【摘要】 为了探究CiMYB33基因在盐胁迫下的功能，以野菊（Chrysanthemum indicum）叶片DNA为模板，克隆了CiMYB33的全长序列，并利用生物信息学软件，与其他植物的MYB33基因进行比对，并构建进化树。通过蘸花法将该基因遗传转化拟南芥（Arabidopsis thaliana），并通过实时荧光定量PCR鉴定，获得转基因植株。对野生型和过表达CiMYB33转基因拟南芥种子进行不同盐浓度(0、75、100和150 mmol·L^-1NaCl)处理，并分析种子发芽率。对幼苗施用200 mmol·L^-1NaCl，观察表型变化和生理指标变化，并分析其耐盐性。系统发育分析结果表明，CiMYB33蛋白与青蒿（Artemisia annua）GAMYB蛋白同源性最高。盐胁迫条件下的研究结果显示，CiMYB33过表达拟南芥表现出显著增强的抗逆性特征，其种子发芽率和幼苗生长状态与野生型相比均呈现明显优势；CiMYB33过表达拟南芥的游离脯氨酸、叶绿素质量分数和相对含水量均高于野生型，相对电导率和丙二醛质量摩尔浓度则低于野生型；CiMYB33过表达拟南芥显示出良好的膜修复能力，与野生型相比，产生较为轻微的细胞膜受损。因此，野菊CiMYB33基因能够增强其异源过表达拟南芥的耐盐能力。更多还原

【Abstract】 To explore the function of CiMYB33 gene under salt stress, the full-length sequence of CiMYB33 was cloned using the DNA from leaves of Chrysanthemum indicumas the template. Bioinformatics software was used to align it with MYB33 genes from other plants and construct a phylogenetic tree. The gene was genetically transformed intoArabidopsis thalianavia the floral dip method, and transgenic plants were obtained and identified by real-time fluorescence quantitative PCR. Seeds of wild-type and CiMYB33-overexpressing transgenic A. thalianawere treated with different salt concentrations(0, 75, 100 and 150 mmol·L^-1NaCl), and the seed germination rate was analyzed. The seedlings were treated with 200 mmol·L^-1NaCl, and the changes in phenotype and physiological indexes were observed to analyze the salt tolerance. Results of phylogenetic analysis showed that CiMYB33 protein had the highest homology with GAMYB protein from Artemisia annua. The results under salt stress showed that CiMYB33-overexpressing A. thaliana exhibited significantly enhanced stress resistance characteristics, and its seed germination rate and seedling growth status showed obvious advantages compared with the wild type. The contents of free proline, chlorophyll and relative water content in CiMYB33-overexpressing A. thaliana were higher than those in the wild type, while the relative conductivity and malondialdehyde molar concentration were lower than those in the wild type.CiMYB33-overexpressing A. thaliana showed good membrane repair ability, with relatively slight cell membrane damage compared with the wild type. Therefore, the CiMYB33 gene from C. indicumcan enhance the salt tolerance of heterologously overexpressed A. thaliana.更多还原