| Preparation |
M. truncatula, Jemalong A17, were sown on an agar medium which prepared in a 500ml sealed glass bottle. The medium contained 1mM potassium phosphate monobasic, 30uM boric acid, 11uM manganese(II) chloride, 1uM zinc chloride, 1mM potassium chloride, 1mM molybdic acid (sodium salt), 3.2uM cupric chloride, 84nM cobalt chloride, 1mM calcium chloride, 50uM Fe-ethylenediaminetetraacetic acid (EDTA), 1mM magnesium sulfate, 10mM 15N-ammonium nitrate (SI Science), 10mM 15N-potassium nitrate (SI Science), and 0.5% glucose. The plants were grown for five weeks under the following conditions: temperature, 25/25°C (day/night); day/night rhythm, 18h/6h. During cultivation, the air was exchanged daily. The shoot (aerial part) and root were separated with a scalpel and they were immediately frozen using liquid nitrogen. After lyophilization, the sample was stored at room temperature. |