IC4R012-Proteomic-2009-19201764

Project Title

Proteomic identification of small, copper-responsive proteins in germinating embryos of Oryza sativa

Although copper (Cu) is an essential micronutrient for plants and algae, excess Cu is toxic to most plants and can cause a wide range of deleterious effects. To investigate the response of rice (Oryza sativa) to Cu stress, a proteomic approach was used to analyse Cu stress-induced changes in the expression of low molecular-weight proteins in germinating rice seed embryos.

Rice (Oryza sativa L. ‘Wuyunjing’ No. 7) seeds were surface-sterilized with 5% (v/v) sodium hypochlorite(NaClO) for 15 min and thoroughly washed in distilled water. Seeds were germinated on moist filter paper. Twenty seeds were randomly placed on the filter paper in 90-mm Petri dishes, and 5 mL of freshly prepared CuSO4.5H2O solution (0–200 mM) was added. Each treatment was performed in triplicate. The seeds were put to germinate for 4–8 d in the dark at 258C with renewal of the Cu solution at 2-d intervals. Seeds were considered to have germinated when the shoots were longer than 2 mm. The radicle length was measured during the exposure period. Embryos, including newly formed shoots, were isolated from the seeds after 6 d for Cu concentration and protein analyses.

College of Life Sciences, Nanjing Agricultural University, Nanjing 210095, China
Asian Natural Environmental Science Center, The University of Tokyo, 1-1-8 Midori-cho, Nishitokyo, Tokyo 188-0002, Japan