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生长素响应因子(auxin response factor,ARF)基因在调控生长素响应基因和生长素信号转导途径以及其它多个生长发育过程具有重要作用。本研究利用Blast P程序比对并获得苹果ARF基因家族,通过DNAMAN 6.0、MEGA 5.0、Web Logo3、Map Inspect和MEME软件对苹果ARF基因进行分析,采用RT-PCR技术研究基因组织表达情况。结果表明,苹果基因组存在29个ARF基因,进化上可分为I、II、III、IV和V组,每组成员数目分别是4、5、10、6和4个。内含子和外显子结构分析表明,该基因家族由2~15个外显子构成。染色体分布结果显示,Md ARF基因在染色体上分布不均匀。分别鉴定出4对和2对Md ARF基因经历了串联复制和片段复制,14个Md ARF基因经历了全基因组复制。保守元件分析表明,苹果ARF基因家族DBD区域、ARF区域、III元件和IV元件结构高度保守。半定量结果表明,大多数Md ARF基因在根、茎、叶、花和果中均有表达。
The auxin response factor (ARF) gene plays an important role in the regulation of auxin response genes and auxin signal transduction pathways and in many other growth and development processes. In this study, we used the Blast P program to compare and obtain the ARF gene family of apple, analyzed the ARF gene in apple by DNAMAN 6.0, MEGA 5.0, Web Logo3, Map Inspect and MEME software, and studied the expression of the gene by RT-PCR. The results showed that there are 29 ARF genes in the apple genome, which can be divided into groups I, II, III, IV and V, with 4, 5, 10, 6 and 4 members in each group. Introns and exons structure analysis showed that the gene family consists of 2 to 15 exons. Chromosome distribution showed that the Md ARF gene was unevenly distributed on chromosomes. Four pairs and two pairs of Md ARF genes were identified as having undergone tandem replication and fragment replication, respectively, and 14 Md ARF genes underwent genome-wide duplication. Analysis of conserved components showed that the structure of the ARF gene family in DBD, ARF, III and IV was highly conserved. Semi-quantitative results showed that most of the Md ARF genes were expressed in roots, stems, leaves, flowers and fruit.