CAJ | 학술논문

【目的】在大田生产条件下研究氮素对灌浆期玉米叶片蛋白质表达的调控。【方法】在大田生产条件下，以紧凑耐密型玉米杂交种登海618为试验材料，研究施氮对玉米穗位叶花后净光合速率、硝酸还原酶（NR）活性、超氧化物歧化酶（SOD）活性、过氧化物酶（POD）活性、丙二醛（MDA）含量和可溶性蛋白含量的影响。采用TCA-丙酮沉淀法提取灌浆期（花后20 d）两个施氮处理下玉米穗位叶总蛋白质，并用双向凝胶电泳技术（2-DE）分离获得蛋白质图谱。采用ImageMaster-2D Elite 7.0图像分析软件对蛋白质图谱进行比较，确定玉米叶片应答灌浆期施氮处理的差异蛋白点。通过MALDI-TOF/TOF MS质谱分析及NCBInr数据库搜索，对差异表达蛋白质进行鉴定并分析其涉及的生物学功能。【结果】开花后，随生育进程的推进，玉米穗位叶净光合速率、叶绿素含量、NR、SOD和POD活性及可溶性蛋白含量均呈下降趋势，而MDA含量则呈上升趋势。相对于不施氮处理，施氮处理下叶片叶绿素含量、NR、SOD和POD活性及可溶性蛋白含量均显著提高，而MDA含量则显著下降。对灌浆期玉米叶片进行双向电泳及图谱分析，分别在施氮和不施氮条件下检测出1086和1170个蛋白点。通过图像分析软件进行成对匹配分析，共得到29个显著差异蛋白点。经质谱鉴定分析，29个显著差异蛋白点中有25个被成功鉴定，鉴定成功的蛋白中除未知蛋白（蛋白点55）和30s核糖体蛋白（蛋白点1089）外，其余蛋白表达量均在施氮条件下上调。通过搜索NCBInr数据库，差异表达的蛋白主要分为8类，包括13个能量相关蛋白，2个防御相关蛋白，2个蛋白合成相关蛋白，2个蛋白目的和储存相关蛋白，1个细胞生长相关蛋白，1个次级代谢相关蛋白，1个转运相关蛋白和3个未知蛋白。【结论】施氮对灌浆期玉米叶片光合能力、碳代谢能力、防御能力、蛋白合成能力、蛋白目的和储存能力、以及次级代谢能力等均有显著提升作用。【目的】在大田生產條件下研究氮素對灌漿期玉米葉片蛋白質錶達的調控。【方法】在大田生產條件下，以緊湊耐密型玉米雜交種登海618為試驗材料，研究施氮對玉米穗位葉花後淨光閤速率、硝痠還原酶（NR）活性、超氧化物歧化酶（SOD）活性、過氧化物酶（POD）活性、丙二醛（MDA）含量和可溶性蛋白含量的影響。採用TCA-丙酮沉澱法提取灌漿期（花後20 d）兩箇施氮處理下玉米穗位葉總蛋白質，併用雙嚮凝膠電泳技術（2-DE）分離穫得蛋白質圖譜。採用ImageMaster-2D Elite 7.0圖像分析軟件對蛋白質圖譜進行比較，確定玉米葉片應答灌漿期施氮處理的差異蛋白點。通過MALDI-TOF/TOF MS質譜分析及NCBInr數據庫搜索，對差異錶達蛋白質進行鑒定併分析其涉及的生物學功能。【結果】開花後，隨生育進程的推進，玉米穗位葉淨光閤速率、葉綠素含量、NR、SOD和POD活性及可溶性蛋白含量均呈下降趨勢，而MDA含量則呈上升趨勢。相對于不施氮處理，施氮處理下葉片葉綠素含量、NR、SOD和POD活性及可溶性蛋白含量均顯著提高，而MDA含量則顯著下降。對灌漿期玉米葉片進行雙嚮電泳及圖譜分析，分彆在施氮和不施氮條件下檢測齣1086和1170箇蛋白點。通過圖像分析軟件進行成對匹配分析，共得到29箇顯著差異蛋白點。經質譜鑒定分析，29箇顯著差異蛋白點中有25箇被成功鑒定，鑒定成功的蛋白中除未知蛋白（蛋白點55）和30s覈糖體蛋白（蛋白點1089）外，其餘蛋白錶達量均在施氮條件下上調。通過搜索NCBInr數據庫，差異錶達的蛋白主要分為8類，包括13箇能量相關蛋白，2箇防禦相關蛋白，2箇蛋白閤成相關蛋白，2箇蛋白目的和儲存相關蛋白，1箇細胞生長相關蛋白，1箇次級代謝相關蛋白，1箇轉運相關蛋白和3箇未知蛋白。【結論】施氮對灌漿期玉米葉片光閤能力、碳代謝能力、防禦能力、蛋白閤成能力、蛋白目的和儲存能力、以及次級代謝能力等均有顯著提升作用。
【목적】재대전생산조건하연구담소대관장기옥미협편단백질표체적조공。【방법】재대전생산조건하，이긴주내밀형옥미잡교충등해618위시험재료，연구시담대옥미수위협화후정광합속솔、초산환원매（NR）활성、초양화물기화매（SOD）활성、과양화물매（POD）활성、병이철（MDA）함량화가용성단백함량적영향。채용TCA-병동침정법제취관장기（화후20 d）량개시담처리하옥미수위협총단백질，병용쌍향응효전영기술（2-DE）분리획득단백질도보。채용ImageMaster-2D Elite 7.0도상분석연건대단백질도보진행비교，학정옥미협편응답관장기시담처리적차이단백점。통과MALDI-TOF/TOF MS질보분석급NCBInr수거고수색，대차이표체단백질진행감정병분석기섭급적생물학공능。【결과】개화후，수생육진정적추진，옥미수위협정광합속솔、협록소함량、NR、SOD화POD활성급가용성단백함량균정하강추세，이MDA함량칙정상승추세。상대우불시담처리，시담처리하협편협록소함량、NR、SOD화POD활성급가용성단백함량균현저제고，이MDA함량칙현저하강。대관장기옥미협편진행쌍향전영급도보분석，분별재시담화불시담조건하검측출1086화1170개단백점。통과도상분석연건진행성대필배분석，공득도29개현저차이단백점。경질보감정분석，29개현저차이단백점중유25개피성공감정，감정성공적단백중제미지단백（단백점55）화30s핵당체단백（단백점1089）외，기여단백표체량균재시담조건하상조。통과수색NCBInr수거고，차이표체적단백주요분위8류，포괄13개능량상관단백，2개방어상관단백，2개단백합성상관단백，2개단백목적화저존상관단백，1개세포생장상관단백，1개차급대사상관단백，1개전운상관단백화3개미지단백。【결론】시담대관장기옥미협편광합능력、탄대사능력、방어능력、단백합성능력、단백목적화저존능력、이급차급대사능력등균유현저제승작용。
Objective]The objective of this experiment is to explore the regulating effect of nitrogen on protein expression of leaf of summer maize under the field production condition.[Method]Denghai 618 was selected as an experimental material, different nitrogen treatments were designed to investigate the photosynthetic rate, the activity of nitrate reductase (NR), superoxide dismutase (SOD) and peroxidase (POD), malondialdehyde (MDA) content and soluble protein content in maize ear leaves after tasseling. The total proteins from harvested sample at the stage of 20 days after tasseling were extracted by the trichloroacetate (TCA)-acetone precipitation method and the protein profiles of maize ear leaves were set up by two-dimensional electrophoresis (2-DE). The proteins in maize leaves responding to nitrogen were identified by ImageMaster-2D Elite 7.0 software. The functions of these differentially expressed proteins were identified by matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry (MALDI-TOF/TOF MS) analysis and NCBInr database searching.[Result]After tasseling, the photosynthetic rate, activities of nitrate reductase (NR), superoxide dismutase (SOD) and peroxidase (POD), and soluble protein content were significantly decreased, and malondialdehyde (MDA) content was significantly increased. The photosynthetic rate, NR, SOD and POD, and soluble protein content were significantly increased, and MDA content was significantly decreased in nitrogen supply treatment when compared to nitrogen limited treatment. There were 1 086 and 1 170 protein spots were found after 2-DE analysis in nitrogen limited and nitrogen supply treatment, respectively. After a comparative proteomics analysis and MALDI-TOF/TOF mass spectrometry (MS), 29 differentially expressed proteins were found, including 25 which were successfully identified. They were all up-regulated in nitrogen supply treatment except spots 55 and 1089. They were mainly involved in energy, defense-related pathways, protein synthesis, metabolism, protein transportation and storage, cell growth, secondary metabolism, transcription and unknown.[Conclusion]Nitrogen supply can significantly improve the photosynthesis, carbohydrate catabolism, defense-related pathways, protein synthesis, protein transportation and storage and secondary metabolism in maize leaves at filling stage.