《植物生理学报》 2013, 49(10): 1089-1098

不同提取液对巴西橡胶树橡胶粒子蛋白质提取和分离的影响

代龙军, 黎瑜, 聂智毅, 康桂娟, 段翠芳, 曾日中*

中国热带农业科学院橡胶研究所, 农业部橡胶树生物学与遗传资源利用重点实验室, 省部共建国家重点实验室培育基地, 海南儋州571737

通信作者：曾日中；E-mail: hnzrz@aliyun.com；Tel: 0898-23300776

摘　要：

橡胶粒子是巴西橡胶树(Hevea brasiliensis)乳管细胞内进行橡胶生物合成的亚细胞结构; 对橡胶粒子的蛋白质组学研究, 可为揭示天然橡胶生物合成机理提供线索。采用5种提取液分别提取橡胶粒子蛋白, 并对橡胶粒子蛋白进行SDS-PAGE和16-BAC/SDS-PAGE电泳分离及MALDI TOF/TOF串联质谱分析, 证明不同提取液抽提的橡胶粒子蛋白具有不同组成, 发现分子量较高的橡胶延伸因子家族蛋白更难从橡胶粒子上被洗脱和提取。通过检索橡胶树转录组数据库, 鉴定了3个新的橡胶粒子蛋白, 即醌氧化还原酶、含蓖麻毒素B链凝集素结构域蛋白及枯萎/脱水相关蛋白。本研究建立的橡胶粒子蛋白质提取和分离方法, 为进一步鉴定低丰度和具有重要功能的橡胶粒子蛋白提供了参照体系。

关键词：巴西橡胶树; 橡胶粒子; 蛋白质; 提取液

收稿：2013-06-04   修定：2013-08-13

资助：国家自然科学基金(31270713)、中国热带农业科学院橡胶研究所基本科研业务费专项(1630022013014)和海南省自然科学基金(312028)

Effects of Different Solutions on Extraction and Separation of Rubber Particle Proteins of Hevea brasiliensis Muell. Arg.

DAI Long-Jun, LI Yu, NIE Zhi-Yi, KANG Gui-Juan, DUAN Cui-Fang, ZENG Ri-Zhong*

State Key Laboratory Incubation Base, Key Laboratory of Biology and Genetic Resources of Rubber Tree, Ministry of Agriculture, Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou, Hainan 571737, China

Corresponding author: ZENG Ri-Zhong; E-mail: hnzrz@aliyun.com; Tel: 0898-23300776

Abstract:

Rubber particles are organelles responsible for the biosynthesis of natural rubber in Hevea brasiliensis (rubber tree). Proteomics investigations into rubber particles may provide clues for elucidating the mechanisms of natural rubber biosynthesis. In this study total proteins extracted from rubber particles using five different buffers were subjected to SDS-PAGE and 16-BAC/SDS-PAGE analysis, respectively, and then analyzed by MALDI TOF/TOF tandem mass spectrometry. The data demonstrated that rubber particle proteins extracted with different buffers had different components, and proteins with higher molecular weight in the rubber elongation factor family were not easily extracted from the rubber particles. Three novel rubber particle proteins, quinone oxidoreductase, protein containing ricin B chain lectin domain, and senescence/dehydration-related protein, were identified through searching a transcriptome database of H. brasiliensis. Our study establishes a reference system which is helpful to identify novel rubber particle proteins with low-abundance and important biological functions.

Key words: Hevea brasiliensis; rubber particle; protein; extraction buffer