细叶百合Catalase基因的克隆及表达分析  

Cloning and Expression Analysis of Catalase Gene in Lilium pumilum

作　　者：宋煜 林文昊 荆一博 董懿 金淑梅 SONG Yu;LIN Wenhao;JING Yibo;DONG Yi;JIN Shumei(Key Laboratory of Saline-alkai Vegetation Ecology Restoration,Ministry of Education,College of Life Sciences,Northeast Forestry University,Harbin 150040;Aulin College,Northeast Forestry University,Harbin 150040;College of Forestry,Northeast Forestry University,Harbin 150040)

机构地区：[1]东北盐碱植被恢复与重建教育部重点实验室,东北林业大学生命科学学院,哈尔滨150040 [2]东北林业大学奥林学院,哈尔滨150040 [3]东北林业大学林学院,哈尔滨150040

出　　处：《植物研究》2023年第5期756-767,共12页Bulletin of Botanical Research

基　　金：东北林业大学大学生创新训练计划项目资助(180602011)。

摘　　要：为探究细叶百合(Lilium pumilum)Catalase(CAT)基因与耐盐碱胁迫的关系,从细叶百合鳞茎中成功克隆出LpCat基因。该基因的ORF区长度为1479 bp,编码492个氨基酸,对其进行序列比对和系统进化树分析,发现细叶百合Catalase蛋白与通江百合(L.sargentiae)、菠萝(Ananas comosus)、油棕(Elaeis guineensis)等植物的Catalase蛋白亲缘关系较近。构建了pQE-LpCat原核表达载体,以1 mol·L^(-1)IPTG为诱导剂进行LpCat蛋白的体外诱导表达与纯化。在添加50 mmol·L^(-1)NaHCO_(3)胁迫下,携带PQE-LpCat蛋白的菌液生长浓度高于对照菌株。在1 mol·L^(-1)NaHCO_(3)、2.5 mol·L^(-1)H_(2)O_(2)胁迫下过表达LpCAT基因烟草(Nicotiana plumbaginifolia)植株与野生型相比,枯萎程度相对较低。通过净光合速率、气孔导度、胞间CO_(2)和蒸腾速率,H_(2)O_(2)含量,丙二醛(MDA)等生理指标检测说明过表达LpCat基因烟草植株比野生型烟草植株更耐盐碱。To explore the relationship between the Catalase(CAT)gene and saline-alkali stress tolerance of Lilium pumilum,the CAT gene was successfully cloned from L.pumilum bulb.The length of region ORF was 1479 bp,encoding 492 amino acids,and sequence alignment and phylogenetic tree analysis were performed,and the Catalase protein of L.pumilum was found to be closely related to the Catalase proteins of L.sargentiae,Ananas comosus,Elaeis guineensis and other plants.The Catalase protein was induced to express,and purified in vitro with 1 mol·L^(-1)IPTG as the inducer after constructing prokaryotic expression vector pQE-LpCat.The results showed that the growth concentration of bacteria solution containing PQE-LpCat protein was higher than that of the control strain under the stress of 50 mmol·L^(-1)NaHCO_(3).Under the stress of 1 mol·L^(-1)NaHCO_(3)and 2.5 mol·L^(-1)H_(2)O_(2),less wilting was observed in plants overexpressing the LpCAT gene compared to wild-type.Determination of the physiological indexes including net photosynthetic rate,stomatal conductance,intercellular CO_(2)and transpiration rate,H_(2)O_(2)concentration and contents of malondialdehyde(MDA)showed that tobacco plants overexpressing the LpCat gene were more tolerant to saline and alkali stress than wild-type tobacco plants.

关 键 词：细叶百合 Catalase基因 盐碱逆境 

分 类 号：Q949.71[生物学—植物学]