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Increasing Phosphatidylinositol (4,5)-Bisphosphate Biosynthesis Affects Basal Signaling and Chloroplast Metabolism in Arabidopsis thaliana

机译:增加磷脂酰肌醇(4,5)-双磷酸酯的生物合成影响拟南芥中的基础信号和叶绿体代谢。

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One challenge in studying the second messenger inositol(1,4,5)-trisphosphate (InsP3) is that it is present in very low amounts and increases only transiently in response to stimuli. To identify events downstream of InsP3, we generated transgenic plants constitutively expressing the high specific activity, human phosphatidylinositol 4-phosphate 5-kinase Iα (HsPIPKIα). PIP5K is the enzyme that synthesizes phosphatidylinositol (4,5)-bisphosphate (PtdIns(4,5)P2); this reaction is flux limiting in InsP3 biosynthesis in plants. Plasma membranes from transgenic Arabidopsis expressing HsPIPKIα had 2–3 fold higher PIP5K specific activity, and basal InsP3 levels in seedlings and leaves were 2-fold higher than wild type. Although there was no significant difference in photosynthetic electron transport, HsPIPKIα plants had significantly higher starch (2–4 fold) and 20% higher anthocyanin compared to controls. Starch content was higher both during the day and at the end of dark period. In addition, transcripts of genes involved in starch metabolism such as SEX1 (glucan water dikinase) and SEX4 (phosphoglucan phosphatase), DBE (debranching enzyme), MEX1 (maltose transporter), APL3 (ADP-glucose pyrophosphorylase) and glucose-6-phosphate transporter (Glc6PT) were up-regulated in the HsPIPKIα plants. Our results reveal that increasing the phosphoinositide (PI) pathway affects chloroplast carbon metabolism and suggest that InsP3 is one component of an inter-organelle signaling network regulating chloroplast metabolism.
机译:研究第二种信使肌醇(1,4,5)-三磷酸酯(InsP 3 )的一个挑战是,它的含量非常低,并且仅在刺激后才短暂增加。为了鉴定InsP 3 下游的事件,我们生成了组成型表达高比活性的转基因植物,即人磷脂酰肌醇4-磷酸5-激酶Iα(HsPIPKIα)。 PIP5K是合成磷脂酰肌醇(4,5)-双磷酸(PtdIns(4,5)P 2 )的酶;该反应是植物中InsP 3 生物合成中通量的限制。表达HsPIPKIα的转基因拟南芥的质膜具有较高的PIP5K比活性2-3倍,而幼苗和叶片中基础InsP 3 的水平则比野生型高2倍以上。尽管光合电子传递没有显着差异,但与对照相比,HsPIPKIα植物的淀粉含量显着更高(2-4倍),花色苷含量高出20%。在白天和黑暗时期结束时,淀粉含量都较高。此外,涉及淀粉代谢的基因的转录物,例如SEX1(葡聚糖水二激酶)和SEX4(磷酸葡聚糖磷酸酶),DBE(去支化酶),MEX1(麦芽糖转运蛋白),APL3(ADP-葡萄糖焦磷酸化酶)和葡萄糖-6-磷酸酯。 HsPIPKIα植物中的转运蛋白(Glc6PT)被上调。我们的结果表明,增加磷酸肌醇(PI)途径会影响叶绿体的碳代谢,并表明InsP 3 是调控叶绿体代谢的细胞间信号网络的一个组成部分。

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