%0 Journal Article %T 菊花转录因子CmMYB59的克隆与表达特性分析 %A 王萃铂 %A 张瓒 %A 张晓雪 %A 展妍丽 %A 亓钰莹 %A 蒋甲福 %J 南京农业大学学报 %D 2016 %R 10.7685/jnau.201503029 %X [目的]研究菊花转录因子CmMYB59的功能,阐释其对冷胁迫的影响。[方法]以菊花品种‘神马’为试验材料,采用RACE和RT-PCR技术克隆得到一个MYB转录因子的cDNA全长及其可变剪切;采用实时荧光定量PCR法研究了CmMYB59在不同组织器官及低温胁迫下的表达,通过酵母单杂交验证了其转录激活的活性,同时通过洋葱表皮细胞瞬时表达系统进行亚细胞定位。[结果]该基因全长904 bp,开放阅读框为768 bp,编码255个氨基酸,蛋白质相对分子质量为61.99×103。生物信息学分析表明,此基因为典型的R2R3型MYB转录因子,具有保守的结构域和调控基序,因与拟南芥的AtMYB59同源性较高,故命名为CmMYB59。亚细胞定位表明CmMYB59蛋白在细胞核上表达。酵母转录激活活性试验表明CmMYB59具有转录激活活性。荧光定量PCR分析其表达模式,发现CmMYB59在根中表达量最高,茎、叶中次之,茎尖和花器官中最低;CmMYB59对低温胁迫有响应。[结论]初步推测,CmMYB59可能与细胞分裂相关,参与根的发育过程,与冷胁迫相关。</br>[Objectives]We had a preliminary study on the function of CmMYB59 and explained its response to the cold stress. [Methods]In this study,a novel MYB and its alternative splicing transcript were isolated from Chrysanthemum morifolium‘Jinba’by RT-PCR and RACE methods. The expression of CmMYB59 in different tissues and cold treatment conditions was studied by qRT-PCR. Furthermore,its transcriptional activation and subcellular localization were analyzed by the yeast one-hybrid assay and transient expression in onion epidermal cells respectively. [Results]The full length of this gene was 904 bp,containing an open reading frame of 768 bp,encoding 255 amino acids,with relative molecular mass of 61.99×103. The multiple alignment of the protein sequences showed the protein had a typical R2R3-MYB domain and was most closed to AtMYB59. Thus,the gene was named CmMYB59. qRT-PCR showed that the expression of CmMYB59 was the highest in roots,higher in stems and leaves,the lowest in shoot tip and floral organ,and it was responsive to the cold stress. The subcellular localization indicated that the protein was located in nucleus. The yeast one-hybrid assay showed that the CmMYB59 protein had a transcriptional activation function. [Conclusions]It is speculated that CmMYB59 might be related to cell division,participating in the development process of root,and being associated with cold stress %K 菊花 %K MYB转录因子 %K 基因克隆 %K 亚细胞定位 %K 荧光定量PCR %K 转录激活< %K /br> %K Chrysanthemum morifolium %K MYB transcription factor %K gene cloning %K subcellular localization %K qRT-PCR %K transcriptional activity %U http://nauxb.njau.edu.cn/oa/darticle.aspx?type=view&id=201601009