Abstract

Durian (Durio zibethinus L.) is a major economic crop native to Southeast Asian countries, including Thailand. Accordingly, understanding durian fruit ripening is an important factor in its market worldwide, owing to the fact that it is a climacteric fruit with a strikingly limited shelf life. However, knowledge regarding the molecular regulation of durian fruit ripening is still limited. Herein, we focused on cytochrome P450, a large enzyme family that regulates many biosynthetic pathways of plant metabolites and phytohormones. Deep mining of the durian genome and transcriptome libraries led to the identification of all P450s that are potentially involved in durian fruit ripening. Gene expression validation by RT-qPCR showed a high correlation with the transcriptome libraries at five fruit ripening stages. In addition to aril-specific and ripening-associated expression patterns, putative P450s that are potentially involved in phytohormone metabolism were selected for further study. Accordingly, the expression of CYP72, CYP83, CYP88, CYP94, CYP707, and CYP714 was significantly modulated by external treatment with ripening regulators, suggesting possible crosstalk between phytohormones during the regulation of fruit ripening. Interestingly, the expression levels of CYP88, CYP94, and CYP707, which are possibly involved in gibberellin, jasmonic acid, and abscisic acid biosynthesis, respectively, were significantly different between fast- and slow-post-harvest ripening cultivars, strongly implying important roles of these hormones in fruit ripening. Taken together, these phytohormone-associated P450s are potentially considered additional molecular regulators controlling ripening processes, besides ethylene and auxin, and are economically important biological traits.

Highlights

  • Durian (Durio zibethinus L., see S1 Fig), well known as the king of fruit, is an important economic fruit crop in Southeast Asia

  • We found that the remarkable expression of P450-dependent monoxygenases (P450s) is potentially involved in the metabolism of phytohormones during the ripening stages of durian fruit, suggesting possible crosstalk between phytohormones that regulate fruit ripening

  • All durian P450s were classified and named based on the protein sequence homology [18]; families share 40% identity, and subfamilies share 55% identity when aligned with the officially classified P450s that are publicly provided on the Cytochrome P450 Homepage [17]

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Summary

Introduction

Durian (Durio zibethinus L., see S1 Fig), well known as the king of fruit, is an important economic fruit crop in Southeast Asia. In terms of global trade, Thailand has been the top durian exporter with 65.9% market share of the 2019 global production, comprising more than 1.47 billion USD for export value (https://www.tridge.com/, search in 2021) This climacteric fruit has a short post-harvest life, posing problems to extending the marketing period and export to distant markets [2]. Biosynthetic pathways and their regulatory mechanisms governing important traits, such as aril flesh softening, the synthesis of pigment and bioactive compounds, sugar metabolism, and phytohormone crosstalk, are still unknown in durian and should be further investigated. We found that the remarkable expression of P450s is potentially involved in the metabolism of phytohormones during the ripening stages of durian fruit, suggesting possible crosstalk between phytohormones that regulate fruit ripening

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