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Acta Agronomica Sinica ›› 2022, Vol. 48 ›› Issue (10): 2567-2574.doi: 10.3724/SP.J.1006.2022.14205

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

Effects of increased nitrogen fertilizer on square Bt protein expression and nitrogen metabolism in cotton

LI Han-Jia(), LI Yuan, LIU Zhen-Yu, ZHANG Chen-Xia, XU Ze, WU Tian-Fan, CHEN Yuan, ZHANG Xiang, CHEN Yuan, CHEN De-Hua()   

  1. Jiangsu Key Laboratory of Crop Genetics and Physiology / Co-Innovation Center for Modern Production Technology of Grain Crops, Yangzhou University, Yangzhou 225009, Jiangsu, China
  • Received:2021-10-30 Accepted:2022-02-25 Online:2022-10-12 Published:2022-03-22
  • Contact: CHEN De-Hua E-mail:1311190875@qq.com;cdh@yzu.edu.cn
  • Supported by:
    National Natural Science Foundation of China(31471435);National Natural Science Foundation of China(31671613);Natural Science Foundation of Jiangsu Province(BK20191439);Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD)

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Abstract:

To investigate the effects of increased nitrogen application on square Bt protein concentration in cotton, the experiments were conducted at Key Laboratory of Genetics and Physiology of Yangzhou University in 2017 and 2018. The conventional cultivar Sikang 1 and hybrid cultivar Sikang 3 were used as the experimental materials. Enhanced nitrogen fertilizer rates of increased 25% to 100% nitrogen (300 (CK), 375, 450, 525, and 600 kg hm-2 as pure nitrogen) were designed to study the effect on square Bt protein content and nitrogen metabolic physiological in cotton. Compared with the control (300 kg hm-2), the squares Bt protein content increased first, but bolstered extent was reduced when increased nitrogen was at 1.50-1.75 times of conventional nitrogen rates (450-525 kg hm-2). Bt protein content in square increased by 4.5%-132.7% with the increase of 25%-100% nitrogen application. The maximum value of square Bt protein content was observed at nitrogen rate of 450-525 kg hm-2. The physiological mechanism of nitrogen metabolism showed that the trend of soluble protein (SP), free amino acid (AA), and key enzymes of protein synthesis (glutamic oxalacetic Transaminase, GOT and glutamine synthetase, GS) in squares were consistent with those of Bt protein. The activities of key enzymes of protein decomposition (protease and peptidase) decreased with the increase of nitrogen application. In conclusion, the increasing nitrogen application at optimum level in soil would promote Bt protein synthesis and benefit the insect resistance.

Key words: Bt cotton, enhanced nitrogen fertilizer application, insecticidal protein, nitrogen metabolism

Table 1

Effects of increased nitrogen application on square Bt protein contents in cotton (ng g-1 FW)"

品种
Variety		 处理
Treatment		 2017 2018
18 d 21 d 24 d 18 d 21 d 24 d
SK-1 CK 103.36 d 153.64 b 195.74 g 133.65 fg 141.38 e 149.29 e
N1 163.00 d 195.61 b 249.73 f 147.53 de 159.51 d 162.81 d
N2 209.64 b 338.91 a 309.45 de 165.83 c 177.05 c 182.88 c
N3 240.48 a 323.90 a 387.85 ab 196.02 a 204.43 a 209.05 a
N4 198.60 bc 160.58 b 279.40 ef 153.26 d 170.06 c 176.32 c
SK-3 CK 94.20 d 175.06 b 318.58 cd 126.76 g 130.16 f 136.54 f
N1 144.72 c 193.67 b 332.95 cd 138.21 ef 142.21 e 149.32 e
N2 179.94 bc 354.54 a 354.32 bc 152.40 d 156.78 d 160.85 d
N3 176.10 bc 374.84 a 410.19 a 183.14 b 187.89 b 192.27 b
N4 164.28 bc 318.11 a 339.67 cd 144.98 de 149.95 de 156.77 d

Table 2

Effects of increased nitrogen application on square soluble protein content in cotton (mg g-1 FW)"

品种
Variety		 处理
Treatment		 2017 2018
18 d 21 d 24 d 18 d 21 d 24 d
SK-1 CK 0.45 e 0.57 cd 0.73 bc 0.25 de 0.22 e 0.43 c
N1 0.98 bc 0.83 bcd 0.81 ab 0.57 bc 0.34 de 0.78 bc
N2 1.18 ab 0.87 bc 0.90 ab 0.82 a 0.54 bcd 0.79 bc
N3 1.44 a 1.36 a 1.02 a 0.69 ab 0.70 bc 1.29 ab
N4 0.60 de 0.77 bcd 0.76 ab 0.63 abc 0.47 cde 0.75 bc
SK-3 CK 0.67 de 0.56 d 0.41 c 0.16 e 0.37 de 0.58 c
N1 0.84 cd 0.74 bcd 0.65 bc 0.32 de 0.53 bcd 0.86 bc
N2 1.26 a 0.87 b 0.97 ab 0.39 cde 1.24 a 1.54 a
N3 1.36 a 1.16 a 1.06 a 0.42 cd 0.80 b 1.87 a
N4 0.79 cd 0.68 bcd 0.63 bc 0.31 de 0.59 bcd 1.27 ab

Table 3

Effect of increased nitrogen application on square free amino acid content in cotton (mg g-1 FW)"

品种
Variety		 处理
Treatment		 2017 2018
18 d 21 d 24 d 18 d 21 d 24 d
SK-1 CK 6.83 e 6.72 g 6.23 d 3.86 b 3.51 f 5.37 e
N1 8.66 d 8.35 f 8.47 c 4.01 b 4.64 ef 6.54 d
N2 10.31 bc 11.37 cd 12.19 ab 6.35 a 6.35 cd 8.00 abc
N3 10.79 b 12.11 bc 12.81 a 6.52 a 6.86 bc 7.22 bcd
N4 9.34 cd 10.24 de 8.94 c 4.59 b 5.77 cde 6.94 cd
SK-3 CK 6.80 e 9.24 ef 5.73 d 2.60 c 5.29 de 6.79 cd
N1 8.72 d 9.78 e 6.69 d 3.54 bc 5.40 de 6.89 cd
N2 10.98 b 13.28 ab 11.01 b 3.81 b 7.72 ab 9.14 a
N3 12.53 a 14.33 a 12.32 ab 6.57 a 8.24 a 8.29 ab
N4 8.72 d 11.36 cd 8.57 c 3.65 bc 5.90 cde 7.74 bcd

Table 4

Effects of increased nitrogen application on square GOT activities in cotton (μmol g-1 h-1)"

品种
Variety		 处理
Treatment		 2017 2018
18 d 21 d 24 d 18 d 21 d 24 d
SK-1 CK 4.18 c 6.50 e 11.70 bc 3.79 d 4.13 c 2.69 c
N1 4.26 c 7.18 de 11.97 b 3.94 cd 4.47 c 3.65 abc
N2 4.84 bc 8.63 c 11.96 b 4.07 bcd 4.88 b 4.17 ab
N3 5.36 bc 13.97 a 10.76 cd 4.19 bcd 5.25 a 4.62 a
N4 4.84 bc 10.07 b 9.28 ef 3.97 cd 4.47 c 3.66 abc
SK-3 CK 4.49 bc 5.76 e 8.39 f 4.30 bcd 3.34 d 3.52 bc
N1 7.13 abc 6.98 e 9.82 de 4.57 abc 3.53 d 3.79 ab
N2 10.15 a 8.50 cd 10.81 cd 5.03 a 3.70 d 4.29 ab
N3 9.83 a 9.18 bc 13.67 a 4.78 ab 4.26 c 4.39 ab
N4 7.63 ab 6.62 e 11.95 b 4.72 ab 3.54 d 3.94 ab

Table 5

Effects of increased nitrogen application on square GS activities in cotton (U g-1 FW min-1)"

品种
Variety		 处理
Treatment		 2017 2018
18 d 21 d 24 d 18 d 21 d 24 d
SK-1 CK 3.92 f 6.46 h 7.00 f 3.47 f 5.78 f 6.94 cd
N1 6.20 e 9.32 f 9.10 d 5.35 d 7.10 d 7.51 cd
N2 8.37 c 11.25 d 11.42 b 6.30 b 8.07 bc 10.27 a
N3 9.22 ab 12.72 b 12.36 a 6.82 a 8.31 ab 10.35 a
N4 6.53 de 9.51 ef 9.61 c 5.98 c 8.05 bc 8.13 bc
SK-3 CK 3.70 f 7.52 g 7.33 e 4.92 e 5.85 f 6.77 d
N1 6.82 d 9.52 ef 9.30 d 5.45 d 6.46 e 7.35 cd
N2 9.10 b 11.68 c 11.61 b 6.84 a 7.82 c 9.13 ab
N3 9.51 a 13.57 a 12.22 a 6.85 a 8.61 a 9.51 a
N4 6.36 e 9.78 e 9.11 d 5.90 c 6.71 e 7.82 cd

Table 6

Effects of increased nitrogen application on square protease activities in cotton (μg g-1 min-1)"

品种
Variety		 处理
Treatment		 2017 2018
18 d 21 d 24 d 18 d 21 d 24 d
SK-1 CK 14.47 a 14.49 a 12.52 a 15.22 bc 16.67 ab 33.96 ab
N1 13.21 bc 14.28 a 11.97 a 14.61 bcd 15.76 abc 28.20 bcd
N2 12.34 de 12.16 cd 9.84 abc 13.88 cd 12.83 abcd 26.81 cde
N3 11.24 f 11.60 de 9.22 bc 10.59 e 11.67 bcd 17.97 fg
N4 11.65 ef 10.13 f 8.81 c 10.00 e 8.40 d 10.55 h
SK-3 CK 14.15 a 13.74 ab 12.40 a 17.81 a 18.35 a 34.48 a
N1 14.14 a 12.80 bc 11.72 ab 16.64 ab 16.21 ab 29.87 abc
N2 13.88 ab 12.93 bc 11.13 abc 12.38 de 14.23 abc 22.32 def
N3 12.72 cd 12.59 bcd 11.00 abc 9.93 e 11.36 bcd 20.66 ef
N4 12.30 de 10.98 ef 10.70 abc 9.92 e 10.21 cd 14.19 gh

Table 7

Effects of increased nitrogen application on square peptidase activities in cotton (U g-1 FW h-1)"

品种
Variety		 处理Treatment 2017 2018
18 d 21 d 24 d 18 d 21 d 24 d
SK-1 CK 13.11 a 13.83 a 19.78 ab 16.45 a 16.04 ab 19.51 a
N1 10.91 b 11.25 bc 16.10 c 14.63 ab 14.45 abc 14.99 bc
N2 8.71 c 9.93 cd 15.62 c 10.11 cd 12.97 bcd 13.73 cd
N3 7.05 d 8.81 de 13.11 d 10.04 cd 10.98 d 13.02 cde
N4 5.52 e 6.09 f 11.06 e 8.60 d 10.79 d 10.49 e
SK-3 CK 13.82 a 14.39 a 20.65 a 15.08 ab 17.03 a 18.22 a
N1 12.52 a 12.06 b 18.65 b 14.66 ab 15.98 ab 16.89 ab
N2 10.26 b 11.05 bc 16.51 c 13.10 abc 12.46 cd 13.76 cd
N3 7.79 cd 9.04 de 13.83 d 12.42 abcd 10.34 d 12.54 cde
N4 5.35 e 8.03 e 11.88 e 12.18 bcd 10.29 d 11.39 de

Fig. 1

Relationship between square Bt insecticidal protein content and main chemical substances, key enzyme activities of nitrogen metabolism SP: soluble protein; AA: free amino acid; GOT: glutamic oxalacetic transaminase; GS: glutamine synthetase. The positive or negative coefficients represent the positive or negative effects and the magnitude of contribution to Bt protein content. **: P < 0.01."

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[1] WANG Yan;QIU Li-Ming;XIE Wen-Juan;HUANG Wei;YE Feng;ZHANG Fu-Chun;MA Ji. Cold Tolerance of Transgenic Tobacco Carrying Gene Encoding Insect Antifreeze Protein[J]. Acta Agron Sin, 2008, 34(03): 397 -402 .
[2] XING Guang-Nan, ZHOU Bin, ZHAO Tuan-Jie, YU De-Yue, XING Han, HEN Shou-Yi, GAI Jun-Yi. Mapping QTLs of Resistance to Megacota cribraria (Fabricius) in Soybean[J]. Acta Agronomica Sinica, 2008, 34(03): 361 -368 .
[3] ZHENG Yong-Mei;DING Yan-Feng;WANG Qiang-Sheng;LI Gang-Hua;WANG Hui-Zhi;WANG Shao-Hua. Effect of Nitrogen Applied before Transplanting on Tillering and Nitrogen Utilization in Rice[J]. Acta Agron Sin, 2008, 34(03): 513 -519 .
[4] LÜ Li-Hua;TAO Hong-Bin;XIA Lai-Kun; HANG Ya-Jie;ZHAO Ming;ZHAO Jiu-Ran;WANG Pu;. Canopy Structure and Photosynthesis Traits of Summer Maize under Different Planting Densities[J]. Acta Agron Sin, 2008, 34(03): 447 -455 .
[5] SHAO Rui-Xin;SHANG-GUAN Zhou-Ping. Effects of Exogenous Nitric Oxide Donor Sodium Nitroprusside on Photosynthetic Pigment Content and Light Use Capability of PS II in Wheat under Water Stress[J]. Acta Agron Sin, 2008, 34(05): 818 -822 .
[6] Huang Ce;Wang Tian-duo. COMPUTER SIMULATION OF BIOMASS PRODUCTION IN RICE COMMUNITY[J]. Acta Agron Sin, 1986, (01): 1 -8 .
[7] CHEN Ji-Bao;JING Rui-Lian;MAO Xin-Guo;CHANG Xiao-Ping;WANG Shu-Min. A Response of PvP5CS2 Gene to Abiotic Stresses in Common Bean[J]. Acta Agron Sin, 2008, 34(07): 1121 -1127 .
[8] LIU Wu-Ge;WANG Feng;JIN Su-Juan;ZHU Xiao-Yuan;LI Jin-Hua;LIU Zhen-Rong;LIAO Yi-Long;ZHU Man-Shan;HUANG Hui-Jun; FU Fu-Hong;LIU Yi-Bai. Improvement of Rice Blast Resistance in TGMS Line by Pyramiding of Pi-1 and Pi-2 through Molecular Marker-Assisted Selection[J]. Acta Agron Sin, 2008, 34(07): 1128 -1136 .
[9] ZHANG Wen-Jing;HU Hong-Biao;CHEN Bing-Lin;WANG You-Hua;ZHOU Zhi-Guo. Difference of Physiological Characteristics of Cotton Bolls in Development of Fiber Thickening and Its Relationship with Fiber Strength[J]. Acta Agron Sin, 2008, 34(05): 859 -869 .
[10]

CHANG Li-Ying;GU Dong-Xiang;ZHANG Wen-Yu;YANG Jie;CAO Wei-Xing;ZHU Yan

. A Simulation Model of Leaf Elongation Process in Rice[J]. Acta Agron Sin, 2008, 34(02): 311 -317 .
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