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以杜梨叶片为试材,采用同源序列和RACE技术克隆了梨ARRO(adventitious rooting related oxygenase)基因,并进行了基因的生物信息学和时空表达分析,以期对梨属植物不定根产生研究提供帮助。结果表明:PbARRO?1基因开放读码框全长831bp,编码276个氨基酸,相对分子量为30.535 9kD,等电点pI为5.53;推导氨基酸序列同源性分析显示该基因与苹果、梅花、毛果杨等具有较高同源性,其中苹果达到92%。荧光实时定量PCR分析表明,该基因在秋子梨根、茎、叶中均有表达,叶片中表达量初期较高,后期较低;茎中表达量则呈现先升高后降低的趋势,在第15天达到峰值,推测PbARRO-1基因与茎上不定根的形成密切相关。
In order to study the adventitious rooting related oxygenase (ARRAR) gene in pear (Pyrus pyrifolia Nakai) cultivars, we used the homologous sequence and RACE technique to clone the gene of adventitious rooting related oxygenase (ARRO) and analyzed its bioinformatics and spatiotemporal expression. . The results showed that the full-length open reading frame (ORF) of PbARRO 1 gene was 831bp, encoding a protein of 276 amino acids with a relative molecular mass of 30.535 9kD and a pI of isoelectric point of 5.53. Homology analysis of deduced amino acid sequence showed that the gene was homologous to apple, Yang and other high homology, of which apple reached 92%. Real-time quantitative PCR analysis showed that the gene was expressed in the roots, stems and leaves of A. thaliana, and the expression level in the leaves was higher at the early stage and lower at the late stage. The expression level in the stem increased first and then decreased, 15 days to reach the peak, presumably PbARRO-1 gene and adventitious root formation is closely related.