Study on biosynthesis pathway and accumulation mechanism of the dihydrochalcones in Lithocarpus litseifolius

Abstract

Dihydrochalcones (DHCs) are highly accumulated in tender leaves of Lithocarpus litseifolius but their biosynthetic pathway and accumulation mechanism remain unclear. In this study, candidate genes including one cinnamoyl-CoA reductase (LlCCR), two double bond reductases (LlDBR1 ~ 2), three aldehyde hydrogenases (LlALDH1 ~ 3), two 4-coumaroyl:CoA ligases (Ll4CL1 ~ 2) and four phloretin glycosyltransferases (LlP4′GT, LlP2′GT1~3) were comprehensively investigated. The substrate specificities and catalytic kinetics of these gene-encoded enzymes were achieved. Through successive catalysis of LlALDH1, Ll4CL2, and chalcone synthase 1 (LlCHS1) or combined action of LlCCR and LlCHS1, phloretin was biosynthesized from direct precursor dihydro-p-coumaraldehyde, which had been converted from initial precursor p-coumaroyl-CoA by LlCCR-mediated carboxylic acid reduction and LlDBR1-catalyzed α,β-double bond saturation. High accumulation of the DHCs in tender leaves of L. litseifolius was mainly driven by efficient catalysis of LlCCR toward p-coumaroyl-CoA and highly expressed genes in the pathway, especially the LlP4′GT and LlP2′GT1 which contributed to biosynthesis of trilobatin and phlorizin, respectively. Antisense oligodeoxyribonucleotide treatments against the LlCCR, LlDBR1, LlALDH1, Ll4CL2, LlP4′GT, and LlP2′GT1 significantly reduced transcripts of the target genes and content of DHCs, confirming these genes might be involved in the pathway. This finding provides insight into the biosynthesis and accumulation mechanism of DHCs in planta

Acknowledgements

This work was financially supported by the National Natural Science Foundation of China (grant 32272763), China Agriculture Research System of MOF and MARA. and the Zhejiang Science and Technology Major Program on Agricultural New Variety Breeding-Tea Plant (grant 2021C02067-6). The authors also want to thank Jingli Cheng for her kind help of instrument analysis and maintenance, and to express our gratitude to Mr. Peihua Zhao from Huzhou Xinya Ecological Agriculture Technology Co., Ltd for kindly providing the L. litseifolius samples.

Author contributions

Jian-Liang Lu (Conceptualization), Yu-Si Yang, Yu-Ke Du (Methodology), Yu-Si Yang (Software), Cun-Yu Li (Validation), Yu-Ke Du, Jia-Li Li (Formal analysis), Yu-Si Yang, Jia-Li Li (Investigation), Yong-Kang Wang (Resources), Yu-Si Yang (Data curation), Yu-Si Yang (Writing-original draft preparation), Jian-Liang Lu, Zhou-Tao Fang (Writing-review & editing), Yu-Si Yang (Visualization), Jian-Hui Ye (Supervision), Xin-Qiang Zheng, Yue-Rong Liang (Project administration), Jian-Liang Lu (Funding acquisition). All authors have read and agreed to the published version of the manuscript.

Data availability

All data related to this research are available in this paper and its supplementary materials published online.

Conflicts of interest statement

The authors declare no competing financial interest.

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