小麦胚乳PCD响应花后干旱胁迫的蛋白质组学研究

The development of wheat endosperm was developed from the viable tissue to nutritional storage organ and companied by the progression of programmed cell death(PCD). The endosperm PCD was programmed by the genes, which controlled the active death process and encoded the protein to implement function. The endosperm PCD was significantly impacted by the abiotic stress. The progression of endosperm PCD was accelerated by the drought condition, which shorted the period of grain filling and led to the low grain quality and yield of wheat in most of XinJiang. In this study, two winter wheat cultuvars were employed to explore the progression of endosperm PCD under the drought condition. The protein characteristic and expression profile of endosperm PCD under drought stress was elucidated. The proteins involved in endosperm PCD were identified of the composition, structure, properties and expression level under drought stress. The progeression of endosperm PCD was investigated by Evans blue staining and transmission electron microscopy. The proteins involved in endosperm PCD were researched by two-dimensional electrophoresis, mass spectrometry and bioinformatics. The objective of this study is to provide more information for using genetic manipulation and improving the wheat grain yield and quality reduced by the drought.

小麦胚乳细胞发育经历了由活细胞到死的营养贮存库的转变，并伴随着胚乳细胞程序性死亡(PCD)过程。小麦胚乳PCD是由基因控制主动的程序性死亡过程，胚乳PCD相关基因其功能最终是通过蛋白质来实现。外界环境胁迫对胚乳PCD发生具有重要的影响。新疆大部分小麦产区灌浆期干旱频发，加速小麦胚乳PCD进程，缩短籽粒灌浆期，严重影响小麦产量与品质。本研究设想通过探明干旱胁迫下胚乳PCD相关蛋白组变化特征，揭示小麦干旱胁迫下胚乳PCD 相关蛋白表达图谱，分析鉴定小麦干旱胁迫下胚乳PCD相关蛋白。因此本项目以新疆两个冬小麦主栽品种为供试材料，利用组织化学染色、透射电镜观察、双向电泳检测、质谱鉴定、生物信息学分析等技术，研究揭示干旱胁迫下胚乳PCD相关蛋白组学变化特征以及PCD相关蛋白组成、结构、性质和表达水平，预测PCD相关蛋白功能。结果可为通过遗传改良方法，来减轻花后干旱胁迫对新疆小麦生产的危害提供依据。

小麦胚乳细胞的发育经历了由活细胞到死的营养贮存库的转变。胚乳细胞程序性死亡（Programmed Cell Death，PCD）是由基因控制的主动的PCD过程。这些胚乳PCD相关基因其功能最终是通过编码的蛋白质来实现。本项目以新疆主栽两个冬小麦品种新冬18号和新冬22号为研究材料，设置田间小区试验，采用小麦花后停止灌水（阴雨天搭遮雨棚，阻止大气降水）形成干旱胁迫条件，研究小麦胚乳PCD响应花后干旱胁迫的蛋白质组学，为通过遗传改良方法，减轻花后干旱胁迫对新疆小麦生产危害提供依据。取得以下重要和新颖结果：.1、探明小麦花后干旱胁迫下胚乳PCD相关蛋白质组变化特征研究.（1）外界干旱胁迫加速胚乳PCD进程，胚乳细胞核由凝聚态变成分散状态遍布胚乳细胞各部；.（2）花后干旱胁迫加速胚乳细胞数目增加，核酸酶活性极显著升高，从而加速胚乳细胞核的分解；.2、揭示了小麦胚乳PCD响应花后干旱胁迫的蛋白质组学规律.（1）通过iTRAQ方法经生物信息学分析鉴定到48个差异蛋白参与了小麦花后干旱胁迫下胚乳PCD进程，主要包括：泛素类蛋白、微管蛋白、类枯草杆菌蛋白酶、丝氨酸蛋白酶家族蛋白、天冬氨酰蛋白酶家族蛋白、26S蛋白酶类蛋白、羧肽酶类蛋白、ATP合成酶类蛋白等26类；.（2）本研究对鉴定得到的小麦花后干旱胁迫下胚乳PCD蛋白的相关基因在转录组水平进行了多组学和基因表达的验证。.上述结果揭示小麦胚乳PCD响应花后干旱胁迫的蛋白质组学规律，为通过遗传改良方法，减轻花后干旱胁迫对新疆小麦生产危害提供依据。.本项目按照计划内容实施，超额完成预期目标和任务。共发表学术论文11篇，其中被SCI收录论文4篇，出版专著1部，授权国家发明专利2项，审定冬小麦新品种2个，培养毕业博士生1人，硕士生4人。