不同气候环境中团棵期烟草叶片蛋白质组学分析

doi:10.3864/j.issn.0578-1752.2013.04.020

摘要/Abstract

摘要： 【目的】应用比较蛋白组学策略，在蛋白表达水平揭示团棵期烟草对不同气候环境响应的分子机理。【方法】在相同土壤和大田栽培管理条件下，分别在昆明市云南农业大学农场和丽江市玉龙县金庄两个不同气候环境的生态试验点盆栽烤烟品种云烟87，在团棵期进行农艺性状、SPAD值和净光合速率测定，并运用双向电泳和MALDI-TOF-MS质谱鉴定技术研究烟草叶片蛋白质表达谱的变化。【结果】金庄点烟草具有较高的SPAD值和净光合速率，较大的株高、节间距、茎围、叶宽和叶面积等农艺性状参数。蛋白质组学分析表明，在两个生态点烟草叶片中共检测到39个表达量相差2倍以上的蛋白质点，通过MALDI-TOF-MS质谱鉴定和数据库检索注释了33个蛋白质点，相对于昆明点，金庄点有3个（9.09%）特异表达，7个（21.12%）表达量上调，19个（57.57%）表达量下调的蛋白质点，而昆明点有4个（12.12%）特异表达蛋白质点；功能分类表明，它们分别参与光合作用、防御/抗胁迫、蛋白质合成、矿质代谢等细胞过程。【结论】金庄点烟草具有较高SPAD值，净光合速率，较大的株高、节间距、茎围、叶宽和叶面积等农艺性状参数，这与该点烟草叶片中积累较多高光合效率相关的蛋白质相一致，而昆明点烟草叶片则富集较多的叶绿体蛋白质合成、防御/胁迫以及氮、硫等矿质元素代谢相关的蛋白质。该结果初步阐明了丽江烟区烟叶总糖、还原糖含量较高，总氮、烟碱含量偏低特征的分子机制。

关键词: 烟草 , 团棵期 , 农艺性状 , 蛋白质组 , 气候环境

Abstract: 【Objective】 The effects of different climatic conditions on the molecular developmental mechanism of rosette stage tobacco were studied. 【Method】 Under the same conditions of soil, cultivation and management, flue-cured tobacco cultivar Yun 87 was pot-planted in the farm of Yunnan Agricultural University in Kunming and Jinzhuang of Yulong county in Lijiang, respectively. When tobacco developed into the rosette stage, the agronomic traits, the SPAD values and the net photosynthetic rate were determined, simultaneously, total proteins of the tobacco leaves were separated by two-dimensional electrophoresis(2-DE) and the more than 2 times differential expression proteins were analyzed through MALDI-TOF-MS. 【Result】 Compared with the Kunming experimental site, the tobaccos planted in Jingzhuang had the higher SPAD value, net-photosynthetic rate and the speeding of developmental process resulted in the increase of the plant height, length of inter stem nodes, stem circumference, leaf width as well as leaf area. The proteomic analysis shown that the leaves of tobaccos planted in both experimental sites had 39 protein spots with more than 2 times differential expression, in which 33 were identified and annotated. Among the annotated ones, compared with Kunming site, tobaccos of Jingzhang site had 3 (9.09%) specifically expressed, 7 (21.21%) up-regulated, 19 (57.57%) down-regulated proteins, whilst the tobaccos planted in Kunming had 4 (12.12%) specific proteins. The functional classification of the differentially expressed proteins belonged to tobacco metabolism of photosynthesis, defense or (and) stress tolerance , chloroplast protein synthesis, mineral metabolism and so on.【Conclusion】 Compared with rosette stage tobaccos of Kunming, the counterparts of Jingzhuang had higher SPAD value, net-photosynthetic rate，and larger agronomic traits, such as plant height, length of inter stem nodes, stem circumference, leaf width as well as leaf area, and those performance was consistent to that the Jingzhuang tobaccos accumulated proteins related to higher photosynthetic efficiency, whilst the Kunming tobaccos enriched the proteins for improving stress tolerance, chloroplast protein biosynthesis and mineral metabolism as well. The results preliminarily illuminated the molecular mechanism of Lijiang tobacco development with characteristics which contains much more total sugars and reduction sugars , while less total nitrogen and nicotine.

Key words: Nicotiana tabacum , rosette stage , agronomic traits , proteome , climatic conditions

蔡永占, 周普雄, 李佛琳, 赵昶灵, 林春, 杨焕文, 毛自朝. 不同气候环境中团棵期烟草叶片蛋白质组学分析[J]. 中国农业科学, 2013, 46(4): 859-870.

CAI Yong-Zhan, ZHOU Pu-Xiong, LI Fu-Lin, ZHAO Chang-Ling, LIN Chun, YANG Huan-Wen, MAO Zi-Chao. Proteomic Analysis of Tobacco Rosette Stage Leaves Under Different Climatic Conditions[J]. Scientia Agricultura Sinica, 2013, 46(4): 859-870.

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.0578-1752.2013.04.020

https://www.chinaagrisci.com/CN/Y2013/V46/I4/859

参考文献

[1]周冀衡, 刘建利, 余砚碧, 杨宏光, 杨春江, 罗华元. 对我国部分替代进口烟叶工作的思考. 烟草农业科学, 2006, 2(3):216-217.

Zhou J H, Liu J L, Yu Y B, Yang H G, Yang C J, Luo H Y. Thoughts on works related to partial substitution for leaf tobacco import in China. Tobacco Agricultural Science, 2006, 2(3):216-217.(in Chinese)

[2]刘杰, 周清明, 彭新辉, 宾柯. 云南烟区烤烟中部烟叶化学成分比较分析. 作物研究, 2009, 23(2):104-107.

Liu J, Zhou Q M, Peng X H, Bin K. Comparative analysis on chemical components in middle leaves of flue-cured tobacco from tobacco areas of Yunnan Province. Crop Research, 2009, 23(2): 104-107. (in Chinese)

[3]胡小曼, 李佛琳, 杨焕文, 方志存, 杨悦章, 李成杰, 陈茂建. 丽江烟区烤烟烟叶致香物质和常规化学成分及其相关性研究. 云南农业大学学报, 2011, 26(S2):41-46.

Hu X M, Li F L, Yang H W, Fang Z C, Yang Y Z, Li C J, Chen M J. Study on aroma component content and regular chemical composition of flue-cured tobacco leaves and their correlation in Lijiang. Journal of Yunnan Agricultural University, 2011, 26(S2):41-46. (in Chinese)

[4]杨兴有, 刘国顺, 伍仁军, 夏林, 邢小军, 张建慧, 杜卫民, 谢良文. 不同生育期降低光强对烟草生长发育和品质的影响. 生态学杂志, 2007,26(7):1014-1020.

Yang X Y, Liu G S, Wu Z J, Xia L, Xing X J, Zhang J H, Du W M, Xie L W. Effects of shading at different growth stages on the growth, development and quality of tobacco. Chinese Journal of Ecology, 2007,26(7):1014-1020. (in Chinese)

[5]黄勇, 周冀衡, 郑明, 杨虹琦, 张苧元. UV-B对烟草生长发育及次生代谢的影响. 中国生态农业学报, 2009, 17(1):140-144.

Huang Y, Zhou J H, Zheng M, Yang H Q, Zhang N Y. Effect of UV-B on growth and development and secondary metabolism of flue-cured tobacco. Chinese Journal of Eco-Agriculture, 2009,17(1):140-144. (in Chinese)

[6]梁太波, 王建伟, 尹启生, 张艳玲, 蔡宪杰, 过伟民, 刘阳. 水分胁迫对烤烟氨同化和生物量的影响. 中国烟草学报, 2012,18(3): 50-54.

Liang T B, Wang J W, Yin Q S, Zhang Y L, Cai X J, Guo W M, Liu Y. Effects of water stress on ammonium assimilation and biomass of flue-cured tobacco. Acta Tabacaria Sinica, 2012,18(3):50-54. (in Chinese)

[7]时鹏, 申国明, 向德恩, 陈明辉, 王正旭, 向必坤, 孟贵星, 张琳, 曹学鸿. 恩施烟区主要气候因子与烤烟烟叶化学成分的关系. 中国烟草科学,2012, 33(4):13-16.

Shi P, Shen G M, Xiang D G, Chen M H, Wang Z X, Xiang B K, Meng G X, Zhang L, Cao X H. Relationships between main clinatic factors and chemical components of flue-cured tobacco leaves in Enshi. Chinese Tobacco Science, 2012,33(4):13-16. (in Chinese)

[8]崔红, 冀浩, 张华, 邵惠芳, 李东宵, 陈亮. 不同生态区烟草叶片蛋白质组学的比较. 生态学报,2008,28(10):4874-4880.

Cui H, Ji H, Zhang H, Shao H F, Li D X, Chen L. Comparative analysis of leaf proteomes between tobacco plants growing in different ecological regions of China. Acta Ecologica Sinca,2008, 28(10):4874-4880. (in Chinese)

[9]陈宗瑜, 毕婷, 吴潇潇. 滤减 UV-B 辐射对烤烟蛋白质组变化的影响. 生态学杂志, 2012, 31(5):1129-1135.

Chen Z Y, Bi T, Wu X X. Effects of reduced UV-B radiation on the variation of flue-cured tobacco proteome. Chinese Journal of Ecology, 2012, 31(5):1129-1135. (in Chinese)

[10]Isaacson T, Damasceno C M B, Saravanan R S, He Y, Catalá C, Saladié M, Rose J K C. Sample extraction techniques for enhanced proteomic analysis of plant tissues. Nature Protocols, 2006, 1(2):769-774.

[11]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(1-2):248-254.

[12]Boguth G, Harder A, Scheibe B, Wildgruber R, Weiss W. The current state of two-dimensional electrophoresis with immobilized pH gradients. Electrophoresis, 2000, 21:1037-1053.

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

[14]梅杨, 李海蓝, 谢晋, 罗红艺. 核酮糖-1, 5-二磷酸羧化酶/加氧酶 (Rubisco). 植物生理学通讯, 2007, 43(2):363-368.

Mei Y, Li H L, Xie J, Luo H Y. Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). Plant Physiology Communications, 2007, 43(2):363-368. (in Chinese)

[15]Spreitzer R J. Role of the small subunit in ribulose-1, 5-bisphosphate carboxylase/oxygenase. Archives of Biochemistry and Biophysics, 2003, 414(2):141-149.

[16]吴沿友, 张红萍, 吴德勇, 刘建, 李国祥. 植物叶片和角果的碳酸酐酶与光合速率的关系研究. 西北植物学报,2007,26:2094-2098.

Wu Y Y, Zhang H P, Wu D Y, Liu J, Li G X. Relations between carbonic anhydrase activities and photosynthetic rates in the leaves and pods of plants. Acta Botanica Boreali-Occidentalia Sinica, 2007,26:2094-2098. (in Chinese)

[17]Rengel Z. Carbonic anhydrase activity in leaves of wheat genotypes differing in Zn efficiency. Journal of Plant Physiology, 1995, 147(2):251-256.

[18]邓秋红, 甘露, 付春华, 栗茂腾, 余龙江. 芸薹属中几个物种碳酸酐酶活性的比较. 植物生理学通讯, 2009 (7):663-666.

Deng Q H, Gan L, Fu C H, Li M T, Yu L J. Comparison of carbonic anhydrase activities of several species in Brassica. Plant Physiology Communications, 2009 (7):663-666. (in Chinese)

[19]Liu J, Liu X, Dai L, Wang G. Recent progress in elucidating the structure, function and evolution of disease resistance genes in plants. Journal of Genetics and Genomics, 2007, 34(9):765-776.

[20]Alam I, Sharmin S A, Kim K H, Yang J K, Choi M S, Lee B H. Proteome analysis of soybean roots subjected to short-term drought stress. Plant and Soil, 2010, 333(1):491-505.

[21]Cantoni G. S-Adenosylmethionine: a new intermediate formed enzymatically from L-methionine and adenosinetriphosphate. Journal of Biological Chemistry, 1953, 204(1):403-416.

[22]Woodson W R, Park K Y, Drory A, Larsen P B, Wang H. Expression of ethylene biosynthetic pathway transcripts in senescing carnation flowers. Plant Physiology, 1992, 99(2): 526-532.

[23]Espartero J, Pintor-Toro J A, Pardo J M. Differential accumulation of S-adenosylmethionine synthetase transcripts in response to salt stress. Plant Molecular Biology, 1994, 25(2): 217-227.

[24]林凡云, 王士强, 胡银岗, 何蓓如. 糜子 SAMS 基因的克隆及其在干旱复水中的表达模式分析. 作物学报, 2008, 34(5): 777-782.

Lin F Y, Wang S Q, Hu Y G, He B R. Cloning of a s-adenosylmethionine synthetase gene from Broomcorn Millet (Panicum miliaceum L.) and its expression during drought and re-watering. Acta Agronomica Sinica, 2008,34(5):777-782. (in Chinese)

[25]Apel K, Hirt H. Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annual Review of Plant Biology, 2004, 55: 373-399.

[26]Davletova S, Schlauch K, Coutu J, Mittler R. The zinc-finger protein Zat12 plays a central role in reactive oxygen and abiotic stress signaling in Arabidopsis. Plant Physiology, 2005, 139(2):847-856.

[27]Koussevitzky S, Suzuki N, Huntington S, Armijo L, Sha W, Cortes D, Shulaev V, Mittler R. Ascorbate peroxidase 1 plays a key role in the response of Arabidopsis thaliana to stress combination. Journal of Biological Chemistry, 2008, 283(49): 34197-34203.

[28]Gökirmak T, Paul A L, Ferl R J. Plant phosphopeptide-binding proteins as signaling mediators. Current Opinion in Plant Biology, 2010,13(5): 527-532.

[29]Yan S, Tang Z, Su W, Sun W. Proteomic analysis of salt stress-responsive proteins in rice root. Proteomics, 2005, 5(1): 235-244.

[30]Jiang Y, Yang B, Harris N S, Deyholos M K. Comparative proteomic analysis of NaCl stress-responsive proteins in Arabidopsis roots. Journal of Experimental Botany, 2007, 58(13):3591-3607.

[31]Stone J M, Walker J C. Plant protein kinase families and signal transduction. Plant Physiology, 1995, 108(2):451-457.

[32]Mizoguchi T, Ichimura K, Shinozaki K. Environmental stress response in plants: the role of mitogen-activated protein kinases. Trends in Biotechnology, 1997, 15(1):15-19.

[33]Mizoguchi T, Irie K, Hirayama T, Hayashida N, Yamaguchi-Shinozaki K, Matsumoto K, Shinozaki K. A gene encoding a mitogen-activated protein kinase kinase kinase is induced simultaneously with genes for a mitogen-activated protein kinase and an S6 ribosomal protein kinase by touch, cold, and water stress in Arabidopsis thaliana. Proceedings of the National Academy of Sciences, 1996, 93(2): 765-769.