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Acta Agronomica Sinica ›› 2019, Vol. 45 ›› Issue (1): 111-117.doi: 10.3724/SP.J.1006.2019.84054

• TILLAGE & CULTIVATION · PHYSIOLOGY & BIOCHEMISTRY • Previous Articles     Next Articles

Relationship between GS isoenzyme activity and nitrogen transportation in flue-cured tobacco leaves

Jian-Fei ZHOU1(),Yun-Jie WU1,Gang XUE1,An-Qian ZHANG1,Pei TIAN1,Yu-Fu PENG2,Tie-Zhao YANG1,*()   

  1. 1 College of Tobacco Science, Henan Agricultural University, Zhengzhou 450002, Henan, China
    2 Technical Center, Henan Tobacco Industry Co., Ltd., Zhengzhou 450000, Henan, China
  • Received:2018-04-12 Accepted:2018-08-20 Online:2018-09-14 Published:2018-09-14
  • Contact: Tie-Zhao YANG E-mail:ndzhoujianfei@163.com;yangtiezhao@126.com
  • Supported by:
    This study was supported by the Henan Tobacco Industry Co., Ltd. Science and Technology Project(HNZY102015004);Henan Tobacco Company Science and Technology Project(2018410000270035)

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Abstract:

A pot experiment was conducted to study the dynamic changes of glutamine synthetase isoenzyme activity and the related physiological indexes in leaves of flue-cured tobacco with different nitrogen efficiencies at maturity stage, and understand the physiological and biochemical mechanism of nitrogen metabolism in flue-cured tobacco leaves. Three different flue-cured tobacco varieties Zhongyan 100, K326, and NC89 with different nitrogen efficiencies were used as materials, and the 12th leaves were taken at different leaf ages. Western blot method was used to determine the content of glutamine synthetase isoenzyme isoforms in leaf mesophyll and main veins. At the same time, the concentrations of NH4 +, total nitrogen, apoplastic NH4 + and ammonia volatilization were determined. The results show, GS2 was the main form of GS isoenzyme in mesophyll of flue-cured tobacco leaves, the protein subunit content gradually decreased with the age of the leaf. The GS1 isoenzyme dominated in the veins, and its protein subunits showed a trend of increasing first and then decreasing. From 45 to 65 days of leaf age, the GS isoenzyme activities of leaf mesophyll and main vein were all expressed as NC89 > K326 > Zhongyan 100, and the difference among varieties was significant. The activities of GS1 in mesophyll and main veins were negatively correlated with total nitrogen and leaf ammonium concentrations, and positively correlated with apoplastic ammonium concentration and ammonia volatilization. The GS2 activity in mesophyll was positively correlated with total nitrogen and leaf ammonium, and negatively correlated with apoplastic ammonium concentration and ammonia volatilization. The GS2 activity of leaf veins was only significantly correlated with the total nitrogen and ammonia volatilization. The activity of two glutamine synthetase isoenzymes in leaves of nitrogen-inefficient flue-cured tobacco varieties at the mature period was lower, and the ability of nitrogen transfer and reuse was poor, resulting in most of the nitrogen absorbed by plants evaporated in the form of ammonia, and the leaves had a faster rate of aging. However, nitrogen efficient varieties had strong nitrogen reutilization capacity, so the ammonia volatilization was small and the aging was postponed.

Key words: flue-cured tobacco, glutamine synthetase isozyme, nitrogen metabolism, ammonia volatilization

Fig. 1

Native-PAGE binding activity staining of mesophyll GS isoenzyme 1-15 represents the 12th leaf mesophyll sample of K326, NC89, and ZY100 at leaf age 55 days with five replicates for each species."

Fig. 2

Native-PAGE binding activity staining of GS isoenzymes in vein 16-30 represents the 12th leaf main vein sample of K326, NC89, and ZY100 at leaf age 55 days with five replicates for each species."

Fig. 3

Dynamic changes of GS isoenzyme activity in leaf mesophyll 1: leaf age 35 days; 2: leaf age 45 days; 3: leaf age 55 days; 4: leaf age 65 days; 5: leaf age 75 days."

Fig. 4

Dynamic changes of GS isoenzyme protein subunit content in leaf mesophyll Bars superscripted by different letters are significantly different at P < 0.05 within the same period."

Fig. 5

Dynamic changes of GS isoenzyme activity in main vein 1: leaf age 35 days; 2: leaf age 45 days; 3: leaf age 55 days; 4: leaf age 65 days; 5: leaf age 75 days."

Fig. 6

Dynamic changes of GS isoenzyme protein subunit content in main vein Bars superscripted by different letters are significantly different at P < 0.05 within the same period."

Fig. 7

Dynamic changes of leaf total nitrogen content and NH4+ concentration Bars superscripted by different letters are significantly different at P < 0.05 within the same period."

Fig. 8

Dynamic changes of apoplastic NH4+ concentration and ammonia volatilization in leaves Bars superscripted by different letters are significantly different at P < 0.05 within the same period."

Table 1

Correlation coefficients of protein subunit content with physiological indexes of GS isoenzyme"

蛋白亚基含量
Protein subunit content		 总氮
Total N		 L-NH4+ A-NH4+ NH3-V
叶肉 GS1 -0.374* -0.415* 0.183* 0.408*
叶肉 GS2 0.527** 0.364* -0.291* -0.651**
叶脉 GS1 -0.478* -0.553** 0.497* 0.457**
叶脉 GS2 0.178* 0.245 0.125 -0.279*
[1] 史宏志, 韩锦峰 . 烤烟碳氮代谢几个问题的探讨. 烟草科技, 1998,129(2):34-36.
Shi H Z, Han J F . Discussion on several problems of carbon and nitrogen metabolism in flue-cured tobacco. Tobacco Sci Technol, 1998,129(2):34-36 (in Chinese).
[2] 严小龙, 张福锁 . 植物营养遗传. 北京: 中国农业出版社, 1997. pp 78-106.
Yan X L, Zhang F S. Genetics of Plant Nutrition. Beijing: China Agriculture Press, 1997. pp 78-106(in Chinese).
[3] Hirel B, Andrieu B, Renard S, Quillere I, Chelle M, Pommel B, Fournier C . Physiology of maize II: Identification of physiological markers representative of the nitrogen status of maize (Zea mays) leaves during grain filling. Physiol Planta, 2005,124:178-188.
doi: 10.1111/j.1399-3054.2005.00511.x
[4] Fei H, Chaillou S, Hirel B, Polowick P, Mahon J, Vessey J . Effects of the overexpression of a soybean cytosolic glutamine synthetase gene (GS15) linked to organ-specific promoters on growth and nitrogen accumulation of pea plants supplied with ammonium. Plant Physiol Biochem, 2006,44:543-550.
doi: 10.1016/j.plaphy.2006.09.007 pmid: 17067806
[5] Li X P, Zhao X Q, He X, Zhao G Y, Li B, Liu D C, Zhang A M, Zhang X Y, Tong Y P, Li Z S . Haplotype analysis of the genes encoding glutamine synthetase plastic isoforms and their association with nitrogen-use and yield-related traits in bread wheat. New Phytol, 2011,189:449-458.
doi: 10.1111/j.1469-8137.2010.03490.x
[6] 邓扬悟 . 甜瓜谷氨酰胺合成酶基因的克隆和功能研究. 上海交通大学博士学位论文, 上海, 2010.
Deng Y W . Cloning and Characterization of Glutamine Synthetase Genes in Melon . PhD Dissertation of Shanghai JiaoTong University, Shanghai, China, 2010 (in Chinese with English abstract).
[7] 卢素萍, 李玉娥, 刘鹏飞, 赵铭钦, 张渝婕, 李志鹏 . 氮用量对南阳不同基因型烤烟烟叶品质的影响. 江西农业大学学报, 2016,38:67-73.
Lu S P, Li Y E, Liu P F, Zhao M Q, Zhang Y J, Li Z P . Effects of nitrogen fertilizer on quality of different genotypes of flue-cured tobacco in Nanyang county. Acta Agric Univ Jiangxiensis, 2016,38:67-73 (in Chinese with English abstract).
[8] 崔昌范, 王书凯, 宋立君 . 烤烟新品种K326和NC89特征特性及配套栽培技术规范. 延边大学农学学报, 1999,21(1):70-75.
Cui C F, Wang S K, Song L J . Characteristics of flue-cured tobacco new varieties K326 and NC89 and their cultivation techniques. J Agric Sci Yanbian Univ, 1999,21(1):70-75 (in Chinese).
[9] 武云杰, 李飞, 杨铁钊, 张小全 . 氮素营养水平对衰老期烟叶氮代谢的影响及品种间差异. 中国烟草学报, 2014,20(4):41-47.
doi: 10.3969/j.issn.1004-5708.2014.04.009
Wu Y J, Li F, Yang T Z, Zhang X Q . Effects of nitrogen nutrition on nitrogen metabolism in flue-cured tobacco leaves and differences among varieties. Acta Tabac Sin, 2014,20(4):41-47 (in Chinese with English abstract).
doi: 10.3969/j.issn.1004-5708.2014.04.009
[10] 黄培堂 . 分子克隆实验指南. 北京: 化学工业出版社, 2007. pp 155-157.
Huang P T. The Condensed Protocols from Molecular Cloning: a Laboratory Manual. Beijing: Chemical Industry Press, 2007. pp 155-157(in Chinese).
[11] Wang X, Wei Y, Shi L, Ma X, Theg S . New isoforms and assembly of glutamine synthetase in the leaf of wheat (Triticum aestivum L.). J Exp Bot, 2015,66:6827-6834.
doi: 10.1093/jxb/erv388 pmid: 4623691
[12] 邹琦 . 植物生理学实验指导. 北京: 中国农业出版社, 2000. pp 56-89.
Zou Q. The Guidance of Plant Physiology Experiment. Beijing: China Agriculture Press, 2000. pp 56-89(in Chinese).
[13] 段旺军, 杨铁钊, 刘化冰, 张小全, 戴亚, 李东亮 . 烟叶氨气补偿点的品种间差异及其与氮素代谢的关系研究. 植物营养与肥料学报, 2011,17:419-424.
doi: 10.11674/zwyf.2011.9440
Duan W J, Yang T Z, Liu H B, Zhang X Q, Dai Y, Li D L . Differences in NH3 compensation point among tobacco (Nicotiana tabacum L.) cultivars and its relationship with nitrogen metabolism. J Plant Nutr Ferti, 2011,17:419-424 (in Chinese with English abstract).
doi: 10.11674/zwyf.2011.9440
[14] 武云杰, 杨铁钊, 张小全, 李飞, 李丽华 . 不同烤烟品种烟叶衰老期氨气挥发及其与氮素代谢的相关性. 中国农业科学, 2013,46:4027-4034.
doi: 10.3864/j.issn.0578-1752.2013.19.008
Wu Y J, Yang T Z, Zhang X Q, Li F, Li L H . Study on correlation between ammonia volatilization and nitrogen metabolism during the tobacco leaves senescence. Sci Agric Sin, 2013,46:4027-4034 (in Chinese with English abstract).
doi: 10.3864/j.issn.0578-1752.2013.19.008
[15] Nielsen K H, Schjoerring J K . Regulation of apoplastic NH4 + concentration in leaves of oilseed rape . Plant Physiol, 1998,118:1361-1368.
doi: 10.1104/pp.118.4.1361
[16] 范江, 杨继洪, 王守旗, 肖志新, 梁梅, 李海平, 邱尧 . 打顶对烤烟生长和各器官烟碱积累分配的影响. 作物研究, 2016,30(1):36-40.
doi: 10.16848/j.cnki.issn.1001-5280.2016.01.011
Fan J, Yang J H, Wang S Q, Xiao Z X, Liang M, Li H P, Qiu Y . Effect of topping on growth of flue-cured tobacco and nicotine accumulation and distribution in different organs. Crop Res, 2016,30(1):36-40 (in Chinese with English abstract).
doi: 10.16848/j.cnki.issn.1001-5280.2016.01.011
[17] 王云华, 欧吉权, 王志强, 周忠新, 欧阳敏, 张楚富 . 黄瓜 (Cucumis sativus L.)子叶发育过程中谷氨酰胺合成酶同工酶的变化. 武汉大学学报(理学版), 2005,51:517-520.
Wang Y H, Ou J Q, Wang Z Q, Zhou Z X, Ou Y M, Zhang C F . Changes of glutamine synthetase isozymes during the development of cucumber (Cucumis sativus L.) cotyledons. J Wuhan Univ (Nat Sci Edn), 2005,51:517-520(in Chinese with English abstract).
[18] Rueda-López M, Crespillo R, Cánovas F M, Avila C . Differential regulation of two glutamine synthetase genes by a single Dof transcription factor. Plant J, 2008,56:73-85.
doi: 10.1111/j.1365-313X.2008.03573.x pmid: 18547397
[19] Stanulović V S, Garcia R M, Labruyère W T, Ruijter J M, Hakvoort T B M, Lamers W H . The 3'-UTR of the glutamine-synthetase gene interacts specifically with upstream regulatory elements, contains mRNA-instbility elements and is involved in glutamine sensing. Biochimie, 2006,88:1255-1264.
doi: 10.1016/j.biochi.2006.05.012
[20] Kumar R, Taware R, Gaur V S, Guru S K, Kumar A . Influence of nitrogen on the expression of TaDof1 transcription factor in wheat and its relationship with photo synthetic and ammonium assimilating efficiency. Mol Biol Rep, 2009,36:2209-2216.
doi: 10.1007/s11033-008-9436-8 pmid: 19123069
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