Cloning, subcellular localization and expression analysis of SmIAA7 gene from Salvia miltiorrhiza
10.16438/j.0513-4870.2024-0789
- VernacularTitle:丹参SmIAA7基因的克隆、亚细胞定位与表达分析
- Author:
Yu-ying HUANG
1
;
Ying CHEN
2
;
Bao-wei WANG
2
;
Fan-yuan GUAN
3
;
Yu-yan ZHENG
3
;
Jing FAN
3
;
Jin-ling WANG
3
;
Xiu-hua HU
4
;
Xiao-hui WANG
3
Author Information
1. Modern Research Center for Traditional Chinese Medicine, Beijing Institute of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing 102488, China; College of Biomedicine and Health, Anhui Science and Technology University, Chuzhou 233100, China
2. School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 102488, China
3. Modern Research Center for Traditional Chinese Medicine, Beijing Institute of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing 102488, China
4. School of Life and Science, Beijing University of Chinese Medicine, Beijing 102488, China
- Publication Type:Research Article
- Keywords:
italic>Salvia miltiorrhiza;
italic>SmIAA7;
prokaryotic expression;
subcellular localization;
expression analysis
- From:
Acta Pharmaceutica Sinica
2025;60(2):514-525
- CountryChina
- Language:Chinese
-
Abstract:
The auxin/indole-3-acetic acid (Aux/IAA) gene family is an important regulator for plant growth hormone signaling, involved in plant growth, development, as well as response to environmental stresses. In the present study, we identified SmIAA7 which is potentially associated with Salvia miltiorrhiza leaf development through comparatively analyzed the transcriptome data from different pinnate leaves. SmIAA7 was successfully isolated from S. miltiorrhiza using the specific primers. Then subsequent bioinformatic analysis, prokaryotic expression and purification, subcellular localization, and induction expression analysis under auxin and abiotic stress were performed. The full-length of SmIAA7 contained an ORF of 684 bp encoding a protein of 227 amino acid with a molecular weight of 25.3 kD. Conserved domain analysis showed that SmIAA7 contains the conserved Aux_IAA domain (pfam02309). Sequence analysis and phylogenetic tree analysis results indicated SmIAA7 was phylogenetically close to IAA7 and IAA14 from other plants, suggesting SmIAA7 involved in plant growth and development as well as response to environmental stresses. The prokaryotic expression vector pET28a-SmIAA7 was constructed and SmIAA7 recombinant protein was successfully expressed in E. coli Rosetta (DE3) strain. Subcellular localization experiment demonstrated that SmIAA7 localized in the nucleus of plant cells. Real-time fluorescence quantitative PCR results showed that the expression level of SmIAA7 was upregulated in response to auxin. Drought, low temperature, and salt stress significantly increased the transcript level of SmIAA7 gene. This study lays a foundation for further elucidating the role of SmIAA7 in leaf development, signal transduction, and stress defense in S. miltiorrhiza.
