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毛竹茎秆快速生长期类囊体膜蛋白复合物BN-PAGE分析
引用本文:傅卢成,卜柯丽,王灵杰,栗青丽,高培军,高岩,张汝民. 毛竹茎秆快速生长期类囊体膜蛋白复合物BN-PAGE分析[J]. 浙江农林大学学报, 2020, 37(4): 664-672. DOI: 10.11833/j.issn.2095-0756.20190398
作者姓名:傅卢成  卜柯丽  王灵杰  栗青丽  高培军  高岩  张汝民
作者单位:浙江农林大学 省部共建亚热带森林培育国家重点实验室,浙江 杭州 311300
基金项目:国家自然科学基金资助项目(31570686,30972397,31470704)
摘    要:  目的  探讨毛竹Phyllostachys edulis笋竹茎秆的光合特性和光系统的发育情况。  方法  以当年生毛竹叶片和笋竹茎秆为材料,采用蓝绿温和胶电泳(BN-PAGE)分析茎秆和叶片类囊体膜蛋白,同时测定了光合色素含量和77 K低温荧光发射光谱。  结果  茎秆叶绿素和类胡萝卜素质量分数显著低于叶片(P<0.01),随着茎秆发育,叶绿素和类胡萝卜素质量分数显著升高。茎秆和叶片类囊体膜PSⅡ核心复合物较完整,捕光色素较多;叶片和茎秆基部PSⅠ核心复合物分离主要得到PsaA/B和PsaD亚基,茎秆中部得到PsaA/B,茎秆顶部未发现PsaA/B。叶片和茎秆77 K低温荧光发射光谱在685和745 nm处有2个明显主峰,四阶导数光谱出现6个极大值,主要是PSⅡ和PSⅠ核心复合物的荧光发射峰以及由PSⅡ外周捕光天线(LHCⅡ)、PSⅡ内周捕光天线(CP47)、PSⅡ内周捕光天线(CP43)、PSⅠ反应中心复合体(RCI)、PSⅠ捕光天线(LHCⅠ)的发射荧光峰引起的肩峰,其中茎秆顶部LHCⅡ和PSⅡ核心复合体的特征发射峰与叶片相比有明显蓝移现象。  结论  毛竹茎秆中PSⅡ核心复合体已形成,随着茎秆发育,笋衣逐渐脱落,色素大量合成,内周天线蛋白CP47和CP43以及外周捕光天线蛋白逐渐形成;同时,茎秆受到光照后PSⅠ核心蛋白PsaA和PsaB开始形成,逐渐组装合成PSⅠ核心复合体。图4表2参45

关 键 词:毛竹   类囊体膜蛋白   光系统   发射荧光
收稿时间:2019-07-04

BN-PAGE analysis of thylakoid membrane protein complex during rapid growth of Phyllostachys edulis
FU Lucheng,BU Keli,WANG Lingjie,LI Qingli,GAO Peijun,GAO Yan,ZHANG Rumin. BN-PAGE analysis of thylakoid membrane protein complex during rapid growth of Phyllostachys edulis[J]. Journal of Zhejiang A&F University, 2020, 37(4): 664-672. DOI: 10.11833/j.issn.2095-0756.20190398
Authors:FU Lucheng  BU Keli  WANG Lingjie  LI Qingli  GAO Peijun  GAO Yan  ZHANG Rumin
Affiliation:State Key Laboratory of Subtropical Silviculture, Zhejiang A&F University, Hangzhou 311300, Zhejiang, China
Abstract:  Objective  The objective of this research is to reveal the photosynthetic characteristics and the development of photosystem of the stem of Phyllostachys edulis.  Method  Blue-greengel electrophoresis (BN-PAGE) was used to analyze the thylakoid membrane proteins in stems and leaves, and the changes in pigment content and the 77 K low temperature fluorescence emission spectrum were measured.  Result  The content of chlorophyll and carotenoid in stems was significantly lower than that in leaves (P<0.01), and with the development of stems, the pigment content increased significantly. The core complex of PSⅡ in the thylakoid membrane of stems and leaves was relatively complete and there were more light-catching pigments. PsaA/B and PsaD subunits were mainly isolated from PSI core complex in leaves and the stem base, and PsaA/B was obtained from the middle of stem, but no PsaA/B was found at the top of stem. There were two obvious main peaks at 685 and 745 nm in the 77 K low temperature fluorescence emission spectrum of leaves and stems, and six maximum values in the fourth derivative spectrum, which were mainly the fluorescence emission peaks of the core complex of PSⅡ and PSⅠ, and the shoulder peak caused by the emission fluorescence peak of the PSⅡ peripheral light catching antenna (LHC Ⅱ), PSⅡ inner peripheral light catching antenna (CP47), PSⅡ inner peripheral light catching antenna (CP43), PSⅠ reaction center complex (RCI), and PSI light catching antenna (LHC Ⅰ). The characteristic emission peak of LHC Ⅱ and PSⅡ core complex at the top of stem had obvious blue shifts compared with that of leaves.  Conclusion  The core complex of PSⅡ in stems of Ph. edulis has been formed. With the development of stems, the bamboo shoot coat gradually falls off, the pigments are synthesized in large amount, the inner antenna proteins CP47 and CP43 and the peripheral light-catching complex are formed gradually. Meanwhile, the PSⅠ core proteins PsaA and PsaB begin to form after the stems are exposed to light, and the core complex of PSⅠ is gradually assembled and synthesized. [Ch, 4 fig. 2 tab. 45 ref.]
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