri
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arisen in very few species due to c
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Chapter 5: BACTERIAL MOLECULAR TYPI
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Fig 5.1 - Photographs of ARDRA gel
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Coefficient: Dice Algorithm: lPGMAA
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The closest published relative for
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identification was made based on 15
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Isolate number Closest phylogenetic
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e included due to its small sequenc
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5.3.2 - Identification of the AFP a
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explains the fluctuation in related
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contamination was lower as the PCR
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seen to a certain extent in the ARD
Page 173 and 174: American Type Culture Collection (A
Page 175 and 176: Phylogenetic cluster alignment of t
Page 177 and 178: et al., 1994) and the Antarctic SIM
Page 179 and 180: Chapter 6: BACTERIAL COMMUNITY ANAL
Page 181 and 182: 6.2.1 - Detection of AFP active iso
Page 183 and 184: using the same PCR protocol as orig
Page 185 and 186: Figure 6.1 - Denaturing gradient ge
Page 187 and 188: 6.2.2 - Temporal and spatial variat
Page 189 and 190: Figure 6.4c - Gel I image showing b
Page 191 and 192: Coefficient: Dice Algorithm: UPGMA
Page 193 and 194: etween Om and 2m was obviously the
Page 195 and 196: All PCR-based rRNA molecular techni
Page 197 and 198: 6.3.2.3 - Cellular lysis and DNA pu
Page 199 and 200: hybrids should be equal. However, S
Page 201 and 202: contain contaminants. If the profil
Page 203 and 204: (Section 6.3.2.4.3 ) could not be r
Page 205 and 206: most dominant species in the commun
Page 207 and 208: study was characterised as M protea
Page 209 and 210: 11 species) showed AFP activity. Th
Page 211 and 212: that AFP activity could have evolve
Page 213 and 214: 7.2 - Future Work The 19 AFP active
Page 215 and 216: REFERENCES ABYZOV, S. S. (1993). Mi
Page 217 and 218: BELL, E. M. and LAYBOURN-PARRY, J.
Page 219 and 220: Symposium on Environmental Biogeoch
Page 221 and 222: Research. 5,477-493. CLESCERI, L. S
Page 223: DRICKAMER, K. (1999). C-type lectin
Page 227 and 228: FRANZMANN, P. D. and DOBSON, S. J.
Page 229 and 230: GRIFFITH, M., ALA, P., YANG. D. S.
Page 231 and 232: IVANOVA, E. P., KIPRIANOVA, E. A.,
Page 233 and 234: Antarctic Ecosystems. G. A. Llano (
Page 235 and 236: Howard-Williams and I. Hawes). Balk
Page 237 and 238: W. (1998). Rep-PCR-mediated genomic
Page 239 and 240: putida strains from Antarctica. Mic
Page 241 and 242: NICHOLS, D., BOWMAN, J., SANDERSON,
Page 243 and 244: PEARCE, D. A. and BUTLER, H. G. (20
Page 245 and 246: (1999). Palaeohydrological modellin
Page 247 and 248: tracers of biogeochemical processes
Page 249 and 250: R. G. Landes Company, Austin, Texas
Page 251 and 252: partial 16S rDNA sequencing. Applie
Page 253 and 254: YABUUCHI, E., WANG, L., ARAKAWA, M.
Page 255 and 256: APPENDICES Al. Media Tryptic Soya A
Page 257 and 258: "D C 'O I. .. r .. r 6 rr E , 4mo .
Page 259 and 260: . + + + I I I I I I + I + + I I I I
Page 261 and 262: Ö ON N M 1 O - N 'zt "o l'" 00 C N
Page 263 and 264: 1 T 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1
Page 265 and 266: 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1
Page 267 and 268: 1 1 1 1 I 1 + I I 1 I 1 1 I 1 1 1 I
Page 269 and 270: 1 1 1 1 1 1 1 I I 1 I 1 I 1 1 I T I
Page 271 and 272: Cl M 'tt tn '. p t- 00 O\ N M O - (
Page 273 and 274: d' O - (N M 'nt V) \O N. 00 O> N M
Page 275 and 276:
1 1 T 1 1 1 T 1 1 1 1 1 1 1 1 1 1 1
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+ + i i i i i i " i " . + + + + " +
Page 279 and 280:
e O --ý N M t Vn 1,0 [-- 00 O> N M
Page 281 and 282:
FT ce + I I r. to \, O t- 00 O\ N M
Page 283 and 284:
kn 110 r- 00 Oý N M O - N M Itt vn
Page 285 and 286:
, It N M "t N M Iýt N M M M M Oý
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gj a - - - - - - - - - - - - - - -
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