Abstract

Red pitaya (Hylocereus polyrhizus) fruit is a high-value, functional food, containing a high level of betalains. Several genes potentially related to betalain biosynthesis, such as cytochrome P450-like (CytP450-like), have been identified in pitaya fruit, while their transcriptional regulation remains unclear. In this work, the potential involvement of a WRKY transcription factor, HpWRKY44, in regulating CytP450-like1 expression in pitaya fruit was examined. HpWRKY44, a member of the Group 1 WRKY family, contains two conserved WRKY motifs and is localized in the nucleus. HpWRKY44 also exhibits trans-activation ability. Gene expression analysis showed that the expression of HpCytP450-like1 and HpWRKY44 increased steadily during pitaya fruit coloration, which corresponded with the production of elevated betalain levels in the fruit. HpWRKY44 was also demonstrated to directly bind to and activate the HpCytP450-like1 promoter via the recognition of the W-box element present in the promoter. Collectively, our findings indicate that HpWRKY44 transcriptionally activates HpCytP450-like1, which perhaps, at least in part, contributes to betalain biosynthesis in pitaya fruit. The information provided in the current study provides novel insights into the regulatory network associated with betalain biosynthesis during pitaya fruit coloration.

Highlights

  • Betalains are naturally occurring, water-soluble pigments produced from amino acid L-tyrosine

  • DNA fragments, including an oligonucleotide harboring two W-box (TTGAC) motifs and the HpCytP450-like[1] promoter with 74 bp containing two consensus W-box motifs were biotin-labeled at the 5′ end

  • To the best of our knowledge, this is the first report of the involvement of WRKY TFs in regulating HpCytP450-like[1] in fruits

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Summary

Introduction

Water-soluble pigments produced from amino acid L-tyrosine. Betalains are widely used as economically important natural food colorants and functional foods.[6,7] understanding the regulatory mechanisms responsible for betalain biosynthesis could provide information to assist the development of biotechnological approaches aimed to improve betalain production in plants

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