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园艺学报 ›› 2020, Vol. 47 ›› Issue (7): 1237-1252.doi: 10.16420/j.issn.0513-353x.2019-0761

• 研究论文 • 上一篇    下一篇

苹果miR396家族鉴定及在不定根发育过程中的表达分析

韦燕红,刘 桢,李 珂,孟 媛,汪 蕙,毛江萍,马豆豆,李少欢,马娟娟,卢 显,张 东*   

  1. 西北农林科技大学园艺学院,国家苹果改良中心杨凌分中心,陕西杨凌 712100
  • 出版日期:2020-07-25 发布日期:2020-07-25
  • 基金资助:
    国家重点研发计划重点项目(2019YFD1001803);陕西省重点研发计划项目(2017ZDXM-NY-019,2019TSLNY02-04);国家现代农业产业技术体系建设专项资金项目(CARS-27);“仲英青年学者”项目

Genome-wide Identification and Expression Analysis of miR396 Family During Adventitious Root Development in Apple

WEI Yanhong,LIU Zhen,LI Ke,MENG Yuan,WANG Hui,MAO Jiangping,MA Doudou,LI Shaohuan,MA Juanjuan,LU Xian,and ZHANG Dong*   

  1. College of Horticulture,Northwest A & F University,National Apple Improvement Center Yangling Sub-center,Yangling,Shaanxi 712100,China
  • Online:2020-07-25 Published:2020-07-25

摘要: 分析了苹果miR396家族进化特性及其在苹果不定根发育过程中的表达模式。结果表明:苹果miR396家族有4条成熟体和7条前体序列(pre-miRNA)。Mfold预测显示Pre-miR396家族7个成员序列均可形成典型稳定的茎环二级结构,最小折叠自由能介于–62.9 kal ? mol-1(pre-miR396b)~–51.9 kal ? mol-1(pre-miR396g)之间。系统发育进化树分析显示,pre-miR396家族亲缘关系可分为3个亚组(G1、G2、G3),每个亚组内基因数量不同,分别含有11、9、19个。靶基因预测显示,苹果miR396靶基因包括MdGRF1、MdGRF2和MdGRF5等,降解组测序进一步验证了miR396对其候选靶基因MdGRF1、MdGRF2和MdGRF5的剪切关系。苹果miR396家族成员在侧根和果实中的表达量显著高于其他组织,其候选靶基因表达量则在花芽和腋芽中显著高于其他组织;不定根发育过程中,miR396家族不同成员表达模式存在显著差异,整体上呈上调表达趋势,其候选靶基因呈下调表达趋势;外源IBA处理显著诱导miR396家族成员的表达,尤其是在不定根诱导期和根系生长期更为显著。

关键词: 苹果, miR396, 不定根发育, 生物信息学分析, 基因表达

Abstract: In this study the evolutionary characteristics of apple miR396 family and their expression pattern during adventitious root development were analyzed. The results demonstrated that the apple miR396 family contained 4 mature sequences and 7 precursor sequences. Mfold analysis showed that 7 precursor sequences of pre-miR396 members could form a typical stable stem loop structures,and their minimal folding free energies(ΔG)ranged from–62.9 to–51.9 kal ? mol-1. Phylogenetic tree analysis showed that the genetic relationship of pre-miR396 family in apple could be classified into 3 subgroups (G1,G2 and G3). According to its homology,each subgroup contained 11,9,19 members respectively. Prediction of target genes for apple miR396 family revealed that their targets were MdGRF1,MdGRF2 and MdGRF5 genes. The members of miR396 family showed significantly higher expression level in lateral roots and fruits as compared to other tissues. In addition,the candidate target genes kept higher expression level in flower buds and axillary buds than other tissues. During the process of adventitious root development,the expression patterns of different miR396 family members had significant differences,and overall expression was up-regulated,however,the candidate target genes were down-regulated during the adventitious root development. Exogenous IBA treatment significantly induced the expression of miR396 family members,especially in induction and root growth phases during adventitious root formation.

Key words: apple, miR396, adventitious root development, bioinformatics analysis, gene expression

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