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Expression of a peroxisome proliferator-activated receptor gene (xPPARα) from Xenopus laevis in tobacco (Nicotiana tabacum) plants

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Abstract

In this work, we have genetically transformed tobacco (Nicotiana tabacum) plants with the peroxisome proliferator-activated receptor cDNA (xPPARα) from Xenopus laevis, which is a transcriptional factor involved in the peroxisomal proliferation and induction of fatty acid ß-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 ß-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 xPPARα protein, in leaves from line T. Taken together, these findings suggest that xPPARα 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 xPPARα.

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Abbreviations

ACOX:

Acyl-CoA oxidase

BSA:

Bovine serum albumin

ACDH:

Acyl-CoA dehydrogenase

CaMV:

Cauliflower mosaic virus

CAT:

Catalase

CTAB:

Hexadecyltrimethylammonium bromide

DAB:

3,3'-diaminobenzidine-HCl

DTT:

Dithiothreitol

EDTA:

Ethylenediaminetetraacetic acid

npt II :

Neomycin phosphotransferase II gene

MS:

Murashige-Skoog plant growth medium

MSF:

Phenylmethanesulphonyl

PVP:

Polyvinylpolypyrrolidone

xPPARα:

Xenopus laevis peroxisome proliferator-activated receptor α

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Acknowledgments

We are indebted to CONACYT for a PhD scholarship to A.G.N.M. We thank Dr. W. Wahli for the kind gift of the cDNA derived from the xPPARα gene from Xenopus laevis. We also thank UNESCO for a travel fellowship to A.G.N.M. The electron microscopy analysis were carried out at the Centre of Scientific Instrumentation of the University of Granada.

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Authors and Affiliations

  1. Departamento de Ingeniería Genética y, CINVESTAV Irapuato, Km 9.6, carretera Irapuato-León, Apartado postal 629, 36500, Irapuato, GTO, México

    Alejandro G. Nila & Miguel A. Gómez-Lim

  2. Biotecnología y Bioquímica, CINVESTAV Irapuato, Km 9.6, carretera Irapuato-León, Apartado postal 629, 36500, Irapuato, GTO, México

    Mercedes G. López

  3. Departamento de Bioquímica, Biología Celular y Molecular de Plantas, Estación Experimental del Zaidín, CSIC, Apartado 419, 18080, Granada, España

    Luisa M. Sandalio & Luis A. del Río

  4. Departamento de Agroecología y Protección Vegetal, Estación Experimental del Zaidín, CSIC, Apartado 419, 18080, Granada, España

    Manuel Gómez

Authors
  1. Alejandro G. Nila
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  2. Luisa M. Sandalio
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  3. Mercedes G. López
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  4. Manuel Gómez
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  5. Luis A. del Río
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  6. Miguel A. Gómez-Lim
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Correspondence to Miguel A. Gómez-Lim.

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Nila, A.G., Sandalio, L.M., López, M.G. et al. Expression of a peroxisome proliferator-activated receptor gene (xPPARα) from Xenopus laevis in tobacco (Nicotiana tabacum) plants. Planta 224, 569–581 (2006). https://doi.org/10.1007/s00425-006-0246-8

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