Abstract

Anthocyanins are a group of secondary metabolites that colour fruit and flowers orange, red, purple or blue depending on a number of factors, such as the basic structure, co-pigmentation, metal ion complexation and vacuolar pH. The biosynthesis of anthocyanin is regulated at the transcriptional level by a group of transcription factors, the MYB–bHLH–WD40 (MBW) complex. In this study, the purple colouration in several kiwifruit (Actinidia) species was identified and characterised as red cyanidin-based and blue delphinidin-based anthocyanins. The differential pigmentation in the skin and flesh can be attributed to the differential ratio of cyanidin and delphinidin derivatives accumulated in the total anthocyanin profile. The expression of anthocyanin biosynthetic genes chalcone synthase (CHS), flavonoid 3-O-glucosyltransferase (F3GT), flavonoid 3′-hydroxylase (F3′H) and flavonoid 3′5′-hydroxylase (F3′5′H) is crucial for anthocyanin accumulation. However, the balance of expression of the F3′H and F3′5′H genes appears responsible for the ratio of cyanidin and delphinidin derivatives, while a lack of CHS, F3GT and MYB110 expression is responsible for a lack of total anthocyanins. The transcriptional regulation of the F3′H and F3′5′H promoters by the R2R3 MYB transcription factor MYB110 is markedly different in tobacco transient assays. When kiwifruit MYB10 or MYB110 are over-expressed in Actinidia chinensis both cyanidin-based and delphinidin-based anthocyanins are elevated, but F3′H and F3′5′H genes are not strongly correlated with MYB expression. These results suggest that the core kiwifruit anthocyanin pathway genes are dependent on characterised MYB transcription factors, while other regulatory proteins are more directly responsible for the expression of the F3′H and F3′5′H genes.

Highlights

  • Anthocyanins are flavonoids that belong to the polyphenols group

  • The results showed that MYB110 strongly activated certain flavonoid 3′ hydroxylase (F3′H) promoters, 33-fold higher in Me and 25-fold higher in arguta var. purpurea (Ap) (Fig. 7a)

  • The data presented here show that the key change during the ripening process of A. melanandra, A. arguta var. purpurea and the two macrosperma × A. melanandra (MaMe) kiwifruits is the increase in total anthocyanin concentration while the composition and the ratio of cyanidin to delphinidin remained the same between the colourchange stage and the ripe stage

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Summary

Introduction

Anthocyanins are flavonoids that belong to the polyphenols group. These secondary metabolites are synthesised in plants and are essential for plant growth and development[1]. In addition to colour pigmentation in plants, anthocyanin accumulation in leaves of plants and skin of fruits may absorb high light energy and scavenge free radicals to offer protection against photo-oxidative effects[3]. F3′H belongs to the CYP75B subfamily of the cytochrome P450-dependent monooxygenase (P450) superfamily and converts dihydrokaempferol to dihydroquercetin which is the precursor of cyanidin-based anthocyanin. F3′5′H belongs to the CYP75A subfamily of the P450 superfamily and converts dihydrokaempferol and dihydroquercetin to dihydromyricetin, which is the

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