• ISSN 1008-505X
  • CN 11-3996/S

钙对苹果果实钙调蛋白含量和Ca2+ -ATPase活性及其基因表达的影响

Effects of calcium on content of calmodulin, activity of Ca2 +-ATPase and their gene expressions in apple (Malus pumila Mill.) fruits

  • 摘要: 研究不同浓度钙(0、1和10 mmo1/L CaCl2 ;5 mmo1/L EGTA)对苹果果实钙调蛋白(CaM)含量和Ca2+-ATPase活性及其基因表达的影响。利用同源克隆方法分离CaM和Ca2+-ATPase基因,采用荧光定量PCR方法研究它们表达特征。结果表明,果实切片外源补钙,可溶性Ca2+及CaM含量在高钙处理12 h达到高峰;高钙处理12 h质膜Ca2+-ATPase活性显著增加,与胞内CaM含量增加时间一致;高钙处理24 h液泡膜Ca2+-ATPase活性显著增加;随着质膜和液泡膜Ca2+-ATPase活性显著增加,可溶性Ca2+含量在加钙处理48 h显著下降。研究基因表达发现,加钙处理6 h苹果CaM基因的表达量显著增加;加钙处理12 h苹果Ca2+-ATPase基因的表达量显著增加,与CaM含量及质膜Ca2 +-ATPase活性变化一致。果实缺钙处理显著增加CaM基因表达量,而苹果Ca2 +-ATPase基因的表达量没有显著变化。上述研究表明,苹果补钙可以提高细胞内可溶性Ca2+和CaM含量以及CaM基因的表达量,有效启动钙信使系统;质膜及液泡膜Ca2+-ATPase是调控胞内Ca2+关键的酶,通过提高质膜及液泡膜Ca2+-ATPase的活性及Ca2+-ATPase基因的表达量,维持胞内Ca2+的稳态水平。

     

    Abstract: Effects of calcium on calmodulin (CaM) content, Ca2 +-ATPase activity and their gene expressions in apple (Malus pumila) fruits were investigated under the hydroponic condition with 0,1,10 mmo1/L CaCl2 and 5 mmo1/L EGTA. CaM and Ca2 +-ATPase gene were isolated by homologous cloning and their expression patterns were analyzed by realtime PCR. Addition of 10 mmo1/LCaCl2 to the culture solutions with 12 h exposure significantly increased soluble Ca2 + and CaM contents in apple fruits. With the CaM content increasing, plasma and tonoplast membrane Ca2 +-ATPase activities were enhanced after 12 and 24 h Ca exposure respectively,resulting in the decrease in the soluble Ca2 + content after 48 h Ca exposure. In molecular level, the abundances of CaM and Ca2 +- ATPase genes were up-regulated after 6 h and 12 h Ca exposure, respectively, in accordance with change of the CaM content and plasma membrane Ca2 +-ATPaseactivity. Addition of 5 mmo1/L EGTA to the culture solutions induced up-regulated the expression of the CaM gene, however, did not markedly change the expression of the Ca2 +-ATPase gene. It was concluded that external calcium might increase soluble Ca2 + and CaM contents and the expression of theCaM gene to stimulate an intracellular Ca2 +/calmodulin signaling system. Plasma and tonoplast membrane Ca2 +-ATPase are of major physiological importance to expel Ca2 + from eukaryotic cells and to maintain overall Ca2 + homoeostasis by enhancing their activities and gene expression.

     

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