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The Choline/Ethanolamine Kinase Family in Arabidopsis: Essential Role of CEK4 in Phospholipid Biosynthesis and Embryo Development

机译:拟南芥中的胆碱/乙醇胺激酶家族:CEK4在磷脂生物合成和胚胎发育中的重要作用

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Phospholipids are highly conserved and essential components of biological membranes. The major phospholipids, phosphatidylethanolamine and phosphatidylcholine (PtdCho), are synthesized by the transfer of the phosphoethanolamine or phosphocholine polar head group, respectively, to the diacylglycerol backbone. The metabolism of the polar head group characterizing each phospholipid class is poorly understood; thus, the biosynthetic pathway of major phospholipids remains elusive in Arabidopsis thaliana. The choline/ethanolamine kinase (CEK) family catalyzes the initial steps of phospholipid biosynthesis. Here, we analyzed the function of the four CEK family members present in Arabidopsis. Knocking out of CEK4 resulted in defective embryo development, which was complemented by transformation of genomic CEK4. Reciprocal genetic crossing suggested that CEK4 knockout causes embryonic lethality, and microscopy analysis of the aborted embryos revealed developmental arrest after the heart stage, with no defect being found in the pollen. CEK4 is preferentially expressed in the vasculature, organ boundaries, and mature embryos, and CEK4 was mainly localized to the plasma membrane. Overexpression of CEK4 in wild-type Arabidopsis increased the levels of PtdCho in seedlings and mature siliques and of major membrane lipids in seedlings and triacylglycerol in mature siliques. CEK4 may be the plasma membrane-localized isoform of the CEK family involved in the rate-limiting step of PtdCho biosynthesis and appears to be required for embryo development in Arabidopsis.
机译:磷脂是高度保守的生物膜必不可少的组成部分。主要的磷脂,磷脂酰乙醇胺和磷脂酰胆碱(PtdCho),是通过将磷酸乙醇胺或磷酸胆碱的极性头基团分别转移到二酰基甘油骨架上来合成的。表征每个磷脂类别的极性首基的代谢了解甚少;因此,拟南芥中主要磷脂的生物合成途径仍然难以捉摸。胆碱/乙醇胺激酶(CEK)家族催化磷脂生物合成的初始步骤。在这里,我们分析了拟南芥中四个CEK家族成员的功能。敲除CEK4会导致胚胎发育缺陷,并辅之以基因组CEK4转化。相互的遗传杂交表明,敲除CEK4会导致胚胎致死,对流产胚胎的显微镜分析表明,心脏阶段后发育停滞,但在花粉中未发现缺陷。 CEK4优先在脉管系统,器官边界和成熟胚胎中表达,而CEK4主要定位在质膜上。 CEK4在野生型拟南芥中的过表达增加了幼苗和成熟角果树中PtdCho的水平,以及成熟角果树中幼苗中的主要膜脂和三酰甘油。 CEK4可能是参与PtdCho生物合成限速步骤的CEK家族的质膜定位同工型,似乎是拟南芥中胚胎发育所必需的。

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