Fiebig A, Mayfield J A, Miley N L, Chau S, Fischer R L, Preuss D
Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, Illinois 60637, USA.
Plant Cell. 2000 Oct;12(10):2001-8. doi: 10.1105/tpc.12.10.2001.
Very long chain lipids contribute to the hydrophobic cuticle on the surface of all land plants and are an essential component of the extracellular pollen coat in the Brassicaceae. Mutations in Arabidopsis CER genes eliminate very long chain lipids from the cuticle surface and, in some cases, from the pollen coat. In Arabidopsis, the loss of pollen coat lipids can disrupt interactions with the stigma, inhibiting pollen hydration and causing sterility. We have positionally cloned CER6 and demonstrate that a wild-type copy complements the cer6-2 defect. In addition, we have identified a fertile, intragenic suppressor, cer6-2R, that partially restores pollen coat lipids but does not rescue the stem wax defect, suggesting an intriguing difference in the requirements for CER6 activity on stems and the pollen coat. Importantly, analysis of this suppressor demonstrates that low amounts of very long chain lipids are sufficient for pollen hydration and germination. The predicted CER6 amino acid sequence resembles that of fatty acid-condensing enzymes, consistent with its role in the production of epicuticular and pollen coat lipids >28 carbons long. DNA sequence analysis revealed the nature of the cer6-1, cer6-2, and cer6-2R mutations, and segregation analysis showed that CER6 is identical to CUT1, a cDNA previously mapped to a different chromosome arm. Instead, we have determined that a new gene, CER60, with a high degree of nucleotide and amino acid similarity to CER6, resides at the original CUT1 locus.
极长链脂质构成了所有陆地植物表面的疏水角质层,并且是十字花科植物细胞外花粉壁的重要组成部分。拟南芥CER基因的突变会消除角质层表面以及某些情况下花粉壁上的极长链脂质。在拟南芥中,花粉壁脂质的缺失会破坏与柱头的相互作用,抑制花粉水合作用并导致不育。我们通过定位克隆得到了CER6,并证明野生型拷贝能够弥补cer6 - 2的缺陷。此外,我们还鉴定出了一个可育的基因内抑制子cer6 - 2R,它能部分恢复花粉壁脂质,但不能挽救茎蜡缺陷,这表明CER6在茎和花粉壁上发挥作用的需求存在有趣的差异。重要的是,对这个抑制子的分析表明,少量的极长链脂质就足以支持花粉水合作用和萌发。预测的CER6氨基酸序列与脂肪酸缩合酶的序列相似,这与其在生成长度大于28个碳的表皮和花粉壁脂质中的作用一致。DNA序列分析揭示了cer6 - 1、cer6 - 2和cer6 - 2R突变的性质,分离分析表明CER6与CUT1相同,CUT1是一个先前被定位到不同染色体臂上的cDNA。相反,我们确定在原来的CUT1位点存在一个新基因CER60,它与CER6在核苷酸和氨基酸水平上具有高度相似性。