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. 2023 Jun 12;11(9):5166-5173.
doi: 10.1002/fsn3.3475. eCollection 2023 Sep.

Identification of two terpenoids that accumulate in Chinese water chestnut in response to fresh-cut processing

Hui Nie  1   2 , Yanghe Luo  1   2 , Shuangquan Huang  1 , Yuwei Mo  1 , Zhenli Huang  1 , Yuemei Liao  1 , Lirui Jiang  1 , Wen Cai  1 , Mubo Song  1
Affiliations

Identification of two terpenoids that accumulate in Chinese water chestnut in response to fresh-cut processing

Hui Nie et al. Food Sci Nutr. .

Abstract

As a form of vegetable in China, freshly cut corms of Chinese water chestnuts (Eleocharis dulcis) are well received by consumers. Few studies have investigated the metabolites present in fresh-cut E. dulcis, particularly during the storage stage. Two compounds, triterpenoids and apocarotenoids, were identified in fresh-cut E. dulcis during the late storage period using thin-layer chromatography (TLC), high-performance liquid chromatography (HPLC), and nuclear magnetic resonance (NMR) spectroscopy. The content of these two compounds gradually increased in the surface tissue of fresh-cut E. dulcis during storage. Moreover, the transcript levels of 10 genes involved in terpenoid backbone biosynthesis and five genes involved in carotenoid precursor biosynthesis were evaluated via quantitative real-time PCR (qRT-PCR). Expression of the rate-limiting enzyme-coding genes CwDXS and CwHMGS was significantly induced by wounding. CwMYC and CwbHLH18, which belong to bHLH transcription factors (TFs) IIIe and VIa subgroup, were isolated from E. dulcis corm. Phylogenetic analysis showed that CwMYC and CwbHLH18 grouped with other terpenoid-regulated bHLHs, and their transcript levels were strongly induced after fresh-cut processing. These results suggested that the biosynthesis of terpenoids and apocarotenoids in fresh-cut E. dulcis strongly depended on the transcriptional regulation of structural genes involved in the methylerythritol 4-phosphate (MEP) and mevalonate (MVA) pathways. However, the complex secondary metabolism of fresh-cut E. dulcis during late storage requires further investigation.

Keywords: Chinese water chestnut; purification; secondary metabolism; structural identification; terpenoid biosynthesis.

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Conflict of interest statement

The authors declare that they have no conflict of interest.

Figures

FIGURE 1
FIGURE 1
Chemical structures of compounds 1 and 2 isolated from fresh‐cut Eleocharis dulcis.
FIGURE 2
FIGURE 2
Time course of the 6β‐Hydroxystigmast‐4‐en‐3‐one and blumenol A accumulation in fresh‐cut Eleocharis dulcis during storage.
FIGURE 3
FIGURE 3
Expression of genes that encode key enzymes involved in terpenoid backbone biosynthesis in Eleocharis dulcis during storage. Total RNA was extracted from fresh‐cut E. dulcis 0, 2, 4, and 6 days after peeling. The expression data were normalized against the expression of 18S rRNA, and gene expression levels were set to 1 in the day 0 samples. Each column represents the mean ± SE of three biological replicates.
FIGURE 4
FIGURE 4
Expression of genes that encode key enzymes involved in carotenoid biosynthesis in Eleocharis dulcis during storage. Total RNA was extracted from fresh‐cut E. dulcis 0, 2, 4, and 6 days after peeling. The expression data were normalized against the expression of 18S rRNA, and gene expression levels were set to 1 in the day 0 samples. Each column represents the mean ± SE of three biological replicates.
FIGURE 5
FIGURE 5
Changes in transcript levels of CwMYC and bHLH18 in Eleocharis dulcis during storage. Total RNA was extracted from fresh‐cut E. dulcis 0, 2, 4, and 6 days after peeling. The expression data were normalized against the expression of 18S rRNA, and gene expression levels were set to 1 in the day 0 samples. Each column represents the mean ± SE of three biological replicates.
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