与水稻根尖细胞程序性死亡相关的线粒体热激蛋白70的研究
|
摘要
前期研究以盐敏感水稻品种武运粳8号为材料,研究了盐胁迫对其根尖细胞程序性死亡(PCD)的诱导。本文通过线粒体蛋白质组双向电泳证明线粒体热激蛋白70(mtHSP70)受PCD诱导,且PCD过程中呼吸链复合物Ⅰ表达量下降。进一步对mtHSP70进行了生物信息学分析,并在转录和翻译水平研究了mtHSP70的表达状况,结果如下：
采用生物信息学的方法对已经在蛋白数据库中注册的由水稻基因组其中的Os02g0774300基因编码的mtHSP70进行系统分析,包括该蛋白的组成成分、拷贝、疏水性／亲水性、跨膜拓扑结构、信号肽、导肽、可溶性、蛋白质二级结构、基序及功能域分。结果表明该蛋白是非跨膜的可溶性蛋白,α螺旋和无规卷曲是其蛋白质二级结构的主要结构元件,β折叠零星散布于整个蛋白质中,包含多个基序位点和功能域,主要作为一种分子伴侣发挥作用,该蛋白在水稻线粒体中存在一个同源蛋白,序列一致性高达92%。
mRNA被转录翻译后形成的初始蛋白一般要经过加工修饰才能成为有功能的蛋白质,因此蛋白在基因水平和蛋白水平的表达状况并不是完全一致的。本研究运用半定量RT-PCR和免疫印迹对其在盐胁迫诱导的水稻根尖细胞PCD过程中mtHSP70基因水平的表达情况进行分析。结果表明,该蛋白及其同源蛋白在盐胁迫诱导的PCD过程中mRNA和蛋白水平的表达量均上调。研究结果提示,mtHSP70可能通过影响呼吸链复合物的稳定性,进而调节线粒体内活性氧的产生。
Salt stress-induced PCD in which mitochondria plays an important role in rice root tip cells was studied by using rice seedling Wuyunjing 8th in early period. In the previous study, it was indicated that 500 mmol/L NaCl could lead to PCD in root tip cells. Mitochondrial proteome was studied by two-dimensional gel electrophoresis in this sduty, the result showed that heat shock protein 70 (mtHSP70) was induced by the PCD and respiratory chain complex I decreased. Then, mtHSP70 was analyzed by bioinformatics. The results are as follows:
The amino acid sequences of heat shock protein 70 from Oryza sativa (japonica cultivar-group), which were registered in Protein databases, were analyzed and predicted by the tools of informatics in the following aspects:the composition amino acid sequences, copies, hydrophobicity or hydrophilicity, trans-membrane topological structure, singnal peptides, target peptides, solubility, secondary structure, motifs and functional domains of protein, etc. The results were as follows:mtHSP70 was a hydrophilic, non-transmembrane and soluble protein; the main motifs of predicted secondary strcture of mtHSP70 were alpha helix and random coil, extended strand were spread to the whole secondary structure of protein; the amino acid sequences of mtHPS70 included many motifs and conserved functional domains; mtHSP70 often function as molecular chaperone; there is a homologous protein sequences of which up to 92% consistency and also located in the mitochondria.
Generally, functional protein only be formated after the initial protein be modified. The expressions of mtHSP70 and its Homologous protein, were analysised by the tool of RT-PCR. The result shows that, their expressions on the gene level are all updated. The results suggest that, mtHSP70 may affect the tability of respiratory chain complex s, and regulate the generation of mitochondrial reactive oxygen species.
采用生物信息学的方法对已经在蛋白数据库中注册的由水稻基因组其中的Os02g0774300基因编码的mtHSP70进行系统分析,包括该蛋白的组成成分、拷贝、疏水性／亲水性、跨膜拓扑结构、信号肽、导肽、可溶性、蛋白质二级结构、基序及功能域分。结果表明该蛋白是非跨膜的可溶性蛋白,α螺旋和无规卷曲是其蛋白质二级结构的主要结构元件,β折叠零星散布于整个蛋白质中,包含多个基序位点和功能域,主要作为一种分子伴侣发挥作用,该蛋白在水稻线粒体中存在一个同源蛋白,序列一致性高达92%。
mRNA被转录翻译后形成的初始蛋白一般要经过加工修饰才能成为有功能的蛋白质,因此蛋白在基因水平和蛋白水平的表达状况并不是完全一致的。本研究运用半定量RT-PCR和免疫印迹对其在盐胁迫诱导的水稻根尖细胞PCD过程中mtHSP70基因水平的表达情况进行分析。结果表明,该蛋白及其同源蛋白在盐胁迫诱导的PCD过程中mRNA和蛋白水平的表达量均上调。研究结果提示,mtHSP70可能通过影响呼吸链复合物的稳定性,进而调节线粒体内活性氧的产生。
Salt stress-induced PCD in which mitochondria plays an important role in rice root tip cells was studied by using rice seedling Wuyunjing 8th in early period. In the previous study, it was indicated that 500 mmol/L NaCl could lead to PCD in root tip cells. Mitochondrial proteome was studied by two-dimensional gel electrophoresis in this sduty, the result showed that heat shock protein 70 (mtHSP70) was induced by the PCD and respiratory chain complex I decreased. Then, mtHSP70 was analyzed by bioinformatics. The results are as follows:
The amino acid sequences of heat shock protein 70 from Oryza sativa (japonica cultivar-group), which were registered in Protein databases, were analyzed and predicted by the tools of informatics in the following aspects:the composition amino acid sequences, copies, hydrophobicity or hydrophilicity, trans-membrane topological structure, singnal peptides, target peptides, solubility, secondary structure, motifs and functional domains of protein, etc. The results were as follows:mtHSP70 was a hydrophilic, non-transmembrane and soluble protein; the main motifs of predicted secondary strcture of mtHSP70 were alpha helix and random coil, extended strand were spread to the whole secondary structure of protein; the amino acid sequences of mtHPS70 included many motifs and conserved functional domains; mtHSP70 often function as molecular chaperone; there is a homologous protein sequences of which up to 92% consistency and also located in the mitochondria.
Generally, functional protein only be formated after the initial protein be modified. The expressions of mtHSP70 and its Homologous protein, were analysised by the tool of RT-PCR. The result shows that, their expressions on the gene level are all updated. The results suggest that, mtHSP70 may affect the tability of respiratory chain complex s, and regulate the generation of mitochondrial reactive oxygen species.
引文
[1]Ameisen JC. (2002) On the origin, evolution, and nature of programmed cell death:a timeline of four billion years. Cell Death Differ 9:367-393.
[2]Van Doom WG, Woltering DJ. (2005) Many ways to exit? Cell death categories in plants. Trends Plant Sci.10:117-122.
[3]Armika HL, Gunawardena AN, Deborah M. (2001) Characterisation of programmed cell death during aerenchyma formation induced by ethylene or hypoxia in roots of maize. Planta 212:205-214.
[4]G. van Loo, X. Saelens, G.M. van, M. MacFarlane, S.J. Martin, P. Vandenabeele, The role of mitochondrial factors in apoptosis:a Russian roulette with more than one bullet, Cell Death Differ.9 (2002) 1031-1042.
[5]X. Wang, The expanding role of mitochondria in apoptosis, Genes Dev.15(2001)2922-2933.
[6]A. Jones, Does the plant mitochondrion integrate cellular stress and regulate programmed cell death? Trends Plant Sci.5 (2000) 225-230.
[7]E. Lam, N. Kato, M. Lawton, Programmed cell death, mitochondria and the plant hypersensi-tive response, Nature 411 (2001) 848-853.
[8]Gupta S, Agrawal A, Agrawal S, Su H, Gollapudi S. (2006) A paradox of immunodeficiency and inflammation in human aging. Immunity and Ageing 3(5):1-8.
[9]Kroemer G, Reed JC. (2000) Mitochondrial control of cell death. Nat Med 6:513-519.
[10]Desagher S, Martinou JC. (2000) Mitochondria as the central control point of apoptosis. Trends Cell Biol 10:369-377.
[11]Van Gurp M, Festjens, N, van Loo, G, Saelens X, Vandenabeele P. (2003) Mitochondrial intermembrane proteins in cell death. Biochem Biophys Res Commun 304:487-497.
[12]Li LY, Luo X, Wang X. (2001) Endonuclease G (endo G) is an apoptotic DNAse when released from mitochondria. Nature 412:95-99.
[13]Kaufmann SH, Hengartner MO. (2001) Programmed cell death:alive and well in the new millennium. Trends Cell Biol 11:526-534.
[14]Yu XH, Perdue TD, Heimer YM, and Jones AM. (2002) Mitochondrial involvement in tracheary element programmed cell death. Cell death differ 9:189-198.
[15]Warmke H, Lee SLJ. (1997) Mitochondrial degeneration in Texas cytoplasmic male-sterile corn anthers. J hered 68:213-222.
[16]Li SQ, Wan CX, Kong J, Zhang ZJ, Li YS, Zhu YG (2004) Programmed cell death during microgenesis in a Honglian CMS line of rice is correlated with oxidative stress in mitochondria. Funct Plant Biol 31:369-376.
[17]Balk J, Leaver CJ, McCabe PF. (1999) Translocation of cytochrome c from the mitochondria to the cytoplasm occurs during heat-induced programmed cell death in cucumber plants. FEBS Lett 463:151-154.
[18]Vacca RA, Valenti D, Bobba A, Merafina RS, Passarella S, Marra E. (2006) Cytochrome c is released in a reactive oxygen species-dependent manner and is degraded via caspase-like proteases in tobacco BY-2 cells en route to heat shock-induced cell death. Plant Physiol 141:208-219.
[19]Balk J, Chew SK, Leaver CJ, McCabe PF. (2003) The intermembrane space of plant mitochondria contains a DNase activity that may be involved in programmed cell death. Plant J 34:573-583.
[20]Jimenez A, Hernandez JA, del Rio LA, Sevilla F. (1997) Evidence for the presence of the ascorbate-glutathione cycle in mitochondria and peroxisomes of pea leaves. Plant Physiol 114:275-284.
[21]Simons BH, Millenaar FF, Mulder L, Van Loon LC, Lambers H. (1999) Enhanced expression and activation of the alternative oxidase during infection of Arabidopsis with Pseudomonas syringae pv. tomato. Plant Physiol 120:529-538.
[22]He SC, Abad AR, Gelvin SB, Mackenzie SA. (1996) A cytoplasmic male sterility-associated mitochondrial protein causes pollen disruption in transgenic tobacco. Proc. Natl. Acad. Sci. USA 93:11763-11768.
[23]陈辉,肖卫民,张阳德.肝细胞生长因子对TN F-α诱导肝细胞凋亡的保护作用及其机制的研究[J].中国现代医学杂志,2002,12(14)：4-7.
[24]方瑗,王玉,童萼塘,等.BFGF对缺血性大鼠脑细胞凋亡及相关基因表达的影响[J].中国药理学通报,2001,17(2)：208-210.
[25]GUSAROVA V, CAPLAN AJ,BRODSKY JL, et al. Apoprotein B degradation is promoted by the molecular chaperones HSP90 and HSP70[J]. Biol Chem,2001,276:24891-24892
[26]NYLANDSTED J, ROHDE M, BRAND K, et al. Selective depletion of heat shock protein 70(HSP70) actives a tumor-specific death program that is independent of caspases and bypasses Bcl-2[J]. Proc Natl Aacad Sci USA,2000,97:7871-7873.
[27]Beere HM, Green DR. Stressmanagement heat shock p rotein-70 and the regulation of apop-tosis [J]. Trends Cell Biol,2001,11 (1):6-10.
[28]Didelot C, Schmitt E, BrunetM, et al. Heat shock p roteins:endogenousmodulators of apop totic cell death [J].Handb Exp Pharmacol,2006,172:171-198.
[29]YenariMA,L IU I, Zheng Z, et al. Antiapop totic and anti-inflammatory mechanisms of heat-shock p rotein p rotection [J]. Ann N Y Acad Sci,2005,1053:74-83.
[30]http://www. ornl gov/TechResources/Human-Genome/home, html [DB].
[31]Sasaki T, Burr B. International rice genome sequencing project:The effort to completely sequence the rice genome[J]. Current Opinion in Plant Biology,2000,3 (2):138-141.
[32]陈润生.生物信息学[J].生物物理学报,1999,15(1)：5-13.
[33]王亚辉.世纪之交生物学发展的主要趋势[J].中国科学基金,2000,14(3)：167-168.
[34]Verbeek F J, Law son KA, Bard J B L. Developmental bioinformatics:L inking genetic data to virtual embryos[J]. International Journal of Developmental Biology,1999,43 (7):761-771.
[35]Trifonov E N. Earliest pages of bioinformatics[J].Bioinformatics,2000,16(1):529.
[36]Collins F S, Patrinos A, Jordan E, et al. New goals for the U.S. Human Genome Project: 1998-2003[J]. Science,1998,282:682-689
[37]陈力学.生物信息学在基因组研究中的应用[J].国外医学临床生物化学与检验学分册,2003,24(6)：3392340.
[38]陈润生.生物信息及其研究进展[J].医学研究通讯,2002,31(12)：2-5.
[39]张成岗,贺福初.生物信息学方法与实践[M].北京：科学出版社,2002.
[40]郑国清.生物信息学的形成与发展[J].河南农业科学,2002,11：427.
[41]李宏；我国生物信息学研究的发展策略[J].重庆工商大学学报(自然科学版),2005,22(2)：105-108.
[42]Galperin M Y. The Molecular Biology Database Collection:2007 update [J]. Nucleic Acids Research,2007,35(Database issue):D3-4.
[43]J.A. Swidzinski, C.J. Leaver, L.J. Sweetlove, A proteomic analysis of plant programmed cell death, Phytochemistry 65 (2004) 1829-1838.
[44]H. Tsunezuka, M. Fujiwara, T. Kawasaki, K. Shimamoto, Proteome analysis of programmed cell death and defense signaling using the rice lesion mimic mutant cdr2, Mol. Plant Microbe Interact.18(2005)52-59.
[45]V. Kruft, H. Eubel, L. Jansch, W. Werhahn, H.P. Braun, Proteomic approach to identify novel mitochondrial proteins in Arabidopsis, Plant Physiol.127 (2001) 1694-1710.
[46]C. Mihr, M. Baumga'rtner, J.H. Dieterich, U.K. Schmitz, H.P. Braun, Proteomic approach for investigation of cytoplasmic male sterility (CMS) in Brassica, J. Plant Physiol.158 (2001) 787-794.
[47]J. Li, A. Jiang, W. Zhang, Primary studies on salt stress-induced programmed cell death in rice root tip cells, J. Integr. Plant Biol.49 (2007) 1-5.
[48]A. Jiang, Y. Cheng, J. Li, W. Zhang, A zinc-dependent nuclear endonuclease is responsible for DNA laddering during salt induced programmed cell death in root tip cells of rice, J. Plant Physiol.165(2008)1134-1141.
[49]Kruft V, Eubel H, Jansch L, Werhahn W, Braun HP. (2001) Proteomic approach to identify novel mitochondrial proteins in Arabidopsis. Plant Physiol 127:1694-1710.
[50]Giege P, Sweetlove LJ, Leaver CJ. (2003) Identification of mitochondrial protein complexes in Arabidopsis using two-dimensional blue-native polyacrylamide gel electrophoresis. Plant Mole Biol Rep 21:133-144.
[51]E. Zerbetto, L. Vergani, F. Dabbeni-Sala, Quantification of muscle mitochondrial oxidative phosphorylation enzymes via histochemical staining of blue native polyacrylamide gels, Electrophoresis 18 (1997) 2059-2064.
[52]Sung DY, Kaplan F, Guy CL. (2001) Plant Hsp70 molecular chaperones:Protein structure, gene family, expression and function. Physiol Plant 113:443-451.
[53]Taylor NL, Heazlewood JL, Day DA, Millar AH. (2005) Differential impact of environmental stresses on the pea mitochondrial proteome. Mol Cell Proteomics 4:1122-1133.
[54]Saleh A, Srinivasula SM, Balkir L, Robbins PD, Alnemri ES. (2000) Negative regulation of the Apaf-1 apoptosome by Hsp70. Nat Cell Biol 2:476-483.
[55]Stankiewicz AR, Lachapelle G, Foo CP, Radicioni SM, Mosser DD. (2005) Hsp70 inhibits heat-induced apoptosis upstream of mitochondria by preventing Bax translocation. J Biol Chem 280:38729-38739.
[56]Krause M, Durner J. (2004) Harpin inactivates mitochondria in Arabidopsis suspension cells. Mol Plant Microbe Interact 17:131-139.
[57]Tsunezuka H, Fujiwara M, Kawasaki T, Shimamoto K. (2005) Proteome analysis of programmed cell death and defense signaling using the rice lesion mimic mutant cdr2. Mol Plant Microbe Interact 18:52-59.
[58]Kuzmin EV, Karpova OV, Elthon TE, Newton KJ. (2004) Mitochondrial respiratory deficiencies signal up-regulation of genes for heat shock proteins. J Biol Chem 279:20672-20677.
[59]Amirsadeghi S, Robson CA, McDonald AE, Vanlerberghe GC. (2006) Changes in plant mitochondrial electron transport alter cellular levels of reactive oxygen species and susceptibility to cell death signaling molecules. Plant Cell Physiol 47:1509-1519.
[60]Moller IM. (2001) PLANT MITOCHONDRIA AND OXIDATIVE STRESS:Electron Transport, NADPH Turnover, and Metabolism of Reactive Oxygen Species. Annu Rev Plant Physiol Plant Mol Biol 52:561-591.
[61]Lam E, Pontier D, del PO. (1999) Die and let live-programmed cell death in plants. Curr Opin Plant Biol 2:502-507.
[62]Katsuhara M, Kawasaki T. (1996) Salt stress induced nuclear and DNA degradation in meristematic cells of barley root. Plant Cell Physiol 37:169-173.
[63]Ning SB, Song YC, Damme PV. (2002) Characterization of the early stages of programmed cell death in maize root cells by using comet assay and the combination of cell electrophoresis with annexin binding. Electrophoresis 23:2096-2102.
[64]Huh GH, Damaz B, Matsumoto T, Reddy MP, Rus AM, Ibeas JI, Narasimhan ML, Bressan RA, Hasegawa PM. (2002) Salt causes ion disequilibrium-induced programmed cell death in yeast and plants. Plant J 29:649-59.
[65]Yu XH, Perdue TD, Heimer YM, and Jones AM. (2002) Mitochondrial involvement in tracheary element programmed cell death. Cell death differ 9:189-198.
[66]Duroc Y, Gaillard C, Hiard S, Defiance MC, Pelletier G, Budar F. (2005) Biochemical and functional characterization of ORF138, a mitochondrial protein responsible for Ogura cytoplasmic male sterility in Brassiceae. Biochimie 87:1089-1100.
[67]Dewey RE, Timothy DH, Levings CS. (1987) A mitochondrial protein associated with cytoplasmic male sterility in the T cytoplasm of maize. Proc. Nati. Acad. Sci. USA. 84:5374-5378.
[68]Gene family, expression and function. Physiol Plant 113:443-451.
[2]Van Doom WG, Woltering DJ. (2005) Many ways to exit? Cell death categories in plants. Trends Plant Sci.10:117-122.
[3]Armika HL, Gunawardena AN, Deborah M. (2001) Characterisation of programmed cell death during aerenchyma formation induced by ethylene or hypoxia in roots of maize. Planta 212:205-214.
[4]G. van Loo, X. Saelens, G.M. van, M. MacFarlane, S.J. Martin, P. Vandenabeele, The role of mitochondrial factors in apoptosis:a Russian roulette with more than one bullet, Cell Death Differ.9 (2002) 1031-1042.
[5]X. Wang, The expanding role of mitochondria in apoptosis, Genes Dev.15(2001)2922-2933.
[6]A. Jones, Does the plant mitochondrion integrate cellular stress and regulate programmed cell death? Trends Plant Sci.5 (2000) 225-230.
[7]E. Lam, N. Kato, M. Lawton, Programmed cell death, mitochondria and the plant hypersensi-tive response, Nature 411 (2001) 848-853.
[8]Gupta S, Agrawal A, Agrawal S, Su H, Gollapudi S. (2006) A paradox of immunodeficiency and inflammation in human aging. Immunity and Ageing 3(5):1-8.
[9]Kroemer G, Reed JC. (2000) Mitochondrial control of cell death. Nat Med 6:513-519.
[10]Desagher S, Martinou JC. (2000) Mitochondria as the central control point of apoptosis. Trends Cell Biol 10:369-377.
[11]Van Gurp M, Festjens, N, van Loo, G, Saelens X, Vandenabeele P. (2003) Mitochondrial intermembrane proteins in cell death. Biochem Biophys Res Commun 304:487-497.
[12]Li LY, Luo X, Wang X. (2001) Endonuclease G (endo G) is an apoptotic DNAse when released from mitochondria. Nature 412:95-99.
[13]Kaufmann SH, Hengartner MO. (2001) Programmed cell death:alive and well in the new millennium. Trends Cell Biol 11:526-534.
[14]Yu XH, Perdue TD, Heimer YM, and Jones AM. (2002) Mitochondrial involvement in tracheary element programmed cell death. Cell death differ 9:189-198.
[15]Warmke H, Lee SLJ. (1997) Mitochondrial degeneration in Texas cytoplasmic male-sterile corn anthers. J hered 68:213-222.
[16]Li SQ, Wan CX, Kong J, Zhang ZJ, Li YS, Zhu YG (2004) Programmed cell death during microgenesis in a Honglian CMS line of rice is correlated with oxidative stress in mitochondria. Funct Plant Biol 31:369-376.
[17]Balk J, Leaver CJ, McCabe PF. (1999) Translocation of cytochrome c from the mitochondria to the cytoplasm occurs during heat-induced programmed cell death in cucumber plants. FEBS Lett 463:151-154.
[18]Vacca RA, Valenti D, Bobba A, Merafina RS, Passarella S, Marra E. (2006) Cytochrome c is released in a reactive oxygen species-dependent manner and is degraded via caspase-like proteases in tobacco BY-2 cells en route to heat shock-induced cell death. Plant Physiol 141:208-219.
[19]Balk J, Chew SK, Leaver CJ, McCabe PF. (2003) The intermembrane space of plant mitochondria contains a DNase activity that may be involved in programmed cell death. Plant J 34:573-583.
[20]Jimenez A, Hernandez JA, del Rio LA, Sevilla F. (1997) Evidence for the presence of the ascorbate-glutathione cycle in mitochondria and peroxisomes of pea leaves. Plant Physiol 114:275-284.
[21]Simons BH, Millenaar FF, Mulder L, Van Loon LC, Lambers H. (1999) Enhanced expression and activation of the alternative oxidase during infection of Arabidopsis with Pseudomonas syringae pv. tomato. Plant Physiol 120:529-538.
[22]He SC, Abad AR, Gelvin SB, Mackenzie SA. (1996) A cytoplasmic male sterility-associated mitochondrial protein causes pollen disruption in transgenic tobacco. Proc. Natl. Acad. Sci. USA 93:11763-11768.
[23]陈辉,肖卫民,张阳德.肝细胞生长因子对TN F-α诱导肝细胞凋亡的保护作用及其机制的研究[J].中国现代医学杂志,2002,12(14)：4-7.
[24]方瑗,王玉,童萼塘,等.BFGF对缺血性大鼠脑细胞凋亡及相关基因表达的影响[J].中国药理学通报,2001,17(2)：208-210.
[25]GUSAROVA V, CAPLAN AJ,BRODSKY JL, et al. Apoprotein B degradation is promoted by the molecular chaperones HSP90 and HSP70[J]. Biol Chem,2001,276:24891-24892
[26]NYLANDSTED J, ROHDE M, BRAND K, et al. Selective depletion of heat shock protein 70(HSP70) actives a tumor-specific death program that is independent of caspases and bypasses Bcl-2[J]. Proc Natl Aacad Sci USA,2000,97:7871-7873.
[27]Beere HM, Green DR. Stressmanagement heat shock p rotein-70 and the regulation of apop-tosis [J]. Trends Cell Biol,2001,11 (1):6-10.
[28]Didelot C, Schmitt E, BrunetM, et al. Heat shock p roteins:endogenousmodulators of apop totic cell death [J].Handb Exp Pharmacol,2006,172:171-198.
[29]YenariMA,L IU I, Zheng Z, et al. Antiapop totic and anti-inflammatory mechanisms of heat-shock p rotein p rotection [J]. Ann N Y Acad Sci,2005,1053:74-83.
[30]http://www. ornl gov/TechResources/Human-Genome/home, html [DB].
[31]Sasaki T, Burr B. International rice genome sequencing project:The effort to completely sequence the rice genome[J]. Current Opinion in Plant Biology,2000,3 (2):138-141.
[32]陈润生.生物信息学[J].生物物理学报,1999,15(1)：5-13.
[33]王亚辉.世纪之交生物学发展的主要趋势[J].中国科学基金,2000,14(3)：167-168.
[34]Verbeek F J, Law son KA, Bard J B L. Developmental bioinformatics:L inking genetic data to virtual embryos[J]. International Journal of Developmental Biology,1999,43 (7):761-771.
[35]Trifonov E N. Earliest pages of bioinformatics[J].Bioinformatics,2000,16(1):529.
[36]Collins F S, Patrinos A, Jordan E, et al. New goals for the U.S. Human Genome Project: 1998-2003[J]. Science,1998,282:682-689
[37]陈力学.生物信息学在基因组研究中的应用[J].国外医学临床生物化学与检验学分册,2003,24(6)：3392340.
[38]陈润生.生物信息及其研究进展[J].医学研究通讯,2002,31(12)：2-5.
[39]张成岗,贺福初.生物信息学方法与实践[M].北京：科学出版社,2002.
[40]郑国清.生物信息学的形成与发展[J].河南农业科学,2002,11：427.
[41]李宏；我国生物信息学研究的发展策略[J].重庆工商大学学报(自然科学版),2005,22(2)：105-108.
[42]Galperin M Y. The Molecular Biology Database Collection:2007 update [J]. Nucleic Acids Research,2007,35(Database issue):D3-4.
[43]J.A. Swidzinski, C.J. Leaver, L.J. Sweetlove, A proteomic analysis of plant programmed cell death, Phytochemistry 65 (2004) 1829-1838.
[44]H. Tsunezuka, M. Fujiwara, T. Kawasaki, K. Shimamoto, Proteome analysis of programmed cell death and defense signaling using the rice lesion mimic mutant cdr2, Mol. Plant Microbe Interact.18(2005)52-59.
[45]V. Kruft, H. Eubel, L. Jansch, W. Werhahn, H.P. Braun, Proteomic approach to identify novel mitochondrial proteins in Arabidopsis, Plant Physiol.127 (2001) 1694-1710.
[46]C. Mihr, M. Baumga'rtner, J.H. Dieterich, U.K. Schmitz, H.P. Braun, Proteomic approach for investigation of cytoplasmic male sterility (CMS) in Brassica, J. Plant Physiol.158 (2001) 787-794.
[47]J. Li, A. Jiang, W. Zhang, Primary studies on salt stress-induced programmed cell death in rice root tip cells, J. Integr. Plant Biol.49 (2007) 1-5.
[48]A. Jiang, Y. Cheng, J. Li, W. Zhang, A zinc-dependent nuclear endonuclease is responsible for DNA laddering during salt induced programmed cell death in root tip cells of rice, J. Plant Physiol.165(2008)1134-1141.
[49]Kruft V, Eubel H, Jansch L, Werhahn W, Braun HP. (2001) Proteomic approach to identify novel mitochondrial proteins in Arabidopsis. Plant Physiol 127:1694-1710.
[50]Giege P, Sweetlove LJ, Leaver CJ. (2003) Identification of mitochondrial protein complexes in Arabidopsis using two-dimensional blue-native polyacrylamide gel electrophoresis. Plant Mole Biol Rep 21:133-144.
[51]E. Zerbetto, L. Vergani, F. Dabbeni-Sala, Quantification of muscle mitochondrial oxidative phosphorylation enzymes via histochemical staining of blue native polyacrylamide gels, Electrophoresis 18 (1997) 2059-2064.
[52]Sung DY, Kaplan F, Guy CL. (2001) Plant Hsp70 molecular chaperones:Protein structure, gene family, expression and function. Physiol Plant 113:443-451.
[53]Taylor NL, Heazlewood JL, Day DA, Millar AH. (2005) Differential impact of environmental stresses on the pea mitochondrial proteome. Mol Cell Proteomics 4:1122-1133.
[54]Saleh A, Srinivasula SM, Balkir L, Robbins PD, Alnemri ES. (2000) Negative regulation of the Apaf-1 apoptosome by Hsp70. Nat Cell Biol 2:476-483.
[55]Stankiewicz AR, Lachapelle G, Foo CP, Radicioni SM, Mosser DD. (2005) Hsp70 inhibits heat-induced apoptosis upstream of mitochondria by preventing Bax translocation. J Biol Chem 280:38729-38739.
[56]Krause M, Durner J. (2004) Harpin inactivates mitochondria in Arabidopsis suspension cells. Mol Plant Microbe Interact 17:131-139.
[57]Tsunezuka H, Fujiwara M, Kawasaki T, Shimamoto K. (2005) Proteome analysis of programmed cell death and defense signaling using the rice lesion mimic mutant cdr2. Mol Plant Microbe Interact 18:52-59.
[58]Kuzmin EV, Karpova OV, Elthon TE, Newton KJ. (2004) Mitochondrial respiratory deficiencies signal up-regulation of genes for heat shock proteins. J Biol Chem 279:20672-20677.
[59]Amirsadeghi S, Robson CA, McDonald AE, Vanlerberghe GC. (2006) Changes in plant mitochondrial electron transport alter cellular levels of reactive oxygen species and susceptibility to cell death signaling molecules. Plant Cell Physiol 47:1509-1519.
[60]Moller IM. (2001) PLANT MITOCHONDRIA AND OXIDATIVE STRESS:Electron Transport, NADPH Turnover, and Metabolism of Reactive Oxygen Species. Annu Rev Plant Physiol Plant Mol Biol 52:561-591.
[61]Lam E, Pontier D, del PO. (1999) Die and let live-programmed cell death in plants. Curr Opin Plant Biol 2:502-507.
[62]Katsuhara M, Kawasaki T. (1996) Salt stress induced nuclear and DNA degradation in meristematic cells of barley root. Plant Cell Physiol 37:169-173.
[63]Ning SB, Song YC, Damme PV. (2002) Characterization of the early stages of programmed cell death in maize root cells by using comet assay and the combination of cell electrophoresis with annexin binding. Electrophoresis 23:2096-2102.
[64]Huh GH, Damaz B, Matsumoto T, Reddy MP, Rus AM, Ibeas JI, Narasimhan ML, Bressan RA, Hasegawa PM. (2002) Salt causes ion disequilibrium-induced programmed cell death in yeast and plants. Plant J 29:649-59.
[65]Yu XH, Perdue TD, Heimer YM, and Jones AM. (2002) Mitochondrial involvement in tracheary element programmed cell death. Cell death differ 9:189-198.
[66]Duroc Y, Gaillard C, Hiard S, Defiance MC, Pelletier G, Budar F. (2005) Biochemical and functional characterization of ORF138, a mitochondrial protein responsible for Ogura cytoplasmic male sterility in Brassiceae. Biochimie 87:1089-1100.
[67]Dewey RE, Timothy DH, Levings CS. (1987) A mitochondrial protein associated with cytoplasmic male sterility in the T cytoplasm of maize. Proc. Nati. Acad. Sci. USA. 84:5374-5378.
[68]Gene family, expression and function. Physiol Plant 113:443-451.