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以梨(Pyrus communis L.)紧凑型矮化砧木‘中矮1号’及其母本‘锦香’新梢韧皮部为试材,根据转录组测序结果设计特异性引物,克隆到1个长1 239 bp的基因序列。该序列在两个品种间不存在差异,均编码412个氨基酸,其氨基酸序列与苹果(np_001280772.1)、梅(xp_008242099.1)、毛果杨(xp_002306421.2)、草莓(xp_00428771.1)的生长素氢转运体基因(AHS)编码的氨基酸序列的相似性在67%~99%之间,命名为Pc AHS。q RT-PCR分析发现,‘中矮1号’新梢韧皮部中Pc AHS的表达量均低于母本‘锦香’,推测其启动子序列存在差异。‘中矮1号’及其母本‘锦香’Pc AHS基因上游启动子长度分别为828 bp和888 bp,二者相似性89.1%。序列分析发现‘中矮1号’Pc AHS基因启动子有一段58 bp(–496bp~–553 bp)的缺失;利用植物顺式作用元件数据库PLACE和PLANTCARE分析表明,‘中矮1号’启动子含有一个‘锦香’没有的BPBF转录因子结合元件P-box。推测‘中矮1号’Pc AHS基因启动子特有的片段缺失和P-box转录因子结合元件可能是导致其表达量低并通过影响生长素的运输最终引起矮化的原因。
Using the specific primers of pear (Pyrus communis L.) compact dwarf rootstock ’Zhongzai 1’ and its female parent ’Jinxiang’ new shoot phloem, the specific primers were designed according to the transcriptome sequencing results and cloned into one long 1 239 bp gene sequence. There was no difference between the two cultivars, which all encoded 412 amino acids. The deduced amino acid sequence was identical with that of apple (np_001280772.1), plum (xp_008242099.1), poplar (xp_002306421.2), strawberry (xp_00428771.1) The amino acid sequence encoded by the auxin hydrogen transporter gene (AHS) has a similarity between 67% and 99% and is named Pc AHS. q RT-PCR analysis showed that Pc AHS expression in phloem of ’Zhongzai 1’ was lower than that of ’Jinxiang’ in the phloem of the ’Zhongzai 1’, and the promoter sequence was presumed to be different. The length of the upstream promoter of Pc AHS gene in ’Zhongkai 1’ and its female parent ’Jinxiang’ were 828 bp and 888 bp, respectively, with a similarity of 89.1%. Sequence analysis showed that there was a 58 bp (-496 bp ~ -553 bp) deletion in the PC AHS gene promoter of Zhongzai 1, and PLACE and PLANTCARE analysis of the plant cis-acting element database showed that the ’Zhongzai 1’ promoter Contains a BPBF transcription factor binding element, P-box, that is not present in ’Jinxiang’. It is speculated that the deletion and P-box transcription factor-binding element specific to the promoter region of Pc AHS gene in ’Zhongkai 1’ may be responsible for its low expression and ultimately dwarfing by affecting the transport of auxin.