香蕉幼苗叶片响应低温胁迫的比较蛋白质组学研究

doi:10.11924/j.issn.1000-6850.casb15040093

摘要/Abstract

摘要： 香蕉是中国热带地区重要的经济作物，香蕉产业受寒害的影响极为严重。明确香蕉植株响应低温胁迫的分子调控机制有可能为培育和筛选出相对抗寒的香蕉品系提供研究基础。应用比较蛋白质组学对7℃低温处理0、3、5和7天的巴西蕉幼苗叶片总蛋白进行分析，发现有85个蛋白质点的表达丰度在低温胁迫后发现明显变化，通过质谱成功地鉴定了其中的82个蛋白，这些蛋白质主要参与蛋白质的加工及翻译后修饰、糖与能量代谢、光合作用等生理学过程。研究结果暗示低温环境下，香蕉幼苗叶片可能通过改变光合作用相关酶、糖类与能量代谢相关酶的表达量来调节能量与碳代谢，保证碳养分供应及吸收，同时通过超氧化物歧化酶清除系统来提高香蕉抵御低温的能力。研究结果将为研究香蕉抗寒的分子机制提供新线索，为香蕉抗寒育种提供新的候选蛋白。

关键词: 日光温室, 日光温室, CO₂浓度, 天气类型, 相关性

Abstract: Banana is one of the very important economic crops in tropical area of China, and the banana industry has been seriously affected by chilling often. Clarifying the molecular mechanism of banana plants in response to low temperature stress could provide the research basis for cultivating and screening relatively cold resistant banana varieties. Comparative proteomics was applied to analyze the differential expression proteins of banana seedling leaves which were exposed at 7℃ for 0, 3, 5, 7 days, and the expression of 85 protein spots was found to be obviously affected by low temperature, finally 82 proteins out of them were identified by mass spectrum. These identified proteins were mainly involved in protein synthesis process, protein posttranslational modification, carbohydrate and energy metabolism, and photosynthesis. The results suggested that the banana seedling leaf might regulate carbon and energy metabolism through the expression pattern change of photosynthesis which was related to enzymes, and carbohydrate which was associated with energy metabolism enzyme to ensure the supply and absorption of carbon nutrients, and also improve the ability of low temperature resistance by superoxide dismutase scavenging system. The results could provide new clues for the study of the molecular mechanism of low temperature resistance of banana, and new candidate proteins for the breeding of cold resistant strain of banana.

中图分类号:

孙勇,王丹,仝征,杨倩,常丽丽,王力敏,何丽平,王旭初. 香蕉幼苗叶片响应低温胁迫的比较蛋白质组学研究[J]. 中国农学通报, 2015, 31(34): 216-228.

Sun Yong,Wang Dan,Tong Zheng,Yang Qian,Chang Lili,Wang Limin,He Liping and Wang Xuchu. Proteomic Analysis of Banana Seedling Leaf Response to Low Temperature[J]. Chinese Agricultural Science Bulletin, 2015, 31(34): 216-228.

参考文献

[1] 陈尚漠,黄寿波,温福光.果树气象学[M].北京:气象出版社,1988:456-486.
[2] 潘嘉念,涂悦贤,李载忠.广东省农业气象灾害及其防灾减灾对策[M].北京:气象出版社,2000:325-343.
[3] 林燕金,黄雄峰,钟秋珍,等.香蕉低温害研究现状及趋势分析[J].福建果树,2010(2):36-39.
[4] Chen W, Provart N J, Glazebrook J, et al. Expression profile matrix of Arabidopsis transcription factor genes suggests their putative functions in response to environmental stresses[J]. The Plant Cell,2002,14(3):559-574.
[5] Fowler S, Thomashow M F. Arabidopsis transcriptome profiling indicates that multiple regulatory pathways are activated during cold acclimation in addition to the CBF cold response pathway[J]. The Plant Cell,2002,14(8):1675-1690.
[6] Greenup A G, Sasani S, Oliver S N, et al. Transcriptome analysis of the vernalization response in barley (Hordeum vulgare) seedlings[J]. PLoS One,2011,6(3):e17900.
[7] Winfield M O, Lu C, Wilson I D, et al. Plant responses to cold: Transcriptome analysis of wheat[J]. Plant Biotechnol J,2010,8(7):749-771.
[8] Santos E, Remy S, Thiry E, et al. Characterization and isolation of a T-DNA tagged banana promoter active during in vitro culture and low temperature stress[J]. BMC Plant Biol,2009,9:77.
[9] Feng D R, Liu B, Li W Y, et al. Over-expression of a cold-induced plasma membrane protein gene (MpRCI) from plantain enhances low temperature-resistance in transgenic tobacco[J]. Environ Exp Bot,2009,65:395-402.
[10] Yan S P, Zhang Q Y, Tang Z C, et al. Comparative proteomic analysis provides new insights into chilling stress responses in rice[J]. Mol Cell Proteomics,2006,5(3):484-496.
[11] Schwanhausser B, Busse D, Li N, et al. Global quantification of mammalian gene expression control[J]. Nature,2011,473(7347):337-342.
[12] Pradet-Balade B, Boulme F, Beug H, et al. Translation control: bridging the gap between genomics and proteomics?[J] Trends Biochem Sci,2001,26(4):225-229.
[13] Kosova K, Vitamvas P, Prasil I T, et al. Plant proteome changes under abiotic stress--contribution of proteomics studies to understanding plant stress response[J]. J Proteomics,2012,74(8):1301-1322.
[14] Yang Q S, Wu J H, Li C Y, et al. Quantitative proteomic analysis reveals that antioxidation mechanisms contribute to cold tolerance in plantain (Musa paradisiaca L.; ABB Group) seedlings[J]. Mol Cell Proteomics,2012,11(12):1853-69.
[15] Wang X C, Li X F, Deng X, et al. A protein extraction method compatible with proteomic analysis for euhalophyte Salicornia europaea[J]. Electrophoresis,2007,28(21):3976-3987
[16] Wang X C, Wang D Y, Wang D, et al. Systematic comparison of technical details in CBB methods and development of a sensitive GAP stain for comparative proteomic analysis[J]. Electrophoresis,2012,33(2):296–306.
[17] 王旭初,范鹏祥,李银心.一种适用于质谱分析的简化胶内酶解方法[J].植物生理与分子生物学学报,2007,33(5):1-7.
[18] Wang X C, Fan P X, Song H M, et al. Comparative proteomic analysis of differentially expressed proteins in shoots of Salicornia europaea under different salinity[J]. J Proteome Res,2009,8:3331-3345.
[19] Ganry J. étude du développement du système foliaire du bananier en fonction de la température[J]. Fruits,1973,28:499-516.
[20] Israeli Y, Lahav E. Injuries to banana caused by adverse climate and weather[M], CAB International, UK.2000.
[21] 黄秉智.香蕉种质资源描述规范和数据标准[M].北京:中国农业出版社,2006:98.
[22] Allen D J, Ort D R. Impacts of chilling temperatures on photosynthesis in warm-climate plants[J]. Trends in Plant Science,2001,6(1):36-42.
[23] Stitt M, Hurry V. A plant for all seasons: alterations in photosynthetic carbon metabolism during cold acclimation in Arabidopsis[J]. Plant Biology,2002,5(3):199-206.
[24] Oquist G. Photosynthesis of overwintering evergreen plants[J]. Annu Rev Plant Biol,2003,54:329-355.
[25] Cui S X, Huang F, Wang J, et al. A proteomic analysis of cold stress responses in rice seedlings[J]. Proteomics,2005,5(12):3162-3172.
[26] Murata N, Mohanty P S, Hayashi H, et al. Glycinebetaine stabilizes the association of extrinsic proteins with the photosynthetic oxygen-evolving complex[J]. FEBS letters,1992,296(2):187-189.
[27] Klosgen R B, Brock I W, Herrmann R G, et al. Proton gradient-driven import of the 16 kDa oxygen-evolving complex protein as the full precursor protein by isolated thylakoids[J]. Plant Molecular Biology,1992,18(5):1031-1034.
[28] Ishida K I, Green B R. Second- and third-hand chloroplasts in dinoflagellates: Phylogeny of oxygen-evolving enhancer 1 (PsbO) protein reveals replacement of a nuclear-encoded plastid gene by that of a haptophyte tertiary endosymbiont[J]. PNAS,2002,99(14):9294-9299.
[29] Ifuku K, Ishihara S, Sato F. Molecular functions of oxygen-evolving complex family Proteins in photosynthetic electron flow[J]. Journal of Integrative Plant Biology, 2010, 52(8): 723-734.
[30] Sproviero E M, McEvoy J P, Gascón J A, et al. Computational insights into the O₂-evolving complex of photosystem II[J]. Photosynth Reseaches,2008,97(1):91-114.
[31] Liang Y, Chen H, Tang M J, et al. Responses of Jatropha curcas seedlings to cold stress: photosynthesis-related proteins and chlorophyll fluorescence characteristics[J]. Physiologia Plantarum,2007,131(3):508-517.
[32] Lee D G, Ahsan N, Lee S H, et al. An approach to identify cold-induced low-abundant proteins in rice leaf[J]. C.R.Biologies,2007,330(3):215-225.
[33] Hashimoto M, Komatsu S. Proteomic analysis of rice seedlings during cold stress[J]. Proteomics,2007,7(8):1293-1302.
[34] Herman E M, Rotter K, Premakumar R, et al. Additional freeze hardiness in wheat acquired by exposure to 23℃ is associated with extensive physiological, morphological and molecular changes[J]. Journal of Experimental Botany,2006,57(14):3601-3618.
[35] Spreitzer R J. Role of the small subunit in ribulose-1,5-bisphosphate carboxylase/oxygenase[J]. Archives of Biochemistry and Biophysics,2003,414(2):141-149.
[36] Peterhansel C, Niessen M, Kebeish R M. Metabolic engineering towards the enhancement of photosynthesis[J]. Photochemistry and Photobiology,2008,84(6):1317-1323.
[37] Komatsu S, Yamada E, Furukawa K. Cold stress changes the concanavalin A-positive glycosylation pattern of proteins expressed in the basal parts of rice leaf sheaths[J]. Amino Acids,2009,36(1):115-123.
[38] Gao F, Zhou Y J, Zhu W P, et al. Proteomic analysis of cold stress-responsive proteins in Thellungiella rosette leaves[J]. Planta,2009,230(5):1033-1046.
[39] Renaut J, Lutts S, Hoffmann L, et al. Responses of poplar to chilling temperatures: proteomic and physiological aspects[J]. Plant Biology,2004,6(1):81-90.
[40] Guo Z, Ou W, Lu S, et al. Differential responses of antioxidative system to chilling and drought in four ricecultivars differing in sensitivity[J]. Plant Physiology and Biochemistry,2006,44(11-12):828-836.
[41] 孙勇,戴绍军.低温对植物叶片蛋白质组的影响[J].现代农业科技,2011(8):29-30.
[42] Goulas E, Schubert M, Kieselbach T, et al. The chloroplast lumen and stromal proteomes of Arabidopsis thaliana show differential sensitivity to short- and long-term exposure to low temperature[J]. The Plant Journal,2006,47(5):720-734.
[43] Apel K, Hirt H. Reactive oxygen species: metabolism, oxidative stress, and signal transduction[J]. Annu Rev Plant Biol,2004,55:373-399.