收稿日期: 2006-12-04
录用日期: 2007-07-11
网络出版日期: 2008-05-30
基金资助
国家自然科学基金(30540056);浙江省自然科学基金(304185);浙江省自然科学基金(303461);浙江省自然科学基金(504135)
PHYTOECOLOGICAL EFFECT OF Al3+ ON THE INDUCTIVITY OF PROGRAMMED CELL DEATH OF BORDER CELLS IN SOYBEAN ROOT
Received date: 2006-12-04
Accepted date: 2007-07-11
Online published: 2008-05-30
设置不同的Al3+浓度(0、25、50、100、200、400 μmol·L -1)和培养时间(12、24 h),研究了边缘细胞活性和大豆(Glycine max)根中过氧化氢酶(CAT)、过氧化物酶(POD)、超氧化物歧化酶(SOD)随Al3+浓度及处理时间变化的规律,并通过Hoechst33342-PI双重荧光染色、梯状DNA(即DNA ladder)分析和末端脱氧核糖核酸转移酶介导的dUTP切口末端标记(即TUNEL原位标记)检测,研究了Al3+对大豆根边缘细胞程序性死亡诱导的生理生态作用。结果表明,Al3+胁迫能诱导边缘细胞的死亡,随着Al3+浓度的升高和处理时间的延长,细胞死亡率增加。通过Hoechst33342-PI双重荧光染色、DNA ladder分析和TUNEL原位标记,检测到Al3+胁迫下发生程序性死亡的边缘细胞。其表现为:在400 μmol·L -1 Al3+诱导大豆根24 h时, 核酸电泳显示细胞DNA发生特异性降解并形成阶梯状电泳条带(DNA ladder),用TUNEL原位标记检测200和400 μmol·L -1 Al3+ 处理12 h后的大豆根边缘细胞,发现DNA的3'-OH端被原位特异标记,二氨基联苯胺(DAB)显色后,细胞核为阳性或强阳性。同时,高浓度Al3+(>100 μmol·L-1)处理下,CAT、POD和SOD活性均有不同程度的下降,CAT和SOD的活性也随处理时间的延长而降低。说明在Al3+胁迫下边缘细胞的死亡可能是一种程序性死亡形式,高浓度Al3+胁迫下,通过诱导活性氧在细胞体内的产生和累积而导致细胞凋亡,此过程是其对逆境胁迫所作出的生理生态防御性应答方式之一。
李荣峰, 蔡妙珍, 刘鹏, 徐根娣, 陈敏燕, 梁和 . Al3+对大豆根边缘细胞程序性死亡诱导的生理生态作用[J]. 植物生态学报, 2008 , 32(3) : 690 -697 . DOI: 10.3773/j.issn.1005-264x.2008.03.019
Aims Programmed cell death (PCD) plays an important role in plant growth and development, which correlates with adaptation of the plant to environment stress. Root border cells have many important biological functions in protecting the plant root tip from biotic and abiotic stress, which is a popular research topic recently. However, few studies have focused on PCD of root border cells and the phytoecological effect of aluminum on it. Our objective was to study the phytoecological and molecular ecological mechanism of root border cells in resisting aluminum (Al) toxicity.
Methods We investigated change in viability of border cells, activities of catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD), and PCD of root border cells by Al3+ induced in soybean (Glycine max) root tips with different Al3+ concentrations (0, 25, 50, 100, 200 and 400 μmol·L -1 Al3+) and treatment times (12 and 24 h). PCD was observed through Hoechst33342-PI fluorescent staining, DNA ladder and TdT-mediated dUTP nick end labeling (TUNEL) analysis.
Important findings Aluminum can induce the death of root border cells. The viability of border cells decreases with increased Al3+ concentration and treatment time. The progressive delineation of fragmented DNA was coincident with the appearance of DNA ladder after being exposed to 400 μmol·L -1 Al3+ for 24 h. TUNEL analysis of border cells revealed that the nuclear DNA strand breaks can be identified by labeling free 3'-OH termini after treatment with 200 and 400 μmol·L -1 Al3+ for 12 h. Diaminobenzidine (DBA) staining indicated that the nucleus was positive and strong positive. Otherwise, CAT and SOD activities declined with increasing Al3+ concentration and treatment time under higher Al3+ concentration (>100 μmol·L-1), and we observed no significant differences in POD activity among different Al3+ concentrations and treatment times. These results indicated that border cell death may be a PCD under Al3+ stress. High Al3+ concentration induced PCD through enhancing reactive oxygen species (ROS), which is one of the ways of resisting adversity in plants.
| [1] | Amako K, Chen GX, Asade K (1994). Separate assays specific for ascorbate peroxidase and guaiacol peroxidase and for the chloroplastic and cytosolic isozymes of ascorbate peroxidase in plants. Plant & Cell Physiology, 35,497-504. |
| [2] | Boscolo PRS, Menossi M, Jorqe RA (2003). Aluminum-induced oxidative stress in maize. Phytochemistry, 62,181-189. |
| [3] | Cakmak I, Horst WJ (1991). Effect of aluminum on lipid peroxidation, superoxide dismutase, catalase, and peroxidase activities in root tips of soybean (Glycine max). Physiologia Plantarum, 834,463-468. |
| [4] | Cai MZ (蔡妙珍), Liu P (刘鹏), Xu GD (徐根娣), Liu HH (刘海华) (2006). Effect of Al3+ toxicity on root border cells in vitro of buckwheat. Journal of Jiangsu University (Natural Science Edition) (江苏大学学报(自然科学版)), 27,293-298. (in Chinese with English abstract) |
| [5] | Cai MZ (蔡妙珍), Liu P (刘鹏), Xu GD (徐根娣), Liu WX (刘文秀), Gong CF (龚春风) (2007). Response of root border cells to Al3+ toxicity in soybean . Scientia Agricultura Sinica (中国农业科学), 40,271-276. (in Chinese with English abstract) |
| [6] | Campbell A, Prasad KN, Bondy SC (1999). Aluminum-induced oxidative events in cell lines: glioma are more responsive than neuroblastoma. Free Radical Biology & Medicine, 26,1166-1171. |
| [7] | de Jong AJ, Hoeberichts FA, Yakimova ET, Maximova E, Woltering EJ (2000). Chemical-induced apoptotic cell death in tomato cells: involvement of caspase-like proteases. Planta, 211,656-662. |
| [8] | de Jong AJ, Elena TY, Veneta MK, Ernist JW (2002). A critical role for ethylene in hydrogen peroxide release during programmed cell death in tomato suspension cells. Planta, 214,537-545. |
| [9] | Du X (杜幸), Liu P (刘鹏), Xu GD (徐根娣), Cai MZ (蔡妙珍) (2006). Response of root border cell in red cowpea to aluminum stress. Plant Nutrition and Fertilizer Science (植物营养与肥料学报), 12,722-726. (in Chinese with English abstract) |
| [10] | Feng YM (冯英明), Yu M (喻敏), Wen HY (温海洋), Zhang YH (张英慧), Xiao HD (萧洪东), Wang HZ (王惠珍), He LL (何丽烂), Liang HD (梁火娣) (2005). Influence of Al on cell viability and mucilage of root border cells of pea (Pisum sativum). Ecological Environment (生态环境), 14,695-699. (in Chinese with English abstract) |
| [11] | Greenberg JT (1996). Programmed cell death: a way of life for plants. Proceedings of the National Academy of Sciences of the United States of America, 93,12094-12097. |
| [12] | Hawes MC, Brigham LA (1992). Impact of root border cells on microbial populations in the rhizosphere. Advances in Plant Pathology, 8,l19-148. |
| [13] | Hawes MC, Gunawardena U, Miyasaka SC, Zhao XW (2000). The role of root border cells in plant defense. Trends in Plant Science, 5,128-133. |
| [14] | Hoeberichts FA, Woltering EJ (2002). Multiple mediators of plant programmed cell death: interplay of conserved cell death mechanisms and plant-specific regulators. Bioessays, 25,47-57. |
| [15] | Jones AW (2001). Programmed cell death in development and defense. Plant Physiology, 125,94-97. |
| [16] | Kochian LV (1995). Cellular mechanisms of aluminum toxicity and resistance in plants. Annual Review of Plant Physiology and Plant Molecular Biology, 46,237-260. |
| [17] | Levine A, Pennell RI, Alvarez ME, Palmer R, Lamb C (1996). Calcium-mediated apoptosis in a plant hypersensitive disease resistance response. Current Biology, 6,427-437. |
| [18] | Lu XD (鲁旭东), Xie ZB (谢志兵) (2003). Programmed cell death in plant growth and development. Journal of Xiaogan University (孝感学院学报), 23 (6),53-56. (in Chinese with English abstract) |
| [19] | Matsumoto H (2000). Cell biology of aluminum toxicity and tolerance in higher plants. International Review of Cytology, 200,41-46. |
| [20] | McCabe PF, Leaver CJ (2000). Programmed cell death in cell cultures. Plant Molecular Biology, 44,359-368. |
| [21] | Mittler R, Lam E (1997). Characterization of nuclease activities and DNA fragmentation induced upon hypersensitive response cell death and mechanical stress. Plant Molecular Biology, 34,209-221. |
| [22] | Miyasaka SC, Hawes MC (2001). Possible role of root border cells in detection and avoidance of aluminum toxicity. Plant Physiology, 125,1978-1987. |
| [23] | Ning SB, Wang L, Song YC (1999). Programmed cell death in plants: a new emerging research field. Development & Reproductive Biology, 8,71-100. |
| [24] | Ormerod MG, Sun XM, Brown D, Snowden RT, Cohen GM (1993). Quantification of apoptosis and necrosis by flow cytometry. Acta Oncologica, 32,417-424. |
| [25] | Orzaez D, Granell A (1997). DNA fragmentation is regulated by ethylene during carpel senescence in Pisum sativum. The Plant Journal, 11,137-144. |
| [26] | Pan JW (潘建伟) (2002). Biological Characters and Mechanisms of Aluminum Toxicity in the Root Tips and Border Cells of Barley (大麦根尖和边缘细胞铝毒生物学特性及其机理研究). PhD dissertation, Zhejiang University, Hangzhou,98-103. (in Chinese) |
| [27] | Pennell RI, Lamb C (1997). Programmed cell death in plants. The Plant Cell, 9,1157-1168. |
| [28] | Richards KD, Schott EJ, Sharma YK, Davis KR, Gardner RC (1998). Aluminum induces oxidative stress genes in Arabidopsis thaliana. Plant Physiology, 116,409-418. |
| [29] | Rosa AV, Maria C, Daniela V, Salvatore P, Ersilia M, Laura DG (2004). Production of reactive oxygen species, alteration of cytosolic ascorbate peroxidase, and impairment of mitochondrial metabolism are early events in heat shock-induced programmed cell death in tobacco bright-yellow cells. Plant Physiology, 134,1100-1112. |
| [30] | Sun YL (孙英丽), Zhao Y (赵允), Liu CX (刘春香), Zhai ZH (翟中和) (1999). Cytochrome c can induce programmed cell death in plant cells. Acta Botanica Sinica (植物学报), 41,379-383. (in Chinese with English abstract) |
| [31] | Tamas L, Budlkova S, Huttova J, Mistrik I, Simonovicovicova M, Siroka B (2005). Aluminum-induced cell death of barley—root border cells is correlated with peroxidase- and oxalate oxidase-mediated hydrogen peroxide production. Plant Cell Reports, 24,189-194. |
| [32] | Wang CB (王淳本) (2003). Applied Biochemistry and Molecular Biology Technology (实用生物化学与分子生物学实验技术) 2nd edn. Hubei Science and Technology Press, Wuhan, 226. (in Chinese) |
| [33] | Wang AG (王爱国), Luo GH (罗广华), Shao CB (邵从本), Wu SJ (吴淑君), Guo JY (郭俊彦) (1983). A study on the superoxide dismutase of soybean seeds. Acta Botanica Sinica (植物学报), 9,77-84. (in Chinese with English abstract) |
| [34] | Wang H, Li J, Bostock RM, Gilchrist DG (1996). Apoptosis: a function paradigm for programmed plant cell death induced by a host-selective phytotoxin and invoked during development. The Plant Cell, 8,375-391. |
| [35] | Xu GD (徐根娣), Liu P (刘鹏), Zhou ZH (周志华) (2004). A progress of studies on development and functions of border cells. Chinese Agricultural Science Bulletin (中国农学通报), 20(5),28-32. (in Chinese with English abstract) |
| [36] | Yamaguchi Y, Yamamoto, Matsumoto H (1999). Cell death process initiated by a combination of aluminum and iron in suspension-cultured tobacco ( Nicotiana tabacum) cells: apoptosis-like cell death mediated by calcium and proteinase . Soil Science & Plant Nutrition, 45,647-657. |
| [37] | Yu M (喻敏), Cui ZX (崔志新), Wen HX (温海祥), Xiao HD (萧洪东), Zhang YH (张英慧), He LL (何丽烂), Liang HD (梁火娣) (2004). Root border cells-A recently defined population of alive cells in rhizophere. Journal of Huazhong Agricultural University (华中农业大学学报), 23,275-280. (in Chinese with English abstract) |
| [38] | Yuan YJ (元英进), Ge ZQ (葛志强), Wang YD (王艳东), Ma ZY (马振毅), Hu ZD (胡宗定) (2001). Ce 4+ induces apoptosis suspension culture of Taxus cuspidata cells . Journal of the Chinese Rare Earth Society (中国稀土学报), 19,357-361. (in Chinese with English abstract) |
| [39] | Zhang ZL (张志良) (1990). The Experimental Guide for Plant Physiology (植物生理学实验指导). Higher Education Press, Beijing, 210-212. (in Chinese) |
| [40] | Zhao YG (赵云罡), Xu JX (徐建兴) (2001). Mitochondrion, AOS and apoptosis. Progress in Biochemistry and Biophysics (生物化学与生物物理进展), 28,168-171. (in Chinese with English abstract) |
| [41] | Zhou N (周楠), Chen WR (陈文荣), Liu P (刘鹏), Xu GD (徐根娣) (2006). Biological characteristic and the response to aluminum toxicity of cucumber border cells. Acta Horticulturae Sinica (园艺学报), 33,1117-1120. (in Chinese with English abstract) |
| [42] | Zhu MY, Ahn S, Matsumoto H (2003). Inhibition of growth and development of root border cells in wheat by Al. Physiologia Plantarum, 117,359-367. |
/
| 〈 |
|
〉 |
