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

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Nitrate (pg L-1) c Nitrate (pg L-1) 0 0 10 20 30 40 50 0 0 10 20 30 40 1 1<strong>ý</strong> 2 2 3 4 3 Depth 5 (m) Depth II s 5 7 I<strong>ý</strong> l 6 8 9 7 10 8 d e tu 60 50 1 40 so so 40 30 r ;r :1 30 20 10 0 06/12/99 25/01/00 15103/00 04/05/00 23/06/00 12/08/00 01/10/00 20111/00 09/01101 Sampling Date Z 20 10 0 06/12/9 25/01/0 15/03/0 04/05/0 23/06/0 12/08/0 0111010 20/11/0 0910110 900000001 Sampling Date Figure 3.5 - Nitrate concentration (µg L-') for the five detailed study lakes. Ace Lake (a), Pendant Lake (b) and Triple Lake (c) were all depth sampled over several sampling dates. Deep Lake (d) and Club Lake (e) were only surface water sampled (0m) over the sampling dates Ace Lake sampling date key: 14/01/00 (-"-), 04/03/00 (- -), 15/07/00 ( ), 07/09/00 () and 15/11/00 ( ). Pendant Lake sampling date key: 23/02/00 (-"-), 17/07/00 (- -), 07/09/00 () and 13/11/00 ( ). Triple Lake sampling date key: 23/01/00 (-"-), 17/07/00 (- -), 07/09/00 () and 10/11/00 ( ). 72
d 14 12 10 14 12 7 10 8 6 4 2 0 00/01/00 01/01/00 02/01/00 03101/00 04/01/00 05/01/00 06/01/00 Sampling Date 6 4 2 0 00/01/00 01101/00 02/01/00 03101100 04101/00 05/01/00 Sampling Date Figure 3.6 - Nitrite concentration (µg L-) for the five detailed study lakes. Ace Lake (a), Pendant Lake (b) and Triple Lake (c) were all depth sampled over several sampling dates. Deep Lake (d) and Club Lake (e) were only surface water sampled (0m) over the sampling dates. Ace Lake sampling date key: 14/01/00 (-"-), 04/03/00 (- -), 15/07/00 ( ), 07/09/00 () and 15/11/00 ( ). Pendant Lake sampling date key: 23/02/00 (-"-), 17/07/00 (- ), 07/09/00 () and 13/11/00 ( ). Triple Lake sampling date key: 23/01/00 (-"-)' 17/07/00 (-N-), 07/09/00 () and 10/11/00 ( ). 73
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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
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
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Page 79 and 80: information regarding the cold adap
Page 81 and 82: 50ppt to 186ppt. Thirteen were sali
Page 83 and 84: M N N 't M M \p \p 00 kf) M kf) bA
Page 85 and 86: as would normally be expected, as b
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: 3.2.2.2.5 - Dissolved organic carbo
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
Page 137 and 138: . ter rý :. 'ý Figure 4.2 -Fryka
Page 139 and 140: -ý- ---- ,. r. __-_-. ___. r. ýý
Page 141 and 142: 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
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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
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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
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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
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FRANZMANN, P. D. and DOBSON, S. J.
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GRIFFITH, M., ALA, P., YANG. D. S.
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IVANOVA, E. P., KIPRIANOVA, E. A.,
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Antarctic Ecosystems. G. A. Llano (
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Howard-Williams and I. Hawes). Balk
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W. (1998). Rep-PCR-mediated genomic
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putida strains from Antarctica. Mic
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NICHOLS, D., BOWMAN, J., SANDERSON,
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PEARCE, D. A. and BUTLER, H. G. (20
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(1999). Palaeohydrological modellin
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tracers of biogeochemical processes
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R. G. Landes Company, Austin, Texas
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partial 16S rDNA sequencing. Applie
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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 " . + + + + " +
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e O --ý N M t Vn 1,0 [-- 00 O> N M
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FT ce + I I r. to \, O t- 00 O\ N M
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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
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gj a - - - - - - - - - - - - - - -
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