Cloning, transcriptional activation, and tissue expression analysis of XsWRI1 from Xanthoceras sorbifolium

doi:10.12302/j.issn.1000-2006.202309001

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2025, Vol. 49 ›› Issue (2): 23-30.doi: 10.12302/j.issn.1000-2006.202309001

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Cloning, transcriptional activation, and tissue expression analysis of XsWRI1 from Xanthoceras sorbifolium

ZHANG Wei(), LI Linkun, LIANG Chongjun, WANG Libing^*()

State Key Laboratory of Tree Genetics and Breeding, Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China

Received:2023-09-01 Accepted:2023-11-29 Online:2025-03-30 Published:2025-03-28
Contact: WANG Libing E-mail:w.zhang35@caf.ac.cn;wlibing@caf.ac.cn

Abstract

Abstract:

【Objective】This study aimed to characterize the sequence identity, transcriptional activation potential, and functional domains of the WRINKLED1 (WRI1) transcription factor in Xanthoceras sorbifolium (yellowhorn), providing insights into its regulatory role in seed oil biosynthesis. 【Method】The full-length cDNA of XsWRI1 was cloned from endosperm tissues of mature yellowhorn using rapid amplification of cDNA ends (RACE). Bioinformatics tools were employed to analyze protein sequence properties. A recombinant pGBKT7-XsWRI1 vector was constructed and transformed into Y2HGold yeast cells to assess transcriptional activation activity. Tissue-specific expression patterns were quantified via quantitative real-time PCR (qRT-PCR) across the root, stem, leaf, petal, stamen and developing embryo tissues. 【Result】The XsWRI1 gene (GenBank accession: OR500287) spans 1 688 bp, encoding a hydrophilic and unstable protein of 414 amino acids. Yeast activity testing confirmed strong transcriptional activation activity of XsWRI1. Tissue-specific expression analysis revealed predominant XsWRI1 expression in developing embryos, with negligible levels in vegetative organs (root, stem, leaf) and reproductive structures (petal, stamen). Structural prediction identified two conserved AP2/EREBP DNA-binding domains (residues 76-148 and 177-238) and a nuclear localization signal. 【Conclusion】This study elucidates the molecular characteristics and tissue-specific regulatory role of XsWRI1 in yellowhorn, highlighting its potential function in lipid biosynthesis pathways. These findings establish a foundation for targeted genetic manipulation to enhance seed oil accumulation in woody oil crops.

Key words: Xanthoceras sorbifolium, XsWRI1, gene cloning, transcriptional activity, tissue-specific expression, seed oil biosynthesis

CLC Number:

S727.32

ZHANG Wei, LI Linkun, LIANG Chongjun, WANG Libing. Cloning, transcriptional activation, and tissue expression analysis of XsWRI1 from Xanthoceras sorbifolium[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2025, 49(2): 23-30.

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引物名称 primer name	引物序列(5'—3') primer sequences 5'-3'	引物用途 primer purpose
3'race outer	GAATAACTCCGTTCTCAGTGC	3'race
3'race inner	GATGCGGGATTTGAGGAGAACATAG
5'race outer	TGTCTCTGTTTTGAGTTTTTCTG	5'race
5'race inner	AGTAACATCAGATAAAGCACAAG
XsWRI1-F	TTTACTACACATTCTGTCTTATT	ORF扩增 ORF amplification
XsWRI1-R	AACAACAAACAACATCAACAA
BD-WRI1-F	aggaggacctgcatatggcc ATGAAGAGGTCTTCTACTAC	pGBKT7载体构建 the pGBKT7 vector construction
BD-WRI1-R	cccgggaattcggcctccat TCAAACAGAATAGATATTAC
qRT WRI1-F	ATAACATACAGAACAAGAAGGGAAG	荧光定量 fluorescence quantitative
qRT WRI1-R	CAGCAAGATCATAAGTACGAGCA
qRT Tip41-F	AAGATGAATTGGCTGATAACGGA	荧光定量内参 fluorescence quantitative internal reference
qRT Tip41-R	AGCTCTCACGAAGAATGACTGGG

SapBase编号	相对表达量relative expression
SapBase编号	44 d	54 d	68 d	81 d
EVM0012075	127.87	112.62	52.33	6.32
EVM0007314	0.33	0.09	0	0
EVM0014879	0	0.02	0	0

Cloning, transcriptional activation, and tissue expression analysis of XsWRI1 from Xanthoceras sorbifolium

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