《植物生理学报》 2014, 50(4): 563-566

毕氏海蓬子光系统II颗粒的分离与鉴定

周峰*, 华春, 郑春梅

南京晓庄学院生物化工与环境工程学院, 南京211171

通信作者：周峰；E-mail: zfibcas@163.com；Tel: 025-86178274

摘　要：

采用去污剂Triton X-100增溶类囊体膜和高速离心的方法, 首次分离和纯化了毕氏海蓬子的光系统II (photosystem II, PSII)颗粒, 通过光谱学和SDS-PAGE对其进行鉴定并与类囊体膜进行比较。室温吸收光谱结果表明, PSII颗粒在蓝区的叶绿素(chlorophyll, Chl) b和胡萝卜素类吸收峰为485 nm, 在红区的Chl b吸收峰为655 nm, 这两个峰值均低于类囊体膜中的。77K荧光发射光谱结果表明, 提取的PSII颗粒基本不含光系统I (photosystem I, PSI)的低温荧光反射峰737 nm。77K荧光激发光谱结果显示, 海蓬子PSII颗粒在470~485 nm之间的Chl b和胡萝卜素类的荧光发射峰明显低于类囊体膜的。这说明在PSII中大部分的PSI已被除去。电泳结果显示, 海蓬子PSII颗粒缺少PSI反应中心蛋白质亚基PsaA和PsaB, 这说明提取到的PSII纯度较高, 这为进一步研究毕氏海蓬子PSII的结构与功能奠定基础。

关键词：海蓬子; 光系统II; 光谱; 电泳

收稿：2013-12-23   修定：2014-02-10

资助：江苏省自然科学基金青年基金(BK2012073)、国家高技术研究发展“863”计划(2012AA021701)和江苏省生态学重点学科建设项目(2012)。

Isolation and Identification of Photosystem II Protein of Salicornia bigelovii Torr.

ZHOU Feng*, HUA Chun, ZHENG Chun-Mei

School of Biochemical and Environmental Engineering, Nanjing Xiaozhuang University, Nanjing 211171, China

Corresponding author: ZHOU Feng; E-mail: zfibcas@163.com; Tel: 025-86178274

Abstract:

The photosystem II (PSII) of Salicornia bigelovii from coastal wetlands was firstly isolated and purified by solubilization of thylakoid membrane using detergent Triton X-100 and high-speed centrifugation. The PSII particles of S. bigelovii were studied and compared with thylakoid membrane by spectroscopy and SDS-PAGE methods. The results of absorption spectra at room temperature showed that the absorption peak of chlorophyll (Chl) b and carotenoid in the blue region was 485 nm, and the absorption peak of Chl b in the red region was 655 nm. Moreover, the two peaks were lower than those in the thylakoid membrane. The results of 77K fluorescence emission spectra showed that PSII particles extracted were free of the low temperature fluorescence peak 737 nm of photosystem I (PSI). It was shown from 77K fluorescence excitation spectra that the emission peak between 470–485 nm was lower than that of the thylakoid membrane. These indicated that most of the PSI has been removed from the extracted PSII. The PSII particles of S. bigelovii were lack of PSI reaction center protein subunits (PsaA and PsaB). It demonstrated that the extracted PSII particles were high purity, which could be used for further study on the structure and function of S. bigelovii.

Key words: Salicornia bigelovii; PSII; spectra; electrophoresis