植物激素处理和环境胁迫对‘金魁’猕猴桃AdRAVs基因表达特性的影响

Effects of phytohormone treatment and environmental stress on expression characteristics of AdRAVs gene from Actinidia deliciosa ‘Jinkui '

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0.1 mmol·L-1水杨酸(SA)、0.05 mmol·L-1茉莉酸甲酯(Me JA)、0.01 mmol·L-11-氨基环丙烷-1-羧酸(ACC)和0.01 mmol·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-1Na Cl胁迫0、4、12和48 h,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、Me JA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、Me JA、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48 h,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。Na Cl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、Me JA、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。

Taking tissue culture seedling and two-year-old cutting seedling of Actinidia deliciosa ‘ Jinkui ’ as experimental materials,relative expression of AdRAVl, 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^-1 methyl jasmonate （ MeJA） , 0. 01 mmol·L^-1 1 -aminocyclopropane-1 -carboxylic acid （ACC） and 0. 01mmol·L^-1 abscisic acid （ABA）, stressed by low temperature （4 ℃）and 0. 2 mmol·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 12 h treated by SA, MeJA and ACC, relative expression of AdRAVl gene increases significantly （P 〈 0 .0 5 ） , 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 decreasessignificantly. After 4, 12 and 48 h treated by four phytohormones, relative expression of AdRAV3 gene decreases significantly as a whole. Under low temperature stress, relative expression of AdRAVl and AdRAV2 genes has no obvious change, but that of AdRAV3 gene increases significantly when stressing for 48 h. When NaCl stressing for 12 h, relative expression of AdRAVl and AdRAV2 genes increases significantly, while that of AdRAV3 gene decreases significantly. When high temperature stressing for 4 h, relative expression of AdRAVl gene decreases significantly, that of AdRAV2 gene increases significantly; when stressing for 2 h, 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 14 d, relative expression of AdRAVl 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 issuggested that different stress conditions have different inductive effects on expression characteristics ofAdRAVs gene from A. deliciosa ‘ Jinkui ’ . According to the experimental results, it is speculated that AdRAVl, 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 AdRAVl gene also may be involved in drought resistance process, while AdRAV3 gene does in cold resistance process.