Jako C, Kumar A, Wei Y, Zou J, Barton D L, Giblin E M, Covello P S, Taylor D C
Seed Oil Biotechnology Group, National Research Council of Canada, Plant Biotechnology Institute, 110 Gymnasium Place, Saskatoon, Saskatchewan S7N 0W9, Canada.
Plant Physiol. 2001 Jun;126(2):861-74. doi: 10.1104/pp.126.2.861.
We recently reported the cloning and characterization of an Arabidopsis (ecotype Columbia) diacylglycerol acyltransferase cDNA (Zou et al., 1999) and found that in Arabidopsis mutant line AS11, an ethyl methanesulfonate-induced mutation at a locus on chromosome II designated as Tag1 consists of a 147-bp insertion in the DNA, which results in a repeat of the 81-bp exon 2 in the Tag1 cDNA. This insertion mutation is correlated with an altered seed fatty acid composition, reduced diacylglycerol acyltransferase (DGAT; EC 2.3.1.20) activity, reduced seed triacylglycerol content, and delayed seed development in the AS11 mutant. The effect of the insertion mutation on microsomal acyl-coenzyme A-dependent DGAT is examined with respect to DGAT activity and its substrate specificity in the AS11 mutant relative to wild type. We demonstrate that transformation of mutant AS11 with a single copy of the wild-type Tag1 DGAT cDNA can complement the fatty acid and reduced oil phenotype of mutant AS11. More importantly, we show for the first time that seed-specific over-expression of the DGAT cDNA in wild-type Arabidopsis enhances oil deposition and average seed weight, which are correlated with DGAT transcript levels. The DGAT activity in developing seed of transgenic lines was enhanced by 10% to 70%. Thus, the current study confirms the important role of DGAT in regulating the quantity of seed triacylglycerols and the sink size in developing seeds.
我们最近报道了拟南芥(生态型哥伦比亚)二酰基甘油酰基转移酶cDNA的克隆和特性分析(邹等人,1999年),并发现拟南芥突变系AS11中,在II号染色体上一个名为Tag1的位点发生的乙基甲磺酸诱导突变,是在DNA中插入了一段147bp的片段,这导致Tag1 cDNA中81bp的外显子2重复。这种插入突变与种子脂肪酸组成改变、二酰基甘油酰基转移酶(DGAT;EC 2.3.1.20)活性降低、种子三酰甘油含量降低以及AS11突变体种子发育延迟相关。相对于野生型,就DGAT活性及其底物特异性而言,研究了AS11突变体中插入突变对微粒体酰基辅酶A依赖性DGAT的影响。我们证明用单拷贝野生型Tag1 DGAT cDNA转化突变体AS11可以互补突变体AS11的脂肪酸和低油表型。更重要的是,我们首次表明在野生型拟南芥中种子特异性过表达DGAT cDNA可增强油脂沉积和平均种子重量,这与DGAT转录水平相关。转基因系发育种子中的DGAT活性提高了10%至70%。因此,当前研究证实了DGAT在调节种子三酰甘油数量和发育种子的库大小方面的重要作用。