<?xml version="1.0" encoding="utf-8" standalone="no"?>
<!DOCTYPE article SYSTEM "http://www.biogeosciences.net/inc/bg/copernicus.dtd">
<article language="en">
	<journal>
		<journal_title>Biogeosciences</journal_title>
		<journal_url>www.biogeosciences.net</journal_url>
		<issn>1726-4170</issn>
		<eissn>1726-4189</eissn>
		<volume_number>6</volume_number>
		<issue_number>7</issue_number>
		<publication_year>2009</publication_year>
	</journal>
	<doi>10.5194/bg-6-1199-2009</doi>
	<article_url>http://www.biogeosciences.net/6/1199/2009/</article_url>
	<abstract_html>http://www.biogeosciences.net/6/1199/2009/bg-6-1199-2009.html</abstract_html>
	<fulltext_pdf>http://www.biogeosciences.net/6/1199/2009/bg-6-1199-2009.pdf</fulltext_pdf>
	<start_page>1199</start_page>
	<end_page>1207</end_page>
	<publication_date>2009-07-17</publication_date>
	<article_title content_type="html">Effects of the pH/&lt;i&gt;p&lt;/i&gt;CO&lt;sub&gt;2&lt;/sub&gt; control method on medium chemistry and phytoplankton growth</article_title>
	<authors>
		<author numeration="1" affiliations="1,2">
			<name>D. Shi</name>
			<email>dshi@princeton.edu</email>
		</author>
		<author numeration="2" affiliations="1,2">
			<name>Y. Xu</name>
		</author>
		<author numeration="3" affiliations="1">
			<name>F. M. M. Morel</name>
		</author>
	</authors>
	<affiliations>
		<affiliation numeration="1" content_type="html">Department of Geosciences, Princeton University, Princeton, New Jersey 08544, USA</affiliation>
		<affiliation numeration="2" content_type="html">These two authors contributed equally to the work.</affiliation>
	</affiliations>
	<abstract content_type="html">The control of key chemical parameters in phytoplankton cultures, such as
&lt;i&gt;p&lt;/i&gt;CO&lt;sub&gt;2&lt;/sub&gt;, pH and Ω (the saturation state of calcium carbonate), is
made difficult by the interdependence of these parameters and by the changes
resulting from the growth of the organisms, such as CO&lt;sub&gt;2&lt;/sub&gt; fixation,
nutrient uptake and, for coccolithophores, calcite precipitation. Even in
cultures where &lt;i&gt;p&lt;/i&gt;CO&lt;sub&gt;2&lt;/sub&gt; or pH is maintained constant, other chemical
parameters change substantially at high cell densities. Experimentally we
observed that various methods of adjustment of &lt;i&gt;p&lt;/i&gt;CO&lt;sub&gt;2&lt;/sub&gt;/pH – acid or base
addition, use of buffers or pH-stats, or bubbling of CO&lt;sub&gt;2&lt;/sub&gt;-enriched air
– can be used, the choice of one or the other depending on the goals of the
experiments. At seawater pH, we measured the same growth rates in cultures
of the diatom &lt;i&gt;Thalassiosira weissflogii&lt;/i&gt; where the &lt;i&gt;p&lt;/i&gt;CO&lt;sub&gt;2&lt;/sub&gt;/pH was controlled by these different
methods. The pH/&lt;i&gt;p&lt;/i&gt;CO&lt;sub&gt;2&lt;/sub&gt; control method also did not affect the rates of
growth or calcification of the coccolithophore &lt;i&gt;Emiliania huxleyi&lt;/i&gt; at seawater pH. At lower
pH/higher &lt;i&gt;p&lt;/i&gt;CO&lt;sub&gt;2&lt;/sub&gt;, in the &lt;i&gt;E. huxleyi&lt;/i&gt; strain PLY M219, we observed increases in rates
of carbon fixation and calcification per cell, along with a slight increase
in growth rate, except in bubbled cultures. In our hands, the bubbling of
cultures seemed to induce more variable results than other methods of
&lt;i&gt;p&lt;/i&gt;CO&lt;sub&gt;2&lt;/sub&gt;/pH control. While highly convenient, the addition of pH buffers to
the medium apparently induces changes in trace metal availability and cannot
be used under trace metal-limiting conditions.</abstract>
	<references>
		<reference numeration="1" content_type="text"> % vor jede Referenz Anderson, D. M. and Morel, F. M. M.: Copper sensitivity of \textitGonyaulax tamarensis, Limnol. Oceanogr., 23, 283–295, 1978. </reference>
		<reference numeration="2" content_type="text"> DOE: Handbook of methods for the analysis of the various parameters of the carbon dioxide system in sea water, Version 2, edited by: Dickson, A. G., and Goyet, C., 1994. </reference>
		<reference numeration="3" content_type="text"> Doney, S. C.: Oceanography: Plankton in a warmer world, Nature, 444, 695–696, 2006. </reference>
		<reference numeration="4" content_type="text"> Feely, R. A., Sabine, C. L., Lee, K., Berelson, W., Kleypas, J., Fabry, V. J., and Millero, F. J.: Impact of anthropogenic CO&lt;sub&gt;2&lt;/sub&gt; on the CaCO&lt;sub&gt;3&lt;/sub&gt; system in the oceans, Science, 305, 362–366, 2004. </reference>
		<reference numeration="5" content_type="text"> Feng, Y., Warner, M. E., Zhang, Y., Sun, J., Fu, F. X., Rose, J. M., and Hutchins, D. A.: Interactive effects of increased $p$CO&lt;sub&gt;2&lt;/sub&gt;, temperature and irradiance on the marine coccolithophore \textitEmiliania huxleyi (Prymnesiophyceae), Eur. J. Phycol., 43, 87–98, 2008. </reference>
		<reference numeration="6" content_type="text"> Ho, T. Y., Quigg, A., Finkel, Z. V., Milligan, A. J., Wyman, K., Falkowski, P. G., and Morel, F. M. M.: The elemental composition of some marine phytoplankton, J. Phycol., 39, 1145–1159, 2003. </reference>
		<reference numeration="7" content_type="text"> Iglesias-Rodriguez, M. D., Buitenhuis, E. T., Raven, J. A., Schofield, O., Poulton, A. J., Gibbs, S., Halloran, P. R., and de Baar, H. J. W.: Response to comment on &quot;Phytoplankton calcification in a High-CO&lt;sub&gt;2&lt;/sub&gt; World&quot;, Science, 322, p 1466, 2008a. </reference>
		<reference numeration="8" content_type="text"> Iglesias-Rodriguez, M. D., Halloran, P. R., Rickaby, R. E. M., Hall, I. R., Colmenero-Hidalgo, E., Gittins, J. R., Green, D. R. H., Tyrrell, T., Gibbs, S. J., von Dassow, P., Rehm, E., Armbrust, E. V., and Boessenkool, K. P.: Phytoplankton calcification in a high-CO&lt;sub&gt;2&lt;/sub&gt; world, Science, 320, 336–340, 2008b. </reference>
		<reference numeration="9" content_type="text"> Langer, G., Geisen, M., Baumann, K. H., Klas, J., Riebesell, U., Thoms, S., and Young, J. R.: Species-specific responses of calcifying algae to changing seawater carbonate chemistry, Geochem. Geophy. Geosy., 7, Q09006, doi:10.1029/2005GC001227, 2006. </reference>
		<reference numeration="10" content_type="text"> Lueker, T. J., Dickson, A. G., and Keeling, C. D.: Ocean $p$CO&lt;sub&gt;2&lt;/sub&gt; calculated from dissolved inorganic carbon, alkalinity, and equations for K-1 and K-2: validation based on laboratory measurements of CO&lt;sub&gt;2&lt;/sub&gt; in gas and seawater at equilibrium, Mar. Chem., 70, 105–119, 2000. </reference>
		<reference numeration="11" content_type="text"> Mash, H. E., Chin, Y. P., Sigg, L., Hari, R., and Xue, H. B.: Complexation of copper by zwitterionic aminosulfonic (good) buffers, Anal. Chem., 75, 671–677, 2003. </reference>
		<reference numeration="12" content_type="text"> Morel, F. M. M. and Hering, J. G.: Principles and applications of aquatic chemistry, John Wiley and Sons, 1993. </reference>
		<reference numeration="13" content_type="text"> Paasche, E. and Brubak, S.: Enhanced calcification in the coccolithophorid \textitEmiliania huxleyi (Haptophyceae) under phosphorus limitation, Phycologia, 33, 324–330, 1994. </reference>
		<reference numeration="14" content_type="text"> Price, N. M., Harrison, G. I., Hering, J. G., Hudson, R. J., Nirel, P. M. V., Palenik, B., and Morel, F. M. M.: Preparation and chemistry of the artificial algal culture medium aquil, Biol. Oceanogr., 6, 443–461, 1988/89. </reference>
		<reference numeration="15" content_type="text"> Riebesell, U., Zondervan, I., Rost, B., Tortell, P. D., Zeebe, R. E., and Morel, F. M. M.: Reduced calcification of marine plankton in response to increased atmospheric CO&lt;sub&gt;2&lt;/sub&gt;, Nature, 407, 364–367, 2000. </reference>
		<reference numeration="16" content_type="text"> Riebesell, U., Schulz, K. G., Bellerby, R. G. J., Botros, M., Fritsche, P., Meyerhofer, M., Neill, C., Nondal, G., Oschlies, A., Wohlers, J., and Zollner, E.: Enhanced biological carbon consumption in a high CO&lt;sub&gt;2&lt;/sub&gt; ocean, Nature, 450, 545–548, 2007. </reference>
		<reference numeration="17" content_type="text"> Rost, B., Zondervan, I., and Wolf-Gladrow, D.: Sensitivity of phytoplankton to future changes in ocean carbonate chemistry: current knowledge, contradictions and research directions, Mar. Ecol.-Prog. Ser., 373, 227–237, 2008. </reference>
		<reference numeration="18" content_type="text"> Sciandra, A., Harlay, J., Lefevre, D., Lemee, R., Rimmelin, P., Denis, M., and Gattuso, J. P.: Response of coccolithophorid \textitEmiliania huxleyi to elevated partial pressure of CO&lt;sub&gt;2&lt;/sub&gt; under nitrogen limitation, Mar. Ecol.-Prog. Ser., 261, 111–122, 2003. </reference>
		<reference numeration="19" content_type="text"> Sunda, W. and Guillard, R. R. L.: Relationship between cupric ion activity and toxicity of copper to phytoplankton, J. Mar. Res., 34, 511–529, 1976. </reference>
		<reference numeration="20" content_type="text"> Sunda, W. G., Price, N. M., and Morel, F. M. M.: Trace metal ion buffers and their use in culture studies, in: Algal culturing techniques, edited by: Andersen, R. A., Elsevier Academic Press, 35–63, 2005. </reference>
		<reference numeration="21" content_type="text"> Tortell, P. D., Payne, C. D., Li, Y. Y., Trimborn, S., Rost, B., Smith, W. O., Riesselman, C., Dunbar, R. B., Sedwick, P., and DiTullio, G. R.: CO&lt;sub&gt;2&lt;/sub&gt; sensitivity of Southern Ocean phytoplankton, Geophys. Res. Lett., 35, L04605, doi:10.1029/2007GL032583, 2008. </reference>
		<reference numeration="22" content_type="text"> Vasconcelos, M. T. S. D. and Leal, M. F. C.: Influence of N-2-hydroxyethylpiperazine-N&apos;-2-ethanesulfonic acid pH buffer on the biological response of marine algae, Environ. Toxicol. Chem., 21, 404–412, 2002. </reference>
		<reference numeration="23" content_type="text"> Xu, Y., Wahlund, T. M., Feng, L., Shaked, Y., and Morel, F. M. M.: A novel alkaline phosphatase in the coccolithophore \textitEmiliania huxleyi (Prymnesiophyceae) and its regulation by phosphorus, J. Phycol., 42, 835–844, 2006. </reference>
		<reference numeration="24" content_type="text"> Zhang, H. N. and Byrne, R. H.: Spectrophotometric pH measurements of surface seawater at in-situ conditions: Absorbance and protonation behavior of thymol blue, Mar. Chem., 52, 17–25, 1996. </reference>
	</references>
</article>

