不同气候环境中团棵期烟草叶片蛋白质组学分析

Proteomic Analysis of Tobacco Rosette Stage Leaves Under Different Climatic Conditions

【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。

【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.