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作物杂志,2021, 第5期: 35–42 doi: 10.16035/j.issn.1001-7283.2021.05.006

• 遗传育种·种质资源·生物技术 • 上一篇    下一篇

不同作物光合蛋白复合物的提取及比较分析

王儒情1,2(), 华玮1, 刘军1()   

  1. 1中国农业科学院油料作物研究所/农业农村部油料作物生物学与遗传育种重点实验室,430062,湖北武汉
    2中国农业科学院研究生院,100081,北京
  • 收稿日期:2020-11-17 修回日期:2021-01-25 出版日期:2021-10-15 发布日期:2021-10-14
  • 通讯作者: 刘军
  • 作者简介:王儒情,研究方向为农艺与种业,E-mail： 365339576@qq.com
  • 基金资助:
    国家自然科学基金(31800196);中国农业科学院农科英才(海外引进工程)择优支持项目(CAAS-ASTIP-OCRI)

Preparations and Comparative Analysis of Photosynthetic Protein Complexes in Different Crops

Wang Ruqing1,2(), Hua Wei1, Liu Jun1()   

  1. 1Oil Crops Research Institute, Chinese Academy of Agricultural Sciences/Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture and Rural Affairs, Wuhan 430062, Hubei, China
    2Graduate School, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2020-11-17 Revised:2021-01-25 Online:2021-10-15 Published:2021-10-14
  • Contact: Liu Jun

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摘要：

以双子叶C3光合模式植物拟南芥和本氏烟草、单子叶C3光合模式作物水稻和C4光合玉米、双子叶C4光合白花菜、C3-C4光合中间型Moricandia suffruticosa（MS）二倍体物种以及多倍体C3作物油菜、大豆和花生为材料,运用蓝绿温和胶[Blue Native (BN)-PAGE]技术手段,在生化分子水平对C3与C4型、单子叶与双子叶、二倍体与多倍体不同作物光合复合物的差异进行解析。首先以拟南芥为材料优化BN-PAGE体系,证明在膜蛋白非离子型去垢剂十二烷基-β-D-麦芽糖苷（n-Dodecyl-β-D-Maltopyranoside,DM）终浓度为1%、处理时间为10min时增溶效果最好。然后用优化后的体系比较不同作物种间光合/类囊体膜复合物的差异。结果表明,光合复合物及其相应代表性亚基在蛋白丰度和组成形式等方面显著不同。与C3、双子叶和二倍体植物相比,C4、单子叶和多倍体作物中光系统II（PSII）核心蛋白D1在复合物中的含量升高;水稻中Cyt b6f含量最高,不同作物之间Cyt b6f核心亚基Cyt f丰度差异较大;玉米中光系统I（PSI）核心亚基PsaA的含量显著高于其他8种作物,且C4作物高于C3作物;单子叶植物中ATP酶复合物核心亚基CF1β含量显著高于双子叶植物。本研究比较了不同植物间光合蛋白复合物差异,为改良作物的光合性状和提高光合效率提供参考。

关键词: 光合作用, 类囊体膜蛋白复合物, 蓝绿温和胶电泳, 光合蛋白, 作物

Abstract:

We performed analysis of the similarities and differences of photosynthetic protein complexes from C3, C4, C3-C4 intermediary plants and polyploid plants by using Blue Native (BN)-PAGE technology. Diploid C3 plants Arabidopsis and tobacco, monocot C3 plant rice, C4 plant maize, white cauliflower, C3-C4 intermediary plant MS, C3 plants rape, soybean and peanut were used as materials. To optimize the BN-PAGE system, we demonstrated that the optimal procedures are as follows. The final solubilization concentration of 1% should be used for the membrane protein non-ionic detergent n-Dodecyl-β-D-Maltopyranoside (DM) and the appropriate treatment time should be 10 minutes in Arabidopsis. This optimized system was then employed to analyze photosynthetic complexes (or thylakoids) in different types of crops. Our datas indicated that the photosynthetic complexes and their key subunits were dramatically distinct in terms of protein abundance and composition. We found that the content of PSII core protein D1 was higher in C4, monocots and polyploid crops compared to C3, dicots and diploid crops, respectively. Oryza sativa had the highest amount of Cyt b6f complex in all the crops. In particular, the core subunit Cyt f of Cyt b6f varied substantially between the different crops. The abundance of PSI core subunit PsaA in Zea mays was remarkably higher relative to the other eight crops, and the contents of C4 plants were higher than that of C3 plants. Monocots contained significantly higher level of ATPase core subunit CF1β than dicots. Taken together, our findings provide reference for further improvement of photosynthetic traits and enhancement of photosynthetic efficiency in crops.

Key words: Photosynthesis, Thylakoid membrane complexes, Blue native polyacrylamide gel electrophoresis (BN-PAGE), Photosynthetic protein, Crops

图1

BN/SDS-PAGE电泳分析模式植物拟南芥类囊体膜蛋白复合物 a：未染色一向BN-PAGE,b：考染一向BN-PAGE,c：二向BN/SDS PAGE分离类囊体膜蛋白

图2

BN-PAGE电泳分析不同作物类囊体膜蛋白复合物 a：9种作物一向BN-PAGE未染色,b：9种作物一向BN-PAGE考染

图3

一向BN-PAGE免疫印迹分析不同作物类囊体膜复合物

图4

免疫印迹分析不同作物光合蛋白

图5

二向BN/SDS PAGE分析不同作物类囊体膜复合物组成亚基

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