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Light affects ascorbate content and ascorbate-related gene expression in tomato leaves more than in fruits

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Abstract

Little is known about the light regulation of vitamin C synthesis in fruits. In contrast, previous studies in leaves revealed that VTC2 (coding for GDP-l-galactose phosphorylase) was one of the key genes up-regulated by light in leaves. Our objective was to determine how the expression of ascorbate (AsA) synthesis genes in tomato (Solanum lycopersicum) was modified according to light irradiance in both leaves and fruits. Seven days of shading strongly decreased total ascorbate (reduced and oxidized form) content in leaves (50%) and to a lesser extent in fruits (10%). Among the last six steps of AsA biosynthesis, only two genes, VTC2 and GPP1 (one of the two unigenes coding for l-galactose-1-P phosphatase in tomato), were down-regulated by long-term shading in red ripe fruits, compared to seven genes regulated in leaves. This underlines that light affects AsA-related gene expression more in leaves than in ripening fruits. Moreover, this study reveals strong daily changes in transcript levels of enzymes of the AsA biosynthetic pathway in leaves (11 of the 12 studied genes showed significant changes in their expression pattern). Among those genes, we found that diurnal variation in transcript levels of VTC2 and GME1 correlated to leaf AsA content measured 8 h later. This study provides a new hypothesis on the role of GME1 in addition to VTC2 in light-regulated AsA biosynthesis.

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Abbreviations

AsA:

Ascorbate

DHA:

Dehydroascorbate

Fv/Fm:

Maximum quantum efficiency of PSII

GMP:

GDP-d-mannose pyrophosphorylase

GME:

GDP-d-mannose 3′,5′-epimerase

GGP:

GDP-l-galactose phosphorylase

GPP:

l-galactose-1-P phosphatase

l-GalDH:

l-galactose dehydrogenase

GLDH:

l-galactono-1,4-lactone dehydrogenase

PAR:

Photosynthetically active radiation

PSII:

Photosystem II

RT-QPCR:

Real-time quantitative polymerase chain reaction

T-AsA:

Total ascorbate (AsA + DHA)

UT:

Universal time

WVA106:

Tomato cultivar West Virginia 106

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Acknowledgments

We are grateful to the greenhouse experimental team and to Yolande Carretero for taking care of the plants. We thank Claude Courbet and Michel Pradier for fruit harvesting, Cécile Garchery and Louise Gilbert for technical support in Q-PCR analyses and Grazielle Faure, Emilie Rubio, Sylvie Sérino and Doriane Bancel for vitamin C and sugar assays. Finally, we thank Valentina Baldazzi for critical reading of the manuscript.

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Authors and Affiliations

  1. INRA UR 1115 Plantes et Systèmes de Culture Horticoles, Domaine St Paul, Site Agroparc, 84914, Avignon, France

    Capucine Massot, Michel Génard, Jean-Jacques Longuenesse & Hélène Gautier

  2. INRA UR 1052 Génétique et Amélioration des Fruits et Légumes, Domaine St Maurice, 84143, Montfavet, France

    Rebecca Stevens

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  1. Capucine Massot
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  2. Rebecca Stevens
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  3. Michel Génard
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  4. Jean-Jacques Longuenesse
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  5. Hélène Gautier
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Corresponding author

Correspondence to Hélène Gautier.

Electronic supplementary material

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The following materials are available in the online version of this article.

Supplemental Fig. S1 Photosynthetically active radiation measured during the two experiments

Supplemental Fig. S2 Relation between T-AsA content and Fv/Fm ratio in leaves in the diurnal experiment

Supplemental Fig. S3 Daily variations of T-AsA content in green mature and red ripe fruits

Supplemental Fig. S4 Relative abundances of AsA-related mRNAs in S. lycopersicum ‘WVa 106’ leaves during a sunny or a cloudy day at midday.

Supplemental Fig. S5 Correlation between relative VTC2 and GME1 mRNA abundance and T-AsA content 8 h later

Supplemental Table S1 PCR primers used to amplify specific regions of genes of interest

Supplementary material 1 (pdf 208 kb)

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Massot, C., Stevens, R., Génard, M. et al. Light affects ascorbate content and ascorbate-related gene expression in tomato leaves more than in fruits. Planta 235, 153–163 (2012). https://doi.org/10.1007/s00425-011-1493-x

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