NIES collection MICROBIAL CULTURE COLLECTION | National Institute for Environmental Studies

<!DOCTYPE html> <html lang="ja"> <head> <title>NIES collection MICROBIAL CULTURE COLLECTION | National Institute for Environmental Studies</title> <meta charset="utf-8"> <meta name="viewport" content="width=1024, user-scalable=1"> <meta name="format-detection" content="telephone=no"> <meta property="og:site_name" content="NIES collection CULTURE COLLECTION | National Institute for Environmental Studies"> <meta property="og:image" content="./OGP_en.png"> <meta property="og:type" content="website"> <meta property="og:locale" content="ja_JP"> <meta name="description" content=""> <meta name="keywords" content=""> <link rel="icon" href="favicon.ico"> <link rel="apple-touch-icon" href="icon.png"> <script src="js/jquery-3.7.1.min.js"></script>  <script src="_common/js/jquery.easing.1.4.min.js"></script> <script src="_common/js/common.js"></script> <script src="_common/js/analytics.js"></script> <script src="js/jquery.pagetop.js"></script> <link rel="stylesheet" href="_common/css/reset.css"> <link rel="stylesheet" href="_common/css/font.css"> <link rel="stylesheet" href="_common/css/layout.css"> <link rel="stylesheet" href="_common/css/style.css"> <script type="text/javascript"> var TRANSLATED_FILE_PATH = '02medium.html';  $(function(){ $("meta").load("_common/_commonEn.jsp"); }); </script> </head> <body id="page" class="english">  <div id="header_wrapper"></div>   <div id="contents_wrapper">  <div id="topic_path"> <ul class="floatclear" itemscope itemtype="http://data-vocabulary.org/Breadcrumb"> <li><a href="./" itemprop="url"><span itemprop="title">HOME</span></a></li> <li>Media list</li> </ul> </div>   <section id="page_contents">  <div id="page_header"> <div id="page_header_wrapper" class="floatclear"> <h1 class="center">Media list</h1> </div> </div>   <nav id="anchor_link"> <ul class="floatclear"> <li><a href="#section01">Media for freshwater, terrestrial, hot spring and salt water algae</a></li> <li><a href="#section02">Media for marine and brackish water microalgae </a></li> <li><a href="#section03">Bacteria-free check media for freshwater algae</a></li> <li><a href="#section04">Bacteria-free check media for marine algae</a></li> <li><a href="#section05">Trace metals, vitamin mixtures and soil extracts</a></li> <li><a href="#section06">Media for protozoa</a></li> <li><a href="#section07">Media for freshwater red algae</a></li> <li><a href="#section08">Media for Charales</a></li> </ul> </nav>   <section id="section00"> <ul class="list_disc"> <li>Please click here to go to <a href="medium/en/media_web_e.html">“ Media list pdf version ”.</a></li> <li>Please click here to go to <a href="method/method_e.html" target="_blank">“ Media preparation, subculture and cryopreservation ”.</a></li> </ul> </section>   <section id="section01"> <h1>1. Media for freshwater, terrestrial, hot spring and salt water algae</h1> <table> <tbody> <tr> <td><a href="#aaf6">AAF–6</a></td> <td><a href="#acid_csi5">Acid–CSi/5</a></td> <td><a href="#af6">AF–6</a></td> </tr> <tr> <td><a href="#af6_2">AF–6/2</a></td> <td><a href="#af6_3">AF–6/3</a></td> <td><a href="#afac">AFAC</a></td> </tr> <tr> <td><a href="#allen">Allen</a></td> <td><a href="#bbm">BBM</a></td> <td><a href="#bg11">BG–11</a></td> </tr> <tr> <td><a href="#c">C</a></td> <td><a href="#c10seawater">C+10% Seawater</a></td> <td><a href="#c6">C/6</a></td> </tr> <tr> <td><a href="#c6g">C/6G</a></td> <td><a href="#c10">C/10</a></td> <td><a href="#ca">CA</a></td> </tr> <tr> <td><a href="#cam">CAM</a></td> <td><a href="#carefoot">Carefoot</a></td> <td><a href="#cb">CB</a></td> </tr> <tr> <td><a href="#cb_v">CB–V</a></td> <td><a href="#cc">CC</a></td> <td><a href="#csi">CSi</a></td> </tr> <tr> <td><a href="#csi_cu">CSi + Cu</a></td> <td><a href="#csi5">CSi/5</a></td> <td><a href="#ct">CT</a></td> </tr> <tr> <td><a href="#cyt">CYT</a></td> <td><a href="#dh_fe">DH + Fe</a></td> <td><a href="#dy_v">DY–V</a></td> </tr> <tr> <td><a href="#hut">HUT</a></td> <td><a href="#m11">M–11</a></td> <td><a href="#ma">MA</a></td> </tr> <tr> <td><a href="#maf6">MAF–6</a></td> <td><a href="#m_allen">M–Allen</a></td> <td><a href="#m_allen_g">M–Allen + Glucose</a></td> </tr> <tr> <td><a href="#m_allen_u">M–Allen + Uracil</a></td> <td height="24"><a href="#mbm">MBM</a></td> <td><a href="#m_chu_no10">M Chu No. 10</a></td> </tr> <tr> <td><a href="#mdm">MDM</a></td> <td><a href="#mg">MG</a></td> <td><a href="#mgm">MGM</a></td> </tr> <tr> <td><a href="#mac">Modified acetate medium (mAC)</a></td> <td><a href="#bg0">Modified BG–11<sub>0</sub></a></td> <td><a href="#jo">Modified Johnson's medium</a></td> </tr> <tr> <td><a href="#mm1">Modified M–1 (mM–1)</a></td> <td><a href="#mw">MW</a></td> <td><a href="#mw_5">MW/5</a></td> </tr> <tr> <td><a href="#n_free">N–Free</a></td> <td><a href="#o">O</a></td> <td><a href="#p35">P 35</a></td> </tr> <tr> <td><a href="#pro">Pro</a></td> <td><a href="#sot">SOT</a></td> <td><a href="#sw">SW</a></td> </tr> <tr> <td><a href="#tap">TAP</a></td> <td><a href="#tre">Tre</a></td> <td><a href="#uro">URO</a></td> </tr> <tr> <td><a href="#uro_h">URO–H</a></td> <td><a href="#uro_soil">URO + Soil</a></td> <td><a href="#uro_t">URO–T</a></td> </tr> <tr> <td><a href="#vt">VT</a></td> <td><a href="#vtac">VTAC</a></td> <td><a href="#vtyt">VTYT</a></td> </tr> <tr> <td><a href="#w">W</a></td> <td><a href="#wc">WC</a></td> <td><a href="#bbm2">9:1 agar medium</a></td> </tr> </tbody> </table> </section>   <section id="section02"> <h1>2. Media for marine and brackish water microalgae</h1> <table> <tbody> <tr> <td><a href="#besm">BESM</a></td> <td><a href="#besm2">BESM 2</a></td> <td><a href="#esm">ESM</a></td> <td><a href="#esm_cbv">ESM + CB–V</a></td> <td><a href="#esm2">ESM2</a></td> <td><a href="#f2">f/2</a></td> <td><a href="#f2_nh4cl">f/2 + NH<sub>4</sub>Cl </a></td> </tr> <tr> <td><a href="#imk">IMK</a></td> <td><a href="#imk10">IMK + NaCl</a></td> <td><a href="#imks">IMK + Soil Extract</a></td> <td><a href="#imk_2">IMK/2</a></td> <td><a href="#k">K</a></td> <td><a href="#k2">K/2</a></td> <td><a href="#k2et">K/2ET</a></td> </tr> <tr> <td><a href="#m_asp7">M–ASP7</a></td> <td><a href="#mb">MB</a></td> <td><a href="#mf">MF</a></td> <td><a href="#mkm">MKM</a></td> <td><a href="#mnk">MNK</a></td> <td><a href="#mimr">Modified IMR ( mIMR )</a></td> <td><a href="#mswm3">Modified SWM–3 (mSWM–3)</a></td> </tr> <tr> <td><a href="#pro99">PRO–99</a></td> <td><a href="#wesm">WESM</a></td> <td> </td> <td> </td> <td> </td> <td> </td> <td> </td> </tr> </tbody> </table> </section>   <section id="section03"> <h1>3. Bacteria-free check media for freshwater algae</h1> <table> <tbody> <tr> <td><a href="#b_i">B–I</a></td> <td><a href="#b_ii">B–II</a></td> <td><a href="#b_iii">B–III</a></td> <td><a href="#b_iv">B–IV</a></td> <td><a href="#b_v">B–V</a></td> <td><a href="#yt">YT</a></td> </tr> </tbody> </table> </section>   <section id="section04"> <h1>4. Bacteria-free check media for marine algae</h1> <table> <tbody> <tr> <td><a href="#bf_2">Bf/2</a></td> <td><a href="#mm23">MM 23</a></td> <td><a href="#stp">STP</a></td> </tr> </tbody> </table> </section>   <section id="section05"> <h1>5. Trace metals, vitamin mixtures and soil extracts</h1> <table> <tbody> <tr> <td><a href="#a2fe">A2 Fe stock solution</a></td> <td><a href="#a2trace">A2 trace element stock solution</a></td> <td><a href="#a5">A<sub>5</sub> solution</a></td> <td><a href="#allen_me">Allen metals</a></td> </tr> <tr> <td><a href="#c_mix">C–Source Mix II</a></td> <td><a href="#d_stock">D stock medium</a></td> <td><a href="#d_trace">D trace mix</a></td> <td><a href="#dy_trace">DY trace metal solution</a></td> </tr> <tr> <td><a href="#fe">Fe ( as EDTA; 1:1 molar )</a></td> <td><a href="#fe_solution">Fe solution</a></td> <td><a href="#f2_metals">f/2 metals</a></td> <td><a href="#hunter">Hutner ’s trace elements</a></td> </tr> <tr> <td><a href="#K_metals">K metals</a></td> <td><a href="#k_2_metals">K/2 metals</a></td> <td><a href="#mimr_trace">mIMR trace metals</a></td> <td><a href="#mm1_trace">mM–1 Trace elements</a></td> </tr> <tr> <td><a href="#mtygm9">mTYGM–9</a></td> <td><a href="#p1_metals">P–1 metals</a></td> <td><a href="#p_ii">P II metals</a></td> <td><a href="#p_iv">P IV metals</a></td> </tr> <tr> <td><a href="#p_n">P<span style="font-size: 50%;">N</span> metals</a></td> <td><a href="#pro99_trace">PRO–99 trace metals</a></td> <td><a href="#soil_extract">Soil extract</a></td> <td><a href="#a5_co">Trace metal mix A<sub>5</sub> + Co</a></td> </tr> <tr> <td><a href="#s3">Vitamin mix S<sub>3</sub></a></td> <td><a href="#w_edta">Waris EDTA</a></td> <td><a href="#wp2">Waris P-2</a></td> <td><a href="#wc_metals">WC trace metals</a></td> </tr> </tbody> </table> </section>   <section id="section06"> <h1>6. Media for protozoa</h1> <table> <tbody> <tr> <td><a href="#af6_wheat">AF–6 + Wheat</a></td> <td><a href="#bley">BLEY</a></td> <td><a href="#dgty">dGTY</a></td> </tr> <tr> <td><a href="#esm_mtygm9_rice">ESM + mTYGM–9 + Rice</a></td> <td><a href="#esm_rice">ESM + Rice</a></td> <td><a href="#f2_mtygm9_rice">f/2 + mTYGM–9 + Rice</a></td> </tr> <tr> <td><a href="#f2_rice">f/2 + Rice</a></td> <td><a href="#f2_wheat">f/2 + Wheat</a></td> <td><a href="#helio">Helio</a></td> </tr> <tr> <td><a href="#hemi">Hemi</a></td> <td><a href="#le">LE</a></td> <td><a href="#mizinko">Mizinko</a></td> </tr> <tr> <td><a href="#whs">Modified WarisH–Si</a></td> <td><a href="#pyg">PYG</a></td> <td><a href="#suy">SUY</a></td> </tr> <tr> <td><a href="#suy1_10">SUY 1/10</a></td> <td><a href="#suy1_10mt">SUY 1/10 + mTYGM–9 + Rice</a></td> <td><a href="#suy1_10_wheat">SUY 1/10 + Wheat</a></td> </tr> <tr> <td><a href="#uro_wheat">URO + Wheat</a></td> <td><a href="#uro_h_wheat">URO–H + Wheat</a></td> <td><a href="#uro_yt_wheat">URO + YT (1/10) + Wheat</a></td> </tr> <tr> <td><a href="#uyts_rice">UYTS + Rice</a></td> <td><a href="#ye100ppm">YE100ppm</a></td> <td> </td> </tr> </tbody> </table> </section>   <section id="section07"> <h1>7. Media for freshwater red algae</h1> <table style="width: 280px;"> <tbody> <tr> <td><a href="#bold3n">Bold 3N</a></td> </tr> </tbody> </table> </section>   <section id="section08"> <h1>8. Media for Charales</h1> <table> <tbody> <tr> <td><a href="#mswc2">mSWC–2 ( Modified SWC–2 )</a></td> <td><a href="#swc1">SWC–1</a></td> <td><a href="#swcn1">SWCN–1</a></td> <td><a href="#swcn2">SWCN–2</a></td> <td><a href="#swcn3">SWCN–3</a></td> <td><a href="#swcn4">SWCN–4</a></td> <td><a href="#1_3asw">1/3 Herbst ASW</a></td> </tr> </tbody> </table> </section>   <section id="section09" class="mediumDetailSection"> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="aaf6" id="aaf6"></a>AAF–6</h1> <div style="text-align: right;"> <a href="./medium/en/aaf6.pdf">Print</a> </div> <p> Prepare as for <a href="02medium.html#af6">AF–6</a> medium but adjust to pH 5.5–5.8. </p> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="acid_csi5" id="acid_csi5"></a>Acid–CSi/5</h1> <div style="text-align: right;"> <a href="./medium/en/acid_csi5.pdf">Print</a> </div> <p> Dilute <a href="02medium.html#csi">CSi</a> <sup>1 )</sup> medium with distilled water to one–fifth. Adjust to pH 3 with sulfuric acid. <br> 1 ) See <a href="02medium.html#csi">CSi</a> </p> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="af6" id="af6"></a>AF–6</h1> <div style="text-align: right;"> <a href="./medium/en/af6.pdf">Print</a> </div> <table> <tbody> <tr> <td>NaNO<sub>3</sub></td> <td>14 mg</td> </tr> <tr> <td>NH<sub>4</sub>NO<sub>3</sub></td> <td>2.2 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>3 mg</td> </tr> <tr> <td>KH<sub>2</sub>PO<sub>4</sub></td> <td>1 mg</td> </tr> <tr> <td>K<sub>2</sub>HPO<sub>4</sub></td> <td>0.5 mg</td> </tr> <tr> <td>CaCl<sub>2</sub> · 2H<sub>2</sub>O </td> <td>1 mg</td> </tr> <tr> <td>CaCO<sub>3</sub><sup> 1 )</sup></td> <td>1 mg</td> </tr> <tr> <td>Fe–citrate</td> <td>0.2 mg</td> </tr> <tr> <td>Citric acid</td> <td>0.2 mg</td> </tr> <tr> <td>Biotin</td> <td>0.2 μg</td> </tr> <tr> <td>Thiamine HCl</td> <td>1 μg</td> </tr> <tr> <td>Vitamin B<sub>6</sub></td> <td>0.1 μg</td> </tr> <tr> <td>Vitamin B<sub>12</sub></td> <td>0.1 μg</td> </tr> <tr> <td>Trace metals<sup> 1 )</sup></td> <td>0.5 mL</td> </tr> <tr> <td>Distilled water</td> <td>99.5 mL</td> </tr> <tr> <td colspan="2">pH 6.6<sup> 2 )</sup></td> </tr> </tbody> </table> <p> 1 ) In the NIES–Collection, CaCO3 is removed and <a href="http://mcc.nies.go.jp/02medium.html#p_iv">PIV metals</a> are used instead of Trace metals. <br> 2 ) In the NIES–Collection, 40 mg MES is added and pH is adjusted to 6.6. </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Kato, S. 1982 Laboratory culture and morphology of <i>Colacium vesiculosum </i> Ehrb. ( Euglenophyceae ). <i>Jpn. J. Phycol.</i>, <b>30</b>, 63-67 ( in Japanese with English summary ). </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="af6_2" id="af6_2"></a>AF–6/2</h1> <div style="text-align: right;"> <a href="./medium/en/af6_2.pdf">Print</a> </div> <p> <a href="http://mcc.nies.go.jp/02medium.html#af6">AF–6</a> <sup>1 )</sup> medium is diluted with distilled water to one-half. <br> 1 ) See <a href="http://mcc.nies.go.jp/02medium.html#af6">AF–6</a> </p> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="af6_3" id="af6_3"></a>AF–6/3</h1> <div style="text-align: right;"> <a href="./medium/en/af6_3.pdf">Print</a> </div> <p> Mix 1 volume of <a href="http://mcc.nies.go.jp/02medium.html#af6">AF–6</a><sup> 1 )</sup> medium and 2 volumes of distilled water.<br> 1 ) See <a href="http://mcc.nies.go.jp/02medium.html#af6">AF–6</a> </p> <h3></h3> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="afac" id="afac"></a>AFAC</h1> <div style="text-align: right;"> <a href="./medium/en/afac.pdf">Print</a> </div> <p> To 100 mL <a href="http://mcc.nies.go.jp/02medium.html#af6">AF–6</a><sup> 1 )</sup> medium add 20 mg sodium acetate.<br> 1 ) See <a href="http://mcc.nies.go.jp/02medium.html#af6">AF–6</a> </p> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="allen" id="allen"></a>Allen</h1> <div style="text-align: right;"> <a href="./medium/en/allen.pdf">Print</a> </div> <table> <tbody> <tr> <td>( NH<sub>4</sub> )<sub>2</sub>SO<sub>4</sub></td> <td>132 mg</td> </tr> <tr> <td>KH<sub>2</sub>PO<sub>4</sub></td> <td>27.2 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>24.6 mg</td> </tr> <tr> <td>CaCl<sub>2</sub> · 2H<sub>2</sub>O </td> <td>7.4 mg</td> </tr> <tr> <td><a href="#allen_me">Allen metals</a><sup> 1 )</sup></td> <td>0.01 mL</td> </tr> <tr> <td>Distilled water</td> <td>99.9 mL</td> </tr> <tr> <td colspan="2">pH 2.5<sup> 2 )</sup></td> </tr> </tbody> </table> <p> 1 ) See <a href="http://mcc.nies.go.jp/02medium.html#allen_me">Allen metals</a> <br> 2 ) pH is adjusted to 2.5 with 0.5 mol / L H2SO4. </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Allen, M. B. 1959 Studies with Cyanidium caldarium, an anomalously pigmented chlorophyte. <i>Arch. Mikrobiol</i>., <b>32</b>, 270-277. </td> </tr> <tr> <td>Starr, R. C., Zeikus, J. A. 1987 UTEX - The culture collection of algae at the University of Texas at Austin. <i>J. Phycol., <b>23, Suppl. to Sept</b> </i>., 1-106. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="bbm" id="bbm"></a>BBM</h1> <div style="text-align: right;"> <a href="./medium/en/bbm.pdf">Print</a> </div> <table> <tbody> <tr> <td>NaNO<sub>3</sub></td> <td>25 mg</td> </tr> <tr> <td>KH<sub>2</sub>PO<sub>4</sub></td> <td>17.5 mg</td> </tr> <tr> <td>K<sub>2</sub>HPO<sub>4</sub></td> <td>10 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>7.5 mg</td> </tr> <tr> <td>CaCl<sub>2</sub> · 2H<sub>2</sub>O </td> <td>2.5 mg</td> </tr> <tr> <td>NaCl</td> <td>2.5 mg</td> </tr> <tr> <td>KOH</td> <td>3.1 mg</td> </tr> <tr> <td>FeSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>0.498 mg</td> </tr> <tr> <td>H<sub>3</sub>BO<sub>3</sub></td> <td>1.142 mg</td> </tr> <tr> <td>ZnSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>0.882 mg</td> </tr> <tr> <td>MnCl<sub>2</sub> · 4H<sub>2</sub>O </td> <td>0.144 mg</td> </tr> <tr> <td>MoO<sub>3</sub></td> <td>0.071 mg</td> </tr> <tr> <td>CuSO<sub>4</sub> · 5H<sub>2</sub>O </td> <td>0.157 mg</td> </tr> <tr> <td>Co( NO<sub>3</sub> )<sub>2</sub> · 6H<sub>2</sub>O </td> <td>0.049 mg</td> </tr> <tr> <td>Na<sub>2</sub>EDTA </td> <td>5 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> </tbody> </table> <p>Add 1.2 g agar to 100 mL of medium to give a solid medium.</p> <table class="media4"> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Bischoff, H. W., Bold, H. C. 1963 Some soil algae from enchanted rock and related algal species. <i>Phycological Studies IV.,Univ. No. 6318</i>, Texas, p. 95. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="bg11" id="bg11"></a>BG–11</h1> <div style="text-align: right;"> <a href="./medium/en/bg11.pdf">Print</a> </div> <table> <tbody> <tr> <td>NaNO<sub>3</sub></td> <td>150 mg</td> </tr> <tr> <td>K<sub>2</sub>HPO<sub>4</sub> · 3H<sub>2</sub>O </td> <td>4 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>7.5 mg</td> </tr> <tr> <td>CaCl<sub>2</sub> · 2H<sub>2</sub>O </td> <td>3.6 mg</td> </tr> <tr> <td>Citric acid</td> <td>0.6 mg</td> </tr> <tr> <td>Ferric ammonium citrate</td> <td>0.6 mg</td> </tr> <tr> <td>Na<sub>2</sub>EDTA – Mg </td> <td>0.1 mg</td> </tr> <tr> <td>Na<sub>2</sub>CO<sub>3</sub></td> <td>2 mg</td> </tr> <tr> <td><a href="#a5_co">Trace metal mix A<sub>5</sub> + Co </a><sup> 1 )</sup></td> <td>0.1 mL</td> </tr> <tr> <td>Distilled water</td> <td>99.9 mL</td> </tr> <tr> <td colspan="2">pH 7.4</td> </tr> </tbody> </table> <p> Add 1.5 g agar to 100 mL of medium to give a solid medium.<br> 1 ) See <a href="#a5_co">Trace metal mix A<sub>5</sub> + Co </a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Waterbury, J. B., Sranier, R. Y. 1981 Isolation and growth of cyanobacteria from marine and hypersaline environments. In <i>The Prokaryotes</i>, Springer Berlin Heidelberg, New York, pp. 221-223. </td> </tr> </tbody> </table> <br/> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="c" id="c"></a>C</h1> <div style="text-align: right;"> <a href="./medium/en/c.pdf">Print</a> </div> <table> <tbody> <tr> <td>Ca( NO<sub>3</sub> )<sub>2</sub> · 4H<sub>2</sub>O </td> <td>15 mg</td> </tr> <tr> <td>KNO<sub>3</sub></td> <td>10 mg</td> </tr> <tr> <td>β–Na<sub>2</sub>glycerophosphate · 5H<sub>2</sub>O </td> <td>5 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>4 mg</td> </tr> <tr> <td>Vitamin B<sub>12</sub></td> <td>0.01 μg</td> </tr> <tr> <td>Biotin</td> <td>0.01 μg</td> </tr> <tr> <td>Thiamine HCl</td> <td>1 μg</td> </tr> <tr> <td><a href="#p_iv">PIV metals</a><sup> 1 )</sup></td> <td>0.3 mL</td> </tr> <tr> <td>Tris ( hydroxymethyl ) aminomethane</td> <td>50 mg</td> </tr> <tr> <td>Distilled water</td> <td>99.7 mL</td> </tr> <tr> <td colspan="2">pH 7.5</td> </tr> </tbody> </table> <p> Add 1.5 g agar to 100 mL of medium to give a solid medium. <br> 1 ) See <a href="#p_iv">PIV metals</a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Ichimura, T. 1971 Sexual cell division and conjugation-papilla formation in sexual reproduction of <i>Closterium strigosum</i>. In <i>Proceedings of the Seventh International Seaweed Symposium</i>, University of Tokyo Press, Tokyo, p. 208-214. </td> </tr> </tbody> </table> <br> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="c10seawater" id="c10seawater"></a>C+10% Seawater ( N. Tezuka, unpubl. )</h1> <div style="text-align: right;"> <a href="./medium/en/c10seawater.pdf">Print</a> </div> <p> <a href="#c">C</a><sup> 1 )</sup> medium with 10% filtered seawater. <br> 1 ) See <a href="#c">C</a> </p> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="c6" id="c6"></a>C/6</h1> <div style="text-align: right;"> <a href="./medium/en/c6.pdf">Print</a> </div> <p> Mix 1 volume of <a href="#c">C</a><sup> 1 )</sup> medium and 5 volumes of distilled water.<br> 1 ) See <a href="#c">C</a> </p> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="c6g" id="c6g"></a>C/6G</h1> <div style="text-align: right;"> <a href="./medium/en/c6g.pdf">Print</a> </div> <p> Mix 1 volume of <a href="#c">C</a><sup> 1 )</sup> medium and 5 volumes of Lake Nojiri water ( sterilized through GF/F filter, and store at 5°C ). <br> 1 ) See <a href="#c">C</a> </p> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="c10" id="c10"></a>C/10</h1> <div style="text-align: right;"> <a href="./medium/en/c10.pdf">Print</a> </div> <p> Mix 1 volume of <a href="#c">C</a><sup> 1 )</sup> medium and 9 volumes of distilled water.<br> 1 ) See <a href="#c">C</a> <br> </p> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="ca" id="ca"></a>CA</h1> <div style="text-align: right;"> <a href="./medium/en/ca.pdf">Print</a> </div> <table> <tbody> <tr> <td>Ca( NO<sub>3</sub> )<sub>2</sub> · 4H<sub>2</sub>O </td> <td>2 mg</td> </tr> <tr> <td>KNO<sub>3</sub></td> <td>10 mg</td> </tr> <tr> <td>NH<sub>4</sub>NO<sub>3</sub></td> <td>5 mg</td> </tr> <tr> <td>β–Na<sub>2</sub>glycerophosphate · 5H<sub>2</sub>O </td> <td>3 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>2 mg</td> </tr> <tr> <td>Vitamin B<sub>12</sub></td> <td>0.01 μg</td> </tr> <tr> <td>Biotin</td> <td>0.01 μg</td> </tr> <tr> <td>Thiamine HCl</td> <td>1 μg</td> </tr> <tr> <td><a href="#p_iv">PIV metals</a><sup> 1 )</sup></td> <td>0.1 mL</td> </tr> <tr> <td><a href="#fe">Fe ( as EDTA; 1:1 molar )</a><sup> 2 )</sup></td> <td>0.1 mL</td> </tr> <tr> <td>HEPES</td> <td>40 mg</td> </tr> <tr> <td>Distilled water</td> <td>99.9 mL</td> </tr> <tr> <td colspan="2">pH 7.2</td> </tr> </tbody> </table> <p> 1 ) See <a href="#p_iv">PIV metals</a><br> 2 ) See <a href="#fe">Fe ( as EDTA; 1:1 molar )</a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Ichimura, T., Watanabe, M. 1974 The <i>Closterium calosporum</i> complex from the Ryukyu Islands - Variation and taxonomical problems. <i>Mem. Natn. Sci. Mus. Tokyo</i>, 7, 89-102, pls. 13-14. </td> </tr> </tbody> </table> <br/> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="cam" id="cam"></a>CAM</h1> <div style="text-align: right;"> <a href="./medium/en/cam.pdf">Print</a> </div> <p> <a href="#ca">CA</a><sup> 1 )</sup> medium with pH adjusted to 6.5 by buffering with MES instead of HEPES. <br> 1 ) See <a href="#ca">CA</a> </p> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="carefoot" id="carefoot"></a>Carefoot</h1> <div style="text-align: right;"> <a href="./medium/en/carefoot.pdf">Print</a> </div> <table> <tbody> <tr> <td>NaNO<sub>3</sub></td> <td>24.7 mg</td> </tr> <tr> <td>CaCl<sub>2</sub> · 2H<sub>2</sub>O </td> <td>1.1 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>4.7 mg</td> </tr> <tr> <td>K<sub>2</sub>HPO<sub>4</sub></td> <td>0.9 mg</td> </tr> <tr> <td>KH<sub>2</sub>PO<sub>4</sub></td> <td>2.3 mg</td> </tr> <tr> <td>NaCl</td> <td>1.5 mg</td> </tr> <tr> <td><a href="#p_iv">PIV metals</a><sup> 1 )</sup></td> <td>0.5 mL</td> </tr> <tr> <td>Distilled water</td> <td>99.5 mL</td> </tr> <tr> <td colspan="2">pH 7.5</td> </tr> </tbody> </table> <p> In the NIES–Collection, 0.02 μg vitamin B<sub>12</sub>, 0.02 μg biotin and 2 μg thiamine HCl are added to this medium. <br> 1 ) See <a href="#p_iv">PIV metals</a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Carefoot, J. R. 1968 Culture and heterotrophy of the freshwater dinoflagellate, <i>Peridinium cinctum fa. ovoplanum Lindeman. J. Phycol</i>., <b>4</b>, 129-131. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="cb" id="cb"></a>CB</h1> <div style="text-align: right;"> <a href="./medium/en/cb.pdf">Print</a> </div> <p> To <a href="#c">C</a><sup> 1 )</sup> medium add Bicine instead of Tris ( hydroxymethyl ) aminomethane, and adjust pH to 9.0. <br> 1 ) See <a href="#c">C</a> </p> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="cb_v" id="cb_v"></a>CB–V</h1> <div style="text-align: right;"> <a href="./medium/en/cb_v.pdf">Print</a> </div> <p> Make <a href="#b_v">B–V</a><sup> 1 )</sup> medium with <a href="#c">C</a><sup> 2 )</sup> medium. <br> 1 ) See <a href="#b_v">B–V</a> <br> 2 ) See <a href="#c">C</a> </p> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="cc" id="cc"></a>CC</h1> <div style="text-align: right;"> <a href="./medium/en/cc.pdf">Print</a> </div> <p> <a href="#c">C</a><sup> 1 )</sup> medium with pH adjusted to 3.0 by buffering with 1,2,3,4–cyclopentane tetracarboxylic acid instead of Tris ( hydroxymethyl ) aminomethane.<br> 1 ) See <a href="#c">C</a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Ichimura, T., Itoh, T. 1977 17. Preservation methods of microalgae ( I ) [17. Bisaisôrui no hozonhô ( I )]. In <i>Preservation methods of microorganisms [ Biseibutsu Hozonhô]</i>, Ed. by Nei, T., University of Tokyo Press, Tokyo, p. 355-373 ( in Japanese without English title ). </td> </tr> </tbody> </table> <p class="mediumCategoryTitle"> <br> 1 Media for freshwater, terrestrial, hot spring and salt water algae </p> <h1><a name="csi" id="csi"></a>CSi</h1> <div style="text-align: right;"> <a href="./medium/en/csi.pdf">Print</a> </div> <p> <a href="#c">C</a><sup> 1 )</sup> medium with pH adjusted to 7.0 by buffering with 50mg HEPES instead of Tris ( hydroxymethyl ) aminomethane. Thereafter, 10 mg Na<sub>2</sub>SiO<sub>3</sub> · 9H<sub>2</sub>O is added. <br> 1 ) See <a href="#c">C</a> </p> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="csi_cu" id="csi_cu"></a>CSi + Cu</h1> <div style="text-align: right;"> <a href="./medium/en/csi_cu.pdf">Print</a> </div> <p> To 100 mL <a href="#csi">CSi</a><sup> 1 )</sup> medium add 0.25 mg CuSO<sub>4</sub> · 5H<sub>2</sub>O and 100 mg agar. <br> 1 ) See <a href="#csi">CSi</a> </p> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="csi5" id="csi5"></a>CSi/5</h1> <div style="text-align: right;"> <a href="./medium/en/csi5.pdf">Print</a> </div> <p> Dilute <a href="#csi">CSi</a><sup> 1 )</sup> medium with distilled water to 1/5.<br> 1 ) See <a href="#csi">CSi</a> <br> </p> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="ct" id="ct"></a>CT</h1> <div style="text-align: right;"> <a href="./medium/en/ct.pdf">Print</a> </div> <p> <a href="#c">C</a><sup> 1 )</sup> medium with pH adjusted to 8.2 by buffering with 40mg TAPS instead of Tris ( hydroxymethyl ) aminomethane.<br> 1 ) See <a href="#c">C</a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Watanabe, M. M., Ichimura, T. 1977 Fresh- and salt-water forms of <i>Spirulina platensis</i> in axenic cultures. <i>Bull. Jpn. Soc. Phycol., <b>25, Suppl. ( Mem. Iss. Yamada )</b> </i>, 371-377. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="cyt" id="cyt"></a>CYT</h1> <div style="text-align: right;"> <a href="./medium/en/cyt.pdf">Print</a> </div> <p> To 100 mL <a href="#c">C</a><sup> 1 )</sup> medium add 100 mg yeast extract and 200 mg tryptone. <br> 1 ) See <a href="#c">C</a> </p> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="dh_fe" id="dh_fe"></a>DH + Fe ( I.I. Brown, unpubl. )</h1> <div style="text-align: right;"> <a href="./medium/en/dh_fe.pdf">Print</a> </div> <table> <tbody> <tr> <td><a href="#d_stock">D stock medium</a><sup> 1 )</sup></td> <td>5 mL</td> </tr> <tr> <td>HEPES</td> <td>0.12 mg</td> </tr> <tr> <td>FeCl<sub>3</sub> · 6H<sub>2</sub>O </td> <td>1.14 mg</td> </tr> <tr> <td>Distilled water</td> <td>95 mL</td> </tr> <tr> <td colspan="2">pH 8.24 – 8.26</td> </tr> </tbody> </table> <p> After autoclaving, keep in room temperature overnight. Next day, adjust pH to 7.5–7.6 and add 1.5 g agar. <br> 1 ) See <a href="#d_stock">D stock medium</a> </p> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="dy_v" id="dy_v"></a>DY–V</h1> <div style="text-align: right;"> <a href="./medium/en/dy_v.pdf">Print</a> </div> <table> <tbody> <tr> <td>MES</td> <td>20 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>5 mg</td> </tr> <tr> <td>KCl</td> <td>0.3 mg</td> </tr> <tr> <td>NH<sub>4</sub>Cl </td> <td>0.27 mg</td> </tr> <tr> <td>NaNO<sub>3</sub></td> <td>2 mg</td> </tr> <tr> <td>β–Na<sub>2</sub>glycerophosphate · 5H<sub>2</sub>O </td> <td>0.22 mg</td> </tr> <tr> <td>H<sub>3</sub>BO<sub>3</sub></td> <td>0.08 mg</td> </tr> <tr> <td>Na<sub>2</sub>EDTA · 2H<sub>2</sub>O </td> <td>0.8 mg</td> </tr> <tr> <td>Na<sub>2</sub>SiO<sub>3</sub> · 9H<sub>2</sub>O </td> <td>1.4 mg</td> </tr> <tr> <td>FeCl<sub>3</sub> · 6H<sub>2</sub>O </td> <td>0.1 mg</td> </tr> <tr> <td>CaCl<sub>2</sub> · 2H<sub>2</sub>O </td> <td>7.5 mg</td> </tr> <tr> <td>Vitamin B<sub>12</sub></td> <td>0.05 μg</td> </tr> <tr> <td>Biotin</td> <td>0.05 μg</td> </tr> <tr> <td>Thiamine HCl</td> <td>10 μg</td> </tr> <tr> <td><a href="#dy_trace">DY trace metal solution</a><sup> 1 )</sup></td> <td>0.1 mL</td> </tr> <tr> <td>Distilled water</td> <td>99.9 mL</td> </tr> <tr> <td colspan="2">pH 6.8</td> </tr> </tbody> </table> <p> 1 ) See <a href="#dy_trace">DY trace metal solution</a><br> </p> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="hut" id="hut"></a>HUT</h1> <div style="text-align: right;"> <a href="./medium/en/hut.pdf">Print</a> </div> <table> <tbody> <tr> <td>KH<sub>2</sub>PO<sub>4</sub></td> <td>2 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>2.5 mg</td> </tr> <tr> <td>Sodium acetate</td> <td>40 mg</td> </tr> <tr> <td>Potassium citrate</td> <td>4 mg</td> </tr> <tr> <td>Polypeptone</td> <td>60 mg</td> </tr> <tr> <td>Yeast extract</td> <td>40 mg</td> </tr> <tr> <td>Vitamin B<sub>12</sub></td> <td>0.05 μg</td> </tr> <tr> <td>Thiamine HCl</td> <td>0.04 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> <tr> <td colspan="2">pH 6.4</td> </tr> </tbody> </table> <p>Add 150 mg agar to 100 mL of medium to give a semi-solid medium.</p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Ichimura, T. 1979 2. Isolation and culture methods of algae. 2.5.B. Freshwater algae [2. Sôrui no bunri to baiyôhô. 2.5.B. Tansui sôrui]. In <i>Methods in Phycological Studies [Sôrui Kenkyûhô],</i> Eds. by Nishizawa, K. & Chihara, M., Kyoritsu Shuppan, Tokyo, p. 301 (in Japanese without English title). </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="m11" id="m11"></a>M–11</h1> <div style="text-align: right;"> <a href="./medium/en/m11.pdf">Print</a> </div> <table> <tbody> <tr> <td>NaNO<sub>3</sub></td> <td>10 mg</td> </tr> <tr> <td>K<sub>2</sub>HPO<sub>4</sub></td> <td>1 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>7.5 mg</td> </tr> <tr> <td>CaCl<sub>2</sub> · 2H<sub>2</sub>O </td> <td>4 mg</td> </tr> <tr> <td>Na<sub>2</sub>CO<sub>3</sub></td> <td>3 mg</td> </tr> <tr> <td>FeSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>0.1 mg</td> </tr> <tr> <td>Na<sub>2</sub>EDTA · 2H<sub>2</sub>O </td> <td>0.1 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> <tr> <td colspan="2">pH 8.0</td> </tr> </tbody> </table> <br> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Hagiwara, T., Yagi, O., Takamura, Y., Sudo, R. 1984 Isolation of bacteria-free <i>Microcystis aeruginosa</i> from Lake Kasumigaura. <i>Jpn. J. Water Poll. Res</i>., <b>7</b>, 437-442 ( in Japanese with English summary ). </td> </tr> <tr> <td>Yagi, O., Okada, M., Sudo, R. 1979 Cultivation of <i>Microcystis</i> and red-tide-organisms. <i>Res. Rep. Natl. Inst. Environ. Stud</i>., <b>No. 6</b>, 223-229 ( in Japanese with English summary ). </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="ma" id="ma"></a>MA</h1> <div style="text-align: right;"> <a href="./medium/en/ma.pdf">Print</a> </div> <table> <tbody> <tr> <td>Ca( NO<sub>3</sub> )<sub>2</sub> · 4H<sub>2</sub>O </td> <td>5 mg</td> </tr> <tr> <td>KNO<sub>3</sub></td> <td>10 mg</td> </tr> <tr> <td>NaNO<sub>3</sub></td> <td>5 mg</td> </tr> <tr> <td>Na<sub>2</sub>SO<sub>4</sub></td> <td>4 mg</td> </tr> <tr> <td>MgCl<sub>2</sub> · 6H<sub>2</sub>O </td> <td>5 mg</td> </tr> <tr> <td>β–Na<sub>2</sub>glycerophosphate · 5H<sub>2</sub>O </td> <td>10 mg</td> </tr> <tr> <td>Na<sub>2</sub>EDTA · 2H<sub>2</sub>O </td> <td>0.5 mg</td> </tr> <tr> <td>FeCl<sub>3</sub> · 6H<sub>2</sub>O </td> <td>0.05 mg</td> </tr> <tr> <td>MnCl<sub>2</sub> · 4H<sub>2</sub>O </td> <td>0.5 mg</td> </tr> <tr> <td>ZnCl<sub>2</sub></td> <td>0.05 mg</td> </tr> <tr> <td>CoCl<sub>2</sub> · 6H<sub>2</sub>O </td> <td>0.5 mg</td> </tr> <tr> <td>Na<sub>2</sub>MoO<sub>4</sub> · 2H<sub>2</sub>O </td> <td>0.08 mg</td> </tr> <tr> <td>H<sub>3</sub>BO<sub>3</sub></td> <td>2 mg</td> </tr> <tr> <td>Bicine</td> <td>50 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> <tr> <td colspan="2">pH 8.6</td> </tr> </tbody> </table> <br> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Ichimura, T. 1979 2. Isolation and culture methods of algae. 2.5.B. Freshwater algae [2. Sôrui no bunri to baiyôhô. 2.5.B. Tansui sôrui]. In <i>Methods in Phycological Studies [Sôrui Kenkyûhô]</i>, Eds. by Nishizawa, K. & Chihara, M., Kyoritsu Shuppan, Tokyo, p. 294-305 ( in Japanese without English title ). </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="maf6" id="maf6"></a>MAF–6</h1> <div style="text-align: right;"> <a href="./medium/en/maf6.pdf">Print</a> </div> <p> To 100 mL of <a href="#af6">AF–6</a><sup> 1 )</sup> medium add 10 mg glucose and 10 mg sodium acetate. <br> 1 ) See <a href="#af6">AF–6</a><br> </p> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="m_allen" id="m_allen"></a>M–Allen</h1> <div style="text-align: right;"> <a href="./medium/en/m_allen.pdf">Print</a> </div> <table> <tbody> <tr> <td>( NH<sub>4</sub> )<sub>2</sub>SO<sub>4</sub></td> <td>262 mg</td> </tr> <tr> <td>KH<sub>2</sub>PO<sub>4</sub></td> <td>54 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>50 mg</td> </tr> <tr> <td>CaCl<sub>2</sub> · 2H<sub>2</sub>O </td> <td>14 mg</td> </tr> <tr> <td><a href="#a2trace">A2 trace element stock solution</a><sup> 1 )</sup></td> <td>0.2 mL</td> </tr> <tr> <td>Distilled water</td> <td>99.4 mL</td> </tr> <tr> <td colspan="2">pH 2.5<sup> 2 )</sup></td> </tr> </tbody> </table> <p> After autoclaving, add 0.4 mL of <a href="#a2fe">A2 Fe stock solution</a><sup> 3 )</sup> ( filter-sterilized ). <br> 1 ) See <a href="#a2trace">A2 trace element stock solution</a> <br> 2 ) pH is adjusted to 2.5 with 0.5 mol / L H<sub>2</sub>SO<sub>4</sub>. <br> 3 ) See <a href="#a2fe">A2 Fe stock solution</a> <br> Indicated as “MA” medium in reference. </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Minoda, A., Sakagami, R., Yagisawa, F., Kuroiwa, T., Tanaka, K. 2004 Improvement of culture conditions and evidence for nuclear transformation by homologous recombination in a red alga, <i>Cyanidioschyzon merolae</i> 10D. <i>Plant Cell Physiol.</i>, <b>45</b>, 667-671.</td> </tr> </tbody> </table> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="m_allen_g" id="m_allen_g"></a>M–Allen + Glucose</h1> <div style="text-align: right;"> <a href="./medium/en/m_allen_g.pdf">Print</a> </div> <p> To 100 mL <a href="#m_allen">M–Allen</a><sup> 1 )</sup> medium add 0.5 –1 g Glucose.<br> 1 ) See <a href="#m_allen">M–Allen</a><br> <br> </p> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="m_allen_u" id="m_allen_u"></a>M–Allen + Uracil</h1> <div style="text-align: right;"> <a href="./medium/en/m_allen_u.pdf">Print</a> </div> <p> To 100 mL <a href="#m_allen">M–Allen</a><sup> 1 )</sup> medium add 50 mg uracil.<br> 1 ) See <a href="#m_allen">M–Allen</a><br> <br> </p> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="mbm" id="mbm"></a>MBM</h1> <div style="text-align: right;"> <a href="./medium/en/mbm.pdf">Print</a> </div> <table> <tbody> <tr> <td>KNO<sub>3</sub></td> <td>25 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>7.5 mg</td> </tr> <tr> <td>K<sub>2</sub>HPO<sub>4</sub></td> <td>7.5 mg</td> </tr> <tr> <td>KH<sub>2</sub>PO<sub>4</sub></td> <td>17.5 mg</td> </tr> <tr> <td>NaCl</td> <td>2.5 mg</td> </tr> <tr> <td>CaCl<sub>2</sub> · 2H<sub>2</sub>O </td> <td>1 mg</td> </tr> <tr> <td><a href="#fe_solution">Fe solution</a><sup> 1 )</sup></td> <td>0.1 mL</td> </tr> <tr> <td><a href="#a5">A<sub>5</sub> solution </a><sup> 2 )</sup></td> <td>0.1 mL</td> </tr> <tr> <td>Agar</td> <td>1.5 g</td> </tr> <tr> <td>Distilled water</td> <td>99.8 mL</td> </tr> <tr> <td colspan="2">pH 6.0</td> </tr> </tbody> </table> <p> 1 ) See <a href="#fe_solution">Fe solution</a>.<br> 2 ) See <a href="#a5">A<sub>5</sub> solution </a>. <br> Indicated as “Modified Bristol medium” in reference. </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Ichimura, T., Itoh, T. 1977 17. Preservation methods of microalgae ( I ) [17. Bisaisôrui no hozonhô ( I )]. In <i>Preservation methods of microorganisms [Biseibutsu Hozonhô]</i>, Ed. by Nei, T., University of Tokyo Press, Tokyo, p. 355-373 ( in Japanese without English title ). </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="m_chu_no10" id="m_chu_no10"></a>M Chu No. 10</h1> <div style="text-align: right;"> <a href="./medium/en/m_chu_no10.pdf">Print</a> </div> <table> <tbody> <tr> <td>Ca( NO<sub>3</sub> )<sub>2</sub> · 4H<sub>2</sub>O </td> <td>2.0 mg</td> </tr> <tr> <td>KH<sub>2</sub>PO<sub>4</sub></td> <td>0.62 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>2.5 mg</td> </tr> <tr> <td>Na<sub>2</sub>CO<sub>3</sub></td> <td>2 mg</td> </tr> <tr> <td>Na<sub>2</sub>SiO<sub>3</sub> · 9H<sub>2</sub>O </td> <td>2.5 mg</td> </tr> <tr> <td>HCl ( 1 mol / L )<sup> 1 )</sup></td> <td>0.025 mL</td> </tr> <tr> <td>Na<sub>2</sub>EDTA · 2H<sub>2</sub>O </td> <td>0.2 mg</td> </tr> <tr> <td>FeCl<sub>3</sub> · 6H<sub>2</sub>O </td> <td>0.1 mg</td> </tr> <tr> <td>H<sub>3</sub>BO<sub>3</sub></td> <td>0.248 mg</td> </tr> <tr> <td>MnCl<sub>2</sub> · 4H<sub>2</sub>O </td> <td>0.139 mg</td> </tr> <tr> <td>( NH<sub>4</sub> )<sub>6</sub>Mo<sub>7</sub>O<sub>24</sub> · 4H<sub>2</sub>O </td> <td>0.1 mg</td> </tr> <tr> <td>Vitamin B<sub>12</sub></td> <td>1 μg</td> </tr> <tr> <td>Thiamine HCl</td> <td>0.1 μg</td> </tr> <tr> <td>Biotin</td> <td>0.1 μg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> </tbody> </table> <p>1 ) In the NIES–Collection, pH is adjusted to 7.6 with 1 mol / L HCl.</p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Chu, S. P. 1942 The influence of the mineral composition of the medium on the growth of planktonic algae. Part I. Methods and culture media. <i>J. Ecol., <b>30</b></i>, 284-325. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="mdm" id="mdm"></a>MDM</h1> <div style="text-align: right;"> <a href="./medium/en/mdm.pdf">Print</a> </div> <table> <tbody> <tr> <td>KNO<sub>3</sub></td> <td>100 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>25 mg</td> </tr> <tr> <td>K<sub>2</sub>HPO<sub>4</sub></td> <td>25 mg</td> </tr> <tr> <td>NaCl</td> <td>10 mg</td> </tr> <tr> <td>CaCl<sub>2</sub> · 2H<sub>2</sub>O </td> <td>1 mg</td> </tr> <tr> <td><a href="#fe_solution">Fe solution</a><sup> 1 )</sup></td> <td>0.1 mL</td> </tr> <tr> <td><a href="#a5">A<sub>5</sub> solution </a><sup> 2 )</sup></td> <td>0.1 mL</td> </tr> <tr> <td>Agar</td> <td>1.5 g</td> </tr> <tr> <td>Distilled water</td> <td>99.8 mL</td> </tr> <tr> <td colspan="2">pH 8.0</td> </tr> </tbody> </table> <p> 1 ) See <a href="#fe_solution">Fe solution</a> <br> 2 ) See <a href="#a5">A<sub>5</sub> solution </a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Watanabe, A. 1960 List of algal strains in collection at the Institute of Applied Microbiology, University of Tokyo. <i>J. Gen. Appl. Microbiol</i>., <b>6</b>, 283-292. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="mg" id="mg"></a>MG</h1> <div style="text-align: right;"> <a href="./medium/en/mg.pdf">Print</a> </div> <table> <tbody> <tr> <td>Ca ( NO<sub>3</sub> )<sub>2</sub>·4H<sub>2</sub>O </td> <td>2 mg</td> </tr> <tr> <td>KNO<sub>3</sub></td> <td>10 mg</td> </tr> <tr> <td>β–Na<sub>2</sub> glycerophosphate·5H<sub>2</sub>O </td> <td>3 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>2 mg</td> </tr> <tr> <td>Vitamin B<sub>12</sub></td> <td>0.01 µg</td> </tr> <tr> <td>Biotin</td> <td>0.01 µg</td> </tr> <tr> <td>Thiamine HCl</td> <td>1 µg</td> </tr> <tr> <td><a href="#p_iv">PIV metals</a> <sup> 1 )</sup></td> <td>0.1  mL</td> </tr> <tr> <td><a href="#fe">Fe ( as EDTA; 1:1 molar )</a><sup> 2 )</sup></td> <td>0.1  mL</td> </tr> <tr> <td>HEPES</td> <td>40 mg</td> </tr> <tr> <td>Distilled water</td> <td>99.8  mL</td> </tr> <tr> <td colspan="2">pH 7.2</td> </tr> </tbody> </table> <p> 1 ) See <a href="#p_iv">PIV metals</a><br> 2 ) See <a href="#fe">Fe ( as EDTA; 1:1 molar )</a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Ichimura, T. 1973 The life cycle and its control in some species of <i>Closterium</i>, with special reference to the biological species problems. <i>Thesis D. Sci</i>., University of Tokyo, 69 pp., 11 tables, 40 figs. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="mgm" id="mgm"></a>MGM</h1> <div style="text-align: right;"> <a href="./medium/en/mgm.pdf">Print</a> </div> <p> <a href="http://mcc.nies.go.jp/02medium.html#mg">MG</a> 1 ) medium with pH adjusted to 6.5 by buffering with MES instead of HEPES. <br> 1 ) See <a href="http://mcc.nies.go.jp/02medium.html#mg">MG</a> </p> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="mac" id="mac"></a>Modified acetate medium ( mAC )</h1> <div style="text-align: right;"> <a href="./medium/en/mac.pdf">Print</a> </div> <p> To 100 mL <a href="#af6">AF–6</a><sup> 1 )</sup> medium, add 40mg glucose, yeast extract, tryptone, and sodium acetate.<br> 1 ) See <a href="#af6">AF–6</a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Nozaki, H., Watanabe, M. M. & Aizawa, K. 1995 Morphology and paedogamous sexual reproduction in <i>Chlorogonium capillatum</i> sp. nov. ( Volvocales, Chlorophyta ). <i>J. Phycol</i>., <b>31</b>, 655-663. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1> <a name="bg0" id="bg0"></a>Modified BG–11<sub>0</sub> </h1> <div style="text-align: right;"> <a href="./medium/en/bg0.pdf">Print</a> </div> <table> <tbody> <tr> <td>1M HEPES–NaOH<sup> 1 )</sup></td> <td>2 mL</td> </tr> <tr> <td>K<sub>2</sub>HPO<sub>4</sub> · 3H<sub>2</sub>O </td> <td>4 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>7.5 mg</td> </tr> <tr> <td>CaCl<sub>2</sub> · 2H<sub>2</sub>O </td> <td>3.6 mg</td> </tr> <tr> <td>Citric acid</td> <td>0.6 mg</td> </tr> <tr> <td>Ferric ammonium citrate</td> <td>0.6 mg</td> </tr> <tr> <td>Na<sub>2</sub>EDTA – Mg </td> <td>0.1 mg</td> </tr> <tr> <td>Na<sub>2</sub>CO<sub>3</sub></td> <td>2 mg</td> </tr> <tr> <td><a href="#a5_co">Trace metal mix A<sub>5</sub> + Co </a><sup> 2 )</sup></td> <td>0.1 mL</td> </tr> <tr> <td>Vitamin B<sub>12</sub></td> <td>0.1 µg</td> </tr> <tr> <td>Biotin</td> <td>0.1 µg</td> </tr> <tr> <td>Thiamine HCl</td> <td>200µg</td> </tr> <tr> <td>Agar</td> <td>1.2 g</td> </tr> <tr> <td>Distilled water</td> <td>97.9 mL</td> </tr> <tr> </tr> </tbody> </table> <p> 1 ) To 60 mL of Distilled water add 23.8 g HEPES, and pH is adjusted to 7.5. Bring to 100 mL with distilled water.v<br> 2 ) See <a href="#a5_co">Trace metal mix A<sub>5</sub> + Co </a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Castenholz, R. W. 1988 Cultureing methods for cyanobacteria. <i>Methods Enzymol</i>., <b>167</b>, 68-93. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="jo" id="jo"></a>Modified Johnson's medium</h1> <div style="text-align: right;"> <a href="./medium/en/du.pdf">print</a> </div> <table> <tbody> <tr> <td>NaCl</td> <td>3 mg</td> </tr> <tr> <td>MgCl<sub>2</sub> · 6H<sub>2</sub>O</td> <td>150 mg</td> </tr> <tr> <td>KNO<sub>3</sub></td> <td>100 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>50 mg</td> </tr> <tr> <td>CaCl<sub>2</sub> · 2H<sub>2</sub>O<sup> 1)</sup> </td> <td>26.5 mg</td> </tr> <tr> <td>KCl</td> <td>20 mg</td> </tr> <tr> <td>NaHCO<sub>3</sub></td> <td>4.3 mg</td> </tr> <tr> <td>KH<sub>2</sub>PO<sub>4</sub></td> <td>3.5 mg</td> </tr> <tr> <td>Tris ( hydroxymethyl ) aminomethane</td> <td>245 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> <tr> <td colspan="2">pH 8.5</td> </tr> </tbody> </table> <p> 1 ) In the NIES-Collection, 20 mg CaCl<sub>2</sub> is replaced by 26.5 mg CaCl<sub>2</sub> · 2H<sub>2</sub>O. </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Johnson, M. K., Johnson, E. J., MacElroy, R. D., Speer, H. L., Bruff, B. S. 1968 Effects of salts on the halophilic alga <i>Dunaliella viridis</i>. <i>J. Bacteriol.</i>, <b>95</b>, 1461-1468. </td> </tr> <tr> <td>Utsunomiya, A., Watanuki, T., Matsushita, K., Tomita, I. 1997 Toxic effects of linear alkylbenzene sulfonate, quaternaryalkylammonium chloride and their complexes on <i>Dunaliella</i>sp. and <i>Chlorella pyrenoidosa</i>. <i>Environ. Toxicol. Chem.</i>, <b>16</b>, 1247-1254. </td> </tr> <tr> <td>Mihirogi, M., Kikuchi, M., Sawai, J. 2012 Development of screening algal growth inhibition test with <i>Dunaliella</i> sp. <i>Jpn. J. Environ. Toxicol.</i>, <b>15</b>, 11-16 (in Japanese with English summary). </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="mm1" id="mm1"></a>Modified M–1 ( mM–1 )</h1> <div style="text-align: right;"> <a href="./medium/en/mm1.pdf">Print</a> </div> <table> <tbody> <tr> <td>CaCl<sub>2</sub> · 2H<sub>2</sub>O </td> <td>0.5 mg</td> </tr> <tr> <td>NaNO<sub>3</sub></td> <td>2.5 mg</td> </tr> <tr> <td>NH<sub>4</sub>Cl </td> <td>0.5 mg</td> </tr> <tr> <td>CaSO<sub>4</sub> · 2H<sub>2</sub>O </td> <td>0.4 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>0.5 mg</td> </tr> <tr> <td>Na<sub>2</sub>SiO<sub>3</sub> · 9H<sub>2</sub>O </td> <td>0.2 mg</td> </tr> <tr> <td><a href="#fe">Fe ( as EDTA; 1:1 molar )</a><sup> 1 )</sup></td> <td>25 mL</td> </tr> <tr> <td><a href="#mm1_trace">mM–1 Trace elements</a><sup> 2 )</sup></td> <td>0.1 mL</td> </tr> <tr> <td>K<sub>2</sub>HPO<sub>4</sub></td> <td>6.96 mg</td> </tr> <tr> <td>KH<sub>2</sub>PO<sub>4</sub></td> <td>266.5 mg</td> </tr> <tr> <td>Distilled water</td> <td>74.9 mL</td> </tr> <tr> <td colspan="2">pH 5.1 – 5.3</td> </tr> </tbody> </table> <p> 1 ) See <a href="#fe">Fe ( as EDTA; 1:1 molar )</a> <br> 2 ) See <a href="#mm1_trace">mM–1 Trace elements</a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Hoham, R. W., Berman, J. D., Rogers, H. S., Felio, J. H., Ryba, J. B., Miller, P. R. 2006 Two new species of green snow algae from Upstate New York, <i>Chloromonas chenangoensis</i> sp. nov. and <i>Chloromonas tughillensis</i> sp. nov. ( Volvocales, Chlorophyceae ) and the effects of light on their life cycle development. <i>Phycologia</i>, <b>45</b>, 319-330. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="mw" id="mw"></a>MW</h1> <div style="text-align: right;"> <a href="./medium/en/mw.pdf">Print</a> </div> <table> <tbody> <tr> <td>Urea</td> <td>0.85 mg</td> </tr> <tr> <td>NaNO<sub>3</sub></td> <td>0.17 mg</td> </tr> <tr> <td>NH<sub>4</sub>Cl </td> <td>0.042 mg</td> </tr> <tr> <td>Ca( NO<sub>3</sub> )<sub>2</sub> · 4H<sub>2</sub>O </td> <td>10 mg</td> </tr> <tr> <td>CaCO<sub>3</sub></td> <td>1 mg</td> </tr> <tr> <td>CaCl<sub>2</sub> · 2H<sub>2</sub>O </td> <td>1.4 mg</td> </tr> <tr> <td>KNO<sub>3</sub></td> <td>1 mg</td> </tr> <tr> <td>KHCO3</td> <td>0.9 mg</td> </tr> <tr> <td>β–Na<sub>2</sub>glycerophosphate · 5H<sub>2</sub>O </td> <td>2 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>1.5 mg</td> </tr> <tr> <td><a href="#p_iv">PIV metals</a><sup> 1 )</sup></td> <td>0.05 mL</td> </tr> <tr> <td>Vitamin B<sub>12</sub></td> <td>0.02 μg</td> </tr> <tr> <td>Thiamine HCl</td> <td>2 μg</td> </tr> <tr> <td>Biotin</td> <td>0.02 μg</td> </tr> <tr> <td>Glycylglycine</td> <td>10 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> <tr> <td colspan="2">pH 7.2</td> </tr> </tbody> </table> <p> 1 ) See <a href="#p_iv">PIV metals</a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Sako, Y., Ishida, Y., Kadota, H., Hata, Y. 1984 Sexual reproduction and cyst formation in the freshwater dinoflagellate <i>Peridinium cunningtonii. Bull. Jpn. Soc. Sci. Fish</i>., <b>50</b>, 743-750. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="mw_5" id="mw_5"></a>MW/5</h1> <div style="text-align: right;"> <a href="./medium/en/mw_5.pdf">Print</a> </div> <p> <a href="#mw">MW</a><sup> 1 )</sup> medium is diluted with distilled water to 1/5. <br> 1 ) See <a href="#mw">MW</a> <br> </p> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="n_free" id="n_free"></a>N–Free</h1> <div style="text-align: right;"> <a href="./medium/en/n_free.pdf">Print</a> </div> <table> <tbody> <tr> <td>NaCl</td> <td>7 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>38 mg</td> </tr> <tr> <td>CaCl<sub>2</sub> · 2H<sub>2</sub>O </td> <td>10.6 mg</td> </tr> <tr> <td>K<sub>2</sub>HPO<sub>4</sub></td> <td>60 mg</td> </tr> <tr> <td>Fe<sub>2</sub>( SO<sub>4</sub> )<sub>3</sub> · 6H<sub>2</sub>O </td> <td>1 mg</td> </tr> <tr> <td>Na<sub>2</sub>EDTA · 2H<sub>2</sub>O </td> <td>2.7 mg</td> </tr> <tr> <td>H<sub>3</sub>BO<sub>3</sub></td> <td>0.3 mg</td> </tr> <tr> <td>MnSO<sub>4</sub> · 4H<sub>2</sub>O<sup> 1 )</sup></td> <td>0.2 mg</td> </tr> <tr> <td>Na<sub>2</sub>MoO<sub>4</sub> · 2H<sub>2</sub>O </td> <td>0.8 mg</td> </tr> <tr> <td>ZnSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>0.03 mg</td> </tr> <tr> <td>CuSO<sub>4</sub> · 5H<sub>2</sub>O </td> <td>8 μg</td> </tr> <tr> <td>CoCl<sub>2</sub> · 6H<sub>2</sub>O </td> <td>3.7 μg</td> </tr> <tr> <td>Agar</td> <td>1.5 g</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> <tr> <td colspan="2">pH 7.5</td> </tr> </tbody> </table> <p> 1 ) In the NIES–Colletion, 0.2 mg MnSO<sub>4</sub> · 4H<sub>2</sub>O is replaced by 0.22 mg MnSO<sub>4</sub> · 5H<sub>2</sub>O. </p> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="o" id="o"></a>O</h1> <div style="text-align: right;"> <a href="./medium/en/o.pdf">Print</a> </div> <table> <tbody> <tr> <td>Glucose</td> <td>100 mg</td> </tr> <tr> <td>Tryptone</td> <td>100 mg</td> </tr> <tr> <td>Yeast extract</td> <td>100 mg</td> </tr> <tr> <td>Beef extract<sup> 1 )</sup></td> <td>50 mg</td> </tr> <tr> <td>Agar</td> <td>150 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 ml</td> </tr> </tbody> </table> <p> 1 ) In the NIES-Collection, beef extract is removed. <br> Indicated as “Ochomonas medium” in reference. </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Ichimura, T., Itoh, T. 1977 17. Preservation methods of microalgae ( I ) [17. Bisaisôrui no hozonhô ( I )]. In <i>Preservation methods of microorganisms [Biseibutsu Hozonhô]</i>, Ed. by Nei, T., University of Tokyo Press, Tokyo, p. 355-373 ( in Japanese without English title ). </td> </tr> <tr> <td>Starr, R. C. 1964 The culture collection of algae at Indiana University. <i>Amer. J. Bot</i>., <b>51</b>, 1013-1044. </td> </tr> </tbody> </table> <br/> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="p35" id="p35"></a>P 35</h1> <div style="text-align: right;"> <a href="./medium/en/p35.pdf">Print</a> </div> <table> <tbody> <tr> <td>NH<sub>4</sub>NO<sub>3</sub></td> <td>10 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>4 mg</td> </tr> <tr> <td>KCl</td> <td>5 mg</td> </tr> <tr> <td>CaCl<sub>2</sub> · 2H<sub>2</sub>O </td> <td>7.4 mg</td> </tr> <tr> <td>β–Na<sub>2</sub>glycerophosphate · 5H<sub>2</sub>O </td> <td>5 mg</td> </tr> <tr> <td>Sodium acetate</td> <td>100 mg</td> </tr> <tr> <td>Vitamin B<sub>12</sub></td> <td>0.01 μg</td> </tr> <tr> <td>Biotin</td> <td>0.01 μg</td> </tr> <tr> <td>Thiamine HCl</td> <td>1 μg</td> </tr> <tr> <td><a href="#p_iv">PIV metals</a><sup> 1 )</sup></td> <td>0.3 mL</td> </tr> <tr> <td>Tris ( hydroxymethyl ) aminomethane</td> <td>50 mg</td> </tr> <tr> <td>Distilled water</td> <td>99.7 mL</td> </tr> <tr> <td colspan="2">pH 8.0</td> </tr> </tbody> </table> <p> 1 ) See <a href="#p_iv">PIV metals</a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Ichimura, T. 1979 2. Isolation and culture methods of algae. 2.5.B. Freshwater algae [2. Sôrui no bunri to baiyôhô. 2.5.B. Tansui sôrui]. In <i>Methods in Phycological Studies [Sôrui Kenkyûhô</i> ], Eds. by Nishizawa, K. & Chihara, M., Kyoritsu Shuppan, Tokyo, p. 294-305 ( in Japanese without English title ). </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="pro" id="pro"></a>Pro</h1> <div style="text-align: right;"> <a href="./medium/en/pro.pdf">Print</a> </div> <p> To 100 mL <a href="#mbm">MBM</a><sup> 1 )</sup> medium add 100 mg proteose peptone. <br> 1 ) See <a href="#mbm">MBM</a> <br> Indicated as “Proteose medium” in reference. </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Ichimura, T., Itoh, T. 1977 17. Preservation methods of microalgae ( I ) [17. Bisaisôrui no hozonhô ( I )]. In <i>Preservation methods of microorganisms [Biseibutsu Hozonhô]</i>, Ed. by Nei, T., University of Tokyo Press, Tokyo, p. 355-373 ( in Japanese without English title ). </td> </tr> <tr> <td>Starr, R. C. 1964 The culture collection of algae at Indiana University. <i>Amer. J. Bot</i>., <b>51</b>, 1013-1044. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="sot" id="sot"></a>SOT</h1> <div style="text-align: right;"> <a href="./medium/en/sot.pdf">Print</a> </div> <table> <tbody> <tr> <td>NaHCO<sub>3</sub></td> <td>1.68 g</td> </tr> <tr> <td>K<sub>2</sub>HPO<sub>4</sub></td> <td>50 mg</td> </tr> <tr> <td>NaNO<sub>3</sub></td> <td>250 mg</td> </tr> <tr> <td>K<sub>2</sub>SO<sub>4</sub></td> <td>100 mg</td> </tr> <tr> <td>NaCl</td> <td>100 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>20 mg</td> </tr> <tr> <td>CaCl<sub>2</sub> · 2H<sub>2</sub>O </td> <td>4 mg</td> </tr> <tr> <td>FeSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>1 mg</td> </tr> <tr> <td>Na<sub>2</sub>EDTA · 2H<sub>2</sub>O </td> <td>8 mg</td> </tr> <tr> <td><a href="#a5">A<sub>5</sub> solution </a><sup> 1 )</sup></td> <td>0.1 mL</td> </tr> <tr> <td>Distilled water</td> <td>99.9 mL</td> </tr> </tbody> </table> <p> 1 ) See <a href="#a5">A<sub>5</sub> solution </a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Ogawa, T., Terui, G. 1970 Studies on the growth of <i>Spirulina platensis</i>. ( I ) On the pure culture of <i>Spirulina platensis. J. Ferment. Technol</i>., <b>48</b>, 361-367. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="sw" id="sw"></a>SW</h1> <div style="text-align: right;"> <a href="./medium/en/sw.pdf">Print</a> </div> <p>Put a small amount of dried soil into a test tube and add 20mL distilled water.</p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Pringsheim, E. G. 1946 The biphasic or soil-water culture method for growing algae and flagellata. <i>J. Ecol</i>., 33, 193-204. </td> </tr> </tbody> </table> <br/> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="tap" id="tap"></a>TAP</h1> <div style="text-align: right;"> <a href="./medium/en/tap.pdf">Print</a> </div> <table> <tbody> <tr> <td>NH<sub>4</sub>Cl </td> <td>40 mg</td> </tr> <tr> <td>CaCl<sub>2</sub> · 2H<sub>2</sub>O </td> <td>5.1 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>10 mg</td> </tr> <tr> <td>K<sub>2</sub>HPO<sub>4</sub></td> <td>11.9 mg</td> </tr> <tr> <td>KH<sub>2</sub>PO<sub>4</sub></td> <td>6.03 mg</td> </tr> <tr> <td><a href="#hunter">Hutner ’s trace elements</a><sup> 1 )</sup></td> <td>0.1 mL</td> </tr> <tr> <td>Acetic acid</td> <td>0.1 mL</td> </tr> <tr> <td>Tris ( hydroxymethyl ) aminomethane</td> <td>242 mg</td> </tr> <tr> <td>Agar</td> <td>1.5 g</td> </tr> <tr> <td>Distilled water</td> <td>99.8 mL</td> </tr> </tbody> </table> <p> 1 ) See <a href="#hunter">Hutner ’s trace elements</a> </p> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="tre" id="tre"></a>Tre</h1> <div style="text-align: right;"> <a href="./medium/en/tre.pdf">Print</a> </div> <p> To 100 mL <a href="#mbm">MBM</a><sup> 1 )</sup> medium add 1 g proteose peptone and 2 g glucose. <br> 1 ) See <a href="#mbm">MBM</a> <br> Indicated as “Trebouxia medium” in reference. </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Ichimura, T., Itoh, T. 1977 17. Preservation methods of microalgae ( I ) [17. Bisaisôrui no hozonhô ( I )]. In <i>Preservation methods of microorganisms [Biseibutsu Hozonhô]</i>, Ed. by Nei, T., University of Tokyo Press, Tokyo, p. 355-373 ( in Japanese without English title ). </td> </tr> <tr> <td>Starr, R. C. 1964 The culture collection of algae at Indiana University. <i>Amer. J. Bot</i>., <b>51</b>, 1013-1044. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="uro" id="uro"></a>URO</h1> <div style="text-align: right;"> <a href="./medium/en/uro.pdf">Print</a> </div> <table> <tbody> <tr> <td>NH<sub>4</sub>NO<sub>3</sub></td> <td>0.5 mg</td> </tr> <tr> <td>β–Na<sub>2</sub>glycerophosphate · 5H<sub>2</sub>O </td> <td>0.4 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>1 mg</td> </tr> <tr> <td>CaCl<sub>2</sub> · 2H<sub>2</sub>O </td> <td>1 mg</td> </tr> <tr> <td>KCl</td> <td>0.1 mg</td> </tr> <tr> <td>Thiamine HCl</td> <td>1 μg</td> </tr> <tr> <td>Vitamin B<sub>12</sub></td> <td>0.01 μg</td> </tr> <tr> <td>Biotin</td> <td>0.01 μg</td> </tr> <tr> <td>Fe-EDTA</td> <td>0.05 mg</td> </tr> <tr> <td><a href="#p_iv">PIV metals</a><sup> 1 )</sup></td> <td>0.1 mL</td> </tr> <tr> <td>Distilled water</td> <td>99.9 mL</td> </tr> <tr> <td colspan="2">pH 7.5<sup> 2 )</sup></td> </tr> </tbody> </table> <p> 1 ) See <a href="#p_iv">PIV metals</a><br> 2 ) pH is adjusted to 7.5 with 0.1 mol / L HCl. </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Kimura, B., Ishida, Y. 1985 Photophagotrophy in <i>Uroglena americana</i>, Chrysophyceae. <i>Jpn. J. Limnol</i>., <b>46</b>, 315-318. </td> </tr> <tr> <td>Nakahara, H., Sako, Y. 1987 2. Life history of freshwater phytoplankton [2. Tansui syokubutsu purankuton no seikatsushi]. In <i>Freshwater red tide</i> [<i>Tansui Akashio</i> ], Ed. by Kadota, H., Kôseisya-Kôseikaku, Tokyo, p. 21-77 ( in Japanese without English title ). </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="uro_h" id="uro_h"></a>URO–H</h1> <div style="text-align: right;"> <a href="./medium/en/uro_h.pdf">Print</a> </div> <p> To 100 mL <a href="#uro">URO</a><sup> 1 )</sup> medium add 40 mg HEPES. <br> 1 ) See <a href="#uro">URO</a><br> </p> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="uro_soil" id="uro_soil"></a>URO + Soil</h1> <div style="text-align: right;"> <a href="./medium/en/uro_soil.pdf">Print</a> </div> <p> Prepare as for 100 mL <a href="#uro">URO</a><sup> 1 )</sup> medium with 89.9 mL instead of 99.9 mL distilled water, add 10 mL <a href="#soil_extract">Soil extract</a><sup> 2 )</sup> and adjust pH 7.5. <br> 1 ) See <a href="#uro">URO</a> <br> 2 ) See <a href="#soil_extract">Soil extract</a> </p> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="uro_t" id="uro_t"></a>URO–T</h1> <div style="text-align: right;"> <a href="./medium/en/uro_t.pdf">Print</a> </div> <p> To 100 mL <a href="#uro">URO</a><sup> 1 )</sup> medium add 50 mg Tris ( hydroxymethyl ) aminomethane. <br> 1 ) See <a href="#uro">URO</a><br> </p> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="vt" id="vt"></a>VT</h1> <div style="text-align: right;"> <a href="./medium/en/vt.pdf">Print</a> </div> <table> <tbody> <tr> <td>Ca( NO<sub>3</sub> )<sub>2</sub> · 4H<sub>2</sub>O </td> <td>11.78 mg</td> </tr> <tr> <td>β-Na<sub>2</sub>glycerophosphate · 5H<sub>2</sub>O </td> <td>5 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>4 mg</td> </tr> <tr> <td>KCl</td> <td>5 mg</td> </tr> <tr> <td>Vitamin B<sub>12</sub></td> <td>0.01 μg</td> </tr> <tr> <td>Biotin</td> <td>0.01 μg</td> </tr> <tr> <td>Thiamine HCl</td> <td>1 μg</td> </tr> <tr> <td><a href="#p_iv">PIV metals</a><sup> 1 )</sup></td> <td>0.3 mL</td> </tr> <tr> <td>Glycylglycine</td> <td>50 mg</td> </tr> <tr> <td>Distilled water</td> <td>99.7 mL</td> </tr> <tr> <td colspan="2">pH 7.5</td> </tr> </tbody> </table> <p> 1 ) See <a href="#p_iv">PIV metals</a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Provasoli, L., Pintner, I. J. 1959 Artificial media for fresh-water algae: problems and suggestions. In <i>The Ecology of Algae. Spec. Pub. No. 2</i>,, Eds. by Tryon, C. A., Jr. & Hartmann, R. T., Pymatuning Laboratory of Field Biology, University of Pittsburgh, Pittsburgh, p. 84-96. </td> </tr> <tr> <td>Starr, R. C. 1973 Special methods-dry soil samples. In <i>Handbook of Phycological Methods. Culture Methods and Growth Measurements</i>, Ed. by Stein, J. R., Cambridge University Press, Cambridge, p. 159-167. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="vtac" id="vtac"></a>VTAC</h1> <div style="text-align: right;"> <a href="./medium/en/vtac.pdf">Print</a> </div> <p> To 100 mL <a href="#vt">VT</a><sup> 1 )</sup> medium add 20 mg sodium acetate.<br> 1 ) See <a href="#vt">VT</a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Nozaki, H., Kuroiwa, H., Mita, T., Kuroiwa, T. 1989 <i>Pleodorina japonica</i> sp. nov. ( Volvocales, Chlorophyta ) with bacteria-like endosymbionts. <i>Phycologia</i>, <b>28</b>, 252-267. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="vtyt" id="vtyt"></a>VTYT</h1> <div style="text-align: right;"> <a href="./medium/en/vtyt.pdf">Print</a> </div> <p> To 100 mL <a href="#vt">VT</a><sup> 1 )</sup> medium add 10 mg yeast extract and 20 mg tryptone. <br> 1 ) See <a href="#vt">VT</a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Ichimura, T., Itoh, T. 1977 17. Preservation methods of microalgae ( I ) [17. Bisaisôrui no hozonhô ( I ) ]. In <i>Preservation methods of microorganisms [Biseibutsu Hozonhô]</i>, Ed. by Nei, T., University of Tokyo Press, Tokyo, p. 355-373 ( in Japanese without English title ). </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="w" id="w"></a>W</h1> <div style="text-align: right;"> <a href="./medium/en/w.pdf">Print</a> </div> <table> <tbody> <tr> <td>Ca( NO<sub>3</sub> )<sub>2</sub> · 4H<sub>2</sub>O </td> <td>10 mg</td> </tr> <tr> <td>KNO<sub>3</sub></td> <td>1 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>1.5 mg</td> </tr> <tr> <td>β–Na<sub>2</sub>glycerophosphate · 5H<sub>2</sub>O </td> <td>2 mg</td> </tr> <tr> <td>Urea</td> <td>1.7 mg</td> </tr> <tr> <td>Thiamine HCl</td> <td>0.2 μg</td> </tr> <tr> <td>Vitamin B<sub>12</sub></td> <td>0.002 μg</td> </tr> <tr> <td>Biotin</td> <td>0.002 μg</td> </tr> <tr> <td><a href="#p_iv">PIV metals</a><sup> 1 )</sup></td> <td>0.05 mL</td> </tr> <tr> <td>Glycylglycine</td> <td>10 mg</td> </tr> <tr> <td>Distilled water</td> <td>99.95 mL</td> </tr> <tr> <td colspan="2">pH 7.5</td> </tr> </tbody> </table> <p> 1 ) See <a href="#p_iv">PIV metals</a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Watanabe, M. M. 1983 Growth characteristics of freshwater red tide alga, <i>Peridinium</i> based on axenic culture. Establishment of synthetic culture medium [Junsuibaiyôhô niyoru tansuiakashio Peridinium no zôsyokutokusei no kaiseki-gôseibaichi no kakuritsu ]. <i>Res. Data Natl. Inst. Environ. Stud</i>., <b>No. 24</b>, 111-121 ( in Japanese without English title ). </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="wc" id="wc"></a>WC</h1> <div style="text-align: right;"> <a href="./medium/en/wc.pdf">print</a> </div> <table> <tbody> <tr> <td height="20">NaNO<sub>3</sub></td> <td>8.5 mg</td> </tr> <tr> <td>CaCl<sub>2</sub> · 2H<sub>2</sub>O</td> <td>3.67 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O</td> <td>3.69 mg</td> </tr> <tr> <td>NaHCO<sub>3</sub></td> <td>1.26 mg</td> </tr> <tr> <td>Na<sub>2</sub>SiO<sub>3</sub> · 9H<sub>2</sub>O</td> <td>2.84 mg</td> </tr> <tr> <td>K<sub>2</sub>HPO<sub>4</sub></td> <td>0.87 mg</td> </tr> <tr> <td><a href="#wc_metals">WC trace metals</a><sup> 1 )</sup></td> <td>0.1 mL</td> </tr> <tr> <td>Vitamin B<sub>12</sub></td> <td>0.05 μg</td> </tr> <tr> <td>Biotin</td> <td>0.05 μg</td> </tr> <tr> <td>Thiamine HCl</td> <td>10 μg</td> </tr> <tr> <td>Distilled water</td> <td>99.9 mL</td> </tr> <tr> <td colspan="2">pH 7.5</td> </tr> </tbody> </table> <p> 1 ) See <a href="#wc_metals">WC trace metals</a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Guillard, R. R. L., Lorenzen, C. J. 1972 Yellow-green algae with chlorohyllide C. <i>J. Phycol.</i>, <b>8</b>, 10-14. </td> </tr> <tr> <td>Andersen, R. A. E, Berges, J. A., Harrison, P. J., Watanabe, M. M. 2005 Appendix A―Recipes for freshwater and seawater media. In <i>Algal culturing techniques</i>, Ed. by Andersen, R. A. E, Elsevier Academic Press, p. 474. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">1 Media for freshwater, terrestrial, hot spring and salt water algae</p> <h1><a name="bbm2" id="bbm2"></a>9:1 agar medium</h1> <div style="text-align: right;"> <a href="./medium/en/bbm2.pdf">Print</a> </div> <table> <tbody> <tr> <td height="20">NaNO<sub>3</sub></td> <td>25 mg</td> </tr> <tr> <td>KH<sub>2</sub>PO<sub>4</sub></td> <td>17.5 mg</td> </tr> <tr> <td>K<sub>2</sub>HPO<sub>4</sub></td> <td>10 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>7.5 mg</td> </tr> <tr> <td>CaCl<sub>2</sub> · 2H<sub>2</sub>O </td> <td>2.5 mg</td> </tr> <tr> <td>NaCl</td> <td>2.5 mg</td> </tr> <tr> <td>KOH</td> <td>3.1 mg</td> </tr> <tr> <td>Na<sub>2</sub>EDTA· 2H<sub>2</sub>O </td> <td>5 mg</td> </tr> <tr> <td>FeSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>0.498 mg</td> </tr> <tr> <td>H<sub>3</sub>BO<sub>3</sub></td> <td>1.142 mg</td> </tr> <tr> <td>ZnSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>0.882 mg</td> </tr> <tr> <td>MnCl<sub>2</sub> · 4H<sub>2</sub>O </td> <td>0.144 mg</td> </tr> <tr> <td>MoO<sub>3</sub></td> <td>0.071 mg</td> </tr> <tr> <td>CuSO<sub>4</sub> · 5H<sub>2</sub>O </td> <td>0.157 mg</td> </tr> <tr> <td>Co( NO<sub>3</sub> )<sub>2</sub> · 6H<sub>2</sub>O </td> <td>0.049 mg</td> </tr> <tr> <td>Vitamin B<sub>12</sub></td> <td>0.01 μg</td> </tr> <tr> <td>Biotin</td> <td>0.01 μg</td> </tr> <tr> <td>Thiamine HCl</td> <td>1 μg</td> </tr> <tr> <td><a href="#soil_extract">Soil extract</a><sup> 1 )</sup></td> <td>10 mL</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> </tbody> </table> <p> Add 1.5 g agar to 100 mL of medium to give a solid medium. <br> 1 ) See <a href="#soil_extract">Soil extract</a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Bischoff, H. W., Bold, H. C. 1963 Some soil algae from enchanted rock and related algal species. <i>Phycological Studies IV.,Univ. No. 6318</i>, Texas, p. 95. </td> </tr> <tr> <td>Watanabe, S., Fučíková, K., Lewis, L. A., Lewis P.O. 2016 Hiding in plain sight: <i>Koshicola spirodelophila</i> gen. et sp. nov. (Chaetopeltidales, Chlorophyceae), a novel green alga associated with the aquatic angiosperm <i>Spirodela polyrhiza</i>. <i>Am. J. Bot.</i>, <b>103</b>, 865-875. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">2 Media for marine and brackish water microalgae</p> <h1><a name="besm" id="besm"></a>BESM</h1> <div style="text-align: right;"> <a href="./medium/en/besm.pdf">Print</a> </div> <p> Make diluted seawater by mixing 27.5 mL seawater and 70 mL distilled water. Make <a href="#esm">ESM</a><sup> 1 )</sup> medium by using this diluted seawater instead of original seawater. <br> 1 ) See <a href="#esm">ESM</a><br> </p> <p class="mediumCategoryTitle">2 Media for marine and brackish water microalgae</p> <h1><a name="besm2" id="besm2"></a>BESM 2</h1> <div style="text-align: right;"> <a href="./medium/en/besm2.pdf">Print</a> </div> <p> Make diluted seawater by mixing 47.5 mL seawater and 50 mL distilled water. Make <a href="#esm">ESM</a><sup> 1 )</sup> medium by using this diluted seawater instead of original seawater. <br> 1 ) See <a href="#esm">ESM</a> </p> <br> <p class="mediumCategoryTitle">2 Media for marine and brackish water microalgae</p> <h1><a name="esm" id="esm"></a>ESM</h1> <div style="text-align: right;"> <a href="./medium/en/esm.pdf">Print</a> </div> <table> <tbody> <tr> <td>NaNO<sub>3</sub></td> <td>12 mg</td> </tr> <tr> <td>K<sub>2</sub>HPO<sub>4</sub></td> <td>0.5 mg</td> </tr> <tr> <td>Vitamin B<sub>12</sub></td> <td>0.1 μg</td> </tr> <tr> <td>Biotin</td> <td>0.1 μg</td> </tr> <tr> <td>Thiamine HCl</td> <td>10 μg</td> </tr> <tr> <td>Fe – EDTA</td> <td>25.9 μg</td> </tr> <tr> <td>Mn – EDTA</td> <td>33.2 μg</td> </tr> <tr> <td>Tris ( hydroxymethyl ) aminomethane</td> <td>100 mg</td> </tr> <tr> <td><a href="#soil_extract">Soil extract</a><sup> 1 )</sup></td> <td>2.5 mL</td> </tr> <tr> <td>Seawater</td> <td>97.5 mL</td> </tr> <tr> <td colspan="2">pH 8.0</td> </tr> </tbody> </table> <p> Add 1.5 g agar to 100 mL of medium to give a solid medium. <br> 1 ) The amount of <a href="#soil_extract">Soil extract</a> depends on the quality of the soil. In the NIES–Collection, <a href="#soil_extract">Soil extract</a> was reduced from 5 mL to 2.5 mL after 2002. See <a href="#soil_extract">Soil extract</a>. </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Okaichi, T., Nishio, S., Imatomi, Y. 1982 Collection and mass culture [Shiryô no saisyu to baiyô]. In <i>Toxic phytoplankton - Occurrence, mode of action, and toxins</i> [<i>Yûdoku Purankuton -Hassei, Sayôkikô, Dokuseibun</i> ], Ed. by Jpn. Fish. Soc., Kôseisya-Kôseikaku, Tokyo, p. 22-34 ( in Japanese without English title ). </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">2 Media for marine and brackish water microalgae</p> <h1><a name="esm_cbv" id="esm_cbv"></a>ESM + CB–V</h1> <div style="text-align: right;"> <a href="./medium/en/esm_cbv.pdf">Print</a> </div> <p> Mix 100 mL of <a href="#esm">ESM</a><sup> 1 )</sup> medium and 0.25 mL of <a href="#cb_v">CB–V</a><sup> 2 )</sup> medium.<br> <br> 1 ) See <a href="#esm">ESM</a> <br> 2 ) See <a href="#cb_v">CB–V</a><br> </p> <p class="mediumCategoryTitle">2 Media for marine and brackish water microalgae</p> <h1><a name="esm2" id="esm2"></a>ESM2</h1> <div style="text-align: right;"> <a href="./medium/en/esm2.pdf">Print</a> </div> <p> Prepare as for 100 mL <a href="#esm">ESM</a><sup> 1 )</sup> medium with 95.5mL instead of 97.5 mL seawater and with 5 mL instead of 2.5 mL <a href="#soil_extract">Soil extract</a><sup> 2 )</sup>. <br> 1 ) See <a href="#esm">ESM</a> <br> 2 ) See <a href="#soil_extract">Soil extract</a><br> </p> <p class="mediumCategoryTitle">2 Media for marine and brackish water microalgae</p> <h1><a name="f2" id="f2"></a>f/2</h1> <div style="text-align: right;"> <a href="./medium/en/f2.pdf">Print</a> </div> <table> <tbody> <tr> <td>NaNO<sub>3</sub></td> <td>7.5 mg</td> </tr> <tr> <td>NaH<sub>2</sub>PO<sub>4</sub> · 2H<sub>2</sub>O </td> <td>0.6 mg</td> </tr> <tr> <td>Vitamin B<sub>12</sub></td> <td>0.05 μg</td> </tr> <tr> <td>Biotin</td> <td>0.05 μg</td> </tr> <tr> <td>Thiamine HCl</td> <td>10 μg</td> </tr> <tr> <td>Na<sub>2</sub>SiO<sub>3</sub> · 9H<sub>2</sub>O </td> <td>1 mg</td> </tr> <tr> <td><a href="#f2_metals">f/2 metals</a><sup> 1 )</sup></td> <td>0.1 mL</td> </tr> <tr> <td>Seawater</td> <td>99.9 mL</td> </tr> <tr> </tr> </tbody> </table> <p> 1 ) See <a href="#f2_metals">f/2 metals</a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Guillard, R. R. L., Ryther, J. H. 1962 Studies of marine planktonic diatoms. I. <i>Cyclotella nana</i> Hustedt, and <i>Detonula confervacea</i> ( Cleve ) Gran. <i>Can. J. Microbiol</i>., <b>8</b>, 229-239. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">2 Media for marine and brackish water microalgae</p> <h1><a name="f2_nh4cl" id="f2_nh4cl"></a>f/2 + NH4Cl</h1> <div style="text-align: right;"> <a href="./medium/en/f2_nh4cl.pdf">Print</a> </div> <p> To 100 mL <a href="#f2">f/2</a><sup> 1 )</sup> medium add 2.67 mg NH<sub>4</sub>Cl. <br> 1 ) See <a href="#f2">f/2</a><br> </p> <p class="mediumCategoryTitle">2 Media for marine and brackish water microalgae</p> <h1><a name="imk" id="imk"></a>IMK</h1> <div style="text-align: right;"> <a href="./medium/en/imk.pdf">Print</a> </div> <p> Into 100 mL seawater dissolve 25.2 mg powder medium of Daigo IMK ( Nihon Pharmaceutical Co., Ltd. ). In the NIES–Collection, IMK medium is used after autoclaving ( 121°C, 20 min ). <br> </p> <p class="mediumCategoryTitle">2 Media for marine and brackish water microalgae</p> <h1><a name="imk10" id="imk10"></a>IMK + NaCl</h1> <div style="text-align: right;"> <a href="./medium/en/imk10.pdf">Print</a> </div> <p> Into 100 mL seawater dissolve 25.2 mg powder medium of Daigo IMK ( Nihon Pharmaceutical Co., Ltd. ). Adjust to salinity 10 percent with NaCl. In the NIES-Collection, IMK + NaCl medium is used after autoclaving ( 121°C, 20 min ). </p> <p class="mediumCategoryTitle">2 Media for marine and brackish water microalgae</p> <h1><a name="imks" id="imks"></a>IMK + Soil Extract</h1> <div style="text-align: right;"> <a href="./medium/en/imks.pdf">Print</a> </div> <p> Into 100 mL seawater dissolve 25.2 mg powder medium of Daigo IMK ( Nihon Pharmaceutical Co., Ltd. ). Thereafter, 2 mL <a href="http://mcc.nies.go.jp/02medium.html#soil_extract">Soil extract</a> 1 ) is added. In the NIES–Collection, IMK + Soil Extract medium is used after autoclaving ( 121°C, 20 min ). <br> 1 ) See <a href="http://mcc.nies.go.jp/02medium.html#soil_extract">Soil extract</a> </p> <p class="mediumCategoryTitle">2 Media for marine and brackish water microalgae</p> <h1><a name="imks" id="imk_2"></a>IMK/2</h1> <div style="text-align: right;"> <a href="./medium/ja/imk_2.pdf">Print</a> </div> <p> Into 100 mL seawater dissolve 12.6 mg powder medium of Daigo IMK ( Nihon Pharmaceutical Co., Ltd. ). In the NIES–Collection, IMK/2 medium is used after autoclaving ( 121°C, 20 min ). <br> </a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Takahashi, K., Lum, W. M., Benico, G., Uchida, H., Ozawa, M., Oikawa, H., Suzuki, T., Nguyen, N. V., Ha, D. V., Iwataki, M. 2021 Harmful algal bloomsToxigenic strains of <i>Azadinium poporum</i> (Amphidomataceae, Dinophyceae) from Japan and Vietnam, with first reports of <i>A. poporum</i> (ribotype A) and <i>A. trinitatum</i> in Asian Pacific. <i>J. Phycol.</i>, <b>69</b>, 175-187.</td> </tr> </tbody> </table> <p class="mediumCategoryTitle">2 Media for marine and brackish water microalgae</p> <h1><a name="k" id="k"></a>K</h1> <div style="text-align: right;"> <a href="./medium/en/k.pdf">Print</a> </div> <table> <tbody> <tr> <td>Tris-base (  pH 7.2) <sup>1 )</sup></td> <td>0.1 mL</td> </tr> <tr> <td>NaNO<sub>3</sub></td> <td>7.5 mg</td> </tr> <tr> <td>NH<sub>4</sub>Cl </td> <td>0.267 mg</td> </tr> <tr> <td>β–Na<sub>2</sub>glycerophosphate · 5H<sub>2</sub>O </td> <td>0.216 mg</td> </tr> <tr> <td>H<sub>2</sub>SeO<sub>3</sub></td> <td>0.129 μg</td> </tr> <tr> <td>Vitamin B<sub>12</sub></td> <td>0.05 μg</td> </tr> <tr> <td>Biotin</td> <td>0.05 μg</td> </tr> <tr> <td>Thiamine HCl</td> <td>0.01 mg</td> </tr> <tr> <td>Na<sub>2</sub>SiO<sub>3</sub> · 9H<sub>2</sub>O<sup>2 )</sup></td> <td>1.535 mg</td> </tr> <tr> <td><a href="#K_metals">K metals</a><sup> 3 )</sup></td> <td>0.1 mL</td> </tr> <tr> <td>Seawater</td> <td>99.8 mL</td> </tr> <tr> </tr> </tbody> </table> <p> 1 ) To 100 mL of Distilled water add 12.11 g Tris-base, and pH is adjusted to 7.2. <br> 2 ) In the NIES-Collection, Na<sub>2</sub>SiO<sub>3</sub> · 9H<sub>2</sub>O is removed. <br> 3 ) See <a href="#K_metals">K metals</a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Keller, M. D., Guillard, R.R.L. 1985 Factors significant to marine diatom culture. pp. 113-6. <i>In</i> Anderson, D.M., White, A.W., Baden, D.G. (eds.) <i>Toxic Dinoflagellates</i>. Elsevier, New York. </td> </tr> <tr> <td>Keller, M.D., Selvin, R.C., Claus, W., Guillard, R.R.L. 1987. Media for the culture of oceanic ultraphytoplankton. <i>J</i>. <i>Phycol</i>., 23: 633-638. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">2 Media for marine and brackish water microalgae</p> <h1><a name="k2" id="k2"></a>K/2 (K/2-modified by Ian Probert)</h1> <div style="text-align: right;"> <a href="./medium/en/k_2.pdf">Print</a> </div> <table> <tbody> <tr> <td>NaNO<sub>3</sub></td> <td>2.44 mg</td> </tr> <tr> <td>NH<sub>4</sub>Cl </td> <td>0.0267 mg</td> </tr> <tr> <td>KH<sub>2</sub>PO<sub>4</sub></td> <td>0.244 mg</td> </tr> <tr> <td>Fe-EDTA</td> <td>0.215 mg</td> </tr> <tr> <td>Vitamin B<sub>12</sub></td> <td>0.05 μg</td> </tr> <tr> <td>Biotin</td> <td>0.0005 μg</td> </tr> <tr> <td>Thiamine HCl</td> <td>0.01 mg</td> </tr> <tr> <td><a href="#k_2_metals">K/2 metals</a><sup> 1 )</sup></td> <td>0.05 mL</td> </tr> <tr> <td>Seawater</td> <td>99.95 mL</td> </tr> <tr> <td colspan="2">pH 8.2</td> </tr> </tbody> </table> <p> K medium is diluted half strength and modified by Ian. This medium should not be autoclaved but filter-sterilized.<br> 1 ) See <a href="#k_2_metals">K/2 metals</a> <br> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Keller, M. D., Guillard, R.R.L. 1985 Factors significant to marine diatom culture. pp. 113-6. <i>In</i> Anderson, D.M., White, A.W., Baden, D.G. (eds.) <i>Toxic Dinoflagellates</i>. Elsevier, New York. </td> </tr> <tr> <td>Keller, M.D., Selvin, R.C., Claus, W., Guillard, R.R.L. 1987. Media for the culture of oceanic ultraphytoplankton. <i>J</i>. <i>Phycol</i>., 23: 633-638. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">2 Media for marine and brackish water microalgae</p> <h1><a name="k2et" id="k2et"></a>K/2ET</h1> <div style="text-align: right;"> <a href="./medium/en/k_2_ETian.pdf">Print</a> </div> <table> <tbody> <tr> <td>NaNO<sub>3</sub></td> <td>2.44 mg</td> </tr> <tr> <td>NH<sub>4</sub>Cl </td> <td>0.0267 mg</td> </tr> <tr> <td>KH<sub>2</sub>PO<sub>4</sub></td> <td>0.244 mg</td> </tr> <tr> <td>Fe-EDTA</td> <td>0.215 mg</td> </tr> <tr> <td>Vitamin B<sub>12</sub></td> <td>0.05 μg</td> </tr> <tr> <td>Biotin</td> <td>0.0005 μg</td> </tr> <tr> <td>Thiamine HCl</td> <td>0.01 mg</td> </tr> <tr> <td><a href="#k_2_metals">K/2 metals</a><sup> 1 )</sup></td> <td>0.05 mL</td> </tr> <tr> <td><a href="#soil_extract">Soil extract</a><sup> 2 )</sup></td> <td>1.5 mL</td> </tr> <tr> <td>Seawater</td> <td>98.45 mL</td> </tr> <tr> <td colspan="2">pH 8.2</td> </tr> </tbody> </table> <p> K medium is diluted half strength and modified by Ian. This medium should not be autoclaved but filter-sterilized.<br> 1 ) See <a href="#k_2_metals">K/2 metals</a> <br> 2 ) See <a href="#soil_extract">Soil extract</a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Keller, M. D., Guillard, R.R.L. 1985 Factors significant to marine diatom culture. pp. 113-6. <i>In</i> Anderson, D.M., White, A.W., Baden, D.G. (eds.) <i>Toxic Dinoflagellates</i>. Elsevier, New York. </td> </tr> <tr> <td>Keller, M.D., Selvin, R.C., Claus, W., Guillard, R.R.L. 1987. Media for the culture of oceanic ultraphytoplankton. <i>J</i>. <i>Phycol</i>., 23: 633-638. </td> </tr> </tbody> </table> <p class="mediumCategoryTitle">2 Media for marine and brackish water microalgae</p> <h1><a name="m_asp7" id="m_asp7"></a>M–ASP7</h1> <div style="text-align: right;"> <a href="./medium/en/m_asp7.pdf">Print</a> </div> <table> <tbody> <tr> <td>NTA</td> <td>7 mg</td> </tr> <tr> <td>NaCl</td> <td>2.5 g</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>900 mg</td> </tr> <tr> <td>KCl</td> <td>70 mg</td> </tr> <tr> <td>CaCl<sub>2</sub> · 2H<sub>2</sub>O </td> <td>30 mg</td> </tr> <tr> <td>NaNO<sub>3</sub></td> <td>5 mg</td> </tr> <tr> <td>NaH<sub>2</sub>PO<sub>4</sub> · 2H<sub>2</sub>O </td> <td>2 mg</td> </tr> <tr> <td>Vitamin B<sub>12</sub></td> <td>0.1 μg</td> </tr> <tr> <td><a href="#s3">Vitamin mix S<sub>3</sub></a><sup> 1 )</sup></td> <td>1 mL</td> </tr> <tr> <td>Na<sub>2</sub>SiO<sub>3</sub> · 9H<sub>2</sub>O </td> <td>1 mg</td> </tr> <tr> <td><a href="#p_n">P<sub>N</sub> metals </a><sup> 2 )</sup></td> <td>3 mL</td> </tr> <tr> <td>Tris ( hydroxymethyl ) aminomethane</td> <td>100 mg</td> </tr> <tr> <td>Distilled water</td> <td>96 mL</td> </tr> <tr> <td colspan="2">pH 8.0</td> </tr> </tbody> </table> <p> 1 ) See <a href="#s3">Vitamin mix S<sub>3</sub></a> <br> 2 ) See <a href="#p_n">P<sub>N</sub> metals </a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Watanabe, M. M., Satake, K. N. ( Eds. ) 1991 <i>NIES-Collection. List of Strains, Third Edition, 1991, Microalgae and Protozoa</i>. Microbial Culture Collection, National Institute for Environmental Studies, Tsukuba, 163 pp.k.pdf </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">2 Media for marine and brackish water microalgae</p> <h1><a name="mf" id="mb"></a>Marine Broth (MB) medium</h1> <div style="text-align: right;"> <a href="./medium/en/mf.pdf">Print</a> </div> <p>Into 100 mL distilled water dissolve 3.74 g powder medium of Difco<sup>TM</sup> Marine broth 2216 (Becton, Dickinson and Company) and 25.2 mg powder medium of Daigo IMK (Nihon Pharmaceutical Co., Ltd.). In the NIES-Collection, MB medium is used after autoclaving (121°C, 20 min). </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Ishii, Y., Maruyama, S., Fujimura-Kamada, K., Kutsuna, N., Takahashi, S., Kawata, M., Minagawa, J. 2018 Isolation of uracil auxotroph mutants of coral symbiont alga for symbiosis studies. <i>Sci Rep</i>, <b>8</b>, 3237.</td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">2 Media for marine and brackish water microalgae</p> <h1><a name="mf" id="mf"></a>MF</h1> <div style="text-align: right;"> <a href="./medium/en/mf.pdf">Print</a> </div> <p> <a href="#f2">f/2</a><sup> 1 )</sup> medium with Na<sub>2</sub>SiO<sub>3</sub> · 9H<sub>2</sub>O replaced by 1 mL <a href="#soil_extract">Soil extract</a><sup> 2 )</sup> and adjusted to pH 8.0 by buffering with 100 mg Tris ( hydroxymethyl ) aminomethane. <br> 1 ) See <a href="#f2">f/2</a> <br> 2 ) See <a href="#soil_extract">Soil extract</a> </p> <br> <p class="mediumCategoryTitle">2 Media for marine and brackish water microalgae</p> <h1><a name="mkm" id="mkm"></a>MKM</h1> <div style="text-align: right;"> <a href="./medium/en/mkm.pdf">Print</a> </div> <table> <tbody> <tr> <td>KNO<sub>3</sub></td> <td>75 mg</td> </tr> <tr> <td>KH<sub>2</sub>PO<sub>4</sub></td> <td>2.5 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>2 mg</td> </tr> <tr> <td>Fe–citrate</td> <td>250 μg</td> </tr> <tr> <td>Agar</td> <td>1.5 g</td> </tr> <tr> <td>Seawater</td> <td>50 mL</td> </tr> <tr> <td>Distilled water</td> <td>50 mL</td> </tr> </tbody> </table> <br> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Watanabe, A. 1960 List of algal strains in collection at the Institute of Applied Micro-biology, University of Tokyo. <i>J. Gen. Appl. Microbiol</i>., <b>6</b>, 283-292. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">2 Media for marine and brackish water microalgae</p> <h1><a name="mnk" id="mnk"></a>MNK</h1> <div style="text-align: right;"> <a href="./medium/en/mnk.pdf">Print</a> </div> <table> <tbody> <tr> <td>NaNO<sub>3</sub></td> <td>2 mg</td> </tr> <tr> <td>K<sub>2</sub>HPO<sub>4</sub></td> <td>0.1 mg</td> </tr> <tr> <td>Na<sub>2</sub>HPO<sub>4</sub> · 12H<sub>2</sub>O </td> <td>0.028 mg</td> </tr> <tr> <td>Vitamin B<sub>12</sub></td> <td>0.015 μg</td> </tr> <tr> <td>Biotin</td> <td>0.015 μg</td> </tr> <tr> <td>Thiamine HCl</td> <td>2 μg</td> </tr> <tr> <td>CoSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>0.12 μg</td> </tr> <tr> <td>ZnSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>0.24 μg</td> </tr> <tr> <td>MnCl<sub>2</sub> · 4H<sub>2</sub>O </td> <td>0.9 μg</td> </tr> <tr> <td>CuSO<sub>4</sub> · 5H<sub>2</sub>O </td> <td>0.006 μg</td> </tr> <tr> <td>Na<sub>2</sub>SeO<sub>3</sub></td> <td>0.003 μg</td> </tr> <tr> <td>Na<sub>2</sub>MoO<sub>4</sub> · 2H<sub>2</sub>O </td> <td>0.07 μg</td> </tr> <tr> <td>Na<sub>2</sub>EDTA · 2H<sub>2</sub>O </td> <td>0.37 μg</td> </tr> <tr> <td>Fe–EDTA</td> <td>2.6 μg</td> </tr> <tr> <td>Mn–EDTA</td> <td>3.3 μg</td> </tr> <tr> <td height="47">Seawater</td> <td>100 mL</td> </tr> </tbody> </table> <p>Vitamins should be added at the end of the preparation. The medium is sterilized by filtering, not autoclaving.<br> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Noël, M.-H., Kawachi, M., Inouye, I. 2004 Induced dimorphic life cycle of a coccolithophorid, <i>Calyptrosphaera sphaeroidea</i> ( Prymnesiophyceae, Haptophyta ). <i>J. Phycol</i>., <b>40</b>, 112-129. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">2 Media for marine and brackish water microalgae</p> <h1><a name="mimr" id="mimr"></a>ModifiedIMR ( mIMR )</h1> <div style="text-align: right;"> <a href="./medium/en/mimr.pdf">Print</a> </div> <table> <tbody> <tr> <td>KNO<sub>3</sub></td> <td>1.26 mg</td> </tr> <tr> <td>K<sub>2</sub>HPO<sub>4</sub></td> <td>0.22 mg</td> </tr> <tr> <td>Na<sub>2</sub>SiO<sub>3</sub><sup> 1 )</sup></td> <td>0.61 mg</td> </tr> <tr> <td><a href="#mimr_trace">mIMR trace metals</a><sup> 2 )</sup></td> <td>0.1 mL</td> </tr> <tr> <td>Thiamine</td> <td>20 μg</td> </tr> <tr> <td>Vitamin B<sub>12</sub></td> <td>1 μg</td> </tr> <tr> <td>Biotin</td> <td>0.1 μg</td> </tr> <tr> <td>Seawater</td> <td>100 mL</td> </tr> </tbody> </table> <p> 1 ) In the NIES–Collection, Na<sub>2</sub>SiO<sub>3</sub> is replaced by Na<sub>2</sub>SiO<sub>3</sub>·9H<sub>2</sub>O. <br> 2 ) See <a href="#mimr_trace">mIMR trace metals</a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Paasche, E., Bruback, S., Skattebol, S., Young, J. R., Green, J. C 1996 Growth and calcification in the coccolithophorid <i>Emiliania huxleyi</i> ( Haptophyceae ) at low salinities. <i>Phycologia</i>, <b>35</b>, 394-403. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">2 Media for marine and brackish water microalgae</p> <h1><a name="mswm3" id="mswm3"></a>Modified SWM–3 ( mSWM–3 )</h1> <div style="text-align: right;"> <a href="./medium/en/mswm_3.pdf">Print</a> </div> <table> <tbody> <tr> <td>NaNO<sub>3</sub></td> <td>17 mg</td> </tr> <tr> <td>NaH<sub>2</sub>PO<sub>4</sub>· 2H<sub>2</sub>O </td> <td>1.56 mg</td> </tr> <tr> <td>Na<sub>2</sub>SiO<sub>3</sub>· 9H<sub>2</sub>O </td> <td>5.68 mg</td> </tr> <tr> <td>Na<sub>2</sub>EDTA· 2H<sub>2</sub>O </td> <td>1.12 mg</td> </tr> <tr> <td>Fe-EDTA</td> <td>0.084 mg</td> </tr> <tr> <td>Na<sub>2</sub>SeO<sub>3</sub></td> <td>0.0346 μg</td> </tr> <tr> <td><a href="#p1_metals">P–1 metals</a></td> <td>1 mL</td> </tr> <tr> <td>Vitamin B<sub>12</sub></td> <td>0.2 μg</td> </tr> <tr> <td><a href="#s3">Vitamin mix S<sub>3</sub></a></td> <td>1 mL</td> </tr> <tr> <td>Tris ( hydroxymethyl ) aminomethane</td> <td>50 mg</td> </tr> <tr> <td>Seawater</td> <td>98.8 mL</td> </tr> <tr> <td colspan="2">pH 7.7 – 7.8</td> </tr> </tbody> </table> <p> 1 ) See <a href="#p1_metals">P–1 metals</a> <br> 2 ) See <a href="#s3">Vitamin mix S<sub>3</sub></a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Imai, I. 2012 High growth rates of the red tide flagellate <i>Chattonella marina</i> (Raphidophyceae) observed in culture. <i>Hokudai Suisan Ihou</i>, <b>62</b>, 71-74 ( in Japanese with English summary ). </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">2 Media for marine and brackish water microalgae</p> <h1><a name="pro99" id="pro99"></a>PRO–99</h1> <div style="text-align: right;"> <a href="./medium/en/pro99.pdf">Print</a> </div> <table> <tbody> <tr> <td>NaH<sub>2</sub>PO<sub>4</sub> · 2H<sub>2</sub>O </td> <td>0.779 mg</td> </tr> <tr> <td>NH<sub>4</sub>Cl </td> <td>4.28 mg</td> </tr> <tr> <td><a href="#pro99_trace">PRO–99 trace metals</a><sup> 1 )</sup></td> <td>0.01 mL</td> </tr> <tr> <td>Seawater</td> <td>100 mL</td> </tr> </tbody> </table> <p>The medium is sterilized by filtering, not autoclaving.<br> 1 ) See <a href="#pro99_trace">PRO–99 trace metals</a><br> </p> <p class="mediumCategoryTitle">2 Media for marine and brackish water microalgae</p> <h1><a name="wesm" id="wesm"></a>WESM</h1> <div style="text-align: right;"> <a href="./medium/en/wesm.pdf">Print</a> </div> <p> Make diluted seawater by mixing 87.5 mL seawater and 10mL distilled water. Make <a href="#esm">ESM</a><sup> 1 )</sup> medium by using this diluted seawater instead of seawater. <br> 1 ) See <a href="#esm">ESM</a> </p> <p class="mediumCategoryTitle">3 Bacteria-free check media for freshwater algae</p> <h1><a name="b_i" id="b_i"></a>B–I</h1> <div style="text-align: right;"> <a href="./medium/en/b_i.pdf">Print</a> </div> <table> <tbody> <tr> <td>Appropriate medium</td> <td>100 mL</td> </tr> <tr> <td>Proteose peptone</td> <td>100 mg</td> </tr> </tbody> </table> <br> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Ichimura, T., Watanabe, M. M. 1977 An axenic clone of <i>Microcystis aeruginosa</i> Kutz. emend. Elenkin from Lake Kasumigaura. <i>Bull. Jpn. Soc. Phycol</i>., <b>25</b>, 177-181. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">3 Bacteria-free check media for freshwater algae</p> <h1><a name="b_ii" id="b_ii"></a>B–II</h1> <div style="text-align: right;"> <a href="./medium/en/b_ii.pdf">Print</a> </div> <table> <tbody> <tr> <td>Appropriate medium</td> <td>100 mL</td> </tr> <tr> <td>Yeast extract</td> <td>500 mg</td> </tr> </tbody> </table> <br> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Ichimura, T., Watanabe, M. M. 1977 An axenic clone of <i>Microcystis aeruginosa</i> Kutz. emend. Elenkin from Lake Kasumigaura. <i>Bull. Jpn. Soc. Phycol</i>., <b>25</b>, 177-181. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">3 Bacteria-free check media for freshwater algae</p> <h1><a name="b_iii" id="b_iii"></a>B–III</h1> <div style="text-align: right;"> <a href="./medium/en/b_iii.pdf">Print</a> </div> <table> <tbody> <tr> <td>Appropriate medium</td> <td>100 mL</td> </tr> <tr> <td>Peptone</td> <td>500 mg</td> </tr> <tr> <td>Beef extract</td> <td>300 mg</td> </tr> </tbody> </table> <br> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Ichimura, T., Watanabe, M. M. 1977 An axenic clone of <i>Microcystis aeruginosa</i> Kutz. emend. Elenkin from Lake Kasumigaura. <i>Bull. Jpn. Soc. Phycol</i>., <b>25</b>, 177-181. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">3 Bacteria-free check media for freshwater algae</p> <h1><a name="b_iv" id="b_iv"></a>B–IV</h1> <div style="text-align: right;"> <a href="./medium/en/_iv.pdf">Print</a> </div> <table> <tbody> <tr> <td>Appropriate medium</td> <td>100 mL</td> </tr> <tr> <td>Glucose</td> <td>100 mg</td> </tr> <tr> <td>Peptone</td> <td>100 mg</td> </tr> </tbody> </table> <br> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Ichimura, T., Watanabe, M. M. 1977 An axenic clone of <i>Microcystis aeruginosa</i> Kutz. emend. Elenkin from Lake Kasumigaura. <i>Bull. Jpn. Soc. Phycol</i>., <b>25</b>, 177-181. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">3 Bacteria-free check media for freshwater algae</p> <h1><a name="b_v" id="b_v"></a>B–V</h1> <div style="text-align: right;"> <a href="./medium/en/b_v.pdf">Print</a> </div> <table> <tbody> <tr> <td>Appropriate medium</td> <td>100 mL</td> </tr> <tr> <td>Sodium acetate</td> <td>50 mg</td> </tr> <tr> <td>Glucose</td> <td>50 mg</td> </tr> <tr> <td>Tryptone</td> <td>50 mg</td> </tr> <tr> <td>Yeast extract</td> <td>30 mg</td> </tr> <tr> </tr> </tbody> </table> <br> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Ichimura, T., Watanabe, M. M. 1977 An axenic clone of <i>Microcystis aeruginosa</i> Kutz. emend. Elenkin from Lake Kasumigaura. <i>Bull. Jpn. Soc. Phycol</i>., <b>25</b>, 177-181. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">3 Bacteria-free check media for freshwater algae</p> <h1><a name="yt" id="yt"></a>YT</h1> <div style="text-align: right;"> <a href="./medium/en/yt.pdf">Print</a> </div> <table> <tbody> <tr> <td>Appropriate medium</td> <td>100 mL</td> </tr> <tr> <td>Yeast extract</td> <td>100 mg</td> </tr> <tr> <td>Tryptone</td> <td>200 mg</td> </tr> </tbody> </table> <br> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Ichimura, T., Itoh, T. 1977 17. Preservation methods of microalgae ( I ) [17. Bisaisôrui no hozonhô ( I ) ]. In <i>Preservation methods of microorganisms [Biseibutsu Hozonhô]</i>, Ed. by Nei, T., University of Tokyo Press, Tokyo, p. 355-373 ( in Japanese without English title ). </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">4 Bacteria-free check media for marine algae</p> <h1><a name="bf_2" id="bf_2"></a>Bf/2</h1> <div style="text-align: right;"> <a href="./medium/en/bf_2.pdf">Print</a> </div> <table> <tbody> <tr> <td>ASP 7<sup> 1 )</sup></td> <td>100 mL</td> </tr> <tr> <td>Trypticase</td> <td>50 mg</td> </tr> <tr> <td>Yeast extract</td> <td>5 mg</td> </tr> </tbody> </table> <p> 1 ) In the NIES-Collection, ASP 7 is replaced by <a href="#f2">f/2</a> medium. See <a href="#f2">f/2</a>. </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Yoshida, Y., Kawaguchi, K. 1983 Buoyancy and phototaxis of <i>Chattonella antiqua</i> ( Hada ) Ono. <i>Bull. Plankton Soc. Jpn</i>., <b>30</b>, 11-19 ( in Japanese with English summary ). </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">4 Bacteria-free check media for marine algae</p> <h1><a name="mm23" id="mm23"></a>MM 23 ( M. Tatewaki, pers. comm. )</h1> <div style="text-align: right;"> <a href="./medium/en/mm23.pdf">Print</a> </div> <table> <tbody> <tr> <td>NaCl</td> <td>1.8 g</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>500 mg</td> </tr> <tr> <td>KCl</td> <td>60 mg</td> </tr> <tr> <td>NaNO<sub>3</sub></td> <td>100 mg</td> </tr> <tr> <td>CaCl<sub>2</sub> · 2H<sub>2</sub>O </td> <td>36.7 mg</td> </tr> <tr> <td>K<sub>2</sub>HPO<sub>4</sub></td> <td>6 mg</td> </tr> <tr> <td>Sucrose</td> <td>400 mg</td> </tr> <tr> <td><a href="#p_ii">PII metals</a><sup> 1 )</sup></td> <td>2 mL</td> </tr> <tr> <td>FeCl<sub>3</sub> · 6H<sub>2</sub>O </td> <td>48 μg</td> </tr> <tr> <td>Thiamine HCl</td> <td>10 μg</td> </tr> <tr> <td>Biotin</td> <td>0.1 μg</td> </tr> <tr> <td>Vitamin B<sub>12</sub></td> <td>0.2 μg</td> </tr> <tr> <td><a href="#c_mix">C–Source Mix II</a><sup> 2 )</sup></td> <td>1 mL</td> </tr> <tr> <td>Tris ( hydroxymethyl ) aminomethane</td> <td>100 mg</td> </tr> <tr> <td>Distilled water</td> <td>97 mL</td> </tr> <tr> <td colspan="2">pH 8.0</td> </tr> </tbody> </table> <p> 1 ) See <a href="#p_ii">PII metals</a> <br> 2 ) See <a href="#c_mix">C–Source Mix II</a> </p> <p class="mediumCategoryTitle">4 Bacteria-free check media for marine algae</p> <h1><a name="stp" id="stp"></a>STP</h1> <div style="text-align: right;"> <a href="./medium/en/stp.pdf">Print</a> </div> <table> <tbody> <tr> <td>NaNO<sub>3</sub></td> <td>20 mg</td> </tr> <tr> <td>K<sub>2</sub>HPO<sub>4</sub></td> <td>1 mg</td> </tr> <tr> <td>Sodium glutamate</td> <td>50 mg</td> </tr> <tr> <td>Glucose</td> <td>20 mg</td> </tr> <tr> <td>Glycine</td> <td>10 mg</td> </tr> <tr> <td>D,L–Alanine</td> <td>10 mg</td> </tr> <tr> <td>Vitamin mix 8<sup> 1 )</sup></td> <td>0.1 mL</td> </tr> <tr> <td>Trypticase</td> <td>20 mg</td> </tr> <tr> <td>Yeast autolysate<sup> 2 )</sup></td> <td>20 mg</td> </tr> <tr> <td>Sucrose</td> <td>100 mg</td> </tr> <tr> <td><a href="#soil_extract">Soil extract</a><sup> 3 )</sup></td> <td>5 mL</td> </tr> <tr> <td>Seawater</td> <td>80 mL</td> </tr> <tr> <td>Distilled water</td> <td>15 mL</td> </tr> <tr> <td colspan="2">pH 7.5</td> </tr> </tbody> </table> <p>1 ) In the NIES–Collection, Vitamin mix 8 is replaced by</p> <p> <a href="#s3">Vitamin mix S<sub>3</sub></a>. See <a href="#s3">Vitamin mix S<sub>3</sub> </a>. <br> 2 ) In the NIES–Collection, yeast autolysate is replaced by yeast extract. <br> 3 ) See <a href="#soil_extract">Soil extract</a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Provasoli, L., McLaughlin, J. J. A., Droop, M. R. 1957 The development of artificial media for marine algae. <i>Arch. Mikrobiol</i>., <b>25</b>, 392-428. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">5 Trace metals, vitamin mixtures and soil extracts</p> <h1><a name="a2fe" id="a2fe"></a>A2 Fe stock solution</h1> <div style="text-align: right;"> <a href="./medium/en/a2fe.pdf">Print</a> </div> <table> <tbody> <tr> <td>EDTA · 2Na</td> <td>700 mg</td> </tr> <tr> <td>FeCl<sub>3</sub> · 6H<sub>2</sub>O </td> <td>400 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> </tbody> </table> <p>Sterilize by passing through a Millipore filter ( 0.22 μm ). </p> <br> <p class="mediumCategoryTitle">5 Trace metals, vitamin mixtures and soil extracts</p> <h1><a name="a2trace" id="a2trace"></a>A2 trace element stock solution</h1> <div style="text-align: right;"> <a href="./medium/en/a2trace.pdf">Print</a> </div> <table> <tbody> <tr> <td>H<sub>3</sub>BO<sub>3</sub></td> <td>285 mg</td> </tr> <tr> <td>MnCl<sub>2</sub> · 4H<sub>2</sub>O<sup> </sup></td> <td>180 mg</td> </tr> <tr> <td>ZnCl<sub>2</sub> </td> <td>10.5 mg</td> </tr> <tr> <td>Na<sub>2</sub>MoO<sub>4</sub> · 2H<sub>2</sub>O </td> <td>39 mg</td> </tr> <tr> <td>CoCl<sub>2</sub> · 6H<sub>2</sub>O </td> <td>4 mg</td> </tr> <tr> <td>CuCl<sub>2</sub> · 2H<sub>2</sub>O </td> <td>4.3 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> </tbody> </table> <p class="mediumCategoryTitle">5 Trace metals, vitamin mixtures and soil extracts</p> <h1> <a name="a5" id="a5"></a>A<sub>5</sub> solution </h1> <div style="text-align: right;"> <a href="./medium/en/a5.pdf">Print</a> </div> <table> <tbody> <tr> <td>H<sub>3</sub>BO<sub>3</sub></td> <td>286 mg</td> </tr> <tr> <td>MnSO<sub>4</sub> · 7H<sub>2</sub>O<sup> 1 )</sup></td> <td>250 mg</td> </tr> <tr> <td>ZnSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>22.2 mg</td> </tr> <tr> <td>CuSO<sub>4</sub> · 5H<sub>2</sub>O </td> <td>7.9 mg</td> </tr> <tr> <td>Na<sub>2</sub>MoO<sub>4</sub> · 2H<sub>2</sub>O </td> <td>2.1 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> </tbody> </table> <p> 1 ) In the NIES–Collection, 250 mg MnSO<sub>4</sub> · 7H<sub>2</sub>O is replaced by 217 mg MnSO<sub>4</sub> · 5H<sub>2</sub>O. </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Holm-Hansen, O., Gerloff, G. C., Skoog, F. 1954 Cobalt as an essential element for blue-green algae. <i>Physiol. Planta</i>., <b>7</b>, 665-675. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">5 Trace metals, vitamin mixtures and soil extracts</p> <h1><a name="allen_me" id="allen_me"></a>Allen metals</h1> <div style="text-align: right;"> <a href="./medium/en/allen_me.pdf">Print</a> </div> <table> <tbody> <tr> <td>Fe–EDTA</td> <td>30.16 g</td> </tr> <tr> <td>MnCl<sub>2</sub> · 4H<sub>2</sub>O </td> <td>1.79 g</td> </tr> <tr> <td>H<sub>3</sub>BO<sub>3</sub></td> <td>2.86 g</td> </tr> <tr> <td>ZnSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>220 mg</td> </tr> <tr> <td>CuSO<sub>4</sub> · 5H<sub>2</sub>O </td> <td>79 mg</td> </tr> <tr> <td>( NH<sub>4</sub> )<sub>6</sub>Mo<sub>7</sub>O<sub>24</sub> · 4H<sub>2</sub>O </td> <td>130 mg</td> </tr> <tr> <td>NH<sub>4</sub>VO<sub>3</sub></td> <td>23 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> </tbody> </table> <p> In the NIES–Collection, <a href="#allen_me">Allen metals</a> are used after dilution with distilled water to 1/1,000. </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Allen, M. B. 1959 Studies with <i>Cyanidium caldarium</i>, an anomalously pigmented chlorophyte. <i>Arch. Mikrobiol</i>., <b>32</b>, 270-277. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">5 Trace metals, vitamin mixtures and soil extracts</p> <h1><a name="c_mix" id="c_mix"></a>C–Source Mix II ( M. Tatewaki, pers. comm. )</h1> <div style="text-align: right;"> <a href="./medium/en/c_mix.pdf">Print</a> </div> <table> <tbody> <tr> <td>Glycine</td> <td>100 mg</td> </tr> <tr> <td>D,L-Alanine</td> <td>100 mg</td> </tr> <tr> <td>L-Asparagine</td> <td>100 mg</td> </tr> <tr> <td>Sodium acetate · 3H<sub>2</sub>O<sup> 1 )</sup></td> <td>200 mg</td> </tr> <tr> <td>Glucose</td> <td>200 mg</td> </tr> <tr> <td>L-Glutamic acid</td> <td>200 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> </tbody> </table> <p> 1 ) In the NIES–Collection, 200 mg sodium acetate · 3H<sub>2</sub>O is replaced by 120 mg sodium acetate, anhydrous. </p> <br> <p class="mediumCategoryTitle">5 Trace metals, vitamin mixtures and soil extracts</p> <h1><a name="d_stock" id="d_stock"></a>D stock medium</h1> <div style="text-align: right;"> <a href="./medium/en/d_stock.pdf">Print</a> </div> <table> <tbody> <tr> <td>NTA</td> <td>0.2 g</td> </tr> <tr> <td><a href="#d_trace">D trace mix</a><sup> 1 )</sup></td> <td>1 mL</td> </tr> <tr> <td>FeCl<sub>3</sub> · 6H<sub>2</sub>O </td> <td>0.58 mg</td> </tr> <tr> <td>CaSO<sub>4</sub> · 2H<sub>2</sub>O </td> <td>120 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>200 mg</td> </tr> <tr> <td>NaCl</td> <td>16 mg</td> </tr> <tr> <td>KNO<sub>3</sub></td> <td>200 mg</td> </tr> <tr> <td>NaNO<sub>3</sub></td> <td>1.4 g</td> </tr> <tr> <td>Na<sub>2</sub>HPO<sub>4</sub><sup> 2 )</sup></td> <td>220 mg</td> </tr> <tr> <td>Distilled water</td> <td>99 mL</td> </tr> </tbody> </table> <p> 1 ) See <a href="#d_trace">D trace mix</a> <br> 2 ) In the NIES-Collection, 220 mg Na<sub>2</sub>HPO<sub>4</sub> is replaced by 550 mg Na<sub>2</sub>HPO<sub>4</sub>· 12H<sub>2</sub>O. </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Castenholz, R. W. 1969 Thermophilic blue-green algae and the thermal environment. <i>Bacteriol. Rev</i>., <b>33</b>, 476-504. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">5 Trace metals, vitamin mixtures and soil extracts</p> <h1><a name="d_trace" id="d_trace"></a>D trace mix</h1> <div style="text-align: right;"> <a href="./medium/en/d_trace.pdf">Print</a> </div> <table> <tbody> <tr> <td>conc · H<sub>2</sub>SO<sub>4</sub></td> <td>0.05 mL</td> </tr> <tr> <td>MnSO<sub>4</sub> · H<sub>2</sub>O<sup> 1 )</sup></td> <td>228 mg</td> </tr> <tr> <td>ZnSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>50 mg</td> </tr> <tr> <td>H<sub>3</sub>BO<sub>3</sub></td> <td>50 mg</td> </tr> <tr> <td>CuSO<sub>4</sub> · 5H<sub>2</sub>O </td> <td>2.5 mg</td> </tr> <tr> <td>Na<sub>2</sub>MoO<sub>4</sub> · 2H<sub>2</sub>O </td> <td>2.5 mg</td> </tr> <tr> <td>CoCl<sub>2</sub> · 6H<sub>2</sub>O </td> <td>4.5 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> </tbody> </table> <p> 1 ) In the NIES–Collection, 228 mg MnSO<sub>4</sub> · H<sub>2</sub>O is replaced by 349 mg MnSO<sub>4</sub> · 5H<sub>2</sub>O. </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Castenholz, R. W. 1969 Thermophilic blue-green algae and the thermal environment. <i>Bacteriol. Rev</i>., <b>33</b>, 476-504. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">5 Trace metals, vitamin mixtures and soil extracts</p> <h1><a name="dy_trace" id="dy_trace"></a>DY trace metal solution</h1> <div style="text-align: right;"> <a href="./medium/en/dy_trace.pdf">Print</a> </div> <table> <tbody> <tr> <td>MnCl<sub>2</sub> · 4H<sub>2</sub>O </td> <td>20 mg</td> </tr> <tr> <td>ZnSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>4 mg</td> </tr> <tr> <td>CoCl<sub>2</sub> · 6H<sub>2</sub>O </td> <td>0.8 mg</td> </tr> <tr> <td>Na<sub>2</sub>MoO<sub>4</sub> · 6H<sub>2</sub>O <sup> 1 )</sup></td> <td>2 mg</td> </tr> <tr> <td>Na<sub>3</sub>VO<sub>4</sub> · 10H<sub>2</sub>O <sup> 2 )</sup></td> <td>0.2 mg</td> </tr> <tr> <td>H<sub>2</sub>SeO<sub>3</sub></td> <td>0.2 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> </tbody> </table> <p> 1 ) In the NIES–Collection, Na<sub>2</sub>MoO<sub>4</sub> · 6H<sub>2</sub>O is replaced by Na<sub>2</sub>MoO<sub>4</sub> · 2H<sub>2</sub>O.<br> 2 ) In the NIES-Collection, 0.2 mg Na<sub>3</sub>VO<sub>4</sub> · 10H<sub>2</sub>O is replaced by 0.1 mg Na<sub>3</sub>VO<sub>4</sub>. </p> <p class="mediumCategoryTitle">5 Trace metals, vitamin mixtures and soil extracts</p> <h1><a name="fe" id="fe"></a>Fe ( as EDTA; 1:1 molar )</h1> <div style="text-align: right;"> <a href="./medium/en/fe.pdf">Print</a> </div> <table> <tbody> <tr> <td>Fe( NH<sub>4</sub> )<sub>2</sub>( SO<sub>4</sub> )<sub>2</sub> · 6H<sub>2</sub>O </td> <td>70.2 mg</td> </tr> <tr> <td>Na<sub>2</sub>EDTA · 2H<sub>2</sub>O </td> <td>66 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> </tbody> </table> <p>1 mL of   this solution contains 0.1 mg Fe.</p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Provasoli, L. 1966 Media and prospects for the cultivation of marine algae. In <i>Cultures and Collections of Algae</i>, Eds. by Watanabe, A. & Hattori, A., Proc. U.S.-Japan Conf., Hakone, Sept. 1966., Jpn. Soc. Plant Physiol., p. 63-75. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">5 Trace metals, vitamin mixtures and soil extracts</p> <h1><a name="fe_solution" id="fe_solution"></a>Fe solution</h1> <div style="text-align: right;"> <a href="./medium/en/fe_solution.pdf">Print</a> </div> <table> <tbody> <tr> <td>FeSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>200 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> <tr> <td>Conc · H<sub>2</sub>SO<sub>4</sub></td> <td>0.026 mL<sup> 1 )</sup></td> </tr> </tbody> </table> <p>1 ) 2 drops/500 mL</p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Ichimura, T., Itoh, T. 1977 17. Preservation methods of microalgae ( I ) [17. Bisaisôrui no hozonhô ( I )]. In <i>Preservation methods of microorganisms [Biseibutsu Hozonhô]</i>, Ed. by Nei, T., University of Tokyo Press, Tokyo, p. 355-373 ( in Japanese without English title ). </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">5 Trace metals, vitamin mixtures and soil extracts</p> <h1><a name="f2_metals" id="f2_metals"></a>f/2 metals</h1> <div style="text-align: right;"> <a href="./medium/en/f2_metals.pdf">Print</a> </div> <table> <tbody> <tr> <td>Na<sub>2</sub>EDTA · 2H<sub>2</sub>O </td> <td>440 mg</td> </tr> <tr> <td>FeCl<sub>3</sub> · 6H<sub>2</sub>O </td> <td>316 mg</td> </tr> <tr> <td>CoSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>1.2 mg</td> </tr> <tr> <td>ZnSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>2.1 mg</td> </tr> <tr> <td>MnCl<sub>2</sub> · 4H<sub>2</sub>O </td> <td>18 mg</td> </tr> <tr> <td>CuSO<sub>4</sub> · 5H<sub>2</sub>O </td> <td>0.7 mg</td> </tr> <tr> <td>Na<sub>2</sub>MoO<sub>4</sub> · 2H<sub>2</sub>O </td> <td>0.7 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> </tbody> </table> <br> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Guillard, R. R. L., Ryther, J. H. 1962 Studies of marine planktonic diatoms. I. <i>Cyclotella nana</i> Hustedt, and <i>Detonula confervacea</i> ( Cleve ) Gran. <i>Can. J. Microbiol</i>., <b>8</b>, 229-239. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">5 Trace metals, vitamin mixtures and soil extracts</p> <h1><a name="hunter" id="hunter"></a>Hutner ’s trace elements</h1> <div style="text-align: right;"> <a href="./medium/en/hunter.pdf">Print</a> </div> <table> <tbody> <tr> <td>Na<sub>2</sub>EDTA · 2H<sub>2</sub>O </td> <td>5 g</td> </tr> <tr> <td>ZnSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>2.2 g</td> </tr> <tr> <td>H<sub>3</sub>BO<sub>3</sub></td> <td>1.14 g</td> </tr> <tr> <td>MnCl<sub>2</sub> · 4H<sub>2</sub>O </td> <td>506 mg</td> </tr> <tr> <td>FeSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>499 mg</td> </tr> <tr> <td>CoCl<sub>2</sub> · 6H<sub>2</sub>O </td> <td>161 mg</td> </tr> <tr> <td>CuSO<sub>4</sub>· 5H<sub>2</sub>O </td> <td>157 mg</td> </tr> <tr> <td>( NH<sub>4</sub> )<sub>6</sub>Mo<sub>7</sub>O<sub>24</sub> · 4H<sub>2</sub>O </td> <td>110 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> </tbody> </table> <p>Adjust the pH to 6.5-6.8 with KOH ( ~1.6 g ). Store the solution in a refrigerator ( 5°C ). The solution should have turned to violet color before use. This process takes a while and is necessary. </p> <p class="mediumCategoryTitle">5 Trace metals, vitamin mixtures and soil extracts</p> <h1><a name="K_metals" id="K_metals"></a>K metals</h1> <div style="text-align: right;"> <a href="./medium/en/k_metals.pdf">Print</a> </div> <table> <tbody> <tr> <td>Na<sub>2</sub>EDTA · 2H<sub>2</sub>O </td> <td>3.722 g</td> </tr> <tr> <td>Fe-EDTA</td> <td>0.493 g</td> </tr> <tr> <td>FeCl<sub>3</sub> · 6H<sub>2</sub>O </td> <td>0.315 g</td> </tr> <tr> <td>MnCl<sub>2</sub> · 4H<sub>2</sub>O </td> <td>17.8 mg</td> </tr> <tr> <td>ZnSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>2.3 mg</td> </tr> <tr> <td>CoSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>1.405 mg</td> </tr> <tr> <td>Na<sub>2</sub>MoO<sub>4</sub> · 2H<sub>2</sub>O </td> <td>0.726 mg</td> </tr> <tr> <td>CuSO<sub>4</sub> · 5H<sub>2</sub>O </td> <td>0.25 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> </tbody> </table> <br> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Keller, M. D., Guillard, R.R.L. 1985 Factors significant to marine diatom culture. pp. 113-6. <i>In</i> Anderson, D.M., White, A.W., Baden, D.G. (eds.) <i>Toxic Dinoflagellates</i>. Elsevier, New York. </td> </tr> <tr> <td>Keller, M.D., Selvin, R.C., Claus, W., Guillard, R.R.L. 1987. Media for the culture of oceanic ultraphytoplankton. <i>J</i>. <i>Phycol</i>., 23: 633-638. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">5 Trace metals, vitamin mixtures and soil extracts</p> <h1><a name="k_2_metals" id="k_2_metals"></a>K/2 metals</h1> <div style="text-align: right;"> <a href="./medium/en/k_2_metals.pdf">Print</a> </div> <p> 1 ) In the NIES–Collection, 0.149 mg NiCl<sub>2</sub> · 6H<sub>2</sub>O is replaced by 0.812 mg NiCl<sub>2</sub>. </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Guillard, R. R. L., Ryther, J. H. 1962 Studies of marine planktonic diatoms. I. <i>Cyclotella nana</i> Hustedt, and <i>Detonula confervacea</i> ( Cleve ) Gran. <i>Can. J. Microbiol</i>., <b>8</b>, 229-239. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">5 Trace metals, vitamin mixtures and soil extracts</p> <h1><a name="mimr_trace" id="mimr_trace"></a>mIMR trace metals</h1> <div style="text-align: right;"> <a href="./medium/en/mimr_trace.pdf">Print</a> </div> <table> <tbody> <tr> <td>FeCl<sub>3</sub> · 6H<sub>2</sub>O </td> <td>100 mg</td> </tr> <tr> <td>MnSO<sub>4</sub> · 4H<sub>2</sub>O </td> <td>85.5 mg</td> </tr> <tr> <td>ZnSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>25 mg</td> </tr> <tr> <td>Na<sub>2</sub>MoO<sub>4</sub> · 2H<sub>2</sub>O </td> <td>13 mg</td> </tr> <tr> <td>CoCl<sub>2</sub> · 6H<sub>2</sub>O </td> <td>0.4 mg</td> </tr> <tr> <td>CuSO<sub>4</sub> · 5H<sub>2</sub>O </td> <td>0.4 mg</td> </tr> <tr> <td>Na<sub>2</sub>SeO<sub>3</sub></td> <td>0.173 mg</td> </tr> <tr> <td>Na<sub>2</sub>EDTA<sup> 1 )</sup></td> <td>600 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> </tbody> </table> <p> 1 ) In the NIES–Collection, Na<sub>2</sub>EDTA is replaced by Na<sub>2</sub>EDTA · 2H<sub>2</sub>O. </p> <br> <p class="mediumCategoryTitle">5 Trace metals, vitamin mixtures and soil extracts</p> <h1><a name="mm1_trace" id="mm1_trace"></a>mM–1 Trace elements</h1> <div style="text-align: right;"> <a href="./medium/en/mm1_trace.pdf">Print</a> </div> <table> <tbody> <tr> <td>CuSO<sub>4</sub> · 5H<sub>2</sub>O </td> <td>10 mg</td> </tr> <tr> <td>MnCl<sub>2</sub> · 4H<sub>2</sub>O </td> <td>10 mg</td> </tr> <tr> <td>Br ( 1mol / L solution )</td> <td>0.01 mL</td> </tr> <tr> <td>ZnSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>10 mg</td> </tr> <tr> <td>CoCl<sub>2</sub> · 6H<sub>2</sub>O </td> <td>5 mg</td> </tr> <tr> <td>BaCl<sub>2</sub> · 2H<sub>2</sub>O </td> <td>1 mg</td> </tr> <tr> <td>H<sub>3</sub>BO<sub>3</sub></td> <td>10 mg</td> </tr> <tr> <td>FeCl<sub>3</sub> · 6H<sub>2</sub>O </td> <td>10 mg</td> </tr> <tr> <td>Na<sub>2</sub>MoO<sub>4</sub> · 2H<sub>2</sub>O </td> <td>5 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> </tbody> </table> <p>Indicated as “Trace elements” in reference.</p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Hoham, R. W., Berman, J. D., Rogers, H. S., Felio, J. H., Ryba, J. B., Miller, P. R. 2006 Two new species of green snow algae from Upstate New York, <i>Chloromonas chenangoensis</i> sp. nov. and <i>Chloromonas tughillensis</i> sp. nov. ( Volvocales, Chlorophyceae ) and the effects of light on their life cycle development. <i>Phycologia</i>, <b>45</b>, 319-330. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">5 Trace metals, vitamin mixtures and soil extracts</p> <h1><a name="mtygm9" id="mtygm9"></a>mTYGM–9</h1> <div style="text-align: right;"> <a href="./medium/en/mtygm9.pdf">Print</a> </div> <h3>Pre–solution</h3> <table> <tbody> <tr> <td>K<sub>2</sub>HPO<sub>4</sub></td> <td>0.28 g</td> </tr> <tr> <td>KH<sub>2</sub>PO<sub>4</sub></td> <td>40 mg</td> </tr> <tr> <td>Casein Digest</td> <td>0.2 g</td> </tr> <tr> <td>Yeast extract</td> <td>0.1 g</td> </tr> <tr> <td>NaCl</td> <td>0.75 g</td> </tr> <tr> <td>Mucin, gastric</td> <td>0.2 g</td> </tr> <tr> <td>Distilled water</td> <td>97 mL</td> </tr> </tbody> </table> <p>Sterilize Pre–solution by autoclaving ( 121°C, 15 min ) , add aseptically 3 mL horse serum and 50 μL 10% Tween 80 dissolved in absolute ethanol ( both filter–sterilized ). Keep in a cool place. </p> <br> <p class="mediumCategoryTitle">5 Trace metals, vitamin mixtures and soil extracts</p> <h1><a name="p_ii" id="p_ii"></a>P II metals</h1> <div style="text-align: right;"> <a href="./medium/en/p_ii.pdf">Print</a> </div> <table> <tbody> <tr> <td>Na<sub>2</sub>EDTA · 2H<sub>2</sub>O </td> <td>100 mg</td> </tr> <tr> <td>H<sub>3</sub>BO<sub>3</sub></td> <td>114 mg</td> </tr> <tr> <td>FeCl<sub>3</sub> · 6H<sub>2</sub>O </td> <td>4.9 mg</td> </tr> <tr> <td>MnSO<sub>4</sub> · 4H<sub>2</sub>O </td> <td>16.4 mg</td> </tr> <tr> <td>ZnSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>2.2 mg</td> </tr> <tr> <td>CoSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>480 μg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> </tbody> </table> <br> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Provasoli, L. 1963 Growing marine seaweeds. In <i>Proceedings of the Fourth International Seaweed Symposium</i>, University of Tokyo Press, Tokyo, p. 9-17. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">5 Trace metals, vitamin mixtures and soil extracts</p> <h1><a name="p1_metals" id="p1_metals"></a>P–1 metals</h1> <div style="text-align: right;"> <a href="./medium/en/p1.pdf">Print</a> </div> <table> <tbody> <tr> <td>H<sub>3</sub>BO<sub>3</sub> </td> <td>618.3 mg</td> </tr> <tr> <td>MnCl<sub>2</sub> · 4H<sub>2</sub>O </td> <td>69.25 mg</td> </tr> <tr> <td>ZnCl<sub>2</sub></td> <td>5.45 mg</td> </tr> <tr> <td>CoCl<sub>2</sub> · 6H<sub>2</sub>O </td> <td>238 μg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> </tbody> </table> <p> 1 ) 1) In the NIES-Collection, CuCl<sub>2</sub> · 2H<sub>2</sub>O is removed. </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Imai, I. 2012 High growth rates of the red tide flagellate <i>Chattonella marina</i> (Raphidophyceae) observed in culture. <i>Hokudai Suisan Ihou</i>, 62, 71-74 (in Japanese with English summary). </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">5 Trace metals, vitamin mixtures and soil extracts</p> <h1><a name="p_iv" id="p_iv"></a>P IV metals</h1> <div style="text-align: right;"> <a href="./medium/en/p_iv.pdf">Print</a> </div> <table> <tbody> <tr> <td>Na<sub>2</sub>EDTA · 2H<sub>2</sub>O </td> <td>100 mg</td> </tr> <tr> <td>FeCl<sub>3</sub> · 6H<sub>2</sub>O </td> <td>19.6 mg</td> </tr> <tr> <td>MnCl<sub>2</sub> · 4H<sub>2</sub>O </td> <td>3.6 mg</td> </tr> <tr> <td>ZnCl<sub>2</sub><sup> 1 )</sup></td> <td>1.04 mg</td> </tr> <tr> <td>CoCl<sub>2</sub> · 6H<sub>2</sub>O </td> <td>0.4 mg</td> </tr> <tr> <td>Na<sub>2</sub>MoO<sub>4</sub> · 2H<sub>2</sub>O </td> <td>0.25 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> </tbody> </table> <p> 1 ) In the NIES–Collection, 1.04mg ZnCl<sub>2</sub> is replaced by 2.2mg ZnSO<sub>4</sub> · 7H<sub>2</sub>O. </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Provasoli, L., Pintner, I. J. 1959 Artificial media for fresh-water algae: problems and suggestions. In <i>The Ecology of Algae. Spec. Pub. No. 2</i>,, Eds. by Tryon, C. A., Jr. & Hartmann, R. T., Pymatuning Laboratory of Field Biology, University of Pittsburgh, Pittsburgh, p. 84-96. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">5 Trace metals, vitamin mixtures and soil extracts</p> <h1> <a name="p_n" id="p_n"></a>P<sub>N</sub> metals </h1> <div style="text-align: right;"> <a href="./medium/en/p_n.pdf">Print</a> </div> <table> <tbody> <tr> <td>Na<sub>2</sub>EDTA · 2H<sub>2</sub>O </td> <td>100 mg</td> </tr> <tr> <td>H<sub>3</sub>BO<sub>3</sub></td> <td>113 mg</td> </tr> <tr> <td>FeCl<sub>3</sub> · 6H<sub>2</sub>O </td> <td>6.3 mg</td> </tr> <tr> <td>CoSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>0.093 mg</td> </tr> <tr> <td>ZnSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>4.66 mg</td> </tr> <tr> <td>MnCl<sub>2</sub> · 4H<sub>2</sub>O </td> <td>3.2 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> </tbody> </table> <br> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Watanabe, M. M., Satake, K. N. ( Eds. ) 1991 <i>NIES-Collection. List of Strains, Third Edition, 1991, Microalgae and Protozoa</i>. Microbial Culture Collection, National Institute for Environmental Studies, Tsukuba, 163 pp. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">5 Trace metals, vitamin mixtures and soil extracts</p> <h1><a name="pro99_trace" id="pro99_trace"></a>PRO–99 trace metals</h1> <div style="text-align: right;"> <a href="./medium/en/pro99_trace.pdf">Print</a> </div> <table> <tbody> <tr> <td>Na<sub>2</sub>EDTA<sup> 1 )</sup></td> <td>0.58 g</td> </tr> <tr> <td>FeCl<sub>3</sub> · 6H<sub>2</sub>O </td> <td>0.422 g</td> </tr> <tr> <td>ZnSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>2.933 mg</td> </tr> <tr> <td>CoCl<sub>2</sub> · 6H<sub>2</sub>O </td> <td>1.332 mg</td> </tr> <tr> <td>MnCl<sub>2</sub> · 4H<sub>2</sub>O </td> <td>24 mg</td> </tr> <tr> <td>Na<sub>2</sub>MoO<sub>4</sub> · 2H<sub>2</sub>O </td> <td>0.839 mg</td> </tr> <tr> <td>Na<sub>2</sub>SeO<sub>3</sub></td> <td>2.306 mg</td> </tr> <tr> <td>NiCl<sub>2</sub> · 6H<sub>2</sub>O </td> <td>0.37 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> </tbody> </table> <p> 1 ) In the NIES–Collection, 0.58 g Na<sub>2</sub>EDTA is replaced by 0.637 g Na<sub>2</sub>EDTA · 2H<sub>2</sub>O. </p> <br> <p class="mediumCategoryTitle">5 Trace metals, vitamin mixtures and soil extracts</p> <h1><a name="soil_extract" id="af110"></a>Soil extract</h1> <div style="text-align: right;"> <a href="./medium/en/soil_extract.pdf">Print</a> </div> <p>To 1000 mL distilled water add 200 mL of soil ( soil from undisturbed deciduous woodland is best ) and heat by autoclaving for 1 h at 105°C. When cool, heat by autoclaving for 1 h at 105°C again. Pass the supernatant through a GF/C filter and Celite, and then pass the filtrate through a GF/F filter. Adjust to 1000 mL by adding distilled water. Dispense 10 mL of the final filtrate into each test tube and sterilize by autoclaving for 20min at 121°C. Keep in a cool place.</p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Provasoli, L., McLaughlin, J. J. A., Droop, M. R. 1957 The development of artificial media for marine algae. <i>Arch. Mikrobiol</i>., <b>25</b>, 392-428. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">5 Trace metals, vitamin mixtures and soil extracts</p> <h1> <a name="a5_co" id="a5_co"></a>Trace metal mix A<sub>5</sub> + Co </h1> <div style="text-align: right;"> <a href="./medium/en/a5_co.pdf">Print</a> </div> <table> <tbody> <tr> <td>H<sub>3</sub>BO<sub>3</sub></td> <td>286 mg</td> </tr> <tr> <td>MnCl<sub>2</sub> · 4H<sub>2</sub>O </td> <td>181 mg</td> </tr> <tr> <td>ZnSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>22.2 mg</td> </tr> <tr> <td>Na<sub>2</sub>MoO<sub>4</sub> · 2H<sub>2</sub>O </td> <td>39 mg</td> </tr> <tr> <td>CuSO<sub>4</sub> · 5H<sub>2</sub>O </td> <td>7.9 mg</td> </tr> <tr> <td>Co( NO<sub>3</sub> )<sub>2</sub>·6H<sub>2</sub>O </td> <td>4.9 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">5 Trace metals, vitamin mixtures and soil extracts</p> <h1> <a name="s3" id="s3"></a>Vitamin mix S<sub>3</sub> </h1> <div style="text-align: right;"> <a href="./medium/en/s3.pdf">Print</a> </div> <table> <tbody> <tr> <td>Thiamine HCl</td> <td>5 mg</td> </tr> <tr> <td>Nicotinic acid</td> <td>1 mg</td> </tr> <tr> <td>Calcium pantothenate</td> <td>1 mg</td> </tr> <tr> <td>p–Aminobenzoic acid</td> <td>0.1 mg</td> </tr> <tr> <td>Biotin</td> <td>0.01 mg</td> </tr> <tr> <td>Inositol</td> <td>50 mg</td> </tr> <tr> <td>Folic acid</td> <td>0.02 mg</td> </tr> <tr> <td>Thymine</td> <td>30 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> </tbody> </table> <br> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Provasoli, L. 1963 Growing marine seaweeds. In <i>Proceedings of the Fourth International Seaweed Symposium</i>, University of Tokyo Press, Tokyo, p. 9-17. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">5 Trace metals, vitamin mixtures and soil extracts</p> <h1><a name="w_edta" id="w_edta"></a>Waris EDTA</h1> <div style="text-align: right;"> <a href="./medium/en/w_edta.pdf">Print</a> </div> <table> <tbody> <tr> <td>EDTA</td> <td>0.522 g</td> </tr> <tr> <td>FeSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>0.498 g</td> </tr> <tr> <td>KOH (1 mol/L solution)</td> <td>5.4 mL</td> </tr> <tr> <td>Distilled water</td> <td>94.6 mL</td> </tr> </tbody> </table> <p> EDTA and FeSO<sub>4</sub> · 7H<sub>2</sub>O is heated for 30 min (100°C ); KOH is added to the cooled mixture.<br> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>McFadden, G. I., Melkonian, M. 1986 Use of Hepes buffer for microalgael culture media and fixation for eledtron microscopy. <i>Phycology</i>, <b>25</b>, 551-557. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">5 Trace metals, vitamin mixtures and soil extracts</p> <h1><a name="wp2" id="wp2"></a>Waris P-2</h1> <div style="text-align: right;"> <a href="./medium/en/wp2.pdf">Print</a> </div> <table> <tbody> <tr> <td>Na<sub>2</sub>EDTA· 2H<sub>2</sub>O </td> <td>0.3 g</td> </tr> <tr> <td>H<sub>3</sub>BO<sub>3</sub></td> <td>0.114 g</td> </tr> <tr> <td>MnCl<sub>2</sub>· 4H<sub>2</sub>O </td> <td>14.4 mg</td> </tr> <tr> <td>ZnSO<sub>4</sub>·7H<sub>2</sub>O </td> <td>2.1 mg</td> </tr> <tr> <td>CoCl<sub>2</sub>·6H<sub>2</sub>O </td> <td>0.4 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> </tbody> </table> <br> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>McFadden, G. I., Melkonian, M. 1986 Use of Hepes buffer for microalgael culture media and fixation for eledtron microscopy. <i>Phycology</i>, <b>25</b>, 551-557. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">5 Trace metals, vitamin mixtures and soil extracts</p> <h1><a name="wc_metals" id="wc_metals"></a>WC trace metals</h1> <div style="text-align: right;"> <a href="./medium/en/wc_metals.pdf">print</a> </div> <table> <tbody> <tr> <td>Na<sub>2</sub>EDTA· 2H<sub>2</sub>O</td> <td>436 mg</td> </tr> <tr> <td>FeCl<sub>3</sub> · 6H<sub>2</sub>O</td> <td>315 mg</td> </tr> <tr> <td>CuSO<sub>4</sub> · 5H<sub>2</sub>O </td> <td>1 mg</td> </tr> <tr> <td>ZnSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>2.2 mg</td> </tr> <tr> <td>CoCl<sub>2</sub> · 6H<sub>2</sub>O </td> <td>1 mg</td> </tr> <tr> <td>MnCl<sub>2</sub> · 4H<sub>2</sub>O </td> <td>18 mg</td> </tr> <tr> <td>Na<sub>2</sub>MoO<sub>4</sub> · 2H<sub>2</sub>O </td> <td>0.6 mg</td> </tr> <tr> <td>H<sub>3</sub>BO<sub>3</sub></td> <td>100 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> </tbody> </table> <br> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Guillard, R. R. L., Lorenzen, C. J. 1972 Yellow-green algae with chlorohyllide C. <i>J. Phycol.</i>, <b>8</b>, 10-14. </td> </tr> <tr> <td>Andersen, R. A. E, Berges, J. A., Harrison, P. J., Watanabe, M. M. 2005 Appendix A — Recipes for freshwater and seawater media. In <i>Algal culturing techniques</i>, Ed. by Andersen, R. A. E, Elsevier Academic Press, p. 474. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">6 Media for protozoa</p> <h1><a name="af6_wheat" id="af6_wheat"></a>AF–6 + Wheat</h1> <div style="text-align: right;"> <a href="./medium/en/af6_wheat.pdf">Print</a> </div> <p> Beforehand, sterilize wheat grains by dry heating ( 150ºC, 30 min ). Keep in a cool place. For use, add a grain of sterile wheat to 10 mL <a href="#af6">AF–6</a><sup> 1 )</sup> medium.<br> 1 ) See <a href="#af6">AF–6</a> </p> <p class="mediumCategoryTitle">6 Media for protozoa</p> <h1><a name="bley" id="bley"></a>BLEY medium ( Barley leaf & Egg-yolk extracted liquid ) </h1> <div style="text-align: right;"> <a href="./medium/ja/bley.pdf">Print</a> </div> <p> <b>BL solution</b>: 1 pack ( 3 g ) of Barley leaf ( Trade name: Oomugi Wakaba Funmatu 100%, Yamamoto Kanpou Seiyaku Co., Ltd. ) is added to 1,000 mL distilled water and boiled for 10 minutes, while stirring. The solution is passed through the qualitative filter paper.<br> <b>EY solution</b>: 9 g of crushed yolk of hard-boiled egg is added to 1,000 mL distilled water and boiled for 10 minutes, while stirring. The solution is passed through the qualitative filter paper.<br> BL and EY Solution are mixed. The mixed solution is dilution with distilled water up to 6,000 mL, and then sterilized by autoclaving ( 121ºC, 15 min ). <br> </p> <p class="mediumCategoryTitle">6 Media for protozoa</p> <h1><a name="dgty" id="dgty"></a>dGTY</h1> <div style="text-align: right;"> <a href="./medium/en/dgty.pdf">Print</a> </div> <table> <tbody> <tr> <td>Glucose</td> <td>0.2 g</td> </tr> <tr> <td>Tryptone</td> <td>0.1 g</td> </tr> <tr> <td>Yeast extract</td> <td>0.05 g</td> </tr> <tr> <td>Seawater</td> <td>100 mL</td> </tr> </tbody> </table> <p class="mediumCategoryTitle">6 Media for protozoa</p> <h1><a name="esm_mtygm9_rice" id="esm_mtygm9_rice"></a>ESM + mTYGM–9 + Rice</h1> <div style="text-align: right;"> <a href="./medium/en/esm_mtygm9_rice.pdf">Print</a> </div> <p> Beforehand, sterilize polished rice by dry heating ( 150ºC, 30 min ). Keep in a cool place. For use, add 500 μL <a href="#mtygm9">mTYGM–9</a><sup> 1 )</sup> and a grain of sterile rice to 10 mL <a href="#esm">ESM</a><sup> 2 )</sup> medium. <br> 1 ) See <a href="#mtygm9">mTYGM–9</a> <br> 2 ) See <a href="#esm">ESM</a><br> </p> <p class="mediumCategoryTitle">6 Media for protozoa</p> <h1><a name="esm_rice" id="esm_rice"></a>ESM + Rice</h1> <div style="text-align: right;"> <a href="./medium/en/esm_rice.pdf">Print</a> </div> <p> Beforehand, sterilize polished rice by dry heating ( 150ºC, 30 min ). Keep in a cool place. For use, add a grain of sterile rice to 10 mL <a href="#esm">ESM</a><sup> 1 )</sup> medium. <br> 1 ) See <a href="#esm">ESM</a> </p> <br> <p class="mediumCategoryTitle">6 Media for protozoa</p> <h1><a name="f2_mtygm9_rice" id="f2_mtygm9_rice"></a>f/2 + mTYGM–9 + Rice</h1> <div style="text-align: right;"> <a href="./medium/en/f2_mtygm9_rice.pdf">Print</a> </div> <p> Beforehand, sterilize polished rice by dry heating ( 150ºC, 30 min ). Keep in a cool place. For use, add 500 μL <a href="#mtygm9">mTYGM–9</a><sup> 1 )</sup> and a grain of sterile rice to 10 mL <a href="#f2">f/2</a><sup> 2 )</sup> medium. <br> 1 ) See <a href="#mtygm9">mTYGM–9</a><br> 2 ) See <a href="#f2">f/2</a> </p> <br> <p class="mediumCategoryTitle">6 Media for protozoa</p> <h1><a name="f2_rice" id="f2_rice"></a>f/2 + Rice</h1> <div style="text-align: right;"> <a href="./medium/en/f2_rice.pdf">Print</a> </div> <p> Beforehand, sterilize polished rice by dry heating ( 150ºC, 30 min ). Keep in a cool place. For use, add a grain of sterile wheat to 10 mL <a href="#f2">f/2</a><sup> 1 )</sup> medium.<br> 1 ) See <a href="#f2">f/2</a><br> </p> <p class="mediumCategoryTitle">6 Media for protozoa</p> <h1><a name="f2_wheat" id="f2_wheat"></a>f/2 + Wheat</h1> <div style="text-align: right;"> <a href="./medium/ja/f2_wheat.pdf">Print</a> </div> <p> Beforehand, sterilize wheat grains by dry heating ( 150ºC, 30 min ). Keep in a cool place. For use, add a grain of sterile wheat to 10 mL <a href="#f2">f/2</a><sup> 1 )</sup> medium.<br> 1 ) See <a href="#f2">f/2</a> </p> <p class="mediumCategoryTitle">6 Media for protozoa</p> <h1><a name="helio" id="helio"></a>Helio</h1> <div style="text-align: right;"> <a href="./medium/en/helio.pdf">Print</a> </div> <table> <tbody> <tr> <td>Sodium acetate</td> <td>10 mg</td> </tr> <tr> <td>Polypeptone</td> <td>10 mg</td> </tr> <tr> <td>Tryptone</td> <td>20 mg</td> </tr> <tr> <td>Yeast extract</td> <td>20 mg</td> </tr> <tr> <td>CaCl<sub>2</sub> · 2H<sub>2</sub>O </td> <td>0.1 mg</td> </tr> <tr> <td>Sea water</td> <td>10 mL</td> </tr> <tr> <td>Distilled water</td> <td>90 mL</td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">6 Media for protozoa</p> <h1><a name="hemi" id="hemi"></a>Hemi</h1> <div style="text-align: right;"> <a href="./medium/ja/hemi.pdf">印刷</a> </div> <p> Into 98.9 mL distilled water 3.6 g powder medium of Daigo's Artificial Seawater SP ( Nihon Pharmaceutical Co., Ltd. ) and 0.1 mL LB broth medium ( BD ). After autoclaving, add aseptically 1 mL horse serum ( Sigma-Aldrich Co. LLC ).<br> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Tashyreva, D., Prokopchuk, G., Yabuki, A., Kaur, B., Faktorová, D., Votýpka, J., Kusaka, C., Fujikura, K., Shiratori, T., Ishida, K., Horák, A., Lukeš, J. 2018 Phylogeny and morphology of new diplonemids from Japan. <i>Protist</i>, <b>169</b>, 158-179. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">6 Media for protozoa</p> <h1><a name="le" id="le"></a>LE</h1> <div style="text-align: right;"> <a href="./medium/en/le.pdf">Print</a> </div> <p> <b>L solution</b>: White part of lettuce is dried at 90ºC for 16–18 h without scorching; 300 mg of the dried lettuce is added to 100 mL boiling water ( 9:1 distilled water to tap water ) and boiled for 30 min, while stirring. The supernatant is passed through cotton wool.<br> <b>E solution</b>: 300 mg of crushed yolk of hardboiled egg is added to 100 mL water ( 9:1 distilled water to tap water ) and boiled for 30 min, while stirring. The supernatant is passed through cotton wool. <br> Equal quantities of L and E solutions are mixed. The pH is adjusted to 6.8–7.0 with 1 mol / L NaOH, and 100 mL of the solution is dispensed into each 200–mL Erlenmeyer flask and sterilized by autoclaving ( 121ºC, 15 min ). <br> </p> <br> <p class="mediumCategoryTitle">6 Media for protozoa</p> <h1><a name="mizinko" id="mizinko"></a>Mizinko</h1> <div style="text-align: right;"> <a href="./medium/en/mizinko.pdf">Print</a> </div> <p> Put 10 - 20 of dried water fleas commercially available for aquarium fish into a test tube and add 10mL distilled water. </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Takahashi, Y., Yoshida, M., Inouye, I., Watanabe, M. M. 2014 <i>Diplophrys mutabilis</i> sp. nov., a new member of Labyrinthulomycetes from freshwater habitats. <i>Protist</i>, <b>165</b>, 50-65. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">6 Media for protozoa</p> <h1><a name="whs" id="whs"></a>Modified WarisH–Si</h1> <div style="text-align: right;"> <a href="./medium/en/1_2WarisH+Si.pdf">Print</a> </div> <table> <tbody> <tr> <td>HEPES</td> <td>11.915 mg</td> </tr> <tr> <td>KNO<sub>3</sub></td> <td>5 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>1 mg</td> </tr> <tr> <td>(NH<sub>4</sub>)<sub>2</sub>HPO<sub>4</sub></td> <td>1 mg</td> </tr> <tr> <td>Ca(NO<sub>3</sub>)<sub>2</sub> · 4H<sub>2</sub>O </td> <td>5 mg</td> </tr> <tr> <td><a href="#wp2">Waris P-2</a></td> <td>0.05 mL</td> </tr> <tr> <td><a href="#w_edta">Waris EDTA</a></td> <td>0.05 mL</td> </tr> <tr> <td>Vitamin B<sub>12</sub></td> <td>0.01 μg</td> </tr> <tr> <td>Biotin</td> <td>0.05 μg</td> </tr> <tr> <td>Thiamine HCl</td> <td>5 μg</td> </tr> <tr> <td>Niacinamide</td> <td>0.005 μg</td> </tr> <tr> <td>Soil extract<sup> 3 )</sup></td> <td>0.5 mL</td> </tr> <tr> <td>Na<sub>2</sub>SiO<sub>3</sub> · 9H<sub>2</sub>O </td> <td>2.842 mg</td> </tr> <tr> <td>Distilled water</td> <td>99.4 mL</td> </tr> <tr> <td colspan="2">pH 7.0</td> </tr> <tr> </tr> </tbody> </table> <p> Waris H is diluted half strength and added Na<sub>2</sub>SiO<sub>3</sub> · 9H<sub>2</sub>O. <br> 1 ) See <a href="#wp2">Waris P-2</a><br> 2 ) See <a href="#w_edta">Waris EDTA</a><br> 3 ) See <a href="#soil_extract">Soil extract</a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>McFadden, G. I., Melkonian, M. 1986 Use of Hepes buffer for microalgael culture media and fixation for eledtron microscopy. <i>Phycology</i>, <b>25</b>, 551-557. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">6 Media for protozoa</p> <h1><a name="pyg" id="pyg"></a>PYG</h1> <div style="text-align: right;"> <a href="./medium/en/pyg.pdf">Print</a> </div> <table> <tbody> <tr> <td>Proteose Peptone</td> <td>0.15 g</td> </tr> <tr> <td>Yeast extract</td> <td>0.1 g</td> </tr> <tr> <td>Glucose</td> <td>0.5 g</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> <tr> <td colspan="2">pH 7.0</td> </tr> </tbody> </table> <p>PPYG medium is diluted half strength and reduced proteose peptone.</p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Mori, K., Yomo, T., Kashiwagi, A. 2011 Single-cell isolation and cloning of tetrahymena thermophila cells with a fluorescence-activated cell sorter. <i>J. Eukaryotic Microbiol.</i>, <b>58</b>(1), 37-42. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">6 Media for protozoa</p> <h1><a name="suy" id="suy"></a>SUY</h1> <div style="text-align: right;"> <a href="./medium/en/suy.pdf">Print</a> </div> <p> Prepare as for 100 mL <a href="#uro">URO</a><sup> 1 )</sup> medium with seawater instead of distilled water. Add 10 mg yeast extract and 20 mg tryptone. Indicated as " URO–YT " in reference. <br> 1 ) See <a href="#uro">URO</a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Moriya, M., Nakayama, T., Inouye, I. 2000 Ultrastructure and 18S rDNA sequence analysis of <i>Wobblia lunata</i> gen. et sp. nov., a new heterotrophic flagellate ( Stramenopiles, <i>incertae sedis</i> ). <i>Protist</i>, <b>151</b>, 41-55. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">6 Media for protozoa</p> <h1><a name="suy1_10" id="suy1_10"></a>SUY 1/10</h1> <div style="text-align: right;"> <a href="./medium/en/suy1_10.pdf">Print</a> </div> <p> Prepare as for 100 mL <a href="#uro">URO</a><sup> 1 )</sup> medium with seawater instead of distilled water. Add 1 mg yeast extract and 2 mg tryptone. Indicated as " URO–1/10 YT " in reference. <br> 1 ) See <a href="#uro">URO</a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Moriya, M., Nakayama, T., Inouye, I. 2002 A new class of the Stramenopiles, Placididea Classis nova: description of <i>Placidia cafeteriopsis</i> gen. et sp. nov. <i>Protist</i>, <b>153</b>, 143-156. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">6 Media for protozoa</p> <h1><a name="suy1_10mt" id="suy1_10mt"></a>SUY 1/10 + mTYGM–9 + Rice</h1> <div style="text-align: right;"> <a href="./medium/en/suy1_10mt.pdf">Print</a> </div> <p> Beforehand, sterilize polished rice by dry heating ( 150ºC, 30 min ). Keep in a cool place. For use, add 500 μL <a href="#mtygm9">mTYGM–9</a><sup> 1 )</sup> and a grain of sterile rice to 10 mL <a href="#suy1_10">SUY 1/10</a><sup> 2 )</sup> medium. <br> 1 ) See <a href="#mtygm9">mTYGM–9</a><br> 2 ) See <a href="#suy1_10">SUY 1/10</a> </p> <br> <p class="mediumCategoryTitle">6 Media for protozoa</p> <h1><a name="suy1_10_wheat" id="suy1_10_wheat"></a>SUY 1/10 + Wheat</h1> <div style="text-align: right;"> <a href="./medium/en/suy1_10_wheat.pdf">Print</a> </div> <p> Beforehand, sterilize wheat grains by dry heating ( 150ºC, 30 min ). Keep in a cool place. For use, add a grain of sterile wheat to 10 mL <a href="#suy1_10">SUY 1/10</a><sup> 1 )</sup> medium. <br> 1 ) See <a href="#suy1_10">SUY 1/10</a> </p> <br> <p class="mediumCategoryTitle">6 Media for protozoa</p> <h1><a name="uro_wheat" id="uro_wheat"></a>URO + Wheat</h1> <div style="text-align: right;"> <a href="./medium/en/uro_wheat.pdf">Print</a> </div> <p> Beforehand, sterilize wheat grains by dry heating ( 150ºC, 30 min ). Keep in a cool place. For use, add a grain of sterile wheat to 10 mL <a href="#uro">URO</a><sup> 1 )</sup> medium. <br> 1 ) See <a href="#uro">URO</a> </p> <br> <p class="mediumCategoryTitle">6 Media for protozoa</p> <h1><a name="uro_h_wheat" id="uro_h_wheat"></a>URO–H + Wheat</h1> <div style="text-align: right;"> <a href="./medium/en/uro_h_wheat.pdf">Print</a> </div> <p> Beforehand, sterilize wheat grains by dry heating ( 150ºC, 30 min ). Keep in a cool place. For use, add a grain of sterile wheat to 10 mL <a href="#uro_h">URO–H</a><sup> 1 )</sup> medium.<br> 1 ) See <a href="#uro_h">URO–H</a> </p> <br> <p class="mediumCategoryTitle">6 Media for protozoa</p> <h1><a name="uro_yt_wheat" id="uro_yt_wheat"></a>URO + YT (1/10) + Wheat</h1> <div style="text-align: right;"> <a href="./medium/ja/uro_yt_wheat.pdf">印刷</a> </div> <p> Prepare as for 100 mL <a href="#uro">URO</a><sup> 1 )</sup> medium. Adjust to pH 7.5 with 0.1 mol / L HCl,　and add 10 mg yeast extract and 20 mg tryptone (URO + YT (1/10) medium). Beforehand, sterilize wheat grains by dry heating ( 150ºC, 30 min ). Keep in a cool place. For use, add a grain of sterile wheat to 10 mL URO + YT (1/10) medium. <br> 1 ) See <a href="#uro">URO</a> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Yabuki, A., Nakayama, T., Yubuki, N., Hashimoto, T., Ishida, K-I. Inagaki, Y. 2011 <i></i>Tsukubamonas globosa</i> n. gen., n. sp., a novel excavate flagellate possibly holding a key for the early evolution in "Discoba". <i>J. Eukaryot. Microbiol.</i>, <b>58</b>, 319-331. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">6 Media for protozoa</p> <h1><a name="uyts_rice" id="uyts_rice"></a>UYTS + Rice</h1> <div style="text-align: right;"> <a href="./medium/en/uyts_rice.pdf">Print</a> </div> <p> Prepare as for 100 mL <a href="#uro">URO</a><sup> 1 )</sup> medium with 99.2 mL instead of 99.9 mL distilled water. Adjust to pH 7.5 with 0.1 mol / L HCl, and add 10 mg yeast extract, 20 mg tryptone and 0.3 ml horse serum ( UYTS medium ). Beforehand, sterilize polished rice by dry heating ( 150ºC, 30 min ). Keep in a cool place. For use, add a grain of sterile rice to 10 mL UYTS medium. <br> 1 ) See <a href="#uro">URO</a> </p> <br> <p class="mediumCategoryTitle">6 Media for protozoa</p> <h1><span class="mediumCategoryTitle"><a name="ye100ppm" id="ye100ppm"></a></span>YE100ppm</h1> <div style="text-align: right;"> <a href="./medium/en/ye100ppm.pdf">Print</a> </div> <p> Into 100 mL distilled water 9 mg powder medium of Daigo's Artificial Seawater SP <br> ( Nihon Pharmaceutical Co., Ltd. ) and 10 mg yeast extract. </p> <br> <p class="mediumCategoryTitle">7 Media for freshwater red algae</p> <h1><a name="bold3n" id="bold3n"></a>Bold 3N</h1> <div style="text-align: right;"> <a href="./medium/en/bold3n.pdf">Print</a> </div> <table> <tbody> <tr> <td>NaNO<sub>3</sub></td> <td>75 mg</td> </tr> <tr> <td>CaCl<sub>2</sub> · 2H<sub>2</sub>O </td> <td>2.5 mg</td> </tr> <tr> <td>MgSO<sub>4</sub> · 7H<sub>2</sub>O </td> <td>7.5 mg</td> </tr> <tr> <td>K<sub>2</sub>HPO<sub>4</sub></td> <td>7.5 mg</td> </tr> <tr> <td>KH<sub>2</sub>PO<sub>4</sub><sup> 1 )</sup></td> <td>17.5 mg</td> </tr> <tr> <td>NaCl</td> <td>2.5 mg</td> </tr> <tr> <td>Vitamin B<sub>12</sub><sup> 2 )</sup></td> <td>0. 015 μg</td> </tr> <tr> <td><a href="#p_iv">PIV metals</a><sup> 3 )</sup></td> <td>0.6 mL</td> </tr> <tr> <td><a href="#soil_extract">Soil extract</a><sup> 4 )</sup></td> <td>4 mL</td> </tr> <tr> <td>Distilled water</td> <td>96.4 mL</td> </tr> </tbody> </table> <p> 1 ) In the NIES–Collection, the amount of KH<sub>2</sub>PO<sub>4</sub> is reduced from 17.5 mg to 10.5 mg.<br> 2 ) In the NIES–Collection, the amount of Vitamin B<sub>12</sub> is increased from 0.015 μg to 0.02 μg. <br> 3 ) See <a href="#p_iv">PIV metals</a> <br> 4 ) See <a href="#soil_extract">Soil extract</a> </p> <br> <p class="mediumCategoryTitle">8 Media for Charales</p> <h1><a name="mswc2" id="mswc2"></a>mSWC–2 ( Modified SWC–2 )</h1> <div style="text-align: right;"> <a href="./medium/en/mswc2.pdf">Print</a> </div> <p> Put leaf mould into a glass vessel to make a thin bottom layer, and add river sand onto the bottom layer up to one–quarter to one–fifth from the bottom. Add a pinch of garden lime to the river sand before use. Dampen the soil with deionized water ( or distilled water ). Cover the glass vessel with a plastic cap or aluminum foil, and autoclave it twice with overnight rest in between ( 121ºC, 20 min ). After cooling the mixture to room temperature, pour sterilized deionized water ( or sterilized distilled water ) into it carefully ( so as not to disturb the soil ). In the case of brackish water strains, deionized water is replaced by about one–third–diluted Herbst artificial seawater ( <a href="#1_3asw">1/3 Herbst ASW</a> ). <br> </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Sakayama, H., Hara Y., Nozaki, H. 2004 Taxonomic re-examination of six species of <i>Nitella</i> ( Charales, Charophyceae ) from Asia, and phylogenetic relationships within the genus based on <i>rbc</i>L and <i>atp</i>B gene sequences. <i>Phycologia</i>, <b>43</b>, 91-104. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">8 Media for Charales</p> <h1><a name="swc1" id="swc1"></a>SWC–1</h1> <div style="text-align: right;"> <a href="./medium/en/swc1.pdf">Print</a> </div> <p> Put leaf mould into a glass vessel to make a thin bottom layer, and add black soil onto the bottom layer up to one-quarter to one–fifth from the bottom. Dampen the soil with deionized water ( or distilled water ). Cover the glass vessel with a plastic cap or aluminum foil, and autoclave it twice with overnight rest in between ( 121ºC, 20 min ). After cooling the mixture to room temperature, pour sterilized deionized water ( or sterilized distilled water ) into it carefully ( so as not to disturb the soil ). </p> <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Sakayama, H., Hara Y., Nozaki, H. 2004 Taxonomic re-examination of six species of <em>Nitella</em> ( Charales, Charophyceae ) from Asia, and phylogenetic relationships within the genus based on <em>rbc</em>L and <em>atp</em>B gene sequences. <em>Phycologia</em>, <strong>43</strong>, 91-104. </td> </tr> </tbody> </table> <br> <p class="mediumCategoryTitle">8 Media for Charales</p> <h1><a name="swcn1" id="swcn1"></a>SWCN–1</h1> <div style="text-align: right;"> <a href="./medium/en/swcn1.pdf">Print</a> </div> <p> Put bottom mud from a paddy field into a glass vessel up to one–quarter to one–fifth from the bottom. Dampen the mud with deionized water ( or distilled water ). Cover the glass vessel or jar with a plastic cap or aluminum foil, and autoclave it twice with overnight rest in between ( 121ºC, 20 min ). After cooling the mixture to room temperature, pour sterilized deionized water ( or sterilized distilled water ) into it carefully ( so as not to disturb the soil ). <br> </p> <p class="mediumCategoryTitle">8 Media for Charales</p> <h1><a name="swcn2" id="swcn2"></a>SWCN–2</h1> <div style="text-align: right;"> <a href="./medium/en/swcn2.pdf">Print</a> </div> <p> Put leaf mould into a glass vessel to make a thin bottom layer, and add a mixture of black soil and river sand onto the bottom layer up to one–quarter to one–fifth from the bottom. Dampen the soil with deionized water ( or distilled water ). Cover the glass vessel or jar with a plastic cap or aluminum foil, and autoclave it twice with overnight rest in between ( 121ºC, 20 min ). After cooling the mixture to room temperature, pour sterilized deionized water ( or sterilized distilled water ) into it carefully ( so as not to disturb the soil ). In the case of brackish water strains, deionized water is replaced by about one–third–diluted Herbst artificial seawater ( <a href="#1_3asw">1/3 Herbst ASW</a> ). </p> <p class="mediumCategoryTitle">8 Media for Charales</p> <h1><a name="swcn3" id="swcn3"></a>SWCN–3</h1> <div style="text-align: right;"> <a href="./medium/en/swcn3.pdf">Print</a> </div> <p>Put a mixture of black soil and bottom mud from a paddy field into a glass vessel up to one–quarter to one–fifth from the bottom. Dampen the soil with deionized water ( or distilled water ). Cover the glass vessel or jar with a plastic cap or aluminum foil, and autoclave it twice with overnight rest in between ( 121ºC, 20 min ). After cooling the mixture to room temperature, pour sterilized deionized water ( or sterilized distilled water ) into it carefully ( so as not to disturb the soil ).</p> <p class="mediumCategoryTitle">8 Media for Charales</p> <h1><a name="swcn4" id="swcn4"></a>SWCN–4</h1> <div style="text-align: right;"> <a href="./medium/en/swcn4.pdf">Print</a> </div> <p>Put a mixture of river sand and bottom mud from a paddy field into a glass vessel up to one–quarter to one–fifth from the bottom. Dampen the soil with deionized water ( or distilled water ). Cover the glass vessel or jar with a plastic cap or aluminum foil, and autoclave it twice with overnight rest in between ( 121ºC, 20 min ). After cooling the mixture to room temperature, pour sterilized deionized water ( or sterilized distilled water ) into it carefully ( so as not to disturb the soil ).</p> <p class="mediumCategoryTitle">8 Media for Charales</p> <h1><a name="1_3asw" id="1_3asw"></a>1/3 Herbst ASW</h1> <div style="text-align: right;"> <a href="./medium/en/1_3asw.pdf">Print</a> </div> <table> <tbody> <tr> <td>NaCl</td> <td>3.0 mg</td> </tr> <tr> <td>KCl<sup> 1 )</sup></td> <td>81.4 mg</td> </tr> <tr> <td>CaCl<sub>2</sub><sup> 1 )</sup></td> <td>132 mg</td> </tr> <tr> <td>MgSO<sub>4</sub><sup> 1 )</sup></td> <td>660 mg</td> </tr> <tr> <td>NaHCO<sub>3</sub><sup> 1 )</sup></td> <td>504 mg</td> </tr> <tr> <td>Distilled water</td> <td>100 mL</td> </tr> </tbody> </table> <p> 1 ) In the NIES–Collection, the amount of KCl is reduced from 81.4 mg to 80.0 mg, 132 mg CaCl<sub>2</sub> is replaced by 172 mg CaCl<sub>2</sub> · 2H<sub>2</sub>O, 660 mg MgSO<sub>4</sub> is replaced by 1.35 g MgSO<sub>4</sub> · 7H<sub>2</sub>O, and the amount of NaHCO<sub>3</sub> is reduced from 504 mg to 49.5 mg. <table> <tbody> <tr> <th>Reference(s)</th> </tr> <tr> <td>Okazaki, Y., Shimmen, T., Tazawa, M. 1984 Turgor regulation in a brackish charophyte, <i>Lamprothamnium succinctum</i> I. Artificial modification of intracellular osmotic pressure. <i>Plant Cell Physiol</i>., <b>25</b>, 565-571. </td> </tr> </tbody> </table> </section>   <div id="other_parts"> <hr> <div id="contents_policy_wrapper"></div> </div>  </section>  </div>  <div id="page-top" class="page_back"> <a href="#contents_wrapper"><img src="images/pagetop.png" alt="page-top" border="0" /></a> </div>  <div id="footer_wrapper"></div>  </body> </html>

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