Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
5906069 | Gene | 2014 | 7 Pages |
Abstract
The WUSCHEL (WUS)-related homeobox (WOX) gene family plays an important role in coordinating gene transcription in the early phases of embryogenesis. In this study, we isolated and characterized WOX5 from common wheat and its relatives Triticum monococcum, Triticum urartu, Aegilops speltoides, Aegilops searsii, Aegilops sharonensis, Aegilops longissima, Aegilops bicornis, Aegilops tauschii, and Triticum turgidum. The size of the characterized WOX5 alleles ranged from 1029 to 1038Â bp and encompassed the complete open reading frame (ORF) as well as 5â² upstream and 3â² downstream sequences. Domain prediction analysis showed that the putative primary structures of wheat WOX5 protein include the highly conserved homeodomain besides the WUS-box domain and the EAR-like domain, which is/are present in some members of the WOX protein family. The full-length ORF was subcloned into a prokaryotic expression vector pET30a, and an approximate 26-kDa protein was successfully expressed in Escherichia coli BL21 (DE3) cells with IPTG induction. The WOX5 genes from wheat-related species exhibit a similar structure to and high sequence similarity with WOX5 genes from common wheat. The degree of divergence and phylogenetic tree analysis among WOX5 alleles suggested the existence of three homoeologous copies in the A, B, or D genome of common wheat. Quantitative PCR results showed that TaWOX5 was primarily expressed in the root and calli induced by auxin and cytokinin, indicating that TaWOX5 may play a role related to root formation or development and is associated with hormone regulation in somatic embryogenesis.
Keywords
RT-qPCRUSDA-ARSthermal asymmetric interlaced PCRWUSWOXTAIL-PCRNECPAGECDA6-BASAMIPTGORFNaACTABSDSIAASNPs2,4-D2,4-dichlorophenoxyacetic acidMurashige and Skoog mediumBLAST, basic local alignment search toolCetyltrimethyl ammonium bromideindole-3-acetic acidExpressionpolyacrylamide gel electrophoresisBlastisopropyl β-D-thiogalactopyranosidephylogenetic analysisRAM, Random Access Memory, Read-Write MemoryEmbryogenic callusreal-time quantitative PCRshoot apical meristemsodium dodecyl sulfateCIAopen reading frameroot apical meristemUTR یا untranslated regions homeodomainWUSCHELSingle nucleotide polymorphismneighbor-joiningWheatEAR
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Authors
Shan Zhao, Qian-Tao Jiang, Jian Ma, Xiao-Wei Zhang, Quan-Zhi Zhao, Xiu-Ying Wang, Chang-Shui Wang, Xue Cao, Zhen-Xiang Lu, You-Liang Zheng, Yu-Ming Wei,