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葡萄根系和叶片对氯化镉处理的生理学与细胞学响应

作者:代写论文  来源:http://www.starlunwen.com/  发布时间:2010-05-03 16:49:59

【中文摘要】 以当年生葡萄扦插苗为试材,通过水培和盆栽试验,研究了镉在葡萄根、茎和叶中的分布,比较了20个葡萄品种对镉的富集与迁移,分析了氯化镉(CdCl2)处理下葡萄根系活力、丙二醛含量、根系与叶片质膜ATPase活性和线粒体特性的变化以及活性氧和一氧化氮(NO)生成等的变化规律;同时,利用流式细胞仪、吖啶橙荧光染色和类caspase3/7活性检测与分析等技术,研究了氯化镉处理下葡萄根系细胞程序性死亡(programmed cell death, PCD),探讨了外源水杨酸和氯化钙对葡萄根系镉伤害的缓解作用。结果表明:1.镉在葡萄根、茎、叶中的分布与累积存在明显差异,但总体呈现根>叶>茎的规律。20个葡萄品种对镉的富集及镉由根系向叶片的迁移存在明显差异。镉在根系的富集系数在39.18%~135.76%(其中紫珍香最高,维多利亚最低)、在茎的富集系数在2.55%~14.32%(其中红地球最高,玫瑰香最低)、在叶片的富集系数在12.06%~54.61%之间(其中红地球最高,克瑞森无核最低);镉由根系向叶片的迁移系数(叶/根)在0.16~1.12之间(其中早艳最高,紫珍香最低)。20个葡萄品...更多种对镉的吸收总量由大到小的顺序是:巨峰>紫珍香>红地球>玫瑰香>矢富罗莎>红宝石无核>泽香>奥迪亚无核>美人指>瑰宝>克瑞森无核>乍娜>森田尼无核>早艳>皇家秋天>早黑宝>黄金指>达米娜>龙眼>维多利亚。2.在0~1mmol·L-1浓度范围内,随着氯化镉处理浓度的增加,泽香葡萄根系活力、根系和叶片NO含量、一氧化氮合酶(NOS)活性、Ca2+-ATPase活性先升高后降低,O2-·产生速率、H2O2含量、丙二醛含量逐渐升高,质膜H+-ATPase活性逐渐降低。其中氯化镉浓度在0.1mmol·L-1时,葡萄根系活力、质膜Ca2+-ATPase活性显著提高;在0.5 mmol·L-1和1 mmol·L-1时质膜ATPase活性、NOS活性、根系活力和NO含量显著下降。3.用0.5 mmol?L-1氯化镉处理乍娜、玫瑰香、达米娜、瑰宝、泽香、红地球、巨峰、奥迪亚无核、皇家秋天等9个葡萄品种,它们的根系活力和质膜ATPase活性均降低,根系O2-·产生速率和H2O2含量、丙二醛含量均升高,除奥迪亚无核外NO含量和NOS活性均升高。巨峰葡萄根系活力比其他品种更易受到氯化镉影响,奥迪亚无核葡萄根系活力最不敏感,受氯化镉影响最小;9个品种根系活力下降幅度由大到小的顺序是:巨峰>皇家秋天>玫瑰香>瑰宝>红地球>乍娜>达米娜>泽香>奥迪亚无核。4.在0~1mmol·L-1浓度范围内,随着氯化镉处理浓度的增加,泽香葡萄根系线粒体膜通透性(MPT)增大、膜电位(Δψm)下降、H2O2含量逐渐升高、细胞色素(ccytochrome c, Cyt c)含量逐渐下降。流式细胞仪检测结果表明,随着氯化镉处理浓度升高根系细胞凋亡率提高。细胞程序性死亡关键酶类caspase3/7的活性也随着CdCl2浓度的增加而提高,研究表明:0.5mmol?L-1和1.0mmol?L-1的氯化镉明显诱导泽香葡萄根系发生细胞程序性死亡,吖啶橙染色检测结果也证实了这一点。5.泽香、龙眼、玫瑰香和巨峰4个葡萄品种一年生扦插苗经0.5mmol?L-1氯化镉处理后,根系、叶片线粒体H2O2含量升高,MPTP开放程度增大,Δψm降低,Cyt c含量下降,根系活力明显降低,类caspase3/7的活性显著提高。其中,根系线粒体H2O2含量和Cyt c含量由高到低的顺序为:巨峰>泽香>玫瑰香>龙眼;根系活力、MPTP和Δψm由高到低的顺序为:龙眼>玫瑰香>泽香>巨峰;细胞凋亡率由高到低为:巨峰>泽香>玫瑰香>龙眼。表明巨峰葡萄根系比其他品种更易受到氯化镉的伤害而发生细胞程序性死亡。6.较低浓度(10μmol·L-1和50μmol·L-1)水杨酸预处理显著降低了1.0mmol·L-1氯化镉处理下泽香葡萄根系O2-·、H2O2和NO的生成,阻止了根系活力和质膜H+-ATPase和Ca2+-ATPase活性下降;而较高浓度(100μmol·L-1和200μmol·L-1)水杨酸促进了氯化镉处理下自由基的生成,进一步降低了根系活力和质膜H+-ATPase和Ca2+-ATPase活性,表明低浓度水杨酸可通过降低自由基而减轻氯化镉对葡萄根系功能的损伤,而高浓度水杨酸则会加重氯化镉的伤害。7.较低浓度(1mmol·L-1和5mmol·L-1)氯化钙显著降低了1.0mmol·L-1氯化镉处理下泽香葡萄根系O2-·、H2O2和NO的生成,阻止了根系活力和质膜H+-ATPase和Ca2+-ATPase活性下降,缓解CdCl2对葡萄根系造成的伤害;而较高浓度(10mmol·L-1)氯化钙则促进了氯化镉处理下根系自由基的生成,进一步降低了根系活力和质膜H+-ATPase和Ca2+-ATPase活性,加重了氯化镉对葡萄根系功能的伤害。 

【英文摘要】 The experiment was carried out with the cutting seedlings of grape under hydroponics or pot culture. In this experiment, efforts have been made to investigate the following aspects; the compartment of cadmium in root, stem and leaf were studied; the cadmium accumulation coefficient and migration coefficient in twenty varieties of grape were compared; root activity, MDA content, the activity of ATPase in PM of root and leaf, the character of mitochondria and the generation of ROS and NO were detected. Meanwhile, with the analytic system of flow cytometry, detective techonogy of acridine orange fluorescent staining and the activity detection and analysis of caspase3/7. The PCD characters of grape roots were studied. Furthermore, the alleviating effect of exogenous SA and CaCl2 on Ze Xiang grape under cadmium were also discussed.The main results were as follows:1. The compartment and accumulation of cadmium in root, stem and leaf were significantly different. The accumulation rule was: ro...ot >leaf > stem. In the twenty varieties of grapes chosen for the experiment, the accumulation coefficient in roots ranged from 39.18% to 135.76% (Zi Zhen-xiang grape being the highest and Victoria the lowest), the accumulation coefficient in stem ranged from 2.55% to 14.32% (Red Globe grape being the highest and Muscat Hamburg grape the lowest), the accumulation coefficient in leaf ranged from 12.06% to 54.61% (Ruby Seedless grape being the highest and Crimson Seedless the lowest) and the migration coefficient of cadmium from root to leaf ranged from 0.16 to 1.12(Zao Yan being the highest and Zi Zhen-xiang the lowest).The total assimilation amount in each of the twenty varieties of grape was in the order as below: Kyoho>Zi Zhen-xiang>Red Globe>Muscat Hamburg>Yatomi Rosa>Ruby Seedless>Ze Xiang>Qtilia Seedless>Manicure Finger>Gui Bao>Crimson Seedless>Zana>Centennial Seedless>Zao Yan>Autumn Royal>Zao He-Bao>Golgen Finger>Tamina>Long Yan>Victoria.2. Within the range of 0~1mmol·L-1, with the increase of CdCl2 concentration, root activity of Ze Xiang, the content of NO in roots and leaves, the activity of NOS and the activity of Ca2+-ATPase rose first and then dropped down; MDA content、O2-·generation rate and H2O2 content had a consistant increase while plasma membrane H+-ATPase had a consistant decrease in roots of Ze Xiang grape. It’s worth noticing that the root activity and plasma membrane Ca2+-ATPase activity increased significantly at 0.1 mmol·L-1 CdCl2 in roots of Ze Xiang and plasma membrane ATPase activity, NOS activity, root activity and NO content decreased significantly at 0.5 mmol·L-1 or 1 mmol·L-1 CdCl2 .3. When Zana, Muscat Hambur, Tamina, Gui Bao, Ze Xiang, Red Globe, Kyoho, Qtilia Seedless and Autumn Royal were treated with 0.5 mmol?L-1 CdCl2, the root activity and plasma membrane ATPase activity decreased and the root MDA content、O2-·generation rate and H2O2 content increased. Apart from Otilia Seedless, the content of NO and the activity of NOS in all the rest varieties increased. The root activity of Kyoho was more easily affected by cadmium than the other varieties. The root activity of Qtilia Seedless was the least sensitive and so was least affected by cadmium. The decrease range of root activities for the nine varieties were discribed as below: Kyoho>Autumn Royal>Muscat Hambur>Gui Bao>Red Globe>Zana>Tamina>Ze Xiang>Qtilia Seedless.4. With the increase of CdCl2 concentration within the range of 0~1mmol·L-1, root MPT increased,Δψm decreased, H2O2 content increased, Cyt c content decreased in the root of Ze Xiang grape. The results of flow cytometry indicated that with the increase in treatment concentration, the root cell PCD rate also increased, and the activity of caspase3/7, which is the key enzyme of PCD, too, increased with the increasing of CdCl2 concentration. The result showed that the PCD of grape root cells were induced significantly by CdCl2, which was also reconfirmed by the detection of acridine orange fluorescent staining.5. After treating the roots with 0.5 mmol?L-1 CdCl2, the mitochondrial hydrogen peroxide (H2O2) content increased, mitochondrial permeability transition pores (MPTP) increased, mitochondrial membrane potential (Δψm) decreased, cytochrome c (Cyt c) in roots and root activity decreased and the activity of caspase3/7 increased in the one-year cutting-seedling of 4 grape cultivars (Kyoho, Muscat Hamburg, Long Yan, and Ze Xiang). The order of H2O2 and Cyt c contents was as below: Kyoho>Ze Xiang>Muscat Hamburg>Loing yan the order of root activity, MPTP andΔψm was: Long Yan>Muscat Hamburg>Ze Xiang>Kyoho, and the order of apoptosis rates was: Kyoho>Ze Xiang>Muscat Hamburg>Long Yan. The results indicated that the roots of Kyoho were more easily damaged by CdCl2 than other varieties and thus induced apoptosis.6. Pretreatment of SA at a lower concentration (10μmol·L-1 and 50μmol·L-1) greatly decreased the formation of O2-·, H2O2 and NO, which retarded the declining of root activity and the activity of H+-ATPase and Ca2+-ATPase in PM under 1.0 mmol·L-1 CdCl2 treatment. On the other hand, pretreatment of SA at a higher concentration (100μmol·L-1 and 200μmol·L-1) promoted the formation of free radicals under CdCl2 treatment and further decreased root activity and the activity of H+-ATPase and Ca2+-ATPase in PM. The results indicate that Salicylic acid at a lower concentration can alleviate the damage of CdCl2 to the function of grape roots by inhibiting the formation of free radicals, while at a higher concentration it will aggravate the damage of CdCl2 to grape roots.7. CaCl2 treatment at a lower concentration (1mmol·L-1 and 5mmol·L-1) greatly decreased the formation of O2-·, H2O2 and NO, which retarded the declining of root activity and the activity of H+-ATPase and Ca2+-ATPase in PM under 1.0 mmol·L-1 CdCl2 treatment, alleviated the damage of CdCl2 to the function of grape roots. On the other hand, CaCl2 treatment at a higher concentration (10 mmol·L-1) promoted the formation of free radicals under CdCl2 treatment and further decreased root activity and the activity of H+-ATPase and Ca2+-ATPase in PM, thus aggravated the damage of CdCl2 to grape roots. 

【中文关键词】 葡萄; 氯化镉; 生理反应; 线粒体; 细胞程序性死亡
【英文关键词】 Grape; Chloride cadmium; Physiological response; Mitochondria; PCD


本文TAGS:葡萄根系和叶片对氯化镉处理的生理学与细胞学响应 ——代写农艺学论文
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