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期 刊 ：植物遗传资源学报 2016年 17卷 5期

关键词：花叶木薯变种；栽培种ZM-Seaside；光合参数；蛋白质组；

Keywords:Mosaic-leaf Mutation, ZM-Seaside, parameters, proteomics,

摘 要 ：摘要：为研究木薯叶片光合效率对块根产量的影响，本研究利用蛋白质组学方法分析花叶木薯变种(低产种质)和栽培种ZM-Seaside(高产种质)光合作用能力的差异，揭示其鲜薯产量差异原因，为选育高产木薯品种提供基础数据。研究采用便携式 LI- 6400 光合作用测定仪测定叶片净光合速率(Pn)、气孔导度(Cs) 、胞间 CO2 浓度(Ci)和蒸腾速率(Tr)；利用Western Blot技术分析蛋白质表达水平；采用苯酚法提取叶片全蛋白质；利用蛋白质双向电泳技术分离叶片全蛋白质；利用Delta2D软件确定差异蛋白质点；利用MALDI-TOF-MS鉴定差异蛋白质，结合KEGG数据库将其按照功能进行分类；利用String软件构建蛋白质互作网络。结果发现栽培种ZM-Seaside和花叶木薯变种叶片Cs、Ci和Tr没有显著差异，但ZM-Seaside的Pn显著高于花叶木薯变种。Western Blot方法揭示ZM-Seaside叶片与光合作用相关的蛋白质Rubisco、OEC和PRXQ的表达水平显著高于花叶木薯变种。以花叶木薯为对照，在ZM-Seaside叶片蛋白质双向电泳图谱上得到20个差异蛋白质点，其中上调表达15个，下调表达5个；经质谱技术成功鉴定到其中16个，涉及光合作用(3)、碳和能量代谢(4)、分子伴侣(2)、结构蛋白(2)、保护蛋白(1)、解毒和抗氧化(2)及未知功能蛋白质(2)。通过差异蛋白质相互作用网络推测Ribulose-5-phosphate-3-epimerase和chloroplast latex aldolase-like protein是影响木薯叶片光合效率的关键蛋白质，由于它们的上调表达，对木薯块根产量提高有一定的促进作用。

Abstract:Abstract: In order to indicate the influences of cassava leaf photosynthetic rate on roots yield in the present study, the proteomic methods was used to analyze the differences of photosynthetic capacity in both cassava genotypes ZM-Seaside (high yield) and mosaic-leaf mutation (low yield) to reveal the different reason of their root yield and provide theoretical basis for the selection of high yield cassava varieties. The leaf net photosynthetic rate (Pn), stomatal conductance (Cs), intercellular CO2 concentration (Ci), transpiration rate (Tr) were measured using a portable analyzer LI-6400. Western blot was used to analysis of protein expression level. Leaf global proteins were extracted by phenol extraction, and then were separated using two-dimensional electrophoresis. Delta 2D software was used to determine the differentially expressed proteins. The differentially expressed proteins were identified using MALDI-TOF-MS in combination with the KEGG database to classify proteins according to their functions. The results showed leaf Pn in ZM-Seaside was significantly increased more than that of mosaic-leaf mutation, while Cs, Ci and Tr in both genotypes were no significant differences. Western blot revealed that the expression levels of Rubisco, OEC and PRXQ, related with photosynthetic pathway, in ZM-Seaside was significantly higher than that of mosaic-leaf mutation. 20 different protein spots were detected in the leaves of ZM-Seaside compared with mosaic-leaf mutation, of which 15 were up-regulated, 5 was down-regulated. Sixteen protein spots were successfully identified, in which they involved in photosynthesis (3), carbohydrate and energy metabolism (4), chaperones (2), structure (2), defense (1), detoxifying and antioxidant (2), and unknown functions (2). Ribulose-5-phosphate-3-epimerase and chloroplast latex aldolase-like protein were theorized as key proteins through the network of differential protein-protein interaction. The root yield of cassava ZM-Seaside was improved probably based on their up-regulation in leaves.

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