De Marchis Francesca, Valeri Maria Cristina, Pompa Andrea, Bouveret Emmanuelle, Alagna Fiammetta, Grisan Simone, Stanzione Vitale, Mariotti Roberto, Cultrera Nicolò, Baldoni Luciana, Bellucci Michele
Institute of Biosciences and Bioresources (IBBR), Research Division of Perugia, CNR, Via Madonna Alta 130, 06128, Perugia, Italy.
PlantLab, Institute of Life Sciences, Scuola Superiore Sant'Anna, 56127, Pisa, Italy.
Transgenic Res. 2016 Feb;25(1):45-61. doi: 10.1007/s11248-015-9919-z. Epub 2015 Nov 11.
Taking into account that fatty acid (FA) biosynthesis plays a crucial role in lipid accumulation in olive (Olea europaea L.) mesocarp, we investigated the effect of olive acyl carrier protein (ACP) on FA composition by overexpressing an olive ACP cDNA in tobacco plants. The OeACP1.1A cDNA was inserted in the nucleus or in the chloroplast DNA of different tobacco plants, resulting in extensive transcription of the transgenes. The transplastomic plants accumulated lower olive ACP levels in comparison to nuclear-transformed plants. Moreover, the phenotype of the former plants was characterized by pale green/white cotyledons with abnormal chloroplasts, delayed germination and reduced growth. We suggest that the transplastomic phenotype was likely caused by inefficient olive ACP mRNA translation in chloroplast stroma. Conversely, total lipids from leaves of nuclear transformants expressing high olive ACP levels showed a significant increase in oleic acid (18:1) and linolenic acid (18:3), and a concomitant significant reduction of hexadecadienoic acid (16:2) and hexadecatrienoic acid (16:3). This implies that in leaves of tobacco transformants, as likely in the mesocarp of olive fruit, olive ACP not only plays a general role in FA synthesis, but seems to be specifically involved in chain length regulation forwarding the elongation to C18 FAs and the subsequent desaturation to 18:1 and 18:3.
考虑到脂肪酸(FA)生物合成在油橄榄(Olea europaea L.)中果皮脂质积累中起关键作用,我们通过在烟草植株中过表达油橄榄酰基载体蛋白(ACP)cDNA来研究其对FA组成的影响。将OeACP1.1A cDNA插入不同烟草植株的细胞核或叶绿体DNA中,导致转基因大量转录。与核转化植株相比,转质体植株积累的油橄榄ACP水平较低。此外,前者植株的表型特征为子叶淡绿/白色、叶绿体异常、发芽延迟和生长受抑制。我们认为转质体表型可能是由于叶绿体基质中油橄榄ACP mRNA翻译效率低下所致。相反,高表达油橄榄ACP的核转化植株叶片中的总脂质显示,油酸(18:1)和亚麻酸(18:3)显著增加,同时十六碳二烯酸(16:2)和十六碳三烯酸(16:3)显著减少。这意味着在烟草转化植株的叶片中,就像在油橄榄果实的中果皮中一样,油橄榄ACP不仅在FA合成中起一般作用,而且似乎特别参与链长调节,促进向C18 FAs的延伸以及随后向18:1和18:3的去饱和。