Cloning and expression analysis of transcription factor gene DoWRKY1 in Dendrobium officinale.
- Author:
Jun ZHAO
;
Shi-wei SUN
;
Can-can MENG
;
Qing JIN
;
Hong-hong FAN
;
Yi LIN
;
Yong-ping CAI
- Publication Type:Journal Article
- MeSH:
Cloning, Molecular;
Dendrobium;
genetics;
Gene Expression Regulation, Plant;
Plant Proteins;
genetics;
Transcription Factors;
genetics
- From:
China Journal of Chinese Materia Medica
2015;40(14):2807-2813
- CountryChina
- Language:Chinese
-
Abstract:
WRKY transcription factors are novel transcriptional regulatory factors, which play an important role in regulating plant development, metabolism and other physiological processes. In this study, a new Dendrobium officinale WRKY transcription factor, designated as DoWRKY1 was cloned by using RT-PCR and RACE (GenBank Accession No. KF953910). Bioinformatic analysis demonstrated that, the full-length cDNA of DoWRKY1 was 1,704 bp. And DoWRKY1 contained a 1,629 bp open reading frame (ORF) that encoding a peptide of 542 amino acid residues. The putative DoWRKY1 protein contained two conserved WRKY domains and it belonged to the group I WRKY family protein. Yeast one-hybrid experiment showed that DoWRKY1 had transcriptional activation ability in yeast, and it could activate the expression of downstream report genes (His3 and Ade2). Semi-quantitative RT-PCR experiment showed that DoWRKY1 expressed in roots, stems, leaves and protocorm-like bodies. Real-time qRT-PCR proved that DoWRKY1 could be induced by methyl jasmonate (MeJA) and chitosan (Chitosan), and the expression level of this gene can reach the expression peak at 2 h and 1 h, respectively. These results are useful for further determination of the regulation function of this gene in secondary metabolism of D. officinale.
