降解组测序技术在玉米雌穗发育相关miRNA靶基因中的应用

Research for using degradome sequencing method to find involved miRNA target genes in the developing maize ear

采用降解组测序技术对昌7-2,郑58 2个玉米自交系雌穗不同发育阶段miRNA的靶基因进行了检测,共获得47个被16个miRNA家族95个成员剪切的靶标。这些靶标多数存在于细胞核内,参与转录调控、细胞分化等生物学过程;分子功能多为DNA结合、特异序列DNA结合转录因子活性、金属离子结合、蛋白结合、ATP结合等.结合荧光实时定量PCR技术构建了昌7-2,郑58雌穗的花序分生组织(IM)分化为成对小穗分生组织(SPM)、成对小穗分生组织分化为小穗分生组织(SM)、小花分生组织(FM)3个时期的miRNA靶标表达谱。研究结果表明,在玉米雌穗发育过程中存在依赖于miRNAs的基因调控网络。

Degradome sequencing technology was used to indentify miRNA target genes in genomewide at the different developmental stages of maize ear of chang 7- 2 and zheng 58. And 47 cleavage targets corresponding to 95 conserved miRNAs belonging to 16 RNA family were found. Bioinformatic analysis showed that most of these targets existed in nucleus participating in transcription regulation,cell differentiation and the molecular function most for DNA binding,sequence-specific DNA binding transcription factor activity,metal ion binding,protein binding,and ATP binding etc. We used Chang 7- 2and Zheng 58 at the inflorescence meristems（ IM） phase,the IM produces spikelet pair meristems（ SPM） period,and floral meristems（ FM） as the experimental materials to construct the expression map through real-time fluorescence quantitative PCR techniques. The results of this study indicated that there was miRNA-mediated regulation network of gene expression at maize ear development stage.