Cloning of AhWRKY33 from Asarum heterotropoides and its genetic transformation in Arabidopsis.
10.19540/j.cnki.cjcmm.20200329.107
- Author:
Yong-Cheng YANG
1
;
Xiao-Han WANG
2
;
Yan-Fang WANG
1
;
Xiu-Yan LIU
1
;
Shi-Hai YANG
1
Author Information
1. Laboratory of Cultivation and Breeding of Medicinal Plants of National Administration of Traditional Chinese Medicine, College of Chinese Medicine Materials, Jilin Agricultural University Changchun 130118, China.
2. School of Chinese Materia Medica, Beijing University of Chinese Medicine Beijing 102488, China.
- Publication Type:Journal Article
- Keywords:
Asarum heterotropoides;
WRKY gene;
genetic transformation;
plant lant expression vector
- MeSH:
Arabidopsis;
genetics;
Arabidopsis Proteins;
genetics;
Asarum;
Cloning, Molecular;
Gene Expression Regulation, Plant;
Plant Leaves;
Plant Proteins;
genetics;
Transcription Factors;
Transformation, Genetic
- From:
China Journal of Chinese Materia Medica
2020;45(13):3112-3119
- CountryChina
- Language:Chinese
-
Abstract:
The WRKY family genes, which play an important role in plant morphogenesis and stress response, were selected based on the data of the full-length transcriptome of Asarum heterotropoides. Using AtWRKY33, which regulates the synthesis of the camalexin in the model plant Arabidopsis to compare homologous genes in A. heterotropoides, primers were designed to amplify the open reading frame(ORF) fragment of AhWRKY33 gene by RT-PCR using total RNA of A. heterotropoides leaves as template. Real-time PCR results showed that there was a significant difference between the aerial part and the underground part of A. heterotropoides, the toxic aristolochic acid content is highly expressed in the leaves higher than the root. After verification, the WRKY33 gene of A. heterotropoides is ORF long 1 686 bp, encoding 561 amino acids.AhWRKY33 had two conserved WRKYGQK domains. According to the classical classification, it belongs to group Ⅰ WRKY transcription factor. A. heterotropoides WRKY33 had some homology with amino acids of other species. The study successfully constructed the plant eukaryotic expression vector PHG-AhWRKY33 and transformed Arabidopsis thaliana, the transgenic Arabidopsis was obtained by PCR detection and hygromycin resistant plate screening. It found that the germination of transgenic Arabidopsis seeds was accelerated and the stress resistance was increased. It laid a foundation for further analysis of WRKY transcription factor in the growth and development of A. heterotropoides and the synthesis of secondary metabolites.
