摘要 | 以‘金魁’猕猴桃(Actinidia deliciosa ‘Jinkui’)组培苗及 2 年生扦插苗为实验材料,采用 qRT-PCR 技术对 0. 1mmol• L-1水杨酸(SA)、0. 05 mmol• L-1茉莉酸甲酯(MeJA)、0. 01 mmol• L-1 1-氨基环丙烷-1-羧酸(ACC)和 0. 01mmol• L-1脱落酸(ABA)4 种植物激素处理后 0、 4、 12 和 48 h,低温(4 ℃ )和 0. 2 mol• L-1 NaCl 胁迫 0、4、12 和 48h,高温(48 ℃ )胁迫 0、2 和 4 h 及恢复培养 6 h,以及干旱胁迫 14 d 后叶片中 AdRAV1、AdRAV2 和 AdRAV3 基因的相对表达量进行了测定。 结果显示:不同处理条件下 3 个 AdRAVs 基因相对表达量的变化存在一定差异。 SA、MeJA和 ACC 处理后 4 和 12 h,AdRAV1 基因的相对表达量显著(P<0. 05)升高,但 ABA 处理后该基因的相对表达量无明显变化;SA、MeJA、ACC 和 ABA 处理后 48 h,AdRAV2 基因的相对表达量显著降低;4 种植物激素处理后 4、12 和 48h,AdRAV3 基因的相对表达量总体上显著降低。 低温胁迫下,AdRAV1 和 AdRAV2 基因的相对表达量无明显变化,但胁迫48 h 时 AdRAV3 基因的相对表达量却显著升高。 NaCl 胁迫 12 h 时,AdRAV1 和 AdRAV2 基因的相对表达量均显著升高, 而 AdRAV3 基因的相对表达量则显著降低。 高温胁迫 4 h 时, AdRAV1 基因的相对表达量显著降低,AdRAV2 基因的相对表达量显著升高;胁迫 2 h 时,AdRAV3 基因的相对表达量显著降低;恢复培养 6 h 时,3 个基因的相对表达量均无法恢复至起始水平。 干旱胁迫 14 d 后,AdRAV1 基因的相对表达量显著高于对照(正常浇水);AdRAV2 的相对表达量高于对照,而 AdRAV3 基因的相对表达量则低于对照,且均与对照无显著差异。 研究结果表明:不同胁迫条件对‘金魁’猕猴桃 AdRAVs 基因的表达特性有不同诱导效应。 根据实验结果,推测 AdRAV1、AdRAV2和 AdRAV3 基因可能参与 SA、MeJA、ACC 和 ABA 信号转导途径以及耐盐和耐高温过程;并且,AdRAV1 基因还可能参与耐旱过程,而 AdRAV3 基因则可能参与耐寒过程。 |
Abstract | Taking tissue culture seedling and two-year-old cutting seedling of Actinidia deliciosa ‘Jinkui’ as experimental materials, relative expression of AdRAV1, AdRAV2 and AdRAV3 genes in leaf after 0, 4,12 and 48 h treated by four phytohormones including 0.1 mmol · L-1 salicylic acid (SA),0.05mmol·L-1methyl jasmonate (MeJA), 0.01 mmol·L-1 1-aminocyclopropane-1-carboxylic acid (ACC) and 0. 01 mmol · L-1 abscisic acid ( ABA), stressed by low temperature (4 ℃ ) and 0. 2 mol· L-1 NaCl for 0, 4, 12 and 48 h, stressed by high temperature (48 ℃ ) for 0, 2 and 4 h and recovery culturing for 6 h, and stressed by drought for 14 d were determined by qRT-PCR technique. The results show that there are some differences in change of relative expression of three AdRAVs genes under different treatment conditions. After 4 and 12h treated by SA, MeJA and ACC, relative expression of AdRAV1 gene increases significantly (P<0. 05), but it does not change obviously after treated by ABA. After 48 h treated by SA, MeJA, ACC and ABA, relative expression of AdRAV2 gene decreases significantly. After 4, 12 and 48h treated by four phytohormones, relative expression of AdRAV3 gene decreases significantly as a whole. Under low temperature stress, relative expression of AdRAV1 and AdRAV2 genes has no obvious change, but that of AdRAV3 gene increases significantly when stressing for 48 h. When NaCl stressing for 12h, relative expression of AdRAV1 and AdRAV2 genes increases significantly, while that of AdRAV3 gene decreases significantly. When high temperature stressing for 4 h, relative expression of AdRAV1 gene decreases significantly, that of AdRAV2 gene increases significantly; when stressing for 2h, that of AdRAV3 gene decreases significantly; when recovery culturing for 6 h, that of three genes all cannot return to the starting level. After drought stressed for 14d, relative expression of AdRAV1 gene is significantly higher than that of the control (normal watering); relative expression of AdRAV2 gene is higher than that of the control, while relative expression of AdRAV3 gene is lower than that of the control, both of them have no significant difference with the control. It is suggested that different stress conditions have different inductive effects on expression characteristics of AdRAVs gene from A. deliciosa ‘Jinkui’. According to the experimental results, it is speculated that AdRAV1, AdRAV2 and AdRAV3 genes may be involved in SA, MeJA, ACC and ABA signal transduction pathway and resistance process of salt and high temperature, and AdRAV1 gene also may be involved in drought resistance process, while AdRAV3 gene does in cold resistance process. |
关键词 | ‘金魁'猕猴桃; AdRAVs 基因; 植物激素; 环境胁迫; 相对表达量; qRT-PCR 技术 |
Key words | Actinidia deliciosa ‘Jinkui’; AdRAVs gene; phytohormone; environmental stress; relative expression; qRT-PCR technique |
作者 | 张计育1, 黄胜男1, 程竞卉2, 王刚1, 潘德林1, 郭忠仁1 |
所在单位 | 1. 江苏省中国科学院植物研究所(南京中山植物园), 江苏 南京 210014; 2. 延边朝鲜族自治州种子管理站, 吉林 延吉 133001 |
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基金项目 | 国家自然科学基金资助项目(31401854); 江苏省自然科学基金资助项目(BK20140760); 江苏省科技计划项目( BE2015350); 泰州市科技支撑计划(农业)项目(TN201511) |