Chen Grace Q, Turner Charlotta, He Xiaohua, Nguyen Tasha, McKeon Thomas A, Laudencia-Chingcuanco Debbie
Western Regional Research Center, Agricultural Research Service, U.S. Department of Agriculture, 800 Buchanan St., Albany, CA 94710, USA.
Lipids. 2007 Apr;42(3):263-74. doi: 10.1007/s11745-007-3022-z. Epub 2007 Feb 6.
Castor seed triacylglycerols (TAGs) contain 90% ricinoleate (12-hydroxy-oleate) which has numerous industrial applications. Due to the presence of the toxin ricin and potent allergenic 2S albumins in the seed, it is desirable to produce ricinoleate from temperate oilseeds. To identify regulatory genes or genes for enzymes that may up-regulate multiple activities or entire pathways leading to the ricinoleate and TAG synthesis, we have analyzed expression profiles of 12 castor genes involved in fatty acid and TAG synthesis using quantitative reverse transcription-polymerase chain reaction technology. A collection of castor seeds with well-defined developmental stages and morphologies was used to determine the levels of mRNA, ricinoleate and TAG. The synthesis of ricinoleate and TAG occurred when seeds progressed to stages of cellular endosperm development. Concomitantly, most of the genes increased their expression levels, but showed various temporal expression patterns and different maximum inductions ranging from 4- to 43,000-fold. Clustering analysis of the expression data indicated five gene groups with distinct temporal patterns. We identified genes involved in fatty acid biosynthesis and transport that fell into two related clusters with moderate flat-rise or concave-rise patterns, and others that were highly expressed during seed development that displayed either linear-rise or bell-shaped patterns. Castor diacylglycerol acyltransferase 1 was the only gene having a higher expression level in leaf and a declining pattern during cellular endosperm development. The relationships among gene expression, cellular endosperm development and ricinoleate/TAG accumulation are discussed.
蓖麻籽三酰甘油(TAGs)含有90%的蓖麻醇酸(12-羟基油酸),具有众多工业应用。由于种子中存在毒素蓖麻毒蛋白和强效变应原性2S白蛋白,因此期望从温带油籽中生产蓖麻醇酸。为了鉴定可能上调导致蓖麻醇酸和TAG合成的多种活性或整个途径的调控基因或酶基因,我们使用定量逆转录-聚合酶链反应技术分析了12个参与脂肪酸和TAG合成的蓖麻基因的表达谱。使用一组具有明确发育阶段和形态的蓖麻籽来确定mRNA、蓖麻醇酸和TAG的水平。当种子进入细胞胚乳发育阶段时,蓖麻醇酸和TAG的合成开始。与此同时,大多数基因的表达水平增加,但呈现出不同的时间表达模式和4至43000倍不等的不同最大诱导倍数。对表达数据的聚类分析表明有五个具有不同时间模式的基因组。我们鉴定出参与脂肪酸生物合成和转运的基因,它们分为两个相关的簇,具有中等的平缓上升或凹形上升模式,以及其他在种子发育过程中高表达的基因,呈现线性上升或钟形模式。蓖麻二酰甘油酰基转移酶1是唯一一个在叶片中表达水平较高且在细胞胚乳发育过程中呈下降模式的基因。文中讨论了基因表达、细胞胚乳发育与蓖麻醇酸/TAG积累之间的关系。