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Differential transcriptional regulation of l-ascorbic acid content in peel and pulp of citrus fruits during development and maturation

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Abstract

Citrus fruits are an important source of ascorbic acid (AsA) for human nutrition, but the main pathways involved in its biosynthesis and their regulation are still not fully characterized. To study the transcriptional regulation of AsA accumulation, expression levels of 13 genes involved in AsA biosynthesis, 5 in recycling and 5 in degradation were analyzed in peel and pulp of fruit of two varieties with different AsA concentration: Navel orange (Citrus sinensis) and Satsuma mandarin (Citrus unshiu). AsA accumulation in peel and pulp correlated with the transcriptional profiling of the l-galactose pathway genes, and the myo-inositol pathway appeared to be also relevant in the peel of immature-green orange. Differences in AsA content between varieties were associated with differential gene expression of GDP-mannose pyrophosphorylase (GMP), GDP-l-galactose phosphorylase (GGP) and l-galactose-1-phosphate phosphatase (GPP), myo-inositol oxygenase in peel, and GGP and GPP in pulp. Relative expressions of monodehydroascorbate reductase 3 (MDHAR3) and dehydroascorbate reductase1 (DHAR1) correlated with AsA accumulation during development and ripening in peel and pulp, respectively, and were more highly expressed in the variety with higher AsA contents. Collectively, results indicated a differential regulation of AsA concentration in peel and pulp of citrus fruits that may change during the different stages of fruit development. The l-galactose pathway appears to be predominant in both tissues, but AsA concentration is regulated by complex mechanisms in which degradation and recycling also play important roles.

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Abbreviations

AsA:

Ascorbic acid

AO:

Ascorbate oxidase

APX:

Ascorbate peroxidase

CP:

Crossing point

DHAR:

Dehydroascorbate reductase

GalLDH:

l-Galactono-1,4-lactone dehydrogenase

GalDH:

l-Galactose dehydrogenase

GalUR:

d-Galacturonic acid reductase

GGP:

GDP-l-Galactose phosphorylase

GLOase:

l-Gulono-1,4-lactone oxidase

GME:

GDP-Mannose-3′,5′-epimerase

GMP:

GDP-Mannose pyrophosphorylase

GPP:

l-Galactose-1-phosphate phosphatase

MDHAR:

Monodehydroascorbate reductase

MIOX:

Myo-inositol oxygenase

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Acknowledgments

This work was supported by research grants FP7-PEOPLE-2011-CIG-2011-303652 (Marie Curie Actions, European Union), Proyectos de I + D para Grupos de Investigación Emergentes GV/2012/036 (Generalitat Valenciana, Spain). Enriqueta Alós was recipient of a JAE-Doc (CSIC) post-doctoral contract which is co-funded by Fondo Social Europeo de Desarrollo Regional (FEDER). Financial support by PROMETEO2010/010 (Generalitat Valenciana, Spain), Fun-C-Food (CSD2007-0063C, Ministerio de Ciencia e Innovación, Spain) and AGL-2009-11558 and AGL-2012-34573 (Ministerio Economía y Competitividad, Spain) are also acknowledged. We thank Dr. Luis Navarro (IVIA, Moncada, Valencia) for the use of the Citrus Germplasm Bank and Dr. Jose Vicente Gil (Universitat de Valencia; IATA-CSIC) for allowing the use of the HPLC equipment. We also gratefully acknowledge the assistance of Arancha Gurrea, Joanna Lado and Amparo Beneyto.

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  1. Instituto de Agroquímica y Tecnología de Alimentos (IATA), Consejo Superior de Investigaciones Científicas (CSIC), Avda. Agustín Escardino 7, Paterna, 46980, Valencia, Spain

    Enriqueta Alós, María J. Rodrigo & Lorenzo Zacarías

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  1. Enriqueta Alós
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  2. María J. Rodrigo
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Correspondence to Lorenzo Zacarías.

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Supplementary material 1 (DOC 247 kb)

425_2014_2044_MOESM2_ESM.pptx

Supplementary Figure S1 Changes in fruit diameter, external color (a/b ratio) and maturity index of the juice (Total soluble solids/total acidity) in fruits of Satsuma Owari (white circles) and Washington Navel (black circles) during fruit development and ripening. Data of fruit diameter and external color are mean ± SD of 20 replicates, and those of juice maturity index are mean ± SD of 3 replicates. JN, JL, A, S, O, N, D indicate June, July, August, September, October, November and December, respectively. (PPTX 72 kb)

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Alós, E., Rodrigo, M.J. & Zacarías, L. Differential transcriptional regulation of l-ascorbic acid content in peel and pulp of citrus fruits during development and maturation. Planta 239, 1113–1128 (2014). https://doi.org/10.1007/s00425-014-2044-z

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