摘要
【目的】基于iTRAQ蛋白质组学筛选不同OsAAP6转基因水稻胚乳的差异蛋白,明确胚乳中的差异蛋白,以期为后续利用OsAAP6基因提升稻米的营养品质提供理论依据。【方法】以水稻OsAAP6超量表达阳性[OX(+)]和阴性对照[OX(-)]材料及OsAAP6互补表达阳性[ZpZc(+)]和阴性对照[ZpZc(-)]为试验材料,采用同位素标记相对与绝对定量(iTRAQ)技术对其进行蛋白质组学检测分析,并利用考马斯亮蓝G-250法对其胚乳中的4类储藏蛋白(谷蛋白、醇溶蛋白、清蛋白和球蛋白)进行含量检测。【结果】与ZpZc(-)和OX(-)转基因阴性对照相比,ZpZc(+)和OX(+)转基因阳性胚乳中OsAAP6基因的表达量分别在P<0.01和P<0.001水平极显著升高。与ZpZc(-)和OX(-)转基因阴性对照相比,ZpZc(+)和OX(+)转基因阳性水稻共有58种差异蛋白共同上调或下调表达,其中有39种蛋白显著增加(P<0.05,下同),有19种蛋白显著降低。在39种共同上调表达的蛋白中有27种蛋白参与水稻种子储藏底物的合成与积累,且有16种上调表达蛋白参与种子储藏蛋白的代谢和积累。与ZpZc(-)和OX(-)转基因阴性对照相比,ZpZc(+)和OX(+)转基因阳性水稻胚乳中的谷蛋白、醇溶蛋白、清蛋白和球蛋白含量均显著增加。【结论】OsAAP6基因上调表达,能抑制与支链淀粉合成相关蛋白的积累,但能促进其胚乳中储藏蛋白的积累,进而提高胚乳中谷蛋白、醇溶蛋白、清蛋白和球蛋白含量,可用于高蛋白水稻新品种的分子育种和遗传改良。
【Objective】The purpose of the study was to screen the differential proteins in the endosperm of different OsAAP6 transgenic rice based on iTRAQ proteomics,and to clarify the differential proteins in the endosperm,in a bid to provide a theoretical basis for the future use of the OsAAP6 gene to enhance the nutritional quality of rice.【Method】Using rice OsAAP6 overexpression positive[OX(+)]and negative control[OX(-)]materials,as well as OsAAP6 complementary expression positive[ZpZc(+)]and negative control[ZpZc(-)]experimental materials,the isotope marker relative and absolute quantification(iTRAQ)technology was used for proteomic detection and analysis.And the contents of four types of storage proteins(gluten,gliadin,albumin and globulin)in its endosperm was detected using the Coomassie brilliant blue G-250 method.【Result】Compared with the ZpZc(-)and OX(-)transgenic negative control,the expression of OsAAP6 gene in ZpZc(+)and OX(+)transgenic positive endosperm was extremely significantly increased at P<0.01 and P<0.001 levels,respectively.Compared with the ZpZc(-)and OX(-)transgenic negative control,a total of 58 differential proteins were simultaneously up-regulated or down-regulated for expression in ZpZc(+)and OX(+)transgenic positive rice,of which 39 proteins significantly increased(P<0.05,the same below),and 19 proteins significantly decreased.Among the 39 co-up-regulated proteins,27 proteins were involved in the synthesis and accumulation of rice seed storage substrates,and 16 up-regulated proteins were involved in the metabolism and accumulation of seed storage proteins.Compared with the ZpZc(-)and OX(-)transgenic negative control,the content of glutelin,gliadin,albumin and globulin in the endosperm of ZpZc(+)and OX(+)transgenic positive rice was significantly increased.【Conclusion】Up-regulation of the OsAAP6 gene expression can inhibit the accumulation of proteins related to amylopectin synthesis,but promote the accumulation of storage proteins in the endosperm,thereby increasing the content of gluten,gliadin,albumin and globulin in the endosperm.It can be used for molecular breeding and genetic improvement of new high protein rice varieties.
作者
彭波
孙晓宇
邱静
彭娟
赵强
孙艳芳
宋晓华
张志国
周伟
汪全秀
PENG Bo;SUN Xiao-yu;QIU Jing;PENG Juan;ZHAO Qiang;SUN Yan-fang;SONG Xiao-hua;ZHANG Zhi-guo;ZHOU Wei;WANG Quan-xiu(College of Life Sciences,Xinyang Normal University/Dabie Mountain Agricultural Biological Resource Protection and Utilization Research Institute,Xinyang,Henan 464000,China;Xinyang Plant Protection and Inspection Station,Xinyang,Henan 464000,China;Henan Scientific Research Platform Service Center,Zhengzhou,Henan 450003,China;Xinyang Academy of Agricultural Sciences,Xinyang,Henan 464000,China;Henan Lingrui Pharmaceutical Company Limited,Xinyang,Henan 465550,China)
出处
《南方农业学报》
CAS
CSCD
北大核心
2024年第4期1060-1069,共10页
Journal of Southern Agriculture
基金
国家自然科学基金项目(U2004141)
河南省科技攻关项目(222102110141,222102110115,212102110249)。
