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作物学报 ›› 2017, Vol. 43 ›› Issue (09): 1410-1414.doi: 10.3724/SP.J.1006.2017.01410

• 研究简报 • 上一篇    

玉米不同组织器官谷氨酰胺合成酶同工酶表达差异及聚合方式

王小纯1,2,3,张浩然3,韦一昊1,贾喜婷3,谷明鑫3,马新明1,*   

  1. 1河南农业大学河南粮食作物协同创新中心,河南郑州 450002;2河南农业大学省部共建小麦玉米作物学国家重点实验室,河南郑州 450002;3河南农业大学生命科学学院生物化学系,河南郑州 450002
  • 收稿日期:2016-12-21 修回日期:2017-04-20 出版日期:2017-09-12 发布日期:2017-05-08
  • 通讯作者: 马新明, E-mail: xinmingma@126.com, Tel:13937100780 E-mail:xiaochun.w@163.com
  • 基金资助:

    本研究由国家重点研发计划项目(2016YFD0300205)和小麦玉米作物学国家重点实验室(39990047)资助。

Differential Expression and Assembly Mode of Glutamine Synthetase Isoen-zymes in Different Tissues and Organs of Maize

WANG Xiao-Chun1,2,3,ZHANG Hao-Ran3,WEI Yi-Hao1,JIA Xi-Ting3,GU Ming-Xin3,MA Xin-Ming1,*   

  1. 1 Collaborative Innovation Center of Henan Grain Crops / Henan AgriculturalUniversity, Zhengzhou 450002, China; 2 State Key Laboratory of Wheat and Maize Crop Science in China / Henan AgriculturalUniversity, Zhengzhou 450002, China; 3 Department of Biochemistry, College of Life Science / Henan AgriculturalUniversity, Zhengzhou 450002, China
  • Received:2016-12-21 Revised:2017-04-20 Online:2017-09-12 Published:2017-05-08
  • Contact: Ma Xinming, E-mail: xinmingma@126.com, Tel:13937100780 E-mail:xiaochun.w@163.com
  • Supported by:

    The study was supported by the National Key Research and Development Program of China (2016YFD0300205) and State Key Laboratory of Wheat and Maize Crop Science (39990047).

摘要:

谷氨酰胺合成酶(GS)是作物氮同化及转移利用的关键酶,本试验研究了玉米灌浆期不同组织器官的GS同工酶表达特性,鉴定了玉米GS同工酶的聚合方式。Westernblot结果表明,玉米不同组织器官的GS同工酶亚基表达存在明显差异,分子量约40kD的GS1亚基在所有组织中均大量表达,39kD的GS1亚基仅在穗位节及穗柄中大量表达,分子量约44kD的GS2亚基在叶片等光合组织中微量表达。通过改进BNE技术,结合胶内转移酶活性的测定,分析了玉米GS同工酶全酶的大小;利用2-D胶结合Westernblot鉴定了GS同工酶相应的亚基组成。结果表明,在玉米组织鉴定出3种分子量不同的GS同工酶,GS2全酶分子量约460kD,为十聚体;GS1全酶有2种聚合状态,一种是分子量约410kD的十聚体,另一种是分子量约240kD的五聚体形式,可见玉米GS同工酶表达存在多种方式。

关键词: Maize, Glutamine synthetase (GS), Expression, Blue native PAGE (BNE), Assembly

Abstract:

Glutamine synthetase (GS) is a key enzyme in nitrogen assimilation and recycling in cereals. In this study, the expression characteristics of GS isoenzymes in different tissues and organs of maize in grain-filling period were analyzed, and the assembly of GS isoenzymes were indentified. The GS isoforms expressed differentially in different organs were shown by Western-blot obviously; GS1 with a molecular weight of about 40 kD expressed highly in all tissues, and GS1 with a molecular weight of about 39 kD was merely expressed in the node of ear position and pedical, and GS2 with a molecular weight of about 44 kD was weakly expressed in the photosynthtic tissue such as leaf. With a modified blue naive PAGE (BNE) technique and in-gel activity analysis, the size of GS holoenzyme was calibrated; combined the 2-D gel with western-blot analysis, the subunits composition of GS isoenzymes were identified. Three GS isoenzymes with different sizes were identified in maize. GS2 holoenzyme was about 460 kD and likely a decamer, GS1 holoenzyme existed two kinds of assembly state, one was about 410 kD and likely a decamer, another was about 240 kD and more likely a pentamer; therefore, the expression of GS isoenzymes exists diversity in maize.

Key words: Maize, Glutamine synthetase (GS), Expression, Blue native PAGE (BNE), Assembly

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