梨‘中矮1号’LUE1基因的克隆及功能分析

发布时间：2018-07-21 13:54

【摘要】：以梨优良矮化砧木‘中矮1号’新梢嫩茎为试材,根据白梨(Pyrus bretschneideri Rehd.)基因组预测的相关基因序列信息设计特异引物,克隆获得1个Katanin蛋白家族成员基因。序列分析显示该基因编码区全长1 566 bp,编码521个氨基酸,预测蛋白分子量约为57.2 kD,等电点为8.11,与其他物种的Katanin P60家族氨基酸序列具有78.01%~98.66%的相似性,将该基因命名为PcLUE1。实时荧光定量RT-PCR(qRT-PCR)分析显示,生长慢的矮化品种新梢快速生长时期PcLUE1的表达量显著高于生长快的乔化品种。石蜡切片显示,矮化品种与半矮化及乔化品种间茎组织细胞的排列方式和细胞大小存在较大差异。过表达PcLUE1的烟草植株与对照相比,株高降低,叶片变小,颜色变深,与‘中矮1号’中PcLUE1高表达效应极为相似,说明PcLUE1可能与‘中矮1号’的矮化有一定的关系。
[Abstract]:The tender stem of the new shoot of 'Zhongai 1', an excellent dwarf rootstock of pear, was used as the test material, according to the results of Pyrus bretschneideri Rehd. A member gene of Katanin protein family was cloned by designing a specific primer for gene sequence information. Sequence analysis showed that the coding region of the gene was 1 566 BP, encoding 521 amino acids. The predicted molecular weight of the predicted protein was about 57.2 KD and the isoelectric point was 8.11, which was similar to the amino acid sequence of Katanin P60 family of other species by 78.01% and 98.66% respectively. The gene was named PcLUE1. The results of real-time quantitative RT-PCR (qRT-PCR) showed that the expression of PcLUE1 was significantly higher in the slow-growing dwarf cultivars than in the fast growing varieties. Paraffin sections showed that there were significant differences in the arrangement of stem tissue cells and cell size among dwarf cultivars, semi-dwarf varieties and arborization varieties. Compared with the control, the over-expressed PcLUE1 in tobacco plants had lower plant height, smaller leaves and darker color, which was similar to the high expression effect of PcLUE1 in 'Zhongai 1', which suggested that PcLUE1 might be related to the dwarfing of 'Zhongai 1'.
【作者单位】：中国农业科学院果树研究所农业部园艺作物种质资源利用重点实验室;北京市农林科学院林业果树研究所;
【基金】：中国农业科学院科技创新工程项目国家科技支撑计划项目(2013D02B01,2013BAD01B04-23) 辽宁省果树产业技术体系专项(LNGSCYTX-15-X) 农业公益性行业科研专项(201203075-05)
【分类号】：S661.2

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