结球甘蓝类钙调蛋白基因CML48和CML50在不同胁迫下的表达分析

Expression Analysis of Calmodulin-like Protein Gene CML48 and CML50 under Different Stresses in Brassica oleracea L.var.capitata

为探究结球甘蓝类钙调蛋白(CMLs)响应不同胁迫时的功能,丰富CMLs参与钙信号网络调控,为CMLs参与植物逆境途径及其功能的研究提供依据。以结球甘蓝ZG为材料,克隆得到CML48和CML50基因,序列分析表明CML48开放阅读框(ORF)为672 bp,含4个内含子,编码223 aa,分子量为25.28 ku;CML50开放阅读框ORF为1095 bp,编码364 aa,分子量为38.06 ku,含3个内含子;均为亲水蛋白,均含有2个EF-hand结构域;系统发育树表明CML48和CML50均与甘蓝处于同一进化枝,与白菜、油菜的亲缘关系最近。qPCR表明CML48和CML50在干旱(PEG6000)、盐胁迫(NaCl)、低温(4℃)、高温(37℃)、H_(2)O_(2)、脱落酸(ABA)、水杨酸(SA)和茉莉酸甲酯(MeJA)等不同胁迫处理下,在叶和根中的表达均有上调,且CML50的表达量均显著高于CML48,CML48在茎尖中高度响应高温胁迫,而在根中不响应高温和H_(2)O_(2)胁迫。初步说明CML48和CML50在根和叶中均可响应上述8种非生物胁迫。

In order to explore the function of calmodulin-like proteins(CMLs)in response to abiotic stress in Brassica oleracea L.,enrich CMLs’participation in calcium signal network regulation,and provide evidences for CMLs’participation in plant stress pathways and study of their functions.In this study,CML48 and CML50 were cloned from Brassica oleracea L.The ORF of CML48 is 672 bp,containing 4 introns,encoding 223 aa with molecular mass of 25.28 ku.And the ORF of CML50 is 1095 bp,encoding 364 aa,with molecular mass of 38.06 ku and three introns.Both of them are hydrophilic proteins,and all of which contain two EF-hand domains.Phylogenetic tree showed that CML48 and CML50 with cabbage were in the same evolutionary branch,and were closely related to Brassica nupus and Brassica rape.qPCR showed that the expressions CML48 and CML50 could up-regulate in leaves and roots under stresses of drought(PEG6000),salt(NaCl),low temperature(4℃),high temperature(37℃),H_(2)O_(2),abscisic acid(ABA),salicylic acid(SA)and methyl jasmonate(MeJA).And the expressions of CML50 was significantly higher than that of CML48,which was highly responsive to high temperature stress in shoot tip,but not responsive to high temperature and H_(2)O_(2) stress in root.The results indicated that CML48 and CML50 responded to 8 kinds of abiotic stress above both in leaves and roots.This study is expected to provide reference for response of CML48 and CML50 to plant stress signal pathways and the research of their functions.