华南8号木薯及其四倍体诱导株系叶片蛋白质组及叶绿素荧光差异分析

doi:10.3864/j.issn.0578-1752.2013.19.003

摘要/Abstract

摘要： 【目的】通过对华南8号木薯及其四倍体诱导株系叶片差异蛋白质及叶绿素荧光参数的分析，在蛋白质水平上揭示两者叶片存在的差异以及其与光合效率间的关系。【方法】通过木薯嫩叶、根尖染色体压片及流式细胞观察对华南8号四倍体诱导株系进行鉴定，采用双向电泳技术分离叶片蛋白质，Delta 2D软件分析差异蛋白质并通过质谱技术鉴定，利用Western blot技术对部分差异蛋白质进行验证；采用95%乙醇直接提取法测定叶绿素含量，并用Imaging-Pam测定叶绿素荧光动力学参数。【结果】染色体压片及流式细胞结果均显示，诱导得到SC8多倍体株系为四倍体株系；得到的13个差异蛋白质点中上调表达12个，下调表达1个；经质谱技术成功鉴定到12个，其功能涉及碳代谢及能量代谢、光合作用、抗氧化、蛋白质代谢调控等；1个下调表达的蛋白质未得到成功匹配，其中参与光合作用的蛋白质占46.2%；四倍体株系叶片叶绿素a、叶绿素b和总叶绿素均显著升高；叶绿素荧光参数，包括Fo、ΦPSII、qP、NPQ和ETR均显著升高；【结论】参与碳代谢及能量代谢、光合作用等途径相关蛋白质表达水平的上调；叶绿素含量及叶绿素荧光参数的升高；表明四倍体株系叶片PSII反应中心捕光能力强、光化学转化效率高，从而提高了叶片的光合速率。

关键词: 木薯 , 四倍体株系 , 差异蛋白质 , 光合效率

Abstract: 【Objective】An investigation aimed at a better understanding of the differences in leaves proteins and its relationship with photosynthetic efficiency of cassava diploid and its tetraploid mutants, leaves proteins and chlorophyll fluorescence parameters were analyzed.【Method】Standard chromosome counting and flow cytometric analyses were used to determine the chromosome numbers and ploidy of each taxon, leaves proteins of cassava tetraploid mutants and diploid genotypes were excised and purified from 2D gels. Delta 2D software were used to analyze the differential proteins and MALDI-TOF-TOF-MS/MS were used to identify it. And western blot was used to verify some differential expressed proteins. 95% ethanol extraction method was used to measure the chlorophyll content and imaging-pam was used to determine the fluorescence parameters.【Result】Induced SC8 polyploid mutants were tetraploid, 13 differential expressed protein spots were identified, in which 12 were up-regulated and 1 was down-regulated, and 12 were successfully matched in database. The function of these 12 proteins which up-regulated were associated with carbohydrate and energy metabolism, photosynthesis, antioxidant and protein metabolism, 1 down-regulated protein was not matched in database, in which 46.2% differential expressed proteins were related with photosynthesis, chlorophyll a, chlorophyll b and total chlorophyll in tetraploid mutants were significantly increased, also the fluorescence parameters including Fo, ΦPSII, qP, NPQ and ETR.【Conclusion】Protein expression levels related with carbohydrate and energy metabolism and photosynthesis were up-regulated, chlorophyll content and chlorophyll fluorescence parameters were increased significantly, these indicate that high light-harvesting ability and high photochemical conversion rate in PSII reaction center of cassava tetraploid mutant leaves, then improving photosynthesis efficiency of SC8 tetraploidy mutants.

Key words: Cassava , tetraploid mutants , differential proteins , photosynthetic efficiency

安飞飞1, 凡杰1, 李庚虎2, 简纯平2, 李开绵1. 华南8号木薯及其四倍体诱导株系叶片蛋白质组及叶绿素荧光差异分析[J]. 中国农业科学, 2013, 46(19): 3978-3987.

AN Fei-Fei-1, FAN Jie-1, LI Geng-Hu-2, JIAN Chun-Ping-2, LI Kai-Mian-1. Comparison of Leaves Proteome and Chlorophyll Fluorescence of Cassava cv. SC8 and Its Tetraploid Mutants[J]. Scientia Agricultura Sinica, 2013, 46(19): 3978-3987.

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