王丹丹,王鹏,韩智阳,刘丹梅.干旱胁迫对赤芍生理特性影响及相关基因的克隆表达[J].干旱地区农业研究,2022,40(5):71~80
干旱胁迫对赤芍生理特性影响及相关基因的克隆表达
Effects of drought stress on physiology characteristics of Radix paeoniae Rubra and clonal expression of related genes
  
DOI:10.7606/j.issn.1000-7601.2022.05.08
中文关键词:  赤芍  干旱胁迫  抗性生理  DREB基因  克隆  基因表达水平
英文关键词:Radix paeoniae Rubra  drought stress  resistance physiology  DREB gene  clone  gene expression level
基金项目:中央财政专项(851800501);辽宁省科学事业公益研究项目(20202681;20180550410)
作者单位
王丹丹 辽东学院农学院辽宁 丹东 118000 
王鹏 辽东学院农学院辽宁 丹东 118000 
韩智阳 辽东学院农学院辽宁 丹东 118000 
刘丹梅 辽东学院农学院辽宁 丹东 118000 
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中文摘要:
      为探究不同干旱程度对赤芍生长发育的影响及其抗旱机制,采用盆栽控水法,设置对照组、轻度干旱和重度干旱3个处理,测定赤芍叶片相对含水量、可溶性糖、可溶性蛋白、脱落酸、叶绿素、类胡萝卜素、类黄酮、丙二醛、游离脯氨酸含量、光合参数及超氧化物酶活性等生理指标,评价赤芍抗旱性;采用RT-PCR结合RACE技术克隆抗旱相关基因(DREB、Psb27、PsaK、MDHAR、SOD和GR),探究不同干旱程度对赤芍相关基因表达水平的影响。结果表明,重度干旱胁迫下,赤芍可溶性糖、可溶性蛋白、类胡萝卜素、类黄酮、脯氨酸、丙二醛、抗坏血酸及谷胱甘肽的含量均呈上升趋势,如在重度干旱和轻度干旱24 d后,赤芍叶片可溶性糖含量分别为对照的2.50倍和1.66倍,重度干旱胁迫第16天时,可溶性蛋白含量为对照组的2.43倍,类胡萝卜素与类黄酮的含量分别约为对照组的1.8倍和1.4倍,约为轻度干旱的1.4倍和1.2倍;而叶片相对含水量、蒸腾速率、气孔导度、净光合速率与叶绿素含量均降低,如重度干旱胁迫下赤芍叶片含水量不足20%,而轻度干旱和对照组叶片含水量均高于55%,叶绿素含量在第16天降到最低值,约为对照组的26%,为轻度干旱的33%;干旱胁迫下,光合作用相关基因Psb27、PsaK的表达量上调,且重度干旱上调水平更高;多个抗氧化基因的表达量(SOD、GR)、ABA含量以及胞间CO2浓度呈先升后降趋势,如SOD在重度干旱第16天表达水平达到峰值,分别为轻度干旱与对照组的1.58倍和1.86倍,而后剧烈下降,到第24天时表达水平降到对照组的1.43倍,其中MDHAR的表达量呈下降趋势,重度干旱第24天时表达水平降到对照组的5.53倍。
英文摘要:
      This study used potted water control method to explore the effects of different drought stresses on growth and drought resistance mechanism of Radix paeoniae. Three treatments of control, mild drought, and severs drought were used to test the leaf relative water content, soluble sugar, soluble protein, net photosynthetic rate, transpiration rate, stomatal conductance, intracellular CO2 concentration, ascorbic acid, chlorophyll, carotenoids, flavonoids, maltaraldehyde, free proline, and superoxidase dismutase to evaluate the drought resistance of Radix paeoniae. Six drought\|related genes (DREB、Psb27、PsaK、MDHAR、SOD and GR) were cloned by RT-PCR with RACE to explore the effects of different drought levels on gene expression levels of Radix paeoniae. The results showed that, under severe drought stress, the soluble sugar, soluble protein, carotenoids, flavonoids, proline, maltaraldehyde, ascorbic acid and glutathione all increased, for example, after 24 d of severe and mild drought, the soluble sugar content of Radix paeoniae was about 2.50 times and 1.66 times that of the control group, respectively. After 16 day of severe drought, the soluble protein content was about 2.43 times that of the control group, and the contents of carotenoid and flavonoid were separately about 1.8 times and 1.4 times that of the control group, and about 1.4 times and 1.2 times that of mild drought; while the leaf relative water content, the transpiration rate, stomatal conductivity, net photosynthetic rate and chlorophyll contentall decreased.For example, the leaf relative water content of Radix paeoniae was less than 20% under severe drought stress, while both mild drought and control groups were above 55%, and the chlorophyll content fell to the lowest value at the 16th day, about 26% of the control group and 33% of mild drought. Under severe drought stress, the expression of photosynthesis associated gene Psb27, PsaK was upregulated, and the severe drought upregulated level was higher than mild drought and control. The expression of most of antioxidant genes (SOD,GR), ABA content, and intracellular CO2 concentration all increased first then decreased, such as SOD expression levels peaked at the 16th day under severe drought, about 1.58 times and 1.86 times in the mild drought and the control group, respectively, and then the expression level decreased to 1.43 times by the 24th day of the control group. The expression level of MDHAR decreased, and at the 24th day under severe drought, its expression level decreased to 5.53 times that of the control group.
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