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

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which were, on average, highest in the hypersaline lakes, and lowest in the freshwater lakes. Therefore, the major limiting factor for metabolic processes in the hypersaline lakes was not nutrient availability but extreme high salinities. Oval Lake, although having a high salinity (34ppt), shows very similar inorganic and organic nutrient concentrations to the other saline lakes, except for a low concentration in ammonium, which is also seen in Lake Abraxas and Lake Williams as well as two hypersaline lakes (Table 3.1 a and b). 86
a 250 200 150 100 50 70 60 50 40 ý+ E 30 U 20 0 10 0 E +o, 4 a, D o p co- -1D4-0"1M c -_ -0 +4 ýo 'o 0' a,, c4=°, c vn aca to VI aa --vý a_ c_ 1n a +v (ý a o _E+ -o äx E 0aa, ä Inc in , ad, IL } Eý =. c c r0 4-j 4fi .: -", +vý. ý cc aa (D-T 0) ä, ß co Q, ßyß H C0 =O- } H 'v v' O 15- ( +1 dü Ü4Ü O Q ö 1 Q> -C J > < Üm wO J U ä E _EN" -V , 4, -Z ä. ýa E QJ 0 Lake Names Figure 3.15a The - correlation between salinity (-"-) and DOC (a ) concentrations in the summer sampled lakes. Note DOC concentration for Cemetery Lake was too high to record using the TOC analyser and is noted here as 9999 mg 1-'. The correlation coefficient for the two variables is r=0.895. 140 120 100 p' 80 E 0 ö 60 40 20 0 Figure 3.15b - DOC levels for the summer-sampled hypersaline (-"-), saline ( ) and freshwater () lakes. The lakes are shown in the order (left to right) in which they were shown in tables la, b and c. The last hypersaline lake (Cemetery Lake) recorded off the readable scale and is noted here as 9999µg I-'. 87
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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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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
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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 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: chemocline (Rankin et al., 1999). M
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
Page 143 and 144: SPLAT score. It has been shown that
Page 145 and 146: 3'E t ý _d M! .PJ r rn 15 3.5 4 3
Page 147 and 148: 4.3 - Discussion Of the 866 bacteri
Page 149 and 150: arisen in very few species due to c
Page 151 and 152: Chapter 5: BACTERIAL MOLECULAR TYPI
Page 153 and 154: Fig 5.1 - Photographs of ARDRA gel
Page 155 and 156: 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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