盐胁迫对盐芥悬浮培养细胞超微结构的影响

摘要/Abstract

摘要： 以盐芥悬浮细胞为实验材料建立“悬浮细胞/盐胁迫”体系,并运用透射电镜技术对盐胁迫下盐芥悬浮细胞超微结构的变化进行了观察。结果表明,未胁迫下,盐芥细胞中各细胞器结构完整; 300 mM NaCl胁迫6 h后,开始出现质壁分离,细胞质具有较高的电子密度; 胁迫12 h后,出现明显的质壁分离现象,线粒体数目增多,多数变成月牙形 ,内部模糊不清,高尔基体失去正常形态,变得松散,“反式”面分泌小泡以“出芽”方式出现,同时发现胞内有“吞噬泡”存在。实验说明盐芥悬浮细胞能短时间耐受高浓度(300 mM)的氯化钠胁迫,当胁迫时间延长为12 h,细胞受损严重,亚细胞水平表现在细胞超微结构的改变,细胞“自我吞噬”可能在盐芥细胞盐胁迫响应过程中发挥重要作用。

关键词: 自我吞噬, 盐胁迫, 悬浮培养细胞, 盐芥

Abstract:

Thellungiella halophila suspension-cultured cells were used to establish a 'cell culture/salt stress' system, and electronic transmission microscope was used to examine ultrastructural changes of Thellungiella halophila cells. Organelles remained normal structures in untreated cells, while the plasma membrane began to detach from the cell wall and the cytoplasm appeared electron-opaque when cells were stressed under 300 mM NaCl for 6 hours. The plasma membrane retracted apparently from the cell wall after salt treatment for 12 hours. The number of mitochondria increased quickly and mitochondria were frequently crescent with illegible cristae at this time. Golgi body lost its normal morphology and became loose with small secretion vesicles budding from the trans end. We also found the presence of autophagic vacuoles in salt stressed cells. These results indicate that Thellungiella halophila suspension-cultured cells can tolerate high salinity stress (300 mM NaCl) for a short time, while severe salt stress (300 mM NaCl for 12 h) leads to the impairment of cell with apparent ultrastructural changes. We suggest that autophagy may play an important role in Thellungiella halophila cells in response to salt stress.

Key words: autophagy, salt stress, suspension-cultured cells, Thellungiella halophila

中图分类号:

Q945.78

王进;李新荣. 盐胁迫对盐芥悬浮培养细胞超微结构的影响[J]. 中国沙漠, 2011, 31(4): 878-883.

WANG Jin;LI Xin-rongShapotou . Effect of Salt Stress on Ultrastructure of Thellungiella halophila Suspension-cultured Cells[J]. JOURNAL OF DESERT RESEARCH, 2011, 31(4): 878-883.

参考文献

[1]Flowers T J,Garcia A,Koyama M,et al.Breeding for salt tolerance in crop plants—the role of molecular biology[J].Acta Physiologiae Plantarum,1997,19:427-433.
[2]Katsuhara M,Kawasaki T.Salt stress induced nuclear and DNA degradation in meristematic cells of barley roots[J].Plant Cell Physiology,1996,37:169-173.
[3]Ashraf M.Breeding for salinity tolerance proteins in plants[J].Critical Reviews in Plant Sciences,1994,13:17-42.
[4]Blumwald E,Grover A.Salt tolerance[M]//Halford N G.Plant biotechnology:current and future uses of genetically modified crops.John Wiley & Sons Ltd.,UK,2006:206-224.
[5]Zhu J K.Genetic analysis of plant salt tolerance using Arabidopsis thaliana[J].Plant Physiology,2000,124:941-948.
[6]张有福,陈拓,费贯清,等.盐度对三种荒漠植物渗透调节物质积累影响的研究[J].中国沙漠,2007,27(5):787-790.
[7]刘建新,胡浩斌,赵国林.NaCl胁迫对骆驼蓬幼苗液泡膜H+-ATPase和H+-PPase活性的影响[J].中国沙漠,2008,28(2):274-279.
[8]刘玉冰,赵昕,谭会娟.荒漠植物红砂愈伤组织诱导与增殖研究[J].中国沙漠,2008,28(2):255-258.
[9]Epimashko S,Schliebs E F,Christian A L,et al.Na+/H+-transporter,H+-pumps and an aquaporin in light and heavy tonoplast membranes from organic acid and NaCl accumulating vacuoles of the annual facultative CAM plant and halophyte Mesembryanthemum crystallinum L[J].Planta,2006,224:944-951.
[10]胡月楠,贺康宁,王占林,等.盐胁迫对霸王水势的影响[J].中国沙漠,2009,29(5):905-910.
[11]赵昕,盛芬玲,赵敏桂,等.NaCl胁迫下盐芥和拟南芥化合物含量与蛋白质结构变化比较——傅立叶红外光谱法[J].应用与环境生物学报,2008,14(3):371-377.
[12]Estrella R V,Barkla B J,Ramírez L G,et al.Salt Stress in Thellungiella halophila activates Na+ transport mechanisms required for salinity tolerance[J].Plant Physiology,2005,139:1507-1517.
[13]Amtmann A,Bohnert H J,Bressan R A.Abiotic stress and plant genome evolution.Search for new models[J].Plant Physiology,2005,138:127-130.
[14]王淑莉,赵昕,谭会娟.荒漠植物花棒耐盐性的傅立叶红外光谱法研究[J].中国沙漠,2008,28(5):874-878.
[15]赵可夫,李法增.中国盐生植物[M].北京:科学出版社,1999:173-177.
[16]李妍.耐盐研究模式植物——盐芥[J].生物学通报,2007,42(2):22-23.
[17]刘爱荣,赵可夫.盐胁迫下盐芥渗透调节物质的积累及其渗透调节作用[J].植物生理与分子生物学报,2005,31(4):389-395.
[18]Volkov V,Wang B,Dominy P J,et al.Thellungiella halophilla,a salt relative of Arabidopsis thaliana,possesses effective mechanisms to discriminate between potassium and sodium[J].Plant Cell & Environment,2004,27:1-14.
[19]Inan G,Zhang Q,Li PH,et al.Salt cress,a halophyte and cryophyte Arabidopsis relative model system and its application to molecular genetic analyses of growth and development of extremophiles[J].Plant Physiology,2004,135:1718-1737.
[20]Wong C E,Li Y,Whitty B R,et al.Expressed sequence tags from the Yukon ecotype of Thellungiella reveal that gene expression in response to cold,drought and salinity shows little overlap[J].Plant Molecular Biology,2005,58:561-574.
[21]Murashige T, Skoog F.A revised medium for rapid growth and bioassays with tobacco tissue culture[J].Plant Physiology,1962,15:463-497.
[22]Orzaez D,de Jong A J,Woltering E J.A tomato homologue of the human protein PIRIN is induced during programmed cell death[J].Plant Molecular Biology,2001,46:459-468.
[23]史密斯 H.植物细胞分子生物学[M].北京:科学出版社,1986:67-147.
[24]Crawford S A,Wildens S.Ultrastructural aspects of damage to leaves of Eucalyptus camaldulengis by the psyllid cardiaspina rotator[J].Micron,1996,27:359-366.
[25]Zuppini A,Baldan B,Millioni R,et al.Chitosan induces Ca2+-mediated programmed cell death in soybean cells[J].New Phytologist,2004,161:557-568.
[26]Kourtis N,Tavernarakis N.Autophagy and cell death in model organisms[J].Cell & Differentiation,2009,16:21-30.
[27]Kroemer G,El-Deiry W S,Golstein P,et al.Classification of cell death:recommendations of the nomenclature committee on cell death[J].Cell Death & Differentiation,2005,12:1463-1467.
[28]Arunika H L,Gunawardena A N,Pearce D M,et al.Characterisation of programmed cell death during aerenchyma formation induced by ethylene or hypoxia in roots of maize (Zea mays L.)[J].Planta,2001,212:205-214.
[29]Liu Y,Xiong Y,Bassham D C.Autophagy is required for tolerance of drought and salt stress in plants[J].Autophagy,2009,5:954-963.
[30]Sanders D.Plant biology:The salty tale of Arabidopsis[J].Current Biology,2000,10:486-488.