Takezawa D, Liu Z H, An G, Poovaiah B W
Department of Horticulture, Washington State University, Pullman 99164, USA.
Plant Mol Biol. 1995 Feb;27(4):693-703. doi: 10.1007/BF00020223.
Eight genomic clones of potato calmodulin (PCM1 to 8) were isolated and characterized. Sequence comparisons of different genes revealed that the deduced amino acid sequence of PCM1 had several unique substitutions, especially in the fourth Ca(2+)-binding area. The expression patterns of different genes were studied by northern analysis using the 3'-untranslated regions as probes. The expression of PCM1, 5, and 8 was highest in the stolon tip and it decreased during tuber development. The expression of PCM6 did not vary much in the tissues tested, except in the leaves, where the expression was lower; whereas, the expression of PCM4 was very low in all the tissues. The expression of PCM2 and PCM3 was not detected in any of the tissues tested. Among these genes, only PCM1 showed increased expression following touch stimulation. To study the regulation of PCM1, transgenic potato plants carrying the PCM1 promoter fused to the beta-glucuronidase (GUS) reporter gene were produced. GUS expression was found to be developmentally regulated and touch-responsive, indicating a positive correlation between the expression of PCM1 and GUS mRNAs. These results suggest that the 5'-flanking region of PCM1 controls developmental and touch-induced expression. X-Gluc staining patterns revealed that GUS localization is high in meristematic tissues such as the stem apex, stolon tip, and vascular regions.
分离并鉴定了8个马铃薯钙调蛋白的基因组克隆(PCM1至8)。不同基因的序列比较显示,PCM1推导的氨基酸序列有几个独特的替换,特别是在第四个钙离子结合区域。使用3'非翻译区作为探针,通过Northern分析研究了不同基因的表达模式。PCM1、5和8在匍匐茎顶端的表达最高,在块茎发育过程中表达下降。PCM6在测试的组织中表达变化不大,除了在叶片中表达较低;而PCM4在所有组织中的表达都非常低。在任何测试组织中均未检测到PCM2和PCM3的表达。在这些基因中,只有PCM1在触摸刺激后表达增加。为了研究PCM1的调控,构建了携带与β-葡萄糖醛酸酶(GUS)报告基因融合的PCM1启动子的转基因马铃薯植株。发现GUS表达受发育调控且对触摸有反应,表明PCM1和GUS mRNA的表达呈正相关。这些结果表明,PCM1的5'侧翼区域控制发育和触摸诱导的表达。X-Gluc染色模式显示,GUS在分生组织如茎尖、匍匐茎顶端和维管区域中的定位较高。