NSFC-31271739
国家自然科学基金面上项目
玉米幼胚胚性愈伤组织诱导过程中重要miRNA及靶转录因子的功能研究，国家自然科学基金面上项目，2013.01-2016.12，主持；
玉米幼胚胚性愈伤组织诱导过程中重要miRNA及靶转录因子的功能研究
执行时间 2013-01 至 2016-12
中文关键词 玉米幼胚;胚性愈伤组织;miRNA;转录因子;功能
英文关键词 Immature maize embryo;Embryonic callus;miRNA;Transcript factor;Function
中文摘要：幼胚胚性愈伤组织诱导是玉米转基因育种的重要技术环节，而我国具有高胚性愈伤诱导率的骨干自交系极少，阻碍了转基因玉米育种的快速发展。前期研究以高胚性愈伤诱导率自交系18-599R为材料，通过 深度测序挖掘了4个在胚性愈伤诱导整个过程中急剧下调的miRNA家族，经靶基因预测结合数字表达谱分析和qRT-PCR验证结果，确定了3个转录因子作为以上4个miRNA家族的候选靶基因，而这3个转录因子基因在染色体上的位置与前期定位的胚性愈伤诱导率QTL位点是一致的。本项目拟首先采取5’RACE技术对miRNA剪切靶转录因子的精确位点进行鉴定；同时分别利用胚特异过表达和RNAi技术对以上miRNA和转录因子进行功能验证，获得控制玉米胚性愈伤诱导过程的miRNA和转录因子基因；并通过ChIP-Seq在全基因组水平上挖掘靶转录因子的互作基因，结合基因功能注释构建由miRNA介导的幼胚胚性愈伤诱导过程的分子调控网络。
英文摘要：Introduction of embryonic callus is an essential process in maize transgenic breeding. Low introduction efficiency for most of maize elite inbred lines prevents maize transgenic breeding from rapid development. In our previous study, the high-introduction effeiciency inbred line 18-599R was conducted to small RNA deep sequencing. As a result, 4 miRNA families were displayed to sharply down-regulated during the embryonic callus introduntion. Three transcript factors were respectively decided to be the candidate target genes of the four miRNAs by bioinformatic prediction and digital gene expression profiling. Further analysis revealed that the loci of the transcript factor genes is consistent with the QTL controlling the introduction of maize embryonic callus identified in the previous research. In the project, the accurate cutting site of target mRNA by the miRNAs will be identified using 5’RACE. The function of the miRNAs and targets involved in embryonic callus induction will be determined by embryo-spesific overexpression of the miRNAs and RNAi of the transcript factor genes. In addition, the interaction genes of the three transcript factors will be genome-widely revealed by ChIP-Seq technique. Then the regulatory network involved in the process of maize embryonic callus introduction will be constructed according to gene annotations.
成果：
- 一种新型的转基因专用茎尖分生组织侵染皿 作者：沈亚欧，杨珊，林海建，刘红军，潘光堂，缪林 成果类型：专利 授权机构：国家知识产权局 授权日期：2013/5/27 授权区域：中国
- Genome-wide analysis of transcription factors involved in maize embryonic callus formation 作者：Lujiang Li|Zhiming Zhang|Guangtang Pan|Yaou Shen| 成果类型：期刊论文 收录情况：无 期刊名称：Physiologia Plantarum 发表日期：2016
- 植物ＤＮＡ甲基化调控研究进展 作者：江舟|黄小波|沈亚欧|潘光堂| 成果类型：期刊论文 收录情况：无 期刊名称：中国生物工程杂志 发表日期：2013
- Combined small RNA and degradome sequencing reveals microRNA regulation during immature maize embryo dedifferentiation 作者：Zhiming Zhang|Maojun Zhao|Tingzhao Rong|Guangtang Pan| 成果类型：期刊论文 收录情况：无 期刊名称：BBRC 发表日期：2013
- Haploid Strategies for Functional Validation of Plant Genes 作者：Yaou Shen|Guantang Pan|Thomas Lubberstedt| 成果类型：期刊论文 收录情况：无 期刊名称：Trends in Biotechnology 发表日期：2015