甘蔗rubisco和pepc基因的克隆及其在不同施氮水平下不同甘蔗品种中的表达生物学植物学专业论文.docxVIP

2．通过对荧光定量和Westem 2．通过对荧光定量和Westem blotting结果进行分析发现，氮利用效率 高的甘蔗品种GXB9的三个光合酶基因在转录和蛋白质水平上表达量均比 较稳定，ROC22次之，而氮利用效率最低的甘蔗品种GTll基因表达波动 最为剧烈。实验还发现施氮处理主要通过影响而毋的表达来影响甘蔗的光 合作用。 3．对叶片中光合酶蛋白的Western blotting检测发现LSU在10月份时部 分被降解为略小于全长的肽段，这可能与甘蔗进入成熟期净光合速率降低 有关。 4．在本研究条件下，施氮处理对甘蔗光合速率有明显的影响，虽然中 氮处理和高氮处理的净光合速率相差不大，但都显著高于低氮处理。荧光 定量PCR和Westem blotting分析结果表明：氮主要通过影响而岱基因的表 达来影响Rubisco全酶含量。 关键词：甘蔗 施氮 光合作用 Rubisco PEPC实时荧光定量PCR Westem blotting II 万方数据 CLONEING CLONEING oF王UBISCO AND PEPC FROM SUGARCANE AND THEIR EXPRES SIoN I胭ER DⅡ7FERENT NITROGEN FERTILIZATION AND SUGARCANE CUITⅣ=ARS Wu Chaoxing (College ofAgriculture，Guangxi University,Nanning 530005，China) ABSTRACT Sugarcane(Saccharrum spp．)is an important Ca crop contributing to over 90％of sugar production in China．In recent years，the sugarcane industry is facing a lot of challenges restricting its development，among which over-fertilization is one of the most important concerns．In China,nitrogen over-fertilization is very serious which has led to not only low nitrogen utilization efficiency and high sugarcane production cost，but also heavier occurrence of pests and diseases，and even environment pollution．The Rubisco and PEPC are the key enzymes catalyzing photosynthesis in C4 plant, accounting for the large amount of soluble protein in leaf．Nitrogen carl affect the photosynthesis by impacting the content and enzymatic characteristics． Photosynthesis is closely related with nitrogen utilization，and it is important to study the mechanism of interaction between photosynthesis and nitrogen utilization to improve the．nitrogen utilization efficiency，。in—sugarcane III 万方数据 production．In production． In this study the full length CDS sequences of three genes encoding two key enzymes catalyzing photosynthesis in sugarcane were cloned from cultivar GT 1 1 using PCR technology,and the prokaryotic expression and protein purification were done．The expression characteristics of the three genes were detected in three sugarcane cultivars(GT 1 1，