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园艺学报 ›› 2017, Vol. 44 ›› Issue (12): 2285-2295.doi: 10.16420/j.issn.0513-353x.2017-0073

• 研究论文 • 上一篇    下一篇

龙眼miR166家族的分子进化特性及时空表达分析

林玉玲,张清林,曾友竞,陈晓慧,张梓浩,陈裕坤,赖钟雄*   

  1. (福建农林大学园艺植物生物工程研究所,福州 350002)
  • 出版日期:2017-12-25 发布日期:2017-12-25

Analysis on Evolutionary Characteristics and the Temporal and Spatial Expression Patterns of miR166 Gene Family in Dimocarpus longan

LIN Yuling,ZHANG Qinglin,ZENG Youjing,CHEN Xiaohui,ZHANG Zihao,CHEN Yukun,and LAI Zhongxiong*   

  1. (Institute of Horticultural Biotechnology,Fujian Agriculture and Forestry University,Fuzhou 350002,China)
  • Online:2017-12-25 Published:2017-12-25

摘要:

为了解miR166基因家族的分子进化特性及其在龙眼不同组织部位的表达规律,对龙眼miR166家族成员的成熟体和前体序列及其进化、前体二级结构预测、靶基因预测以及时空表达模式进行了分析。结果表明:龙眼miR166家族含有12条成熟体序列和7个前体序列(pre-miRNA)。Mfold预测显示pre-miR166家族7个成员的序列均能形成典型稳定的茎环二级结构,成熟序列的碱基保守性高,其他位置的则保守性减弱;它们的最小折叠自由能(ΔG)在–35.30 ~–83.30 kal ? mol-1。系统发育进化树分析显示,龙眼pre-miR166家族与葡萄、碧桃的pre-miR166亲缘关系更为接近。靶基因预测显示,龙眼miR166基因家族靶基因包括同源异型域—亮氨酸拉链蛋白、三角状五肽重复结构蛋白、假定蛋白、DNA指导的RNA聚合酶Ⅲ亚单位RPC2、紫外线B受体等。实时荧光定量PCR显示,pre-miR166家族不同成员在龙眼生长发育进程中均有表达且表达模式不尽相同,提示其可能广泛参与龙眼各个器官的调控,且在功能上可能具有分工。

关键词: 龙眼, miR166家族, 进化特性, 实时荧光定量PCR

Abstract:

In order to understand the evolution characteristics of miR166 gene family and its expression pattern in different tissues of Dimocarpus longan,we analyzed the mature sequences,and the phylogenetic of miR166 in longan and some other fruit plants,predicted the secondary structure of precursor miR166(pre-miR166)family members,analyzed their targets,as well as their expression patterns. The results showed that the longan miR166 family contained 12 mature sequences and 7 precursor sequences. Mfold analysis showed that these pre-miR166 members could form a typical stable stem loop structures,and their minimal folding free energies(ΔG)ranged from–35.30 to–83.30 kal ? mol-1. Multiple sequence alignment of these pre-miR166 members showed that the regions generating mature miRNA were highly conservative. Phylogenetic tree analysis showed that the genetic relationship of pre-miR166 family in longan was closer to those from grape and peach. Target gene prediction of longan miR166 family revealed that their targets were homeobox-leucine zipper protein,pentatricopeptide repeat-containing protein,hypothetical protein,DNA-directed RNA polymerase Ⅲ subunit,and Ultraviolet-B receptor UVR8. Real-time quantitative PCR showed that diffierent pre-miR166 family members,had different expression patterns,and more or less expressed at different development stages on vegetative and reproductive growth of longan,suggesting that they may be widely participate in the regulation of longan organs development,and may have function division in longan.

Key words: Dimocarpus longan, miR166 family, evolution, real-time quantitative PCR

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