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


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)


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"

Protein subunit content
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*
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