Cd~(2+)对BY-2细胞的毒性机制及水杨酸的缓解作用
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摘要
镉对生态环境的危害表现出对植物这种定生生物的毒害性。以绿色荧光蛋白(GFP)膜泡标记的烟草BY-2细胞为实验材料,分别在荧光显微镜和激光共聚焦显微镜下观察了Cd2+对植物细胞的毒害作用和机制,并研究了水杨酸(SA)处理对Cd2+植物细胞毒害的缓解作用。研究发现激光共聚焦显微镜可以观察到Cd2+处理3 h后细胞荧光亮度减弱,6 h时细胞皱缩和液泡缩小,9 h细胞大多死亡。在Cd2+胁迫同时加入SA,可显著提高细胞成活时间,荧光亮度增强、液泡化程度加大,同时能够观察到荧光标记的细胞膜包裹Cd2+的高荧光颗粒。SA处理的细胞还通过高度液泡化来缓解重金属Cd2+的毒性。结果表明具有生物活性的SA诱导细胞的液泡化,并通过膜成分对重金属的包裹和结合进一步降低了重金属对植物细胞的毒害。因此水杨酸缓解重金属对植物细胞的毒性,是通过诱导细胞的液泡化和膜对重金属离子的包裹束缚而实现的。
The toxicity to immobilized plants is one of the most harmful effects of heavy mental cadmium to the ecological environment. In this paper the toxicity and mechanism of Cd2 +to plants cells were analyzed by fluorescence microscope and confocal laser scanning microscope observation in GFP membrane labeled BY-2 cell. The relief effects of salicylic acid( SA) treatment to the Cd2 +toxicity were also investigated. The results showed that the GFP fluorescence of the labeled cells was getting dimming after 3 h Cd2 +treatment. The cells and vacuoles were apparently shrunk after 6 h. All the cells were dead after 9 h treatment. When SA was also added during the Cd2 +treatment,the survival time of the treated cells were prolonged significantly. The SA and Cd2 +treated cells also showed stronger fluorescence and better vacuolization compared to the Cd2 +treatment. High fluorescence particles attached to the cell membrane were observed,indicating that there were wrapped Cd2 +microcapsules formed in SA treatment cells. The results suggest that the SA treatment can relieve the toxicity of heavy metals both by high vacuolization inthe cells and by membrane components retention of the heavy metals ion. The conclusion is that the microcapsule retention and vacuolization of the cells induced by SA is the mechanism of the SA toxicity relief.
The toxicity to immobilized plants is one of the most harmful effects of heavy mental cadmium to the ecological environment. In this paper the toxicity and mechanism of Cd2 +to plants cells were analyzed by fluorescence microscope and confocal laser scanning microscope observation in GFP membrane labeled BY-2 cell. The relief effects of salicylic acid( SA) treatment to the Cd2 +toxicity were also investigated. The results showed that the GFP fluorescence of the labeled cells was getting dimming after 3 h Cd2 +treatment. The cells and vacuoles were apparently shrunk after 6 h. All the cells were dead after 9 h treatment. When SA was also added during the Cd2 +treatment,the survival time of the treated cells were prolonged significantly. The SA and Cd2 +treated cells also showed stronger fluorescence and better vacuolization compared to the Cd2 +treatment. High fluorescence particles attached to the cell membrane were observed,indicating that there were wrapped Cd2 +microcapsules formed in SA treatment cells. The results suggest that the SA treatment can relieve the toxicity of heavy metals both by high vacuolization inthe cells and by membrane components retention of the heavy metals ion. The conclusion is that the microcapsule retention and vacuolization of the cells induced by SA is the mechanism of the SA toxicity relief.
引文
[1]和莉莉,李冬梅,吴钢.我国城市土壤重金属污染研究现状和展望[J].土壤通报,2008,39(5):1211-1216He L L,Li D M,Wu G.Heavy metal contamination of urban soils in China:State and prospect[J].Chinese Journal of Soil Science,2008,39(5):1211-1216(in Chinese)
[2]王宏镔,束文圣,蓝崇钰.重金属污染生态学研究现状与展望[J].生态学报,2005,25(3):597-605Wang H B,Shu W S,Lan C Y.Ecology for heavy metal pollution:Recent advances and future prospects[J].Acta Ecologica Sinica,2005,25(3):597-605(in Chinese)
[3]郑小东,荣湘民,罗尊长,等.土壤重金属污染及修复方法研究进展[J].农学学报,2011,37-43Zheng X D,Rong X M,Luo Z Z,et al.Research progress of heavy metal contamination in soil and remediation method[J].Journal of Agriculture,2011,37-43(in Chinese)
[4]代全林.重金属对植物毒害机理的研究进展[J].亚热带农业研究,2006,2(2):49-53Dai Q L.A review of toxicant mechanism s of heavy metals against plants[J].Subtropical Agriculture Research,2006,2(2):49-53(in Chinese)
[5]孔祥海.重金属离子对植物的毒害及其机理[J].龙岩学院学报,2005,23(3):83-87Kong X H.Toxcity of heavy metal ions to plants and their toxicological mechanism[J].Journal of Longyan University,2005,23(3):83-87(in Chinese)
[6]孟雪娇,邸昆,丁国华.水杨酸在植物体内的生理作用研究进展[J].中国农学通报,2010,26(15):207-214Meng X J,Di K,Ding G H.Progress of study on the physiological role of salicylic acid in plant[J].Chinese Agricultural Science Bulletin,2010,26(15):207-214(in Chinese)
[7]Ananieva E A,Christov K N,Popova L P.Exogenous treatment with salicylic acid leads to increased antioxidant capacity in leaves of barley plants exposed to paraquat[J].Plant Physiology,2004,161:319-328(in Chinese)
[8]印莉萍,罗思羽,闫莉婕,等.缺铁水稻根细胞膜泡相关基因群的分析与电镜观察[J].电子显微学报,2005,24(5):489-493Yin L P,Luo S Y,Yan L J,et al.Gene groups analysis relevant to membrane vesicles and electron microscopic observation of rice roots under Fe-deficiency condition[J].Journal of Chinese Electron Microscopy Society,2005,24(5):489-493
[9]Mc Cabe P F,Leave C J.Programmed cell death in cell cultures[J].Plant Molecular Biology,2000,44:359-368
[10]李春烨,丁国华,刘保东.重金属影响植物细胞超微结构和功能的研究进展[J].中国农学通报,2013,29(18):114-118Li C Y,Ding G H,Liu B D.Research progress of heavy metal affecting plant cell ultrastructure and function[J].Chinese Agricultural Science Bulletin,2013,29(18):114-118(in Chinese)
[11]Liu D H,Kottke I.Subcellular localization of Cd in the root cells of Allium sativum by electron energy loss spectroscopy[J].Biosciences,2003,28(4):471-478
[12]Lidon F C,Henriques F S.Role of rice shoot vacuoles in copper toxicity regulation[J].Environmental Experiment Botany,1998,39:197-202
[13]郝怀庆,施国新,杜开和,等.镉污染对水鳖的毒害影响[J].西北植物学报,2001,21(6):1237-1240Hao H Q,Shi G X,Du K H,et al.Toxic effect of cadmium on hydrocharis dubia[J].Acta Botanica Boreali-Occidentalia Sinica,2001,21(6):1237-1240(in Chinese)
[14]宇克莉,孟庆敏,邹金华.镉对玉米幼苗生长、叶绿素含量及细胞超微结构的影响[J].华北农学报,2010,25(3):118-123Yu K L,Meng Q M,Zou J H,Effects of Cd2+on seedling growth,chlorophyll contents and ultrastructures in maize[J].Acta Agriculturae Boreali-Sinica,2010,25(3):118-123(in Chinese)
[15]魏志琴,陈志勇,秦蓉,等.Cu2+对拟南芥根的局部毒性及诱导DNA损伤和细胞死亡[J].植物学报,2013,48(3):303-312Wei Z Q,Chen Z Y,Qin R,et al.Cu2+induced local toxicity and DNA damage,cell death in roots of arabidopsis thaliana[J].Chinese Bulletin of Botany,2013,48(3):303-312(in Chinese)
[16]关伟,张金珠,王占全,等.镉胁迫对桃树根尖细胞超微结构的影响[J].北京农学院学报,2010,25(3):18-25Guan W,Zhang J Z,Wang Z Q,et al.Effect of cadmium stress on the ultrastructure of cell in peach root apex[J].Journal of Beijing University of Agriculture,2010,25(3):18-25(in Chinese)
[17]赵新月.水杨酸和钙对镉胁迫下玉米毒害修复的生理生态基础研究[D].西安建筑科技大学硕士学位论文,2013Zhao X Y.Study on the physiological and ecological base of exogenous salicylic acid and calcium in repairing maize by cadmium[D].Dissertation for Doctor Degree of Xi`an University of Architecture and Technology,2013(in Chinese)
[18]常云霞,徐克东,张小芳,等.外源水杨酸对Hg2+胁迫下小麦幼苗生长及生理的影响[J].麦类作物学报,2014,34(4):540-545Chang Y X,Xu K D,Zhang X F,et al.Effects of salicylic acid on the growth and physiological property of wheat seedlings under Hg2+stress[J].Journal of Triticeae Crops,2014,34(4):540-545(in Chinese)
[19]王林华,梁书荣,吕淑敏,等.外源水杨酸与植物非生物胁迫抗性的关系及其作用机制[J].河南农业科学,2010,8:160-164
[20]Popova L P,Maslenkova L T,Yordanova R Y,et al.Exogenous treatment with salicylic acid attenuates cadmium toxicity in pea seedlings[J].Plant Physiology Biochemistry,2009,47(3):224-231
[21]李丰涛.红麻对重金属的吸收特征及外源GSH缓解镉毒的机理研究[D].福建农业大学博士学位论文,2013Li F T.The study on accumulation characteristics of heavy metals and alleviating mechanism of exogenous GSH in kenaf[D].Dissertation for Doctor Degree of Fujian Agriculture and Forestry University,2013(in Chinese)
[2]王宏镔,束文圣,蓝崇钰.重金属污染生态学研究现状与展望[J].生态学报,2005,25(3):597-605Wang H B,Shu W S,Lan C Y.Ecology for heavy metal pollution:Recent advances and future prospects[J].Acta Ecologica Sinica,2005,25(3):597-605(in Chinese)
[3]郑小东,荣湘民,罗尊长,等.土壤重金属污染及修复方法研究进展[J].农学学报,2011,37-43Zheng X D,Rong X M,Luo Z Z,et al.Research progress of heavy metal contamination in soil and remediation method[J].Journal of Agriculture,2011,37-43(in Chinese)
[4]代全林.重金属对植物毒害机理的研究进展[J].亚热带农业研究,2006,2(2):49-53Dai Q L.A review of toxicant mechanism s of heavy metals against plants[J].Subtropical Agriculture Research,2006,2(2):49-53(in Chinese)
[5]孔祥海.重金属离子对植物的毒害及其机理[J].龙岩学院学报,2005,23(3):83-87Kong X H.Toxcity of heavy metal ions to plants and their toxicological mechanism[J].Journal of Longyan University,2005,23(3):83-87(in Chinese)
[6]孟雪娇,邸昆,丁国华.水杨酸在植物体内的生理作用研究进展[J].中国农学通报,2010,26(15):207-214Meng X J,Di K,Ding G H.Progress of study on the physiological role of salicylic acid in plant[J].Chinese Agricultural Science Bulletin,2010,26(15):207-214(in Chinese)
[7]Ananieva E A,Christov K N,Popova L P.Exogenous treatment with salicylic acid leads to increased antioxidant capacity in leaves of barley plants exposed to paraquat[J].Plant Physiology,2004,161:319-328(in Chinese)
[8]印莉萍,罗思羽,闫莉婕,等.缺铁水稻根细胞膜泡相关基因群的分析与电镜观察[J].电子显微学报,2005,24(5):489-493Yin L P,Luo S Y,Yan L J,et al.Gene groups analysis relevant to membrane vesicles and electron microscopic observation of rice roots under Fe-deficiency condition[J].Journal of Chinese Electron Microscopy Society,2005,24(5):489-493
[9]Mc Cabe P F,Leave C J.Programmed cell death in cell cultures[J].Plant Molecular Biology,2000,44:359-368
[10]李春烨,丁国华,刘保东.重金属影响植物细胞超微结构和功能的研究进展[J].中国农学通报,2013,29(18):114-118Li C Y,Ding G H,Liu B D.Research progress of heavy metal affecting plant cell ultrastructure and function[J].Chinese Agricultural Science Bulletin,2013,29(18):114-118(in Chinese)
[11]Liu D H,Kottke I.Subcellular localization of Cd in the root cells of Allium sativum by electron energy loss spectroscopy[J].Biosciences,2003,28(4):471-478
[12]Lidon F C,Henriques F S.Role of rice shoot vacuoles in copper toxicity regulation[J].Environmental Experiment Botany,1998,39:197-202
[13]郝怀庆,施国新,杜开和,等.镉污染对水鳖的毒害影响[J].西北植物学报,2001,21(6):1237-1240Hao H Q,Shi G X,Du K H,et al.Toxic effect of cadmium on hydrocharis dubia[J].Acta Botanica Boreali-Occidentalia Sinica,2001,21(6):1237-1240(in Chinese)
[14]宇克莉,孟庆敏,邹金华.镉对玉米幼苗生长、叶绿素含量及细胞超微结构的影响[J].华北农学报,2010,25(3):118-123Yu K L,Meng Q M,Zou J H,Effects of Cd2+on seedling growth,chlorophyll contents and ultrastructures in maize[J].Acta Agriculturae Boreali-Sinica,2010,25(3):118-123(in Chinese)
[15]魏志琴,陈志勇,秦蓉,等.Cu2+对拟南芥根的局部毒性及诱导DNA损伤和细胞死亡[J].植物学报,2013,48(3):303-312Wei Z Q,Chen Z Y,Qin R,et al.Cu2+induced local toxicity and DNA damage,cell death in roots of arabidopsis thaliana[J].Chinese Bulletin of Botany,2013,48(3):303-312(in Chinese)
[16]关伟,张金珠,王占全,等.镉胁迫对桃树根尖细胞超微结构的影响[J].北京农学院学报,2010,25(3):18-25Guan W,Zhang J Z,Wang Z Q,et al.Effect of cadmium stress on the ultrastructure of cell in peach root apex[J].Journal of Beijing University of Agriculture,2010,25(3):18-25(in Chinese)
[17]赵新月.水杨酸和钙对镉胁迫下玉米毒害修复的生理生态基础研究[D].西安建筑科技大学硕士学位论文,2013Zhao X Y.Study on the physiological and ecological base of exogenous salicylic acid and calcium in repairing maize by cadmium[D].Dissertation for Doctor Degree of Xi`an University of Architecture and Technology,2013(in Chinese)
[18]常云霞,徐克东,张小芳,等.外源水杨酸对Hg2+胁迫下小麦幼苗生长及生理的影响[J].麦类作物学报,2014,34(4):540-545Chang Y X,Xu K D,Zhang X F,et al.Effects of salicylic acid on the growth and physiological property of wheat seedlings under Hg2+stress[J].Journal of Triticeae Crops,2014,34(4):540-545(in Chinese)
[19]王林华,梁书荣,吕淑敏,等.外源水杨酸与植物非生物胁迫抗性的关系及其作用机制[J].河南农业科学,2010,8:160-164
[20]Popova L P,Maslenkova L T,Yordanova R Y,et al.Exogenous treatment with salicylic acid attenuates cadmium toxicity in pea seedlings[J].Plant Physiology Biochemistry,2009,47(3):224-231
[21]李丰涛.红麻对重金属的吸收特征及外源GSH缓解镉毒的机理研究[D].福建农业大学博士学位论文,2013Li F T.The study on accumulation characteristics of heavy metals and alleviating mechanism of exogenous GSH in kenaf[D].Dissertation for Doctor Degree of Fujian Agriculture and Forestry University,2013(in Chinese)