Estación Experimental de Aula Dei (EEAD-CSIC), Avda. Montañana 1005, 50059 Zaragoza, Spain; Instituto de la Grasa (IG-CSIC), Campus Universidad Pablo de Olavide, Building 46, Ctra. Utrera km. 1, 41013 Seville, Spain.
Instituto de la Grasa (IG-CSIC), Campus Universidad Pablo de Olavide, Building 46, Ctra. Utrera km. 1, 41013 Seville, Spain.
Mol Plant. 2015 Nov 2;8(11):1599-611. doi: 10.1016/j.molp.2015.06.004. Epub 2015 Jun 14.
Plastidial ω-3 desaturase FAD7 is a major contributor to trienoic fatty acid biosynthesis in the leaves of Arabidopsis plants. However, the precise contribution of the other plastidial ω-3 desaturase, FAD8, is poorly understood. Fatty acid and lipid analysis of several ω-3 desaturase mutants, including two insertion lines of AtFAD7 and AtFAD8, showed that FAD8 partially compensated the disruption of the AtFAD7 gene at 22 °C, indicating that FAD8 was active at this growth temperature, contrasting to previous observations that circumscribed the FAD8 activity at low temperatures. Our data revealed that FAD8 had a higher selectivity for 18:2 acyl-lipid substrates and a higher preference for lipids other than galactolipids, particularly phosphatidylglycerol, at any of the temperatures studied. Differences in the mechanism controlling AtFAD7 and AtFAD8 gene expression at different temperatures were also detected. Confocal microscopy and biochemical analysis of FAD8-YFP over-expressing lines confirmed the chloroplast envelope localization of FAD8. Co-localization experiments suggested that FAD8 and FAD7 might be located in close vicinity in the envelope membrane. FAD8-YFP over-expressing lines showed a specific increase in 18:3 fatty acids at 22 °C. Together, these results indicate that the function of both plastidial ω-3 desaturases is coordinated in a non-redundant manner.
质体 ω-3 去饱和酶 FAD7 是拟南芥叶片中三烯脂肪酸生物合成的主要贡献者。然而,其他质体 ω-3 去饱和酶 FAD8 的精确贡献仍知之甚少。几种 ω-3 去饱和酶突变体的脂肪酸和脂质分析,包括 AtFAD7 和 AtFAD8 的两个插入系,表明 FAD8 在 22°C 时部分补偿了 AtFAD7 基因的破坏,表明 FAD8 在这个生长温度下是活跃的,与之前的观察结果形成对比,之前的观察结果将 FAD8 的活性局限在低温下。我们的数据表明,FAD8 对 18:2 酰基脂底物具有更高的选择性,并且对除半乳糖脂以外的脂质,特别是磷脂酰甘油,具有更高的偏好,在研究的任何温度下都是如此。在不同温度下控制 AtFAD7 和 AtFAD8 基因表达的机制也存在差异。共聚焦显微镜和 FAD8-YFP 过表达系的生化分析证实了 FAD8 的质体包膜定位。共定位实验表明,FAD8 和 FAD7 可能位于包膜膜的附近。FAD8-YFP 过表达系在 22°C 时表现出 18:3 脂肪酸的特异性增加。总之,这些结果表明两种质体 ω-3 去饱和酶的功能以非冗余的方式协调。