大花红景天盐胁迫调控基因RcNAC22克隆及功能分析

doi:10.7525/j.issn.1673-5102.2026.02.005

摘要/Abstract

摘要：

本研究克隆大花红景天（Rhodiola crenulata）转录因子NAC家族成员之一RcNAC22基因，并对其进行生物信息学分析和功能验证。结果表明：RcNAC22基因全长为1 049 bp，含2个内含子、3个外显子，编码293个氨基酸。其蛋白质相对分子质量为33 625.02，属亲水蛋白，性质较稳定，不具有跨膜结构域和信号肽且亚细胞定位于细胞核，含有32个可磷酸化位点。RcNAC22基因包含1个NAM结构域（no apical meristem），第4结构域为转录抑制结构域NARD区，与狭叶红景天（R. kirilowii）亲缘关系最近。RcNAC22基因在6个器官（根、成熟茎、幼茎、叶、顶芽和花）中均有表达但主要存在于绿色器官（茎、叶、顶芽）中，同时受多种非生物胁迫和植物激素脱落酸（ABA）、水杨酸（SA）的诱导表达。RcNAC22蛋白对酵母细胞没有毒性，其全长基因和截短C端均具有自激活活性。此外，以丹参（Salviamiltiorrhiza）为转基因受体研究RcNAC22基因过表达对植物生长的影响，结果显示：提高RcNAC22基因表达量不会影响丹参的生长发育过程，但降低其对盐胁迫的耐受性。本研究揭示了RcNAC22基因的生物学功能，为进一步开展大花红景天分子生物学研究提供理论基础。

关键词: 大花红景天, 转录因子, 基因克隆, 盐胁迫

Abstract:

This study cloned the RcNAC22 gene of R. crenulata and conducted bioinformatics analysis and functional verification. The results showed that the full length of RcNAC22 gene was 1 049 bp， containing two introns and three exons， and encoding 293 amino acids. Its protein relative molecular weight was 33 625.02， belonging to a hydrophilic protein， with relatively stable properties， no transmembrane domain or signal peptide， and subcellular localization in the nucleus， containing 32 phosphorylation sites. RcNAC22 contains one NAM domain （no apical meristem）， and the fourth domain is the transcriptional repression domain NARD region， and it was most closely related to that of R.kirilowii. RcNAC22 gene was expressed in all six tested organs （root， mature stem， young stem， leaf， apical bud， flower） but mainly in green organs （stem， leaf， apical bud）， and its expression could be induced by various abiotic stresses and plant hormones ABA and SA. RcNAC22 gene had no cytotoxicity to yeast cells， and both the full-length gene and truncated C-terminal had self-activation activity. In addition， using Salvia miltiorrhiza as the transgenic receptor to study the RcNAC22 gene effect on plant growth， the results showed that overexpression of RcNAC22 gene did not affect the growth and development of Salviamiltiorrhiza， but reduced the tolerance to salt stress. This study revealed the biological function of the RcNAC gene， providing a theoretical basis for further molecular biological research on R. crenulata.

Key words: Rhodiolacrenulata, transcription factor, gene cloning, salt stress

中图分类号:

S714.7

张力鹏, 武美, 王宏鹏, 李天宇. 大花红景天盐胁迫调控基因RcNAC22克隆及功能分析[J]. 植物研究, 2026, 46(2): 246-258.

Lipeng ZHANG, Mei WU, Hongpeng WANG, Tianyu LI. Salt Stress-related RcNAC22 Gene Cloning and Function Analysis in Rhodiolacrenulata[J]. Bulletin of Botanical Research, 2026, 46(2): 246-258.

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引物名称 Primer name	正向引物（5′→3′） Forward primer （5′→3′）	反向引物（5′→3′） Reverse primer （5′→3′）
RcNAC22基因全长扩增	ATGTCGGCTACGAAGTCC	TCAGAGATTAAGCACGTTGAC
植物表达载体构建	cctaggATGTCGGCTACGAAGTCC	ggtcaccTCAGAGATTAAGCACGTTGAC
酵母表达载体构建	gaattcATGTCGGCTACGAAGTCC	ggatccTCAGAGATTAAGCACGTTGAC
基因N端序列酵母载体构建	gaattcATGTCGGCTACGAAGTCC	ggatccTCGCCGCTCTCTGCTCCAAC
基因C端序列酵母载体构建	gaattcATGTGGATCACGACGACGAAG	ggatccTCAGAGATTAAGCACGTTGAC
大花红景天内参基因	TACCCTTCATAGCACCCTCAGA	GAATGGCTTTCCGTGTCCC
丹参内参基因	GGTGCCCTGAGGTCCTGTT	AGGAACCACCGATCCAGACA