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During July SRP increased, along with chlorophyll a concentration (Fig 3.8c & Fig 3. l Oc). However, SRP was still lower than in Ace Lake or Pendant Lake while, overall, chlorophyll a was higher. By September SRP concentrations dropped considerably', but the mean chlorophyll a concentration in September was approximately 50% that of July and remained low through November (Fig 3. l Oc). It is possible that bacteriochlorophv II could be responsible for the decrease in SRP. As bacterial abundance increases, phosphate decreases, despite the decrease in chlorophyll a. Therefore it is suggested that bacterioplankton, not the PNAN, have the biggest impact on phosphate concentrations in Triple Lake. DOC concentrations (Fig 3.9c) were significantly higher than in Ace or Pendant Lake (the correlation between high salinity and high DOC levels is shown in section 3.3.1, figure 3.15a). It is believed this is because of the lack of biological activity in hypersaline lakes such as Triple Lake, which causes a reduction in the utilization of DOC leading to accumulation (Hand, 1980). Chlorophyll a shows an increase in concentration with depth throughout the year (Fig 3.22a). This indicates a shade adapted system. With similar inorganic nutrient concentrations throughout the lake, it is more likely that the phototrophic organisms are functioning with greater efficiency at depth due to reduced light intensities (Fig 3.1 Oc). Bacterial abundance correlates well with chlorophyll a concentrations during July and November, 2000, with r values of 0.96 and 0.95 respectively (Fig 3.22a). However, during November there is no correlation between the two variables, although they still show a similar relationship, i. e. an increase with depth. HNAN and PNAN (Fig 3.12c & 3.13c) abundances were similar to those found in Pendant Lake and Ace Lake. HNAN abundance correlated well with the bacterial abundance (September r=0.97, November r=0.99), which was expected, as HNAN use bacteria as a food source (Hand & Burton, 1981). 104
a Bacterial abundance (x 106 1.1) 05 10 0 0 Bacterial Abundance (x106I. 1) 0 10 20 C 0 Bacterial Abundance (x 106 I. 1) 05 10 II 2 2 2 Depth Depth (m) Depth (m) 4 4 ö 8 05 10 15 0246 05 10 Chlorophyll a (ug 1.1) Chlorophyll a (ug I-1) Chlorophyll a (ug 1.1) Figure 3.22 - Correlation between bacterial abundance (- -) and chlorophyll a concentration (-"-) for three sampling dates in 2000, July (a), September (b) and November (c), in Triple Lake. Correlation coefficients are as follows, (a) r=0.96, (b) r=0.95 and (c) r=0.61. 105
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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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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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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 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: salinity decreased from July to Sep
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
Page 157 and 158: The closest published relative for
Page 159 and 160: identification was made based on 15
Page 161 and 162: Isolate number Closest phylogenetic
Page 163 and 164: e included due to its small sequenc
Page 165 and 166: 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
Page 171 and 172: seen to a certain extent in the ARD
Page 173 and 174: 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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