匍匐翦股颖和多年生黑麦草对镉、铅胁迫的响应及部分基因表达调控机理研究
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摘要
土壤被重金属离子污染造成对植物生长发育不良影响，严重时会威胁到农、牧业的生产安全。在污染地区利用各种方法修复被污染的土壤，降低重金属污染物质对生物代谢和生长发育的威胁，已经成为环境科学、生态学研究的重点。由于部分草坪草能够在被污染的土壤中生长发育，通过对草坪草的反复刈割修剪，可以逐步减少土壤中的重金属离子含量，达到植物修复被污染土壤的目的。探索重金属污染物对草坪草的生态毒理作用、重金属离子胁迫时草坪草对胁迫的响应机理，发挥草坪草在未来污染地区植被恢复、污染土壤的植物修复中的作用，以及为将来培育草坪草抗性品种提供理论基础，进行本论文研究具有重要意义。
论文研究通过将匍匐翦股颖(Agrostis stolonifera)、多年生黑麦草(Lolium perenne)幼苗在Hoagland培养液中的培养，并将硝酸镉、醋酸铅溶液按一定浓度梯度混入培养液中，对培养的草坪草幼苗进行重金属离子胁迫处理，然后分别采用恒重法测定供试样品生物量的积累量；采用原子吸收分光光度法测定供试植物根系与地上茎叶样品中重金属离子积累量；采用荧光分光光度法测定供试样品叶片中二氯荧光黄双乙酸(DCF)的荧光值来间接反映叶片中产生的过氧化氢量的变化，来研究胁迫时活性氧(ROS)物质产生与积累情况；分别采用甲基氮蓝四唑(NBT)光密度测定法、高锰酸钾滴定法和愈创木酚法分别测定供试样品中超氧化物歧化酶(SODs)、过氧化氢酶(CAT)、过氧化物酶(POD)等细胞抗氧化酶活性变化；采用RNA和cDNA探针放射性标记对供试样品中的总RNA和poly(A)~+mRNA进行Northern印迹和杂交检验，来研究胁迫时相关基因表达谱的特征。从而系统探讨重金属离子胁迫对两种草坪草幼苗的生态毒理效应，草坪草在重金属胁迫响应时叶片中ROS形成和积累规律，以及ROS对细胞主要抗氧化酶活性的调节作用和ROS物质中的部分信使分子对主要抗氧化酶基因表达的调控作用。研究结果表明：
1．重金属离子胁迫可造成两种草坪草生物量积累的显著下降，其中当匍匐翦股
Heavy-metal ions stress was a one of adverse condition for plant grow, the metal ions accumulated by heavy metal pollutant in soil had been badly menaced agricultural environment. How to reduce the harm of the heavy metal ions on plant growth and metabolize, it had been a study focus at environment science and ecology. Because of some herbage plants and turfgrass could abate the heavy-metal ions and could grow in heavy-metals pollutant soil, it was significance to study of the toxicological effect of the metal ions on these plants, and the response mechanism and effect on some expression of the metal ions in these plants.
The infancy creeping bent (Agrostis stolonifers ) and perennial ryegrass (Lolium perenne) had been cultivated in Hoagland culture medium by using tank farming. When dealing the infancy lawn with heavy metal ions stress, the different concentration cadmium nitrate and lead acetate were mixed into the Hoagland culture medium. The identical biomass method was used to weigh in the biomass-production of the two species of plant. The fluorescence spectrophotometry method with testing dichlorofluorescein (DCF) in the leaf blade was conducted to determine the effect of the opposite contents of hydrogen peroxide (H_2O_2), a special reactive oxygen species (ROS) in leaf cells of the turfgrass. The photochemical NBT method, the potassium permanganate stactometer and guaiene
论文研究通过将匍匐翦股颖(Agrostis stolonifera)、多年生黑麦草(Lolium perenne)幼苗在Hoagland培养液中的培养，并将硝酸镉、醋酸铅溶液按一定浓度梯度混入培养液中，对培养的草坪草幼苗进行重金属离子胁迫处理，然后分别采用恒重法测定供试样品生物量的积累量；采用原子吸收分光光度法测定供试植物根系与地上茎叶样品中重金属离子积累量；采用荧光分光光度法测定供试样品叶片中二氯荧光黄双乙酸(DCF)的荧光值来间接反映叶片中产生的过氧化氢量的变化，来研究胁迫时活性氧(ROS)物质产生与积累情况；分别采用甲基氮蓝四唑(NBT)光密度测定法、高锰酸钾滴定法和愈创木酚法分别测定供试样品中超氧化物歧化酶(SODs)、过氧化氢酶(CAT)、过氧化物酶(POD)等细胞抗氧化酶活性变化；采用RNA和cDNA探针放射性标记对供试样品中的总RNA和poly(A)~+mRNA进行Northern印迹和杂交检验，来研究胁迫时相关基因表达谱的特征。从而系统探讨重金属离子胁迫对两种草坪草幼苗的生态毒理效应，草坪草在重金属胁迫响应时叶片中ROS形成和积累规律，以及ROS对细胞主要抗氧化酶活性的调节作用和ROS物质中的部分信使分子对主要抗氧化酶基因表达的调控作用。研究结果表明：
1．重金属离子胁迫可造成两种草坪草生物量积累的显著下降，其中当匍匐翦股
Heavy-metal ions stress was a one of adverse condition for plant grow, the metal ions accumulated by heavy metal pollutant in soil had been badly menaced agricultural environment. How to reduce the harm of the heavy metal ions on plant growth and metabolize, it had been a study focus at environment science and ecology. Because of some herbage plants and turfgrass could abate the heavy-metal ions and could grow in heavy-metals pollutant soil, it was significance to study of the toxicological effect of the metal ions on these plants, and the response mechanism and effect on some expression of the metal ions in these plants.
The infancy creeping bent (Agrostis stolonifers ) and perennial ryegrass (Lolium perenne) had been cultivated in Hoagland culture medium by using tank farming. When dealing the infancy lawn with heavy metal ions stress, the different concentration cadmium nitrate and lead acetate were mixed into the Hoagland culture medium. The identical biomass method was used to weigh in the biomass-production of the two species of plant. The fluorescence spectrophotometry method with testing dichlorofluorescein (DCF) in the leaf blade was conducted to determine the effect of the opposite contents of hydrogen peroxide (H_2O_2), a special reactive oxygen species (ROS) in leaf cells of the turfgrass. The photochemical NBT method, the potassium permanganate stactometer and guaiene
引文
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