《植物生理学报》 2018, 54(5): 904-916

基于蛋白质组学对螺旋藻在高温胁迫下响应机制的初步研究

吕冰心^#, 常蓉^#, 李博生^*

北京林业大学生物科学与技术学院食品科学与工程系, 林业食品加工与安全北京市重点实验室(北京林业大学), 北京林业大学螺旋藻研究所, 北京100083

通信作者：李博生；E-mail: libosheng1119@163.com

摘　要：

本实验以螺旋藻为材料, 通过iTRAQ对螺旋藻细胞在高温胁迫下的全蛋白进行定量分析。结果表明: 40ºC是螺旋藻可恢复的最大耐受胁迫温度, 并在此温度胁迫条件下启动响应机制。差异表达蛋白的筛选结果确定了18 523个独特的肽和2 085个蛋白质, 此外, 在UniProtKB/Swiss-Prot数据库中注释了142种独特的蛋白质。GO功能注释中, 共有207条蛋白序列被793条GO功能条目注释, 平均GO层次为6.545。KEGG通路注释中, 检测到呈现显著差异性表达的注释蛋白117个, 涉及光合作用、能量代谢、RNA的转录和翻译等方面。荧光定量PCR结果显示, 测序结果与iTRAQ实验相一致, 光合系统I P700叶绿素脱辅基蛋白A1、果糖1,6-二磷酸酶、核酮糖二磷酸羧化酶大侧链、光合系统I反应中心亚基XI下调; 顺反异构酶、热激蛋白70、热激蛋白90、磷酸甘油酸激酶、二磷酸核苷酸激酶上调。由此可知, 螺旋藻经高温胁迫后, 与光合作用和遗传信息相关的蛋白是影响螺旋藻热应激的关键。

关键词：螺旋藻; 高温胁迫; 蛋白质组学; 双重机制; 光合作用

修定：2018-04-13

资助：国家林业局重点(2017-01)。

Study on the response mechanism of Spirulina platensis to high temperature stress based on proteomics

LÜ Bing-Xin^#, CHANG Rong^#, LI Bo-Sheng^*

Beijing Forestry University, Department of Food Science and Engineering, College of Biological Sciences and Technology, Beijing Key Laboratory of Forest Food Processing and Safety, Institute of Spirulina, Beijing 100083, China

Corresponding author: LI Bo-Sheng; E-mail: libosheng1119@163.com

Abstract:

In this experiment, Spirulina was used as a material to quantitatively analyze whole proteins of Spirulina cells under high temperature stress by iTRAQ. The results showed that 40ºC was the maximum tolerable stress of Spirulina, and the response mechanism was initiated under this stress condition. The screening results for differentially expressed proteins identified 18 523 unique peptides and 2 085 proteins. Besides, 142 unique proteins were annotated in the UniProtKB/Swiss-Prot database. In the GO functional annotation, a total of 207 protein sequences were annotated with 793 GO functional entries, with an average GO level of 6.545. In the KEGG pathway annotation, 117 annotated proteins with significantly different expressions were detected and involved in photosynthesis, energy metabolism, RNA transcription and translation, and other aspects. The results of RT-qPCR showed that the sequencing results were consistent with the iTRAQ experiment. Photosystem I P700 chlorophyll a apoprotein A1, D-fructose 1,6-bisphosphatase, Ribulose bisphosphate carboxylase large chain, Photosystem I reaction center subunit XI were down-regulated; PpiC-type peptidyl-prolyl cis-trans isomerase, Heat shock protein Hsp70, Heat shock protein 90, Phosphoglycerate kinase, Nucleoside diphosphate kinase were up-regulated. Therefore, it is known that the proteins associated with photosynthesis and genetic information were the key factors affecting the heat stress of Spirulina.

Key words: Spirulina; heat stress; proteomics; double mechanism; photosynthesis