【作者】 李兆伟；

【导师】 林文雄；

【作者基本信息】 福建农林大学 ， 草业科学与技术， 2008， 硕士

【摘要】 稻草作为世界主要的农副产品之一,可以作为畜牧业新的饲草资源,对世界草业科学的发展具有重要的意义。本文以超大穗型水稻“金恢809”为实验材料,采用差异蛋白质组学方法研究其在籽粒灌浆期间,水稻秸杆对籽粒灌浆的影响,旨在揭示超大穗型水稻的源库流灌浆机理,为有效地挖掘水稻高产潜力、促进粮食增产、以及谷杆两用稻的选育,提供重要的理论依据。结果显示,水稻叶片在籽粒灌浆期间,共有17个蛋白质点发生显著变化,经MALDI-TOF/MS检测及肽指纹图谱(PMF)分析,其中12个蛋白质的功能得到鉴定,分别涉及物质的合成与降解,碳水化合物运输,植株抗氧化反应,激素代谢,以及细胞骨架的构建和组织成熟。具体表现为:核糖/半乳糖/甲基半乳糖苷运输ATP结合蛋白1在灌浆前中期参与叶片物质向籽粒的运输;生长素响应蛋白IAA27通过调节ATPase活性影响叶片物质运输;N-乙酰谷氨酸半醛脱氢酶在灌浆末期参与叶片体内的多胺代谢;谷胱甘肽S-转移酶和Cu/Zn-超氧化物岐化酶在籽粒灌浆后期的植物解毒和防御活性氧伤害中起着重要作用。此结果表明,大穗型水稻在灌浆前期和中期(0到14天),叶片光合同化物质在ATPase的作用下,供给籽粒灌浆需要;在灌浆前期到中后期(0到21天),生长素响应蛋白IAA27对ATPase的调节作用逐渐加强;籽粒灌浆后期(第28d),多胺代谢活动加强,叶片的抗氧化酶代谢和泛素化作用也相应加强,共同维持叶片的正常生理机能。由此可见,在水稻生育后期,叶片的功能性状具有复杂的阶段性转换现象。叶鞘在灌浆不同时期,发现23个差异表达蛋白,其中11个蛋白得到鉴定,分别参与叶鞘的光合作用,激素调节,物质转运,植株衰老的抗性反应以及细胞信号转导。表现为:α-亚基草酰乙酸脱羧酶、二磷酸核酮糖羧化酶小链、二磷酸核酮糖羧化酶/加氧酶活化酶和核酮糖α亚基结合蛋白参与叶鞘光合作用;生长素响应因子和栊牛儿基牻牛儿基二磷酸合酶与激素代谢相关;ADP-核糖基化因子1和液泡H+-ATPase与物质运输相关;金属硫蛋白Ⅱ相似蛋白-1A与植株抗性反应相关;锌指-C3HC4型家族蛋白参与细胞内信号转导;蛋白激酶在细胞新陈代谢和信号转换进程中起着重要作用。此结果表明,灌浆初期,叶鞘接收叶片的营养物质,具备“库”的生理作用;灌浆前中期(0～21 d),叶鞘又可以合成部分同化物质,执行“源”的生理功能;在整个灌浆期,叶鞘又作为物质向籽粒运输的通道(流)——转运同化物质。叶鞘作为连接叶片和籽粒的通道,是叶片营养物质向籽粒运输的重要保障。籽粒在灌浆期间,有32个差异蛋白,其中27个得到鉴定,分别参与蛋白质代谢,碳水化合物与能量代谢,信号转导,防御代谢,基因转录调节,细胞结构组成,以及生殖代谢。籽粒作为主要的库器官,接收叶、鞘中的营养物质,在体内淀粉合成酶系和蛋白质酶系的作用下,将其转化为贮藏性多糖和贮藏性蛋白,累积于体内,提高稻米产量和品质。更多还原

【Abstract】 Straw is one of the main agricultural byproducts and has become important forage resources of animal production, which play an important role in the development of Pratacultural science in the world.The investigation was conducted to address the influence of rice straw on grain filling of rice Jinhui 809 cultivar with large panicles and high setting percentage during grain filling by the method of differential proteomics. The aim is to reveal the metabolic mechanism of source-sink-stream in large panicles rice during grain filling period and provide important theoretical references to dig the rice high yield potential, accelerate grain production increase and breeding on grain-straw-dual-use-rice.The results show that a total of 17 protein spots in rice flag leaf had significant expression difference; these proteins were identified using MALDI-TOF/MS analysis and database searching. Among them, twelve were knowen proteins in function. These proteins were involved in substance synthesis and degradation, carbohydrate transportation, plant antioxidation reaction, hormone metabolism, cytoskeleton construction and tissues maturation respectively. The detailed performances are as follows: ribose/galactose/methyl galactoside import ATP-binding protein 1 associated with substance transportation from leaf to grain at grain filling prometaphase, auxin-responsive protein IAA27 responsible for the substance transportation by regulating the activity of ATPase, N-acetyl-glutamate semialdehyde dehydrogenase participating in polyamine metabolism at grain filling terminal phase, and glutathione S-transferase as well as superoxide dismutase playing an important role in plant detoxification and recovery oxygen species damage at late grain filling stage. It concluded that, the photosynthesis assimilation substance of rice leaf was provided to grains powered by ATPase at the prophase and metaphase of grain filling (0 to 14 day), and the regulation function of auxin-responsive protein IAA27 on ATPase was strengthened gradually from prophase to middle-late phase of grain filling (from 0 to 21 days). At the anaphase of grain filling (28 day), the activities of polyamine metabolism was enhanced, besides, the metabolism of antioxidant enzymes and ubiquitin pathway were also strengthended correspondingly, which maintain the regular physiological function of leaf. Thus it could be found, in the later period of rice growth, the function characters of leaves had a complex periodical transformation phenomenon.Twenty-three proteins with obviously different abundances were detected in leaf sheath during grain filling period, and eleven of them were identified in function, which were associated with photosynthesis, hormone regulation, substance transportation, resistance response to plant senescence and cell signal transduction in leaf sheath respectively. The performances are: oxaloacetate decarboxylase alpha subunit, ribulose bisphosphate carboxylase small chain, Rubisco activase and Rubisco binding-protein alpha subunit participating in the photosynthesis of leaf sheath. Auxin response factor (ARF) and geranylgeranyl diphosphate synthase (GGPP synthase) related to hormones metabolism. ADP-ribosylation factor 1 (ARF-1), and vacuolar H~+-ATPase (H~+-ATPase) correlative with substance transportation, metallothionein II -like protein 1A (MT II-1 A) involved in resistant response of leaf sheath, Zinc finger C3HC4 type family protein participating in intracellular signal transduction, and protein kinases playing a role in the regulation of cellular metabolism and signal transduction processes. It concluded that, at the early stage of grain filling, leaf sheath performed the physiological function as sink, which received nutrient substance from leaves. And at the prometaphase of grain filling (0～21 d), leaf sheath also performed the physiological function as source, which produced assimilation substance. In the whole grain filling periods, leaf sheath performed function as flow organs which participated in assimilation products transportation from leaves to grains. Leaf sheathes which connected leaves with grains was the important guarantee of nutrient substance transportation from leaves to grains.In addition, thirty-two proteins with obvious different abundances were detected in grains, and twenty-seven of them were identified in function, which were involved in protein metabolism, carbohydrate and energy metabolism, signal transduction, defence metabolism, gene transcriptional regulation, cell structure forming, and reproduction metabolism. The grains were the main sink organ receiving nutrition substances from leaves and sheathes, in which the nutrition substances were transformed to reserved polysouharide and storage proteins catalyzed by the system of starch and protein synthases, consequently accumulated them to improve the production and quality of rice grains.更多还原