白木香转录因子AsMYB1与AsMYB2克隆及表达分析.docVIP

白木香转录因子AsMYB1与AsMYB2克隆及表达分析 　　[摘要] MYB类转录因子是数量最多、功能最多样化的转录因子家族之一。该研究根据白木香转录组高通量测序结果设计引物扩增白木香中MYB基因，利用RTPCR技术从白木香中首次克隆得到2个MYB基因cDNA 全长，分别命名为AsMYB1，AsMYB2，并对MYB蛋白质进行跨膜结构分析和聚类分析，预测其生物学信息；利用实时荧光定量PCR技术检测 AsMYB基因在不同组织， 及在NaCl、低温（4 ℃）、重金属（CdCl2）、干旱（甘露醇）4种非生物胁迫和脱落酸、水杨酸、赤霉素、茉莉酸甲酯4种外源激素处理下的表达差异。该研究中克隆获得的AsMYB1基因全长1 063 bp，编码353个氨基酸；AsMYB2基因全长1 081 bp，编码359个氨基酸。2个基因具有组织表达特异性；AsMYB1，AsMYB2 基因的转录水平受到不同生物胁迫和外源激素的影响，说明MYB基因对白木香防御反应和激素信号传导具有重要的作用。 　　[关键词] 白木香； MYB基因； 表达分析； 非生物胁迫； 激素处理 　　Cloning and expression analysis of transcription factor AsMYB1 and 　　AsMYB2 from Aquilaria sinensis 　　FENG Yingying， DONG Xianjuan， LIU Xiao， YAN Yaru， WANG Jinling， SHI Shepo， WANG Xiaohui*， TU Pengfei* 　　（Modern Research Center for Traditional Chinese Medicine， School of Chinese Materia Medica， 　　Beijing University of Chinese Medicine， Beijing 100029， China） 　　[Abstract] The MYB gene family comprises one of the richest groups of transcription factors in plants. The full length of two MYB genes were isolated through heterologous screening of Aquilaria sinensis calli transcriptome data， and the reverse transcription PCR was performed to obstain the corrected MYB clones， named AsMYB1， AsMYB2. The MYB transmembrane domain and phylogenetic analysis were predicted by different software to analyze the bioinformatics of MYB proteins. The transcript level of AsMYB1， AsMYB2 was performed by realtime quantitative RTPCR in different tissues and in responds to abiotic stresses including salt， cold， metal and drought stress， and hormone treatments including abscisic acid （ABA）， salicylic acid （SA）， gibberellins （GA3） and methyl jasmonate （MeJA） treatment. The AsMYB1 cDNA sequence had an ORF of 1 063 nucleotides， encoding a protein of 353 amino acids. The largest AsMYB2 ORF was 1 081 nucleotides， and its predicted translation products consisted of 359 amino acids. Two MYB genes had a tissuesspecific pattern in A. sinensis. Moreover， the expression level of AsMYB1 and AsMYB2 was regulated by different a