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LAYBOURN-PARRY, J., QUAYLE, W. C.. HENSHAW, T.. RUDDELL. A. and MARCHANT, H. J. (2001b). Life on the edge: the plankton and chemistry of Beaver Lake, an ultra-oligotrophic epishelf lake, Antarctica. Freshwai r- Biology. 46,1205-1217. LAYBOURN-PARRY, J., QUAYLE, W. and HENSHAW. T. (2002). The functional dynamics and evolution of Antarctic saline lakes in relation to salinity and trophy. Polar Biology. 25,542-552.. LEE, S. J., XIE, A., JIANG, W., ETCHEGARAY, J-P., JONES. P. G. and INOUYE. M. (1994). Family of major cold-shock protein, CspA (CS7.4), of Escherichia coli, whose members show a high sequence similarity with the eukaryotic Y-box binding proteins. Molecular Microbiology. 11,833-839. LIESACK, L., JANSSEN, P. H., RAINEY, F. A., WARD-RAINEY. N. L. and STACKERBRANDT, E. (1997). Microbial diversity in soil: the need for a combined approach using molecular and cultivation techniques. In: ltlodt'rn Soil Microbiology. (Eds. J. D. van Elsas, J. T. Trevors, E. M. H. Wellington). Marcel Dekker Inc., New York, pp 375-439. LIN, N. T. and TSENG, Y. H. (1997). Sequence and copy number of the Xanthomonas campestris pv. Campestris gene encoding 16S rRNA. Biochemical and Biophysical Research Communications. 235,2,276-280. LINDH, E., KJAELDGAARD, P., FREDERIKSEN, W. and URSUG, J. (1991). Phenotypical properties of Enterobacter agglomerans (Pantoea agglomerans) from human, animal and plant sources. APMIS: Acta Pathologica Microbiologica at limnological Scandinavica. 99.4,347-352. LOGSDON, J. M. and DOOLITTLE, W. F. (1997). Origin of antifreeze protein genes: A cool tale in molecular evolution. Proceedings of the National Academy of Science. USA. 94,3485-3487. LOPEZ-GARCIA, P., LOPEZ-LOPEZ, A.. MOREIRA, D. and RODRIGUEZ- VALERA, F. (2001). Diversity of free-living prokaryotes from a deep-sea site at the Antarctic Polar Front. FEMS Microbiology Ecology. 36.193-202. LOUWS, F. J., BELLS, J., MEDINA-MORA, C. M., SMART. C. D.. OPGENORTH, D.. ISHIMARU, C. A.. HAUSBECK, M. K.. De BRUIJN. F. J. and FULBRIGHT. D. 216
W. (1998). Rep-PCR-mediated genomic fingerprinting: a rapid and effective method to identify Clavibacter michiganensis. Phytopathologv. 88.8.862-868. MACKERETH, F. J. H., HERON, J. & FALLING. J. I. (1988). Water Analysis. Freshwater Biol. Assoc. Sci. Publ. n° 36. MADURA, J. D., TAYLOR, M. S., WIERZBICKI, A.. HARRINGTON. J. P.. SIKES. C. S. and SÖNNICHSEN, F. (1996). The dynamics and binding of a type III antifreeze protein in water and on ice. Journal of Molecular Structure (Theochem). 388,65-77. MANCUSO, C. A., FRANZMANN, P. D., BURTON, H. R. and NICHOLS, P. D. (1990). Microbial community structure and biomass estimates of a methanogenic Antarctic lake ecosystem as determines by phosphlipid analysis. Aficrobial Ecology. 19,73-95. MARSHALL, W. A. (1996). Biological particles over Antarctica. Nature. 383,680. MARUYAMA, A., HONDA, D., YAMAMOTO, H., KITAMURA, K. and HIGASHIHARA, T. (2000). Phylogenetic analysis of psychrophilic bacteria isolated from the Japan Trench, including a description of the deep-sea species Psychrobacter pacificensis sp. nov. International Journal of Systematic and evolutionary Microbiology. 50,2,835-846. MATTHEWS, B. W. (1987). Genetic and structural analysis of the protein stability problem. Biochemistry. 26,6885-6888. MAZUR, P. (1980). Limits to life at low temperatures and reduced water contents and water activities. Origins of Life. 10,137-159. McCAMMON, S. A., INNES, B. H., BOWMAN. J. P., FRANZMANN. P. D.. DOBSON, S. J., HOLLOWAY, P. E., SKERRATT, J. H.. NICHOLS, P. D. and RANKIN, L. M. (1998). Flavobacterium hibernum sp. nov., a lactose-utilizing bacterium from a freshwater Antarctic lake. International Journal of'Sy. stematic Bacteriology. 48,4.1405-1412. McCAMMON, S. A. and BOWMAN. J. P. (2000). Taxonomy of Antarctic Flavobacterium species: description of Flavobacterium gillisiae sp. nov.. Flavobacterium tegetincola sp. nov.. and Flaw bacterium xanthum sp. nov.. nom. Rev. and reclassification of [Flavobacterium] salegens as Salcgentibacter 217
Page 1 and 2:
COLD ADAPTATION STRATEGIES AND DIVE
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, 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
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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
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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
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 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: Howard-Williams and I. Hawes). Balk
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
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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