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为研究蝴蝶兰Rubisco活化酶(RCA)基因对低温胁迫的响应机制,以蝴蝶兰“满天红”为材料,分析了蝴蝶兰Rubisco活化酶基因PhRCAβ的全长序列(Gen Bank登录号为KU954548)及其表达特性。结果表明,PhRCAβ基因全长1 695 bp,编码440个氨基酸;其编码的蛋白具有RCA的特异位点,含叶绿体转运肽,定位于叶绿体中,其成熟蛋白的分子量为42.52 k D,等电点为蛋白5.54,属于RCA小亚基基因;该基因在绿色组织中转录表达;13℃/8℃(昼/夜)的低温胁迫抑制Ph RCAβ基因的转录表达,并随着低温胁迫时间的延长,PhRCAβ基因的表达水平逐渐降低,在温度恢复正常时其表达水平升高;4℃低温条件下,Ph RCAβ基因的表达水平随着低温处理时间的延长逐渐降低,并一直维持在较低水平。由此推测,PhRCAβ直接参与了蝴蝶兰叶片光合作用的调控。
In order to study the mechanism of Rubisco activating enzyme (RCA) gene response to low temperature stress, the full-length sequence of the gene Phosphatidylinositol Rubisco Activase (PhRCAβ) from Phalaenopsis “Mantianhong” KU954548) and its expression characteristics. The results showed that the full-length PhRCAβ gene was 1 695 bp encoding a protein of 440 amino acids. The encoded protein had a specific RCA site and contained a chloroplast transit peptide, which located in the chloroplast. The molecular weight of the mature protein was 42.52 kD. Was 5.54 and belonged to RCA small subunit gene. The gene was transcribed in green tissue. Under low temperature stress at 13 ℃ / 8 ℃ (day / night), the transcriptional expression of Ph RCAβ gene was inhibited. With the prolongation of low temperature stress, The expression level of PhRCAβgene gradually decreased when the temperature returned to normal, and the expression level of Ph RCAβgene decreased gradually with the prolongation of low temperature treatment at 4 ℃, and remained at a low level. It is speculated that PhRCAβ is directly involved in the regulation of photosynthesis in Phalaenopsis.