摘要
为了探究梨花芽休眠进程中miRNA的表达模式和靶基因,利用Solexa测序技术、生物信息学分析和实时荧光定量PCR(qPCR)技术,对内休眠、内休眠解除和生态休眠解除3个时期梨花芽的miRNA进行高通量测序、筛选和鉴定。结果表明,内休眠、内休眠解除和生态休眠解除时期3个样本文库中分别有12 276 226、10 135 952、11 453 981条Unique序列。miRNA主要分布在21～24 nt之间,其中长度为24 nt的数量最多。共检测到151个已知的miRNA,属于39个不同的家族,并利用生物信息学软件预测到了209个新的miRNAs。比较分析从内休眠进入到生态休眠解除的整个休眠转换时期差异表达的miRNA,筛选出8个miRNA(ahy-miR156b-5p、cpa-miR319、aly-miR172c-3p、aau-miR396、mdm-miR858、aly-miR171b-3p、bdi-miR160f和hbr-miR166a),其靶基因主要参与转录调控、信号传导等过程。利用qPCR验证了8个miRNA及其8个靶基因的表达。
In order to explore the expression pattern and target genes of microRNAs associated with dormancy of pear flower buds,we used Solexa sequencing,bioinformatics analysis and qPCR to screen and identify the small RNAs of pear flower buds at stages of endodormancy,endodormancy release and ecodormancy release. The results showed that there were 12 276 226,10 135 952 and 11 453 981 unique reads in endodormancy,endodormancy release and ecodormancy release libraries,respectively. The size distribution of small RNAs mainly ranged from 21–24 nt,with 24 nt small RNAs being the most abundant. A total of 151 known miRNAs were identified from three samples,which belong to 39 different families,and 209 novel miRNAs were predicted using bioinformatics software. Through comparative expression profiling of the miRNAs during the transition period from endodormancy to ecodormancy release,we found 8 differentially expressed miRNAs(ahy-miR156b-5p,cpa-miR319,aly-miR172c-3p, aau-miR396,mdm-miR858,aly-miR171b-3p,bdi-miR160f and hbr-miR166a). The predicted target genes of the 8 miRNAs were mainly involved in transcription regulation,signal transduction and so on. Finally,we confirmed the differential expression of the 8 miRNAs and 8 target genes by qPCR.
作者
马鑫瑞
李亮
刘瑾航
杨梦洁
陈洁
梁沁
吴少华
李永裕
MA Xinrui;LI Liang;LIU Jinhang;YANG Mengjie;CHEN Jie;LIANG Qin;wu Shaohua;LI Yongyu(College of Horticulture,Fujian Agriculture and Forestry University,Institute of Natural Products of Horticultural Plants,Fujian Agriculture and Forestry University,Fuzhou 350002,China)
出处
《园艺学报》
CAS
CSCD
北大核心
2018年第11期2089-2105,共17页
Acta Horticulturae Sinica
基金
国家自然科学基金项目(31501694)
高等学校博士学科点专项科研基金项目(2011351511011).
