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. 2023 Mar 23:14:1151645.
doi: 10.3389/fgene.2023.1151645. eCollection 2023.

Genomic profiling of WRKY transcription factors and functional analysis of CcWRKY7, CcWRKY29, and CcWRKY32 related to protoberberine alkaloids biosynthesis in Coptis chinensis Franch

Xiaoqiang Huang  1 , An Jia  1 , Tao Huang  1 , Li Wang  1 , Guohua Yang  2 , Wanli Zhao  1   3
Affiliations

Genomic profiling of WRKY transcription factors and functional analysis of CcWRKY7, CcWRKY29, and CcWRKY32 related to protoberberine alkaloids biosynthesis in Coptis chinensis Franch

Xiaoqiang Huang et al. Front Genet. .

Abstract

Coptis chinensis Franch. (Huanglian in Chinese) is an important economic crop with medicinal value. Its rhizome has been used as a traditional herbal medicine for thousands of years in Asia. Protoberberine alkaloids, as the main bioactive component of Coptis chinensis, have a series of pharmacological activities. However, the protoberberine alkaloids content of C. chinensis is relatively low. Understanding the molecular mechanisms affecting the transcriptional regulation of protoberberine alkaloids would be crucial to increase their production via metabolic engineering. WRKY, one of the largest plant-specific gene families, regulates plant defense responses via the biosynthesis of specialized metabolites such as alkaloids. Totally, 41 WRKY transcription factors (TFs) related to protoberberine alkaloid biosynthesis were identified in the C. chinensis genome and classified into three groups based on phylogenetic and conserved motif analyses. Three WRKY genes (CcWRKY7, CcWRKY29, and CcWRKY32) may regulate protoberberine alkaloid biosynthesis, as suggested by gene-specific expression patterns, metabolic pathways, phylogenetic, and dual-luciferase analysis. Furthermore, the CcWRKY7, CcWRKY29, and CcWRKY32 proteins were specifically detected in the nucleus via subcellular localization. This study provides a basis for understanding the regulatory mechanisms of protoberberine alkaloid biosynthesis and valuable information for breeding C. chinensis varieties.

Keywords: Coptis chinensis; WRKY family; berberine biosynthesis; protoberberine alkaloids; transcription factors.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
Schematic diagram of CcWRKYs conserved motifs composition.
FIGURE 2
FIGURE 2
Phylogenetic relationships of CcWRKY proteins in Coptis chinensis and A. thaliana.
FIGURE 3
FIGURE 3
Chromosomal location of CcWRKY gene family in C. chinensis.
FIGURE 4
FIGURE 4
The main protoberberine alkaloids content of C. chinensis in different tissues and the expression profile of CcWRKY genes. (A) The main protoberberine alkaloid content of Coptis chinensis in different tissues. (B) Expression profile of CcWRKY genes and their candidate target genes involved in protoberberine alkaloids biosynthesis in different tissues including, root, petiole, leaf, and rhizome.
FIGURE 5
FIGURE 5
Subcellular localization of CcWRKY7, CcWRKY29 and CcWRKY32 proteins in Nicotiana benthamiana leaves. Laser scanning confocal microscope with wavelengths of 488 and 406 nm for GFP and DAPI, respectively.
FIGURE 6
FIGURE 6
CcWRKY7, CcWRKY29, and CcWRKY32 activate the promoter of CcCNMT.
See this image and copyright information in PMC

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