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1281-1285. WILKINS, P. 0. (1973). Psychrotrophic Gram-positive bacteria: temperature effects on growth and solute uptake. Canadian Journal of Microbiology. 19,8,909-915. WILLIAMS, J. G., KUBELIK, A. R., LIVAK, K. J.. RAFALSKI. J. A. and TINGEY". S. V. (1990). DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Research. 18,22.6531-6535. WILLIAMS, W. D. and SHERWOOD, J. E. (1994). Definition and measurement of salinity in salt lakes. International Journal of Salt Lake Research. 3,53-63 . WILLEMSKY, G., BANG, H., FISCHER, G. and MARAHIEL, M. A. (I 992). Characterisation of csp B, Bacillus subtilis inducible cold shock gene affecting cell viability at low temperatures. Journal of Bacteriology. 174.6326-6335. WILSON, K. (1987). Preparation of genomic DNA from bacteria. In: Current Protocols in Molecular Biology. (Eds. F. M. Ausubel, R. Brent, R. E. Kingston, D. D. Moore, J. A. Smith, J. G. Seidman, and K. Struhl). Wiley, N. Y. WILSON, P. W., BEAGLEHOLE, D. and DeVRIES, A. L. (1993). Antifreeze glycopeptide adsorption on a single crystal ice surfaces using ellipsometry. Biophysical Journal. 64,1878-1884. WILSON, P. W. and LEADER, J. P. (1995). Stabilization of supercooled fluids by thermal hysteresis proteins. Biophysical Journal. 68.2098-2107. WIRSEN, C. 0., JANNASCH, H. W., WAKEHAM, S. G. and CANUEL, E. A. (1987). Membrane lipids of a psychrophilic and barophilic deep-sea bacterium. Current Microbiology. 14,319-322. WOESE, C. R. (1987). Bacterial evolution. Microbiological Review. 51,221-271. WRIGHT, S. W. and BURTON, H. R. (1981). The biology of Antarctic saline lakes. Hydrobiologia, 82,319-338. WU, Y. and FLETCHER, G. L. (2000). Efficacy of antifreeze protein types in protecting liposome membrane integrity depends on phospholipid class. Biochimica ci Biophysica Acta. 1524,11-16. YABUUCHI. E., YANO, I., OYAIZU, H., HASHIMOTO, Y.. EZAKI. T. and YAMAMOTO. H. (1990). Proposal of Sphingomonas paucumoblis gen. no\. and comb. nov., Sphingomonas parapaucimoblis sp. nov., Sphingomonas I"unc)ikUUi'uc ýtý.,
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Page 1 and 2:
COLD ADAPTATION STRATEGIES AND DIVE
Page 3 and 4:
, 71 "If you march your Winter Jour
Page 5 and 6:
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
Page 153 and 154:
Fig 5.1 - Photographs of ARDRA gel
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Coefficient: Dice Algorithm: lPGMAA
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The closest published relative for
Page 159 and 160:
identification was made based on 15
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Isolate number Closest phylogenetic
Page 163 and 164:
e included due to its small sequenc
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5.3.2 - Identification of the AFP a
Page 167 and 168:
explains the fluctuation in related
Page 169 and 170:
contamination was lower as the PCR
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seen to a certain extent in the ARD
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American Type Culture Collection (A
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Phylogenetic cluster alignment of t
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et al., 1994) and the Antarctic SIM
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Chapter 6: BACTERIAL COMMUNITY ANAL
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6.2.1 - Detection of AFP active iso
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using the same PCR protocol as orig
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Figure 6.1 - Denaturing gradient ge
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6.2.2 - Temporal and spatial variat
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Figure 6.4c - Gel I image showing b
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Coefficient: Dice Algorithm: UPGMA
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etween Om and 2m was obviously the
Page 195 and 196:
All PCR-based rRNA molecular techni
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6.3.2.3 - Cellular lysis and DNA pu
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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 and 224: DRICKAMER, K. (1999). C-type lectin
Page 225 and 226: antifreeze proteins from Atlantic s
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: partial 16S rDNA sequencing. Applie
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
Page 277 and 278: + + 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
Page 287 and 288: gj a - - - - - - - - - - - - - - -
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