关键词： 1-naphthoxyacetic acid 2,4-D 2,4-dichlorophenoxyacetic acid 6-BA 6-benzyl-adenine AP BLAST CDA CIA CTAB EAR EC ERF-associated amphiphilic repression Expression HD IAA IPTG MS NAA NEC NJ ORF PAGE Phylogenetic analysis QHB RAM RT-qPCR Real-time quantitative PCR SAM SDS SNPs TAIL-PCR USDA-ARS UTR United States Department of Agriculture—Agricultural Research Service WOX WOX5 WUS WUS-related homeobox Wheat Wuschel adaptor primer basic local alignment search tool callus in differentiation medium after days callus in induced medium after days cetyltrimethyl ammonium bromide embryogenic callus homeodomain indole-3-acetic acid isopropyl β-d-thiogalactopyranoside murashige and skoog medium neighbor-joining non-embryogenic callus open reading frame polyacrylamide gel electrophoresis quiescent center-specific homeobox real-time quantitative PCR root apical meristem shoot apical meristem single nucleotide polymorphism sodium dodecyl sulfate thermal asymmetric interlaced PCR untranslated transcribed region

Mesh ： Amino Acid Sequence Escherichia coli / genetics Gene Expression Regulation, Plant Genes, Homeobox Indoleacetic Acids / metabolism Molecular Sequence Data Multigene Family Open Reading Frames Phylogeny Plant Proteins / genetics metabolism Plant Roots / genetics Poaceae / genetics Protein Structure, Tertiary Triticum / genetics

来  源：   DOI：10.1016/j.gene.2013.12.022   PDF(Sci-hub)

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 1038 bp and encompassed the complete open reading frame (ORF) as well as 5\' upstream and 3\' 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.
摘要：
WUSCHEL（WUS）相关的同源盒（WOX）基因家族在协调胚胎发育早期的基因转录中起着重要作用。在这项研究中,我们从普通小麦及其近缘小麦中分离并表征了WOX5，普通小麦,埃希洛普斯，Aegilopssearsii,夏龙士，龙骨埃希洛普斯，Aegilopsbicornis,Aegilopstauschii,和小麦。表征的WOX5等位基因的大小范围为1029至1038bp，包括完整的开放阅读框（ORF）以及5'上游和3'下游序列。结构域预测分析表明，小麦WOX5蛋白的一级结构除了WUS盒结构域和EAR样结构域外，还包括高度保守的同源结构域。存在于WOX蛋白家族的一些成员中。将全长ORF亚克隆到原核表达载体pET30a中，通过IPTG诱导，在大肠杆菌BL21（DE3）细胞中成功表达了约26kDa的蛋白。来自小麦相关物种的WOX5基因表现出与普通小麦的WOX5基因相似的结构和高度的序列相似性。WOX5等位基因之间的分歧程度和系统发育树分析表明，在A中存在三个同源拷贝，B,或普通小麦的D基因组。定量PCR结果表明，TaWOX5主要在生长素和细胞分裂素诱导的根和愈伤组织中表达，这表明TaWOX5可能与根的形成或发育有关，并且与体细胞胚胎发生中的激素调节有关。
公众号