ý.,,: V. ý ýý . - Nottingham eTheses - University of Nottingham

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as would normally be expected, as biologically available nutrients are locked up in the increased biomass. 3.2.2 - Focus study lakes Only five lakes were chosen for the detailed study from January to November 2000, these were Ace Lake, Pendant Lake, Club Lake, Triple Lake and Deep Lake. They were chosen because they revealed the highest number of AFP positive isolates (refer to chapter 4). 3.2.2.1 - Physico-chemical characteristics 3.2.2.1.1 - Ice cover Ice cover on Ace Lake and Pendant Lake thickened during winter. with some fluctuation. This continued into November, with no observed melt (Fig 3.1 a& b). Triple showed total melt-out during the summer months and the early winter months but ice thickness increased to 0.4m during the winter, followed by a rapid thinning in November (Fig 3.1 c). Deep and Club had no ice cover throughout the year, apart from occasional greasy ice on extremely cold and still days (
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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
Page 33 and 34: dominant, role in the ice-growth in
Page 35 and 36: explanation for this diversity is t
Page 37 and 38: 1.5.3.5 - Membrane bound solute tra
Page 39 and 40: een well established (Russell & Nic
Page 41 and 42: & Bowman, 2001; Labrenz & Hirsch, 2
Page 43 and 44: The reaction mix contains both deox
Page 45 and 46: gene mixtures and hence can be used
Page 47 and 48: displaying a high number of common
Page 49 and 50: AACGTCGAAA AACCTCGAAA (Organism A)
Page 51 and 52: particular set of environmental par
Page 53 and 54: Lake Maximum Depth (m) Approximate
Page 55 and 56: Larsemann Hills __ rp3rtrnent of th
Page 57 and 58: Sampling trips took two days during
Page 59 and 60: Sigma, Sigma-Aldrich Company Ltd.,
Page 61 and 62: oxidized and the sample then conver
Page 63 and 64: sample. An appropriate volume was l
Page 65 and 66: min to ensure complete extension of
Page 67 and 68: Figure 2.3 - Lane delineation image
Page 69 and 70: Figure 2.6b - Molecular weights are
Page 71 and 72: RNA 41F (5'-GCT CAG ATT GAA CGC TGG
Page 73 and 74: I min. The spin column was removed
Page 75 and 76: compiled using Seqman and any erron
Page 77 and 78: ecorded and related to the level of
Page 79 and 80: information regarding the cold adap
Page 81 and 82: 50ppt to 186ppt. Thirteen were sali
Page 83: M N N 't M M \p \p 00 kf) M kf) bA
Page 87 and 88: 1444 6ý L CC vý O Q O O NO u 'c z
Page 89 and 90: 5 - 250 3 245 240 G (, -3 235 a 0-
Page 91 and 92: 3.2.2.2.5 - Dissolved organic carbo
Page 93 and 94: d 14 12 10 14 12 7 10 8 6 4 2 0 00/
Page 95 and 96: SRP ('g L- 05 10 15 20 2 3 Depth4 5
Page 97 and 98: 3.2.2.3 - Biological characteristic
Page 99 and 100: a 0 Bacterial Abundance (x 106 ml-1
Page 101 and 102: 3.3 - Discussion 3.3.1 - Summer sam
Page 103 and 104: to high surface water temperatures
Page 105 and 106: chemocline (Rankin et al., 1999). M
Page 107 and 108: a 250 200 150 100 50 70 60 50 40 ý
Page 109 and 110: The temperature (Fig 3.2a) through
Page 111 and 112: et al., 1999). No melt out was obse
Page 113 and 114: increase in ammonium is probably du
Page 115 and 116: 10 7 9 6 8 E ö 7 5 Co u c m c 40 w
Page 117 and 118: main pigments were chlorophylls a a
Page 119 and 120: Inorganic nitrogen (NO2, NO3 and NH
Page 121 and 122: 744 -- 7 644 6 19 ü SE 400-- 4 2s
Page 123 and 124: salinity decreased from July to Sep
Page 125 and 126: a Bacterial abundance (x 106 1.1) 0
Page 127 and 128: 8 ö a N Ö oý L _> Q O O 4 0 rn O
Page 129 and 130: undoubtedly true of Club Lake. The
Page 131 and 132: suggested that input of bacteria an
Page 133 and 134: Pendant Lake is an unstratified, sa
Page 135 and 136:
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
Page 179 and 180:
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
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contain contaminants. If the profil
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(Section 6.3.2.4.3 ) could not be r
Page 205 and 206:
most dominant species in the commun
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study was characterised as M protea
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11 species) showed AFP activity. Th
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that AFP activity could have evolve
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7.2 - Future Work The 19 AFP active
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REFERENCES ABYZOV, S. S. (1993). Mi
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BELL, E. M. and LAYBOURN-PARRY, J.
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Symposium on Environmental Biogeoch
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Research. 5,477-493. CLESCERI, L. S
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DRICKAMER, K. (1999). C-type lectin
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antifreeze proteins from Atlantic s
Page 227 and 228:
FRANZMANN, P. D. and DOBSON, S. J.
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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 (
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Howard-Williams and I. Hawes). Balk
Page 237 and 238:
W. (1998). Rep-PCR-mediated genomic
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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
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partial 16S rDNA sequencing. Applie
Page 253 and 254:
YABUUCHI, E., WANG, L., ARAKAWA, M.
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APPENDICES Al. Media Tryptic Soya A
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"D C 'O I. .. r .. r 6 rr E , 4mo .
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. + + + I I I I I I + I + + I I I I
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Ö ON N M 1 O - N 'zt "o l'" 00 C N
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1 T 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1
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1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1
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1 1 1 1 I 1 + I I 1 I 1 1 I 1 1 1 I
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1 1 1 1 1 1 1 I I 1 I 1 I 1 1 I T I
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Cl M 'tt tn '. p t- 00 O\ N M O - (
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d' O - (N M 'nt V) \O N. 00 O> N M
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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
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, It N M "t N M Iýt N M M M M Oý
Page 287 and 288:
gj a - - - - - - - - - - - - - - -
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