摘要： 为解析苹果钙调蛋白基因MdCaM在非生物胁迫过程中的功能。以秦冠苹果为材料，克隆了苹果MdCaM基因，并利用MEGA 5.0软件对其编码的蛋白进行了聚类分析，利用qPCR分析了MdCaM在釆后损伤、低氧、高温等胁迫条件下的表达模式。结果表明，不同物种间钙调蛋白具有较高的同源性，MdCaM与拟南芥的钙调蛋白 CaM₁基因关系较近；qPCR结果表明，MdCaM在损伤、低氧胁迫（0 O₂或3% O₂）后均呈先升高后恢复正常水平的表达模式，损伤后6 h达到峰值，低氧（0 O₂）处理1 h后达到峰值；高温（20℃/40℃）诱导后其表达量高于同期对照组，并在处理后12 h达到峰值，表明该基因可能参与了高温胁迫下的全程响应。因此推测，MdCaM基因可能在苹果适应损伤、高温、低氧等逆境胁迫过程中发挥重要作用，可能参与了苹果生长发育过程中的逆境调控。为后继研究MdCaM的功能和作用机制奠定基础。

关键词: 苹果, 采后逆境胁迫, 荧光定量PCR, MdCaM, 基因克隆

Abstract: In order to identify the function of MdCaM gene in apple under different abiotic stresses. MdCaM gene was cloned by RT-PCR from apple, and phylogenetic tree was clustered based on amino acid of it. Also, the relative expression levels of MdCaM gene was analyzed by Real-time quantitative PCR (qPCR) method under wound, low-oxygen, high-temperature stress during apple fruit storage. The results showed that Calmodulin gene had high homology in different species, and MdCaM gene had a closer relationship with Arabidopsis thaliana CaM₁. qPCR analysis showed that the transcription level of MdCaM was up-regulated in the early stage, then returned to the normal level after wound and low-oxygen (0 O₂ and 3%O₂) stress, the difference was wound stress peak at 6 h, while low-oxygen (0 O₂) stress peak at 1 h. Also, MdCaM was up-regulated under high-temperature (20℃ and 40℃) stress compared to control,and the expression level was the highest at 12 h under stress,which indicated that MdCaM gene might be involved in the whole process of high-temperature stress-responsive. These results suggested that MdCaM might function as an stress-responsive gene under wound,high-temperature,low-oxygen stresses during apple storage. This research provided a theoretical basis for further study of the function and molecular regulation mechanism of MdCaM.

Key words: Apple, Postharvest stress, Real-time quantitative PCR, MdCaM, Gene clone

中图分类号: 

• Q78:S661.03