芦笋三种花蕾性别分化相关差异蛋白表达分析

doi:10.3864/j.issn.0578-1752.2016.21.013

摘要/Abstract

摘要： 【目的】探究与芦笋性别分化相关的蛋白，为揭示性别分化的分子机制奠定基础。【方法】以芦笋雌株、雄株、雄性两性株两性分化期的花蕾为材料，采用双向电泳、质谱鉴定和生物信息学相结合的方法，对其进行蛋白质差异表达分析。【结果】通过双向电泳3次重复试验发现，雌、雄花蕾相比，雄花蕾特异蛋白点25个，上调蛋白点5个；雌花蕾特异蛋白点13个，上调蛋白点12个；雄性两性花蕾与雄花蕾相比，特异蛋白点19个，上调蛋白点8个。经过质谱鉴定及生物信息学分析，鉴定出雄花蕾特异或上调表达同源蛋白6个，包括1个促进α淀粉酶合成的luminal binding protein（BiP）；3个参与糖代谢的蛋白，分别为β淀粉酶、3-磷酸甘油醛脱氢酶和胞质磷酸甘油酸激酶；1个与质体相关的脂连接蛋白PAP fibrillin和1个未知功能的Os02g0634900蛋白；雄性两性花蕾和雌花蕾特异或上调表达同源蛋白16个，包括2个参与糖酵解过程的蛋白，即烯醇化酶1和胞质磷酸甘油酸激酶；2个维持细胞结构的蛋白，即肌动蛋白亚型B和肌动蛋白；2个参与能量代谢的蛋白，即ATP合成酶CF1α亚基和ATP合成酶β亚基；2个参与细胞内物质运输的蛋白，即小GTP结合蛋白和GTP结合蛋白，1个抑制蛋白质合成的核糖体失活蛋白，1个参与物质运输和信号转导的蛋白，即ADP核糖激化因子相似蛋白，1个催化磷酸基团转移的蛋白，即核苷二磷酸激酶，1个脂质相关蛋白，1个与光合作用（光系统Ⅱ）有关的蛋白，即放氧增强蛋白1，1个允许离子、糖和氨基酸被动转运穿过外膜的蛋白，即细胞外膜孔道蛋白，2个未知功能的蛋白，即细胞内病程相关蛋白亚型4和假想蛋白。【结论】β淀粉酶、3-磷酸甘油醛脱氢酶、胞质磷酸甘油酸激酶、luminal binding protein (BiP)、PAP fibrillin和Os02g0634900在芦笋上的同源蛋白与芦笋雄性器官发育相关；烯醇化酶1、胞质磷酸甘油酸激酶、肌动蛋白亚型B、肌动蛋白、ATP合成酶CF1α亚基、小GTP结合蛋白、GTP结合蛋白、核糖体失活蛋白、ADP核糖激化因子相似蛋白、核苷二磷酸激酶、脂质相关蛋白、放氧增强蛋白1、细胞外膜孔道蛋白和假想蛋白在芦笋上的同源蛋白，ATP合成酶β亚基、细胞内病程相关蛋白亚型4与芦笋雌性器官发育相关。放氧增强蛋白1和细胞外膜孔道蛋白在芦笋上的同源蛋白可能为雌性器官发育的关键蛋白。

关键词: 芦笋, 雌花蕾, 雄花蕾, 雄性两性花蕾, 性别分化, 差异蛋白

Abstract: 【Objective】 To investigate the proteins which are related to the sex differentiation, and lay a foundation for revealing the molecular mechanism of sex differentiation in Asparagus officinalis L. 【Method】 Differential proteins were analyzed in female flower buds, male flower buds and hermaphroditic flower buds at hermaphroditic differentiation stage by 2-D electrophoresis, mass spectrometry and bioinformatics method. 【Result】 Compared male and female flower buds, 25 specific protein spots and 5 up-regulated protein spots were detected in male flower buds, 13 specific protein spots and 12 up-regulated protein spots were detected in female flower buds. Compared with male flower buds, 19 specific protein spots and 8 up-regulated protein spots were detected in hermaphroditic flower buds. The proteins were identified by LC-MS-MS and analyzed through bioinformatics. Six specific or up-regulated homologous proteins of Asparagus were identified in male flower buds, including luminal binding protein (BiP) which promoted the synthesis of alpha amylase; beta-amylase, glyceraldehyde-3-phosphate and cytosolic phosphoglycerate kinase which are all involved in the glycolytic pathway, PAP fibrillin which is associated with liposomes,Os02g0634900 which its function is unknown. There were 16 specific or up-regulated homologous proteins of Asparagus were identified in hermaphroditic and female flower buds, including enolase 1 and cytosolic phospho-glycerate kinase which are involved in the glycolytic pathway, actin isoform B and actin which maintained cell structure, ATP synthase CF1 alpha subunit and ATP synthase beta subunit which participated in energy metabolism; small GTP-binding protein and GTP-binding protein which participated in material transportation, ribosome inactivating protein RIPm which depressed protein synthesis, ADP-ribosylation factor which participated in material transportation and signal transduction, nucleoside diphosphate kinase which catalyzed phosphate group transfer, plastid-lipid-associated protein which participated in lipid metabolism, oxygen-evolving enhancer protein 1 which was related to photosynthesis, porin which allowed ions, sugars and amino acids across the outer membrane, intracellular pathogenesis-related protein isoform 4 and hypothetical protein which their functions are unknown.【Conclusion】Beta-amylase, glyceraldehyde-3-phosphate dehydrogenase, cytosolic phosphoglycerate kinase, luminal binding protein (BiP), PAP fibrillin and Os02g0634900 which are homologous proteins of Asparagus are related to the male organ development. Enolase 1, cytosolic phosphoglycerate kinase, actin isoform B, actin, ATP synthase CF1 alpha subunit, small GTP-binding protein, GTP-binding protein, ribosome inactivating protein RIPm, with strong similarity to ADP-ribosylation factor, nucleoside diphosphate kinase, plastid-lipid-associated protein, oxygen-evolving enhancer protein 1, porin and hypothetical protein which are homologous proteins of Asparagus, ATP synthase beta subunit and intracellular pathogenesis-related protein isoform 4 are related to the female organ development. Oxygen-evolving enhancer protein 1 and porin which are homologous proteins of Asparagus might be the key proteins of female organ’s development.

Key words: Asparagus officinalis, female flower buds, male flower buds, hermaphroditic flower buds, sex differentiation, differential proteins

刘孟，乜兰春，王珊珊，胡淑明. 芦笋三种花蕾性别分化相关差异蛋白表达分析[J]. 中国农业科学, 2016, 49(21): 4192-4202.

LIU Meng, NIE Lan-chun, WANG Shan-shan, HU Shu-ming . Expression Analysis of Differential Proteins in Three Kinds of Flower Buds with Sex Differentiation of Asparagus[J]. Scientia Agricultura Sinica, 2016, 49(21): 4192-4202.

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