Cloning and functional analysis of the psbD gene in Pseudosasa japonica f. akebonosuji
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摘要: 植物叶绿体光系统Ⅱ是植物进行光能转化、水的裂解和释放氧气的重要蛋白复合物,psbD基因编码光系统Ⅱ(PSⅡ)作用中心蛋白D2,D2和D1构成的异源二聚体上结合着与电子传递有关的大部分辅助因子和色素分子。以花叶矢竹Pseudosasa japonica f. akebonosuji为材料,提取总DNA,聚合酶链式反应(PCR)扩增psbD基因全长序列(Pj-psbD)。序列分析表明:其开放阅读框(ORF)为1 059 bp,编码352个氨基酸,相对分子质量为39.59 kD,等电点为5.34,有6个跨膜结构,有别于高等植物中psbD蛋白普遍存在的5个跨膜区。psbD还具备有多个与光合作用有关的结构域和功能位点。本实验用荧光定量PCR技术检测了花叶矢竹叶片3个不同生长阶段和3种叶色中psbD基因的表达量。分析表明:在花叶矢竹叶片生长发育时期,为了满足叶片生长发育和叶绿体的形成的需要,psbD基因表达量逐步提高。研究还发现:psbD在白叶中的表达量显著高于绿叶,这可能与植物的光保护机制有关,降低强光对白色和部分白叶的光合系统的损伤。图8表1参25Abstract: Photosystem Ⅱ (PSⅡ) of a plant chloroplast is an important protein complex in light-dependent reactions of oxygenic photosynthesis with the psbD gene encoding the center protein D2. Until now, psbD gene in Pseudosasa japonica f. akebonosuji still would not been sequenced and reported. In order to analysis the psbD gene structure and the potential relation of psbD gene function and leaf color variation, in this study the genomic DNA of Pseudosasa japonica f. akebonosuji was extracted and the full-length psbD gene sequence (Pj-psbD) was amplified by polymerase chain reaction (PCR). The expression pattern for Pj-psbD was detected by the fluorescence quantitative PCR technique in three different growth (S1, S2 and S3) stages using green, stripe and albino leaves of P. japonica f. akebonosuji leaves with three material replicates and three expriements replicates. Results showed that the Open Reading Frame (ORF) of Pj-psbD was 1 059 bp long encoding 352 amino acids with a relative molecular weight of 39.59 kD and an isoelectric point of 5.34. In the Pj-psbD polypeptide chain, there were six transmembrane domains, plus multiple photosynthesis domains and other function sites. Among the three colors of leaves the psbD expression level in the three growth stages of the albino leaves was highest with more than 2-4 times than those of the according three growth stages of the green leaves. The continuously increasing expression levels of the gene along with leaf growth and development for the three types of leaves could meet the formation needs of the chloroplast, and the high psbD expression level in the albino leaf could be the protection mechanism that inhibits light damage in the photosynthetic system. [Ch, 8 fig. 1 tab. 25 ref.]
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链接本文:
https://zlxb.zafu.edu.cn/article/doi/10.11833/j.issn.2095-0756.2015.04.010

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