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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。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),表明栽培种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;利用Western Blot技术分析叶片蛋白质表达水平,结果显示ZM-Seaside叶片与光合作用相关蛋白质Rubisco、OEC和PRXQ的表达水平显著高于花叶木薯变种;采用苯酚法提取叶片全蛋白质,并进行双向电泳分离,及Delta2D软件确定差异蛋白质点,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;通过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,成功鉴定到其中16个涉及光合作用、碳和能量代谢、分子伴侣、结构蛋白、保护蛋白、解毒和抗氧化及未知功能蛋白质;利用String在线软件构建蛋白质互作网络,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促进作用。
In order to study the effects of cassava leaf photosynthetic efficiency on root tuber yield, this study analyzed the differences in photosynthetic ability of cassava varieties (low yielding germplasm) and cultivar ZM-Seaside (high yielding germplasm) by proteomics, The reason for the difference in yield provides the basic data for breeding high-yielding cassava varieties. The net photosynthetic rate (Pn), stomatal conductance (Cs), intercellular CO2 concentration (Ci) and transpiration rate (Tr) of leaves were measured by portable LI-6400 photosynthesis analyzer. The results showed that the cultivars ZM-Seaside and Mucous- There was no significant difference in Cs, Ci and Tr among leaves, but Pn in ZM-Seaside was significantly higher than that in Mucous var. Moui. Western Blot analysis of leaf protein expression showed that ZM-Seaside leaves were related to photosynthesis-related proteins Rubisco, OEC and PRXQ The protein level in ZM-Seaside leaves was significantly higher than that in Mucous cassava varieties. The total protein was extracted by phenol method and separated by two-dimensional gel electrophoresis. The differential protein spots were determined by Delta2D software. Twenty different protein spots were obtained, of which 15 were up-regulated and 5 were down-regulated. The differential proteins were identified by MALDI-TOF-MS and classified according to KEGG database. Sixteen of them were successfully identified as photosynthesis, Energy metabolism, molecular chaperones, structural proteins, protective proteins, detoxification and antioxidant and unknown function proteins; construction of protein-protein interaction networks using String online software It is speculated that Ribulose-5-phosphate-3-epimerase and chloroplast latex aldolase-like protein are the key proteins that affect the photosynthetic efficiency of cassava leaves. The upregulation of them contributes to the improvement of tuberous root yield.