Nila Alejandro G, Sandalio Luisa M, López Mercedes G, Gómez Manuel, del Rio Luis A, Gómez-Lim Miguel A
Departamento de Ingeniera Genética y CINVESTAV Irapuato, Km. 9.6, carretera Irapuato-León, Apartado postal 629, 36500 Irapouato, GTP, Mexico.
Planta. 2006 Aug;224(3):569-81. doi: 10.1007/s00425-006-0246-8.
In this work, we have genetically transformed tobacco (Nicotiana tabacum) plants with the peroxisome proliferator-activated receptor cDNA (xPPARalpha) from Xenopus laevis, which is a transcriptional factor involved in the peroxisomal proliferation and induction of fatty acid beta-oxidation in animal cells. Several transgenic lines were generated and one representative line (T) from the R2 generation was selected for further studies. Analysis of free fatty acids revealed that unsaturated fatty acids such as C16:2 and C16:3 were deficient in line T, whereas saturated fatty acids like C16:0, C18:0, and C20:0 were more abundant than in non-transformed plants. Acyl-CoA oxidase (ACOX) activity was assayed as a marker enzyme of beta-oxidation in crude leaf extracts and it was found that in line T there was a threefold increase in enzyme activity. We also found that the peroxisome population was increased and that catalase (CAT) activity was induced by clofibrate, a known activator of xPPARalpha protein, in leaves from line T. Taken together, these findings suggest that xPPARalpha is functional in plants and that its expression in tobacco leads to changes in general lipid metabolism and peroxisomal proliferation as reported in animal cells. Furthermore, it indicates that there is an endogenous ligand in tobacco cells able to activate xPPARalpha.
在这项研究中,我们用非洲爪蟾的过氧化物酶体增殖物激活受体cDNA(xPPARalpha)对烟草(Nicotiana tabacum)植株进行了遗传转化,xPPARalpha是一种转录因子,参与动物细胞中的过氧化物酶体增殖和脂肪酸β氧化的诱导。我们获得了多个转基因株系,并从R2代中挑选了一个代表性株系(T)进行进一步研究。游离脂肪酸分析表明,T株系中C16:2和C16:3等不饱和脂肪酸含量不足,而C16:0、C18:0和C20:0等饱和脂肪酸比未转化植株更为丰富。我们测定了粗叶提取物中作为β氧化标记酶的酰基辅酶A氧化酶(ACOX)活性,发现T株系中的酶活性增加了三倍。我们还发现,T株系叶片中的过氧化物酶体数量增加,并且已知的xPPARalpha蛋白激活剂氯贝丁酯可诱导过氧化氢酶(CAT)活性。综上所述,这些发现表明xPPARalpha在植物中具有功能,并且其在烟草中的表达会导致如动物细胞中所报道的一般脂质代谢和过氧化物酶体增殖的变化。此外,这表明烟草细胞中存在能够激活xPPARalpha的内源性配体。