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Calmodulin Gene Family in Potato: Developmental and Touch-Induced Expression of the mRNA Encoding a Novel Isoform

机译:钙调蛋白基因家族在马铃薯中:编码新型同工型的mRNA的发育和触摸诱导表达。

摘要

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.
机译:分离并鉴定了八个马铃薯钙调蛋白基因组克隆(PCM1至8)。不同基因的序列比较显示,推导的PCM1氨基酸序列具有几个独特的取代,尤其是在第四个Ca(2+)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的表达在茎尖端最高,在块茎发育过程中下降。 PCM6的表达在受试组织中变化不大,但在叶子中表达较低。 PCM4在所有组织中的表达都非常低。在任何测试的组织中均未检测到PCM2和PCM3的表达。在这些基因中,只有PCM1在触摸刺激后显示出增加的表达。为了研究PCM1的调控,生产了带有与β-葡萄糖醛酸苷酶(GUS)报告基因融合的PCM1启动子的转基因马铃薯植株。发现GUS表达受到发育调节和触觉响应,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染色模式表明,在分生组织(例如茎尖,茎尖端和血管区域)中,GUS的定位较高。

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