Department of Plant Biology, Michigan State University, East Lansing, Michigan 48824, USA.
Plant Cell. 2009 Dec;21(12):3885-901. doi: 10.1105/tpc.109.071795. Epub 2009 Dec 29.
Triacylglycerol (TAG) biosynthesis is a principal metabolic pathway in most organisms, and TAG is the major form of carbon storage in many plant seeds. Acyl-CoA:diacylglycerol acyltransferase 1 (DGAT1) is the only acyltransferase enzyme that has been confirmed to contribute to TAG biosynthesis in Arabidopsis thaliana seeds. However, dgat1 null mutants display only a 20 to 40% decrease in seed oil content. To determine whether other enzymes contribute to TAG synthesis, candidate genes were expressed in TAG-deficient yeast, candidate mutants were crossed with the dgat1-1 mutant, and target genes were suppressed by RNA interference (RNAi). An in vivo role for phospholipid:diacylglycerol acyltransferase 1 (PDAT1; At5g13640) in TAG synthesis was revealed in this study. After failing to obtain double homozygous plants from crossing dgat1-1 and pdat1-2, further investigation showed that the dgat1-1 pdat1-2 double mutation resulted in sterile pollen that lacked visible oil bodies. RNAi silencing of PDAT1 in a dgat1-1 background or DGAT1 in pdat1-1 background resulted in 70 to 80% decreases in oil content per seed and in disruptions of embryo development. These results establish in vivo involvement of PDAT1 in TAG biosynthesis, rule out major contributions by other candidate enzymes, and indicate that PDAT1 and DGAT1 have overlapping functions that are essential for normal pollen and seed development of Arabidopsis.
三酰基甘油(TAG)的生物合成是大多数生物的主要代谢途径,而TAG 是许多植物种子中碳储存的主要形式。酰基辅酶 A:二酰基甘油酰基转移酶 1 (DGAT1)是唯一被证实参与拟南芥种子 TAG 生物合成的酰基转移酶。然而,dgat1 缺失突变体的种子油含量仅降低 20%至 40%。为了确定是否有其他酶参与 TAG 合成,候选基因在 TAG 缺陷型酵母中表达,候选突变体与 dgat1-1 突变体杂交,并通过 RNA 干扰 (RNAi) 抑制靶基因。本研究揭示了磷脂:二酰基甘油酰基转移酶 1 (PDAT1; At5g13640) 在 TAG 合成中的作用。在未能从 dgat1-1 和 pdat1-2 的杂交中获得双纯合子植物后,进一步的研究表明,dgat1-1 pdat1-2 双突变导致花粉不育,缺乏可见的油体。在 dgat1-1 背景下沉默 PDAT1 或在 pdat1-1 背景下沉默 DGAT1,导致种子中油含量降低 70%至 80%,并破坏胚胎发育。这些结果确立了 PDAT1 在 TAG 生物合成中的体内参与,排除了其他候选酶的主要贡献,并表明 PDAT1 和 DGAT1 具有重叠的功能,对于拟南芥正常花粉和种子发育是必不可少的。