Enhanced production of resveratrol derivatives in tobacco plants by improving the metabolic flux of intermediates in the phenylpropanoid pathway

Abstract

The biosynthesis of flavonoids such as anthocyanin and stilbenes has attracted increasing attention because of their potential health benefits. Anthocyanins and stilbenes share common phenylpropanoid precursor pathways. We previously reported that the overexpression of sweetpotato IbMYB1a induced anthocyanin pigmentation in transgenic tobacco (Nicotiana tabacum) plants. In the present study, transgenic tobacco (Nicotiana tabacum SR1) plants (STS-OX and ROST-OX) expressing the RpSTS gene encoding stilbene synthase from rhubarb (Rheum palmatum L. cv. Jangyeop) and the RpSTS and VrROMT genes encoding resveratrol O-methyltransferase from frost grape (Vitis riparia) were generated under the control of 35S promoter. Phenotypic alterations in floral organs, such as a reduction in floral pigments and male sterility, were observed in STS-OX transgenic tobacco plants. However, we failed to obtain STS-OX and ROST-OX plants with high levels of resveratrol compounds. Therefore, to improve the production of resveratrol derivatives in plants, we cross-pollinated flowers of STS-OX or ROST-OX and IbMYB1a-OX transgenic lines (SM and RSM). Phenotypic changes in vegetative and reproductive development of SM and RSM plants were observed. Furthermore, by HPLC and LC-MS analyses, we found enhanced production of resveratrol derivatives such as piceid, piceid methyl ether, resveratrol methyl ether O-hexoside, and 5-methyl resveratrol-3,4'-O-β-D-diglucopyranoside in SM and RSM cross-pollinated lines. Here, total contents of trans- and cis-piceids ranged from approximately 104-240 µg/g fresh weight in SM (F2). Collectively, we suggest that coexpression of RpSTS and IbMYB1a via cross-pollination can induce enhanced production of resveratrol compounds in plants by increasing metabolic flux into stilbenoid biosynthesis.

Next-Generation Plant Metabolic Engineering, Inspired by an Ancient Chinese Irrigation System

“…The effects of manipulation of flux on engineering plant metabolites has been extensively studied in phenylpropanoid biosynthesis, because of the degree of the understanding on the pathways and their transcriptional regulators (Besseau et al, 2007;Fornalé et al, 2010;Vogt, 2010;Liu et al, 2011Liu et al, , 2015aLiu et al, , 2017aJeong et al, 2016;Li et al, 2016). Could flux control be applicable to engineering other metabolic pathways such as terpenoid or alkaloid production?…”

Induced extracellular production of viniferins in grapevine cell culture medium by elicitation with methyl jasmonate and stevioside

“…Resveratrol oligomers also have numerous impressive bioactivities, but few studies have been performed due to their trace amounts in natural resources (Shen et al 2009). Thus, there have been many efforts to enhance production of resveratrol and its derivatives in plants and microorganisms through metabolic engineering (Beekwilder et al 2006;Delaunois et al 2009;Donnez et al 2009;Jeong et al 2014;Jeong et al 2015;Jeong et al 2016;Nivelle et al 2017;Shin et al 2011;Yu et al 2006). Currently, plant cell suspension cultures are a promising approach for resveratrol production under controlled aseptic conditions.…”

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“…[56] Anthocyanin and stilbenes have potential health benefits, and both share common phenylpropanoid precursor pathway. [13] Anthocyanin also found accumulated in seedlings. [57] Hydroxycinnamic, flavonoids, and anthocyanins are added during finishing of Tempranillo wines to provide specific color and phenolic attributes to it.…”

“…[12] Anthocyanins are phenolic pigments show cell protective effects [8] and show cytotoxicity to MCF-7 tumor cells. [13] Pink colored pigments found in onion and leeks are also anticancer in nature. [14] Hypocrellin A is a well-known natural pigment and bioactive agent that is used to treat skin diseases.…”

“…[13] There are different sources of pigments as anthocyanin is produced in transgenic tobacco (Nicotiana tabacum) plants. [13] Nor-abietanoid is isolated from Perovskia atriplicifolia Benth, [15] while green pigment from intact garlic (Allium sativum L.) cloves [16] and tanshinones from Salvia miltiorrhiza, a diterpenoid pigments. [17] Bee pollen and honey is also good source of plant pigment.…”