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.
Subject(s)
Cloning, Molecular , Dendrobium , Genetics , Gene Expression Regulation, Plant , Plant Proteins , Genetics , Transcription Factors , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To study the genetic diversity of medicinal Dendrobium by SRAP.</p><p><b>METHOD</b>The genetic diversity of 9 spices Dendrobium was studied by using the optimized SRAP reaction system. The NTSYS software was used to analyze the markers.</p><p><b>RESULT</b>Forty primer pairs were selected from 88 amplified 1 782 polymorphic bands with an average of 44.55 polymorphic bands per primer pair. Cluster analysis using UPGMA method based on the data of SRAP amplified bands by 40 primer pairs showed that 9 spices of could be distinguished into two main groups. Jaccard's similarity coefficient ranged from 0.330 2-0.789 2.</p><p><b>CONCLUSION</b>The results of this research indicate that SRAP molecular marker is efficient to study the medical Dendrobium genetic diversity.</p>