影响静止中心（QC）细胞分裂的EMS突变体库构建与筛选分析.docVIP

影响静止中心（QC）细胞分裂的EMS突变体库构建与筛选分析 　　摘要：采用正向遗传学方法，以转基因拟南芥（Arabidopsis）SCRGVG UAS∶∶axr3-1为材料来进行EMS诱变。通过外源施加激素DEX诱导GVG/UAS系统启动axr3-1在静止中心表达，导致静止中心发生分裂。通过构建EMS突变体库，得到2 616份M2代候选突变体家系。对M2代种子进行初筛得到43个候选家系。经过复筛，获得22个具有稳定表型的突变体家系。根据表型相似性将候选家系分为 5类，共6 组。将候选家系进行组内杂交，根据 F1代植株表型是否与亲本相似，获得2对等位突变体。这些突变体将为研究干细胞小生境的调控机制研究提供新材料。 　　关键词：拟南芥（Arabidopsis）；根的发育；静止中心；EMS突变体库 　　中图分类号：Q291 文献标识码：A 文章编号：0439-8114（2016）05-1307-04 　　DOI：10.14088/j.cnki.issn0439-8114.2016.05.056 　　Construction and Screening of Arabidopsis EMS Mutant Library for the Effection of Divding Quiescent Center Cells 　　LIU Wen-gui，XU Jian 　　（National Key Laboratory of Crop Genetic Improvement，Huazhong Agricultural University，Wuhan 430070， China） 　　Abstract： A forward genetics approach were utilized to transgenic Arabidopsis SCRGVG UAS∶∶axr3-1 for EMS mutagenesis. The transgenic Arabidopsis SCRGVG UAS∶∶axr3-1 was expressed of axr3-1 specifically in the SCR-expressing cells in the presence of DEX， leading to cell divisions in the QC. An EMS mutant library was generated and a total of 2 616 M2 lines was obtained. Through a preliminarily screening of M2 lines，43 candidate lines were identified. A second round of screening was then performed and led to the identification of 22 candidate mutant lines that displayed stable phenotypes over generations. According to the phenotypic similarity， these candidate lines were divided into 5 categories，6 groups.And the candidate lines were hybridized in each groups. Based on the phenotypic similarity between the parental lines and F1 progeny，two pairs of allelic mutants were identified. The mutant materials would provide new materials for the study of mechanisms underlying the quiescence of the QC. 　　Key words： Arabidopsis； root development； quiescent center； EMS mutant library 　　在植物根冠与分生组织区交界处有一个根的组织中心（Organizing center），该处的细胞基本不发生分裂和分化，因此又称为静止中心（Quiescent center QC）。1954年Clowes[1]在玉米根尖中发现，顶端分生组织中最中央区域细胞分裂频率很低或不分裂。利用放射自显影技术，在玉米、蚕豆、洋葱根尖中分生组织区中，有一个区域细胞几乎没有发生DNA的合成，因而确定无细胞的分裂，被命