白叶1号白化过程中叶绿体蛋白质组差异分析

LI Qin1; 2; 3; CHENG Xiaomei1; LI Yongdi1; YANG Peidi4; HUANG Jian'an1; 3*; LIU Zhonghua1; 2; 3*

doi:10.13305/j.cnki.jts.2019.03.010

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茶叶科学 ›› 2019, Vol. 39 ›› Issue (3) : 325-334. DOI: 10.13305/j.cnki.jts.2019.03.010

白叶1号白化过程中叶绿体蛋白质组差异分析

李勤^1,2,3，程晓梅¹，李永迪¹，杨培迪⁴，黄建安^1,3*，刘仲华^1,2,3*

作者信息 +

Analysis of the Chloroplast Proteome Difference of ‘Baiye 1’ [Camellia sinensis (L.) O Kuntze] during Periodic Albinism

LI Qin^1,2,3, CHENG Xiaomei¹, LI Yongdi¹, YANG Peidi⁴, HUANG Jian'an^1,3*, LIU Zhonghua^1,2,3*

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文章历史 +

摘要

白叶1号是一种温度敏感型白化茶树品种，叶绿体的变化是其产生阶段性白化现象的关键因素。本研究以白叶1号鲜叶叶绿体为研究对象，采用双向电泳、质谱鉴定结合生物信息学分析，研究阶段性白化过程中叶绿体蛋白的表达差异，探讨白叶1号阶段性白化现象的分子机制。结果表明，在白叶1号白化前期、白化期和复绿期叶绿体中分别识别726、748、718个蛋白质，其中差异表达的蛋白59个，质谱成功鉴定22个差异表达蛋白。生物信息学分析表明，差异表达蛋白直接或间接参与了光合作用、应激响应、核酸代谢、物质代谢和未知功能等，其中与光合作用相关的差异表达蛋白最多，占31.82%，表明阶段性白化现象可能与这些生理功能相关。通过荧光定量PCR分析发现，差异蛋白的基因表达与蛋白表达存在一定差异，这可能是由于蛋白质翻译后加工及修饰造成的。上述研究为进一步揭示白叶1号阶段性白化现象产生的分子机制奠定了理论基础。

Abstract

‘Baiye 1’ is a kind of temperature sensitive tea cultivar. The change of chloroplast is the key factor for the periodic albinism of ‘Baiye 1’. To understand the mechanism of periodic albinism of ‘Baiye 1’, two dimensional electrophoresis (2-DE) and mass spectrometry was adopted to separate and identify the chloroplast proteins, which were significantly changed during the three developmental periods. The results show that 726, 748 and 718 protein spots were separated at the pre-albinistic, albinistic and regreen stages, respectively. The expression levels of 59 protein spots varied markedly during the three development stages. A total of 22 protein spots were successfully identified by MS, which were involved in photosynthesis, stress response, metabolism of nucleic acid, substance metabolism and unknown function. Photosynthetic proteins were the most affected proteins, which account for 31.82% in the significantly changed proteins. These results indicate that these physiological processes might play crucial roles in the periodic albinism. The gene expression profiles of the differentially expressed proteins were also verified by real-time PCR analysis. The results show that the expressions of genes and proteins were not consistent, which might be related to the protein processing and post-modification. These results provide a theoretical basis for understanding the molecular mechanism of periodic albinism in ‘Baiye 1’.

导出引用

李勤, 程晓梅, 李永迪, 杨培迪, 黄建安, 刘仲华. 白叶1号白化过程中叶绿体蛋白质组差异分析[J]. 茶叶科学. 2019, 39(3): 325-334 https://doi.org/10.13305/j.cnki.jts.2019.03.010

LI Qin, CHENG Xiaomei, LI Yongdi, YANG Peid, HUANG Jian'an, LIU Zhonghua. Analysis of the Chloroplast Proteome Difference of ‘Baiye 1’ [Camellia sinensis (L.) O Kuntze] during Periodic Albinism[J]. Journal of Tea Science. 2019, 39(3): 325-334 https://doi.org/10.13305/j.cnki.jts.2019.03.010

中图分类号： S571.1

参考文献

[1] 李素芳. 安吉白茶返白机理的研究[J]. 中国计量学院学报, 2002, 13(3): 214-217. [2] 杨普香, 李文金, 聂樟清, 等. 安吉白茶茶多酚和氨基酸含量初探[J]. 蚕桑茶叶通讯, 2007(5): 33-34. [3] 成浩, 李素芳, 陈明, 等. 安吉白茶特异性状的生理生化本质[J]. 茶叶科学, 1999, 19(2): 87-92. [4] 曾超珍, 刘仲华. 安吉白茶阶段性白化机理的研究进展[J]. 分子植物育种, 2015, 13(12): 2905-2911. [5] 卢翠, 沈程文. 茶树白化变异研究进展[J]. 茶叶科学, 2016, 36(5): 445-451. [6] Li Q, Huang J, Liu S, et al. Proteomic analysis of young leaves at three developmental stages in an albino tea cultivar [J]. Proteome science, 2011, 9(1): 44. https://doi.org/10.118 6/1477-5956-9-44. [7] 贺庭琪, 郭安平, 杜伟, 等. 植物叶绿体蛋白质组学研究进展[J]. 热带作物学报, 2011, 32(11): 2196-2203. [8] 张立明, 刘亚军, 王云生, 等. 茶树叶绿体及其蛋白的分离研究[J]. 激光生物学报, 2011, 20(6): 802-808. [9] 肖龙, 张彩霞, 宗泽冉, 等. 苹果叶片应答轮纹病菌胁迫的叶绿体蛋白质组学分析[J]. 果树学报, 2016, 33(11): 1357-1366. [10] 肖龙, 张彩霞, 张利义, 等. 苹果叶片叶绿体分离及其蛋白提取、双向电泳方法的优化[J]. 果树学报, 2016, 33(6): 752-761. [11] Lydia Macias-Rubalcava M, Claudia Garcia-Mendez M, King-Diaz B, et al. Effect of phytotoxic secondary metabolites and semisynthetic compounds from endophytic fungus Xylaria feejeensis strain SM3e-1b on spinach chloroplast photosynthesis [J]. Journal of Photochemistry and Photobiology B: Biology, 2017, 166: 35-43. [12] Kley J, Heil M, Muck A, et al. Isolating intact chloroplasts from small Arabidopsis samples for proteomic studies [J]. Analytical Biochemistry, 2010, 398(2): 198-202. [13] 李勤, 黄建安, 刘硕谦, 等. 茶树蛋白质双向电泳样品制备技术研究[J]. 茶叶科学, 2011, 31(3): 173-178. [14] Chomczynski P, Mackey K. Short technical reports. Modification of the TRI reagent procedure for isolation of RNA from polysaccharide- and proteoglycan-rich sources [J]. BioTechniques, 1995, 19(6): 942-945. [15] Baginsky S, Gruissem W. Chloroplast proteomics: potentials and challenges [J]. Journal of Experimental Botany, 2004, 55(400): 1213-1220. [16] Spreitzer RJ, Salvucci ME. Rubisco: Structure, regulatory interactions, and possibilities for a better enzyme [J]. Annual Review of Plant Biology, 2002, 53: 449-475. [17] 苏小静, 汪沛洪, 王永吉. 小麦突变体返白系返白机理研究-Ⅲ.返白阶段叶蛋白质、游离氨基酸变化规律的分析[J]. 西北农林科技大学学报(自然科学版), 1990, 18(4): 16-22. [18] 翁晓燕, 蒋德安, 陆庆. 水稻转绿型白化突变系W25转绿过程中Rubisco、Rubisco活化酶活性与光合速率的变化[J]. 植物生理学报, 2000, 26(3): 213-225. [19] Igamberdiev AU, Kleczkowski LA. Optimization of ATP synthase function in mitochondria and chloroplasts via the adenylate kinase equilibrium [J]. Frontiers in Plant Science, 2015, 6: 10. https://doi.org/10.3389/fpls.2015.00010. [20] Rast A, Heinz S, Nickelsen J. Biogenesis of thylakoid membranes [J]. Biochimica Et Biophysica Acta-Bioenergetics, 2015, 1847(9): 821-830. [21] 董慧. 叶绿体ATP合酶的活性调节与亚基的定点突变[D]. 上海: 中国科学院研究生院（上海生命科学研究院）, 2005. [22] 毛娟, 迟伟, 马今方, 等. 拟南芥PAB1蛋白参与叶绿体ATP合酶复合物调控机制的研究[J]. 生物物理学报, 2009, 25(S1): 242. [23] 胡光珍. 香蒲细胞色素b5基因的克隆和功能研究[D]. 上海: 华东师范大学, 2005. [24] 乌凤章, 王柏臣, 杨传平. 白桦叶绿体RNA结合蛋白的蛋白质组学分析(英文)[J]. 中国生物化学与分子生物学报, 2013, 29(2): 144-150. [25] Dupont FM. Metabolic pathways of the wheat (Triticum aestivum) endosperm amyloplast revealed by proteomics [J]. Bmc Plant Biology, 2008, 8(1): 39. https://doi.org/10.1186/ 1471-2229-8-39. [26] 张在宝, 李婉杰, 李九丽, 等. 植物RNA结合蛋白研究进展[J]. 中国农业科学, 2018, 51(21): 4007-4019. [27] 程晓梅. 安吉白茶阶段性返白过程中叶绿体蛋白质组学研究[D]. 长沙: 湖南农业大学, 2015. [28] 栗振义, 龙瑞才, 张铁军, 等. 植物热激蛋白研究进展[J]. 生物技术通报, 2016, 32(2): 7-13. [29] 王明强, 张道远. 植物热激蛋白70基因家族及其生物学功能研究进展[J]. 基因组学与应用生物学, 2015, 34(2): 421-428. [30] 侯典云. 小麦返白系叶绿体基因组分析及叶绿体超微结构和差异表达蛋白质研究[D]. 杨凌: 西北农林科技大学, 2009. [31] 孙卫红, 王伟青, 孟庆伟. 植物抗坏血酸过氧化物酶的作用机制、酶学及分子特性[J]. 植物生理学通讯, 2005, 41(2): 143-147. [32] 孙云. 茶叶抗坏血酸过氧化物酶（APX）的生理学与分子生物学研究[D]. 福州: 福建农林大学, 2009. [33] 董红霞. 水稻苗期叶色突变体的蛋白质组分析[D]. 武汉: 华中农业大学, 2010. [34] Langer T. AAA proteases: cellular machines for degrading membrane proteins [J]. Trends in Biochemical Sciences, 2000, 25(5): 247-251. [35] Sjogren LL, Stanne TM, Zheng B, et al. Structural and functional insights into the chloroplast ATP-dependent Clp protease in Arabidopsis [J]. The Plant cell, 2006, 18(10): 2635-2649. [36] Lu Y-J, Zhang Y-M, Rock CO. Product diversity and regulation of type II fatty acid synthases [J]. Biochemistry and Cell Biology-Biochimie et Biologie Cellulaire, 2004, 82(1): 145-155. [37] Ferro M, Salvi D, Brugiere S, et al. Proteomics of the chloroplast envelope membranes from Arabidopsis thaliana [J]. Molecular & Cellular Proteomics: MCP, 2003, 2(5): 325-345.