玉米杂交种与亲本苗期叶片差异表达蛋白谱分析

doi:10.3864/j.issn.0578-1752.2013.14.021

摘要/Abstract

摘要： 【目的】建立玉米杂交种与亲本苗期叶片差异表达蛋白谱，探讨叶片大小杂种优势形成的分子机理。【方法】以玉米强优势杂交种Mo17/B73及其亲本发芽后第5天的第3片叶为材料，采用双向电泳技术（2-DE），结合MALDI TOF MS质谱技术，建立叶片细胞分裂和生长关键区域的差异表达蛋白质谱，并对差异表达蛋白进行质谱鉴定。【结果】在检测到的630个蛋白质点中，有52个蛋白质点在杂交种与亲本之间的表达差异达到显著水平，表现为单亲沉默（15个）、偏高亲（13个）、偏低亲（8个）、杂种上调（6个）、杂种下调（7个）和杂种特异表达模式（3个）。另外，还成功鉴定出了其中的28个差异表达蛋白质点，涉及到代谢、胁迫响应、糖酵解、转录调控、蛋白折叠和降解、三羧酸循环、细胞骨架、发育及未知蛋白质等9个功能类别。【结论】玉米杂交种与亲本在蛋白丰度上存在明显的差异，并且差异表达蛋白涉及到多个功能类别，可能与玉米叶片大小杂种优势的形成有关。

关键词: 玉米 , 叶片大小 , 杂种优势 , 差异表达蛋白 , 分子机理

Abstract: 【Objective】To construct protein expression profile of seedling leaves between maize hybrid and its parental lines, with the purpose to give an insight into the molecular basis of leaf size heterosis. 【Method】 Differentially expressed protein profile of the third leaf on 5th day after germination between maize hybrid and its parental lines used for 2-DE analysis and differentially expressed proteins were identified by using MALDI TOF MS method. 【Result】 A total of 630 protein spots were detected, among which 52 protein spots were found to be differentially expressed between maize hybrid and its parental lines. Differentially expressed protein spots could be grouped into six models, that is UPF1 (expression in hybrid and uniparent but not in another parent), HDH(hybrid is equal to the highly expressed parent), LDH (hybrid is equal to the lowly expressed parent), URH (up-regulated in hybrid), DRH (down-regulated in hybrid), F1nBP (hybrid-specific expressed protein spots), and the numbers were 15, 13, 8, 6, 7 and 3, respectively. Moreover, 28 of the 52 differentially expressed protein spots were identified by using MALDI TOF MS, which were grouped into nine functional categories, including metabolism, development, stress, glycolysis, transcription regulation, protein folding and degradation, tricarboxylic acid cycle, cytoskeleton and unknown protein. 【Conclusion】 Significant alterations in protein expression occurred in seedling leaves of hybrids and parental lines and differentially expressed proteins implicated in some functional categories might contribute to heterosis related to leaf size heterosis.

Key words: maize , leaf size , heterosis , differentially expressed protein , molecular mechanism

郭宝健, 隋志鹏, 李洋洋, 冯万军, 闫文文, 李慧敏, 孙其信, 倪中福. 玉米杂交种与亲本苗期叶片差异表达蛋白谱分析[J]. 中国农业科学, 2013, 46(14): 3046-3054.

GUO Bao-Jian, SUI Zhi-Peng, LI Yang-Yang, FENG Wan-Jun, YAN Wen-Wen, LI Hui-Min, SUN Qi-Xin, NI Zhong-Fu. Differentially Expressed Protein Profile of Maize Seedling Leaves Between Hybrid and Its Parental Lines[J]. Scientia Agricultura Sinica, 2013, 46(14): 3046-3054.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2013.14.021

https://www.chinaagrisci.com/CN/Y2013/V46/I14/3046

参考文献

[1]Swanson-Wagner R A, Jia Y, DeCook R, Borsuk L A, Nettleton D, Schnable P S. All possible modes of gene action are observed in a global comparison of gene expression in a maize F1 hybrid and its inbred parents. Proceedings of the National Academy of Sciences of the USA, 2006, 103(18): 6805-6810.

[2]Song S H, Qu H Z, Chen C, Hu S N, Yu J. Differential gene expression in an elite hybrid rice cultivar (Oryza sativa L.) and its parental lines based on SAGE data. BMC Plant Biology, 2007, 7: 49.

[3]Meyer R C, Kusterer B, Lisec J, Steinfath M, Becher M, Scharr H, Melchinger A E, Selbig J, Schurr U, Willmitzer L, Altmann T. QTL analysis of early stage heterosis for biomass in Arabidopsis. Theoretical and Applied Genetics, 2010, 120(2): 227-237.

[4]孙其信. 农作物杂种优势机理研究及展望. 作物杂志, 1998, 4: 31.

Sun Q X. Crop heterosis mechanism and prospects. Crop Journal, 1998, 4: 31. (in Chinese)

[5]Romagnoli S, Maddaloni M, Livini C, Motto M. Relationship between gene expression and hybrid vigor in primary root tips of young maize (Zea mays L.) plantlets. Theoretical and Applied Genetics, 1990, 80(6): 769-775.

[6]Marcon C, Schützenmeister A, Schütz W, Madlung J, Piepho H P, Hochholdinger F. Nonadditive protein accumulation patterns in maize (Zea mays L.) hybrids during embryo development. Journal of Proteome Research, 2010, 9(12): 6511-6522.

[7]张义荣, 姚颖垠, 彭惠茹, 张庆波, 倪中福, 孙其信. 植物杂种优势形成的分子遗传机理研究进展. 自然科学进展, 2009, 19(7): 697-703.

Zhang Y R, Yao Y Y, Peng H R, Zhang Q B, Ni Z F, Sun Q X. Progress in molecular genetic mechanism of plant heterosis. Progress in Natural Science, 2009, 19(7): 697-703. (in Chinese)

[8]Hoecker N, Keller B, Muthreich N, Chollet D, Descombes P, Piepho H P, Hochholdinger F. Comparison of maize (Zea mays L.) F1-hybrid and parental inbred line primary root transcriptomes suggests organ-specific patterns of nonadditive gene expression and conserved expression trends. Genetics, 2008, 179(3): 1275-1283.

[9]Fu Z Y, Jin X N, Ding D, Li Y L, Fu Z J, Tang J H. Proteomic analysis of heterosis during maize seed germination. Proteomics, 2011, 11: 1462-1472.

[10]Paschold A, Jia Y, Marcon C, Lund S, Larson N B, Yeh C T, Ossowski S, Lanz C, Nettleton D, Schnable P S, Hochholdinger F. Complementation contributes to transcriptome complexity in maize (Zea mays L.) hybrids relative to their inbred parents. Genome Research, 2012, 22(12): 2445-2454.

[11]Hoecker N, Lamkemeyer T, Sarholz B, Paschold A, Fladerer C, Madlung J, Wurster K, Stahl M, Piepho H P, Nordheim A, Hochholdinger F. Analysis of nonadditive protein accumulation in young primary roots of a maize (Zea mays L.) F1-hybrid compared to its parental inbred lines. Proteomics, 2008, 8(18): 3882-3894.

[12]Huang Y, Zhang L D, Zhang J W, Yuan D J, Xu C G , Li X H , Zhou D X , Wang S P, Zhang Q F. Heterosis and polymorphisms of gene expression in an elite rice hybrid as revealed by a microarray analysis of 9198 unique ESTs. Plant Molecular Biology, 2006, 62(4/5): 579-591.

[13]Meyer R C, Witucka-Wall H, Becher M, Blacha A, Boudichevskaia A, Dörmann P, Fiehn O, Friedel S, von Korff M, Lisec J, Melzer M, Repsilber D, Schmidt R, Scholz M, Selbig J, Willmitzer L, Altmann T. Heterosis manifestation during early Arabidopsis seedling development is characterized by intermediate gene expression and enhanced metabolic activity in the hybrids. The Plant Journal, 2012, 71(4): 669-683.

[14]Dahal D, Mooney B P, Newton K J. Specific changes in total and mitochondrial proteomes are associated with higher levels of heterosis in maize hybrids. The Plant Journal, 2012, 72(1): 70-83.

[15]Song X, Ni Z F, Yao Y Y, Zhang Y H, Sun Q X. Identification of differentially expressed proteins between hybrid and parents in wheat (Triticum aestivum L.) seedling leaves. Theoretical and Applied Genetics, 2009, 118(2): 213-225.

[16]Wang W, Meng B, Ge X, Song S, Yang Y, Yu X, Wang L, Hu S, Liu S, Yu J. Proteomic profiling of rice embryos from a hybrid rice cultivar and its parental lines. Proteomics, 2008, 8(22): 4808-4821.

[17]Song X, Ni Z F, Yao Y Y, Xie C J, Li Z X, Wu H Y, Zhang Y H, Sun Q X. Wheat (Triticum aestivum L.) root proteome and differentially expressed root proteins between hybrid and parents. Proteomics, 2007, 7(19): 3538-3557.

[18]Ni Z F, Kim E D, Ha M S, Lackey E, Liu J X, Zhang Y R, Sun Q X, Chen Z J. Altered circadian rhythms regulate growth vigour in hybrids and allopolyploids. Nature, 2009, 457: 327-331.

[19]Guo M, Rupe M A, Dieter J A, Zou J, Spielbauer D, Duncan K E, Howard R J, Hou Z, Simmons C R. Cell number regulator1 affects plant and organ size in maize: Implications for crop yield enhancement and heterosis. The Plant Cell, 2010, 22(4): 1057-1073.

[20]Li P, Ponnala L, Gandotra N, Wang L, Si Y, Tausta S L, Kebrom T H, Provart N, Patel R, Myers C R, Reidel E J, Turgeon R, Liu P, Sun Q, Nelson T, Brutnell T P. The developmental dynamics of the maize leaf transcriptome. Nature Genetic, 2010, 42(12): 1060-1067.

[21]Palmer S J. Davies W J. An analysis of relative elemental growth rate, epidermal cell size and xyloglucan endotransglycosylase activity through the growing zone of ageing maize leaves. Journal of Experimental Botany, 1996, 47(296): 339-347.

[22]Bradford M M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 1976, 72: 248-254.

[23]宋方威, 彭惠茹, 张义荣, 杨小红, 刘婷, 孙其信, 倪中福. 利用三重测交群体剖析玉米苗期性状杂种优势的遗传学基础。农业生物技术学报, 2011, 19(5): 785-792.

Song F W, Peng H R, Zhang Y R, Yang X H, Liu T, Sun Q X, Ni Z F. Heterosis for seeding traits in maize (Zea mays L.) revealed by quantitative trait loci analysis of the triple testcross design. Journal of Agricultural Biotechnology, 2011, 19(5): 785-792. (in Chinese)

[24]Frascaroli E, Canè M A, Landi P, Pea G, Gianfranceschi L, Villa M, Morgante M, Pè M E. Classical genetic and quantitative trait loci analyses of heterosis in a maize hybrid between two elite inbred lines. Genetics, 2007, 176: 625-644.

[25]Lange P R, Geserick C, Tischendorf G, Zrenner R. Functions of chloroplastic adenylate kinases in Arabidopsis. Plant Physiology, 2008, 146(2): 492-504.

[26]Regierer B, Fernie A R, Springer F, Perez-Melis A, Leisse A, Koehl K, Willmitzer L, Geigenberger P, Kossmann J. Starch content and yield increase as a result of altering adenylate pools in transgenic plants. Nature Biotechnology, 2002, 20(12): 1256-1260.

[27]Staiger C J. Signaling to the actin cytoskeleton in plants. Annual Review of Plant Physiology and Plant Molecular Biology, 2000, 51: 257-288.

[28]An Y Q, Huang S, McDowell J M, McKinney E C, Meagher R B. Conserved expression of the Arabidopsis ACT1 and ACT3 actin subclass in organ primordia and mature pollen. The Plant Cell, 1996, 8(1): 15-30.

[29]McDowell J M, An Y Q, Huang S, McKinney E C, Meagher R B. The Arabidopsis ACT7 actin gene is expressed in rapidly developing tissues and responds to several external stimuli. Plant Physiology, 1996, 111: 699-711.

[30]McDowell J M, Huang S, McKinney E C, An Y Q, Meagher R B. Structure and evolution of the actin gene family in Arabidopsis thaliana. Genetics, 1996, 142(2): 587-602.

[31]Kandasamy M K, McKinney E C, Meagher R B. A single vegetative actin isovariant overexpressed under the control of multiple regulatory sequences is sufficient for normal Arabidopsis development. The Plant Cell, 2009, 21(3): 701-718.

[32]Winter D, Vinegar B, Nahal H, Ammar R, Wilson G V, Provart N J. An “electronic fluorescent pictograph” browser for exploring and analyzing large-scale biological data sets. PloS ONE, 2007, 2(8): e718.

[33]Shen H S, He H, Li J G, Chen W, Wang X C, Guo L, Peng Z Y, He G M, Zhong S W, Qi Y J, Terzaghi W, Deng X W. Genome-wide analysis of DNA methylation and gene expression changes in two Arabidopsis ecotypes and their reciprocal hybrids. The Plant Cell, 2012, 24: 875-892.

[34]Johnston A J, Matveeva E, Kirioukhova O, Grossniklaus U, Gruissem W. A dynamic reciprocal RBR-PRC2 regulatory circuit controls Arabidopsis gametophyte development. Current Biology, 2008, 18(21): 1680-1686.