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

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3 250 2 1 200 O 1 150 E3 -2 1°O -4 5 50 jÖ -6 a. 2CCD 2i 2 ý- z^ ý_ (V (h vÖÖONRÖNp0 G LL . LL LL_ (6 LL C . 19 0 NN =0 o -mai Qa Lakes and Sampling Dates Figure 3.14a- Air temperature (°C - ), surface water temperature (°C -"-) and salinity (ppt ) for the 12 summer sampled hypersaline lakes. 6 I~ v s 03 E y d a 17 0 -6 1 r-i rr r--- ri° mmm 'ryc memönnn "r ä NN 1)0 N (") ÜQN LL 0 in N E0xmm OOO ' >Q "' Wüö3 0 NyU rn ry Lakes and Dates of Sampling Figure 3.14b- Air temperature (°C surface water temperature (°C -"-) and salinity (ppt ) for the 13 summer sampled saline lakes. 6 25 4 2 2 0 15 -2 I -4 05 -6 -8 EnBZ ý' LL LL LL_ ll NOO týI OOOO NNN LL"' ry % c U OT ZLDOOVO ndv0N jOyOOOO In aNJ I- v Lake and Sampling Dates Figure 3.14c Air -- temperature (°C -), surface water temperature (°C -"-) and salinity (ppt the 11 summer sampled freshwater lakes. ) for 84
chemocline (Rankin et al., 1999). Meromixis has been observed in the following sampled Lakes: Organic, Williams, Abraxas, Ekho, Shield, Oval, Scale, Anderson, Oblong. Laternula and Angel `2' (Gibson, 1999; Labrenz & Hirsch, 2001). Most of the lakes do demonstrate some form of thermal stratification during the year (Table 2.1), generally monomixis. For example the majority of the hypersaline lakes are monomictic (Burton & Barker, 1979; Hand, 1980; Hand & Burton, 1981; Burton, 1981), being stratified during the summer, due to salinity and temperature gradients and then mixing in the winter when the upper warm water is cooled to the temperature of the lower waters. The freshwater lake, Crooked Lake. becomes inversely stratified when ice covered and only mixes in summer if the ice breaks out (Laybourn-Parry et al., 1992); it is, therefore, variably monomictic or amictic. Inorganic nutrients showed variability between the lakes (Tables 3.1 a, b and c). Dissolved organic carbon showed a correlation coefficient of r=0.895 with salinity (Fig 3.15a) indicating lakes with high salinities generally had a higher DOC concentration (Fig 3.15b). However, the correlation between salinity and DOC concentration was stronger at lower salinities. The correlation coefficient (r value) was only 0.70 for the hypersaline lakes. DOC is exuded by the phytoplankton during photosynthesis (Chrost & Faust, 1983), therefore high PNAN populations may cause DOC levels to increase (PNAN abundance was not recorded for the summer lakes). Autotrophic bacterioplankton may also exude DOC and contribute to the DOC pool. The heterotrophic bacterioplankton are the main consumers of DOC, but HNAN have also been known to exploit it (Laybourn-Parry, 1997), although the majority of HNAN are consumers of bacterioplankton. The oligotrophic nature of these lakes coupled with low temperatures results in low microbial biomass and low turnover of the DOC pool. The higher DOC concentrations recorded in the saline and hypersaline lakes could be due to accumulation of photosynthate in the absence of significant microbial decomposition (Hand, 1980; Barker, 1981; Burton, 1981). Viral lysis of bacteria and phytoplankton short circuits the carbon cycle in aquatic systems, preventing transfer of carbon to higher trophic levels (Fuhrman, 1999). Viruses have been identified in the lakes of the Vestfold Hills (Laybourn-Parry et al., 2001 a) but their role in DOC dynamics has yet to be determined. However, in microbially dominated communities like those of Antarctic lakes, viruses are likely to have a significant impact. This same trend can be seen for the major inorganic nutrients (Tables 3.1a, b and c), nitrate, nitrite, ammonium. and SRP, all of 85
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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
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 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: to high surface water temperatures
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
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
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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 - - - - - - - - - - - - - - -
show all

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