Komatsu Setsuko, Yang Guangxiao, Khan Monowar, Onodera Haruko, Toki Seiichi, Yamaguchi Masayuki
National Institute of Crop Science, 2-1-18 Kannondai, Tsukuba 305-8518, Japan.
Mol Genet Genomics. 2007 Jun;277(6):713-23. doi: 10.1007/s00438-007-0220-6. Epub 2007 Feb 21.
Calcium is a ubiquitous signaling molecule and changes in cytosolic calcium concentration are involved in plant responses to various stimuli. The rice calcium-dependent protein kinase 13 (CDPK13) and calreticulin interacting protein 1 (CRTintP1) have previously been reported to be involved in cold stress response in rice. In this study, rice lines transformed with sense CDPK13 or CRTintP1 constructs were produced and used to investigate the function of these proteins. When the plants were incubated at 5 degrees C for 3 days, leaf blades of both the sense transgenic and vector control rice plants became wilted and curled. When the plants were transferred back to non-stress conditions after cold treatment, the leaf blades died, but the sheaths remained green in the sense transgenic rice plants. Expression of CDPK13 or CRTintP1 was further examined in several rice varieties including cold-tolerant rice varieties. Accumulation of these proteins in the cold-tolerant rice variety was higher than that in rice varieties that are intermediate in their cold tolerance. To examine whether over-expression of CDPK13 and CRTintP1 would have any effect on the proteins or not, sense transgenic rice plants were analyzed using proteomics. The 2D-PAGE profiles of proteins from the vector control were compared with those of the sense transgenic rice plants. Two of the proteins that differed between these lines were calreticulins. The results suggest that CDPK13, calreticulin and CRTintP1 might be important signaling components for response to cold stress in rice.
钙是一种普遍存在的信号分子,胞质钙浓度的变化参与植物对各种刺激的反应。先前有报道称,水稻钙依赖蛋白激酶13(CDPK13)和钙网蛋白相互作用蛋白1(CRTintP1)参与水稻的冷胁迫反应。在本研究中,构建了过表达CDPK13或CRTintP1的水稻株系,并用于研究这些蛋白的功能。当植株在5℃下培养3天时,过表达转基因水稻植株和载体对照水稻植株的叶片均出现萎蔫和卷曲。冷处理后将植株转移回非胁迫条件下,过表达转基因水稻植株的叶片死亡,但叶鞘仍保持绿色。在包括耐冷水稻品种在内的几个水稻品种中进一步检测了CDPK13或CRTintP1的表达。这些蛋白在耐冷水稻品种中的积累高于耐冷性中等的水稻品种。为了检测CDPK13和CRTintP1的过表达是否会对蛋白质产生影响,利用蛋白质组学对过表达转基因水稻植株进行了分析。将载体对照的蛋白质二维电泳图谱与过表达转基因水稻植株的图谱进行了比较。这两个株系之间差异的两种蛋白质是钙网蛋白。结果表明,CDPK13、钙网蛋白和CRTintP1可能是水稻响应冷胁迫的重要信号成分。