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Acta Agronomica Sinica ›› 2023, Vol. 49 ›› Issue (9): 2505-2516.doi: 10.3724/SP.J.1006.2023.24217

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

Effects of increased nitrogen on Bt protein expression and nitrogen metabolism in the leaf subtending to cotton boll

LI Yi-Yang(), LI Yuan, ZHAO Zi-Xu, ZHANG Ding-Shun, DU Jia-Ning, WU Shu-Juan, SUN Si-Qi, CHEN Yuan, ZHANG Xiang, CHEN De-Hua(), LIU Zhen-Yu()   

  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:2022-09-30 Accepted:2023-02-10 Online:2023-09-12 Published:2023-02-23
  • Supported by:
    National Natural Science Foundation of China(31901462);National Natural Science Foundation of China(31671613);Natural Science Foundation of Jiangsu Province(BK20191439);Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD);Higher School in Jiangsu Province College Students’ Innovation and Entrepreneurship Training Programs(202111117074Y)

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Abstract:

The objective of this study is to explore the effect of soil nitrogen increase on the expression of insecticidal proteins in the leaf subtending to cotton boll and the related physiological mechanism of nitrogen metabolism and to provide the theoretical and technical support for agronomic regulation of Bt cotton boll stage insect resistance. A split-plot experiment was conducted. 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 Bt protein content and nitrogen metabolic physiological in the leaf subtending to cotton boll. The results showed that the content of Bt protein in the leaf subtending to cotton boll of two types of cultivars showed a constant increase with the increase of nitrogen application rate. Compared with the control (300 kg hm-2), Bt protein content in the leaf subtending to cotton boll increased by 6.1%-96.9% with the increase of 25%-100% nitrogen application. 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 the leaf subtending to cotton boll 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. The leaf area index (LAI) increased with the increase of nitrogen application, while the yield increased first and then decreased with the increase of soil nitrogen. The optimum LAI and the maximum yield were both 1.25 times (375 kg hm-2) that of conventional nitrogen application. In conclusion, on the basis of conventional nitrogen application, an appropriate amount of nitrogen fertilizer was beneficial to the synthesis of Bt protein, the formation of the optimal LAI, and the increase of yield in the counter-position leaves of cotton boll, which was conductive to the synergistic expression of high yield and insect resistance of Bt cotton.

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

Table 1

Effects of increased nitrogen application on Bt protein contents in the leaf subtending to cotton boll (ng g-1 FW)"

品种
Cultivar		 处理
Treatment		 2017年花后天数Days after flowering in 2017 2018年花后天数Days after flowering in 2018
15 d 20 d 25 d 15 d 20 d 25 d
SK-1 CK 169.54±5.96 c 117.61±5.59 g 133.67±2.91 e 136.30±2.70 ef 138.88±1.36 e 135.93±1.09 d
N1 184.31±4.54 c 162.29±5.79 ef 151.92±0.40 d 156.53±3.78 c 158.37±2.86 d 145.17±2.79 bcd
N2 257.71±2.31 b 179.08±0.68 e 174.42±0.44 c 163.06±1.30 bc 171.06±1.31 bc 152.36±4.02 bc
N3 277.43±21.15 ab 201.19±11.67 d 197.97±11.41 b 172.19±2.84 b 175.14±2.28 b 155.25±1.23 b
N4 317.85±12.69 a 231.54±17.47 bc 202.76±5.72 b 192.62±2.28 a 198.64±5.12 a 174.89±1.56 a
SK-3 CK 171.21±5.89 c 151.66±5.44 f 128.29±5.80 e 125.30±3.58 f 134.86±2.53 e 133.76±2.41 d
N1 265.59±3.91 b 162.09±5.84 ef 137.19±2.96 e 141.18±2.72 de 155.79±1.77 d 141.86±3.41 cd
N2 272.65±30.34 ab 215.44±3.01 cd 153.26±0.58 d 151.48±3.47 cd 157.99±2.99 d 144.76±0.64 bcd
N3 288.06±19.46 ab 250.09±5.92 ab 183.66±2.82 c 161.54±4.20 bc 163.77±2.23 cd 151.22±2.10 bc
N4 315.68±29.14 a 263.03±0.15 a 232.21±5.81 a 187.59±1.92 a 192.42±5.03 a 170.83±2.64 a
方差分析ANOVA
品种Variety NS NS NS NS * NS
处理Treatment ** ** ** ** ** **
品种×处理Variety×treatment NS ** NS NS NS NS

Table 2

Effects of increased nitrogen application on soluble protein contents in the leaf subtending to cotton boll (mg g-1 FW)"

品种
Cultivar		 处理
Treatment		 2017年花后天数Days after flowering in 2017 2018年花后天数Days after flowering in 2018
15 d 20 d 25 d 15 d 20 d 25 d
SK-1 CK 0.46±0.06 d 0.43±0.01 d 0.40±0.01 e 0.42±0.00 f 0.39±0.08 e 0.44±0.07 e
N1 0.58±0.12 cd 0.65±0.24 d 0.60±0.01 de 0.49±0.04 ef 0.45±0.07 e 0.54±0.03 de
N2 0.76±0.18 bc 0.93±0.04 c 0.72±0.01 cd 0.65±0.03 ef 0.75±0.26 cd 0.69±0.10 cd
N3 0.94±0.01 ab 1.03±0.07 bc 0.89±0.01 bc 1.27±0.16 b 0.79±0.17 cd 0.68±0.24 cd
N4 1.14±0.00 a 1.27±0.04 b 1.01±0.02 ab 1.41±0.04 b 0.91±0.21 c 1.10±0.13 ab
SK-3 CK 0.44±0.00 d 0.56±0.03 d 0.38±0.01 e 0.51±0.03 ef 0.44±0.21 e 0.55±0.11 de
N1 0.58±0.03 cd 0.93±0.01 c 0.67±0.06 cd 0.74±0.01 de 0.66±0.15 d 0.72±0.20 c
N2 0.74±0.09 bc 1.13±0.18 bc 0.80±0.06 bcd 0.97±0.01 cd 1.13±0.19 b 0.79±0.14 c
N3 0.78±0.09 bc 1.19±0.01 b 1.01±0.12 ab 1.18±0.13 bc 0.93±0.16 bc 0.98±0.25 b
N4 1.03±0.00 a 1.59±0.07 a 1.19±0.07 a 1.75±0.09 a 2.00±0.09 a 1.18±0.21 a
方差分析ANOVA
品种 Variety NS NS NS NS NS NS
处理 Treatment ** ** ** ** ** **
品种×处理 Variety×treatment NS NS NS * NS NS

Table 3

Effects of increased nitrogen application on free amino acid contents in the leaf subtending to cot ton boll (mg g-1 FW)"

品种
Cultivar		 处理
Treatment		 2017年花后天数 Days after flowering in 2017 2018年花后天数Days after flowering in 2018
15 d 20 d 25 d 15 d 20 d 25 d
SK-1 CK 1.27±0.02 g 1.13±0.02 g 1.23±0.02 g 1.32±0.02 d 1.66±0.02 c 1.64±0.02 f
N1 1.43±0.02 e 1.18±0.02 fg 1.35±0.04 de 1.33±0.02 d 1.90±0.02 b 1.86±0.04 e
N2 1.53±0.05 cd 1.25±0.03 ef 1.38±0.04 cd 1.66±0.05 c 1.67±0.03 c 2.21±0.04 c
N3 1.65±0.03 b 1.34±0.02 d 1.39±0.02 c 1.80±0.03 b 1.76±0.02 bc 2.44±0.02 b
N4 1.75±0.03 a 1.44±0.07 b 1.48±0.04 b 1.94±0.03 a 1.94±0.07 ab 2.59±0.04 a
SK-3 CK 1.27±0.02 g 1.28±0.02 de 1.26±0.02 g 0.79±0.02 e 1.30±0.02 d 1.51±0.02 g
N1 1.34±0.02 f 1.35±0.02 cd 1.31±0.02 f 1.23±0.02 d 1.83±0.02 bc 1.72±0.02 f
N2 1.51±0.03 d 1.42±0.09 bc 1.32±0.04 ef 1.69±0.03 c 1.77±0.09 bc 1.99±0.04 d
N3 1.58±0.07 c 1.49±0.04 b 1.38±0.06 cd 1.79±0.07 b 1.87±0.04 b 2.27±0.06 c
N4 1.73±0.04 a 1.66±0.03 a 1.53±0.03 a 1.95±0.04 a 2.12±0.03 a 2.37±0.03 b
方差分析ANOVA
品种 Variety NS NS NS NS NS NS
处理 Treatment ** ** ** ** ** **
品种×处理 Variety×treatment NS NS NS NS NS NS

Table 4

Effects of increased nitrogen application on GPT activities in the leaf subtending to cotton boll (μmol g-1 h-1)"

品种
Cultivar		 处理
Treatment		 2017年花后天数 Days after flowering in 2017 2018年花后天数Days after flowering in 2018
15 d 20 d 25 d 15 d 20 d 25 d
SK-1 CK 4.63±0.56 f 4.43±0.56 e 4.90±0.50 f 5.45±0.03 cd 6.27±0.96 b 3.02±0.30 f
N1 6.02±0.57 de 5.67±0.57 de 5.96±0.34 e 5.52±0.33 cd 6.27±0.82 b 4.71±0.22 de
N2 7.05±1.10 bcd 7.21±0.58 bc 8.19±0.42 c 6.13±0.54 bcd 6.74±0.84 b 4.99±0.54 cde
N3 8.18±0.59 b 7.14±0.76 bc 8.94±0.43 b 7.64±0.78 bc 7.99±0.31 ab 6.47±0.23 bc
N4 9.96±0.59 a 8.17±0.59 ab 9.53±0.37 ab 8.08±1.04 b 9.07±0.11 ab 8.85±0.93 a
SK-3 CK 5.50±0.57 ef 4.84±0.56 de 4.79±0.33 f 5.49±0.45 d 6.45±0.33 b 3.76±0.53 ef
N1 6.64±0.75 cde 6.08±0.57 cd 6.34±0.35 e 5.01±0.08 cd 6.48±0.12 b 4.39±0.53 ef
N2 7.41±0.58 bc 7.28±0.18 bc 7.37±0.18 d 6.61±0.37 bcd 6.55±0.03 b 6.26±0.17 bcd
N3 9.54±0.15 a 7.99±0.16 ab 9.43±0.20 ab 8.25±0.75 b 6.98±0.03 b 7.42±0.25 ab
N4 9.58±0.07 a 8.92±0.12 a 9.89±0.95 a 9.14±0.84 a 9.89±0.88 a 8.15±0.71 a
方差分析ANOVA
品种 Variety NS NS * NS NS NS
处理 Treatment ** ** ** ** ** **
品种×处理 Variety×treatment NS NS NS ** NS NS

Table 5

Effects of increased nitrogen application on GS activities in the leaf subtending to cotton boll (U g-1 FW min-1)"

品种
Cultivar		 处理
Treatment		 2017年花后天数 Days after flowering in 2017 2018年花后天数Days after flowering in 2018
15 d 20 d 25 d 15 d 20 d 25 d
SK-1 CK 0.66±0.06 f 0.66±0.01 d 0.72±0.01 c 0.48±0.05 e 0.82±0.05 cd 0.96±0.07 cd
N1 0.90±0.12 e 0.90±0.24 cd 0.84±0.01 bc 0.57±0.03 de 1.07±0.09 bc 1.38±0.01 b
N2 1.08±0.01 cd 0.96±0.01 bc 0.90±0.01 b 0.69±0.04 cde 1.08±0.06 bc 1.42±0.17 ab
N3 1.14±0.02 bcd 1.14±0.01 abc 1.26±0.07 a 0.93±0.06 bcd 1.13±0.09 b 1.63±0.13 ab
N4 1.26±0.03 ab 1.20±0.13 ab 1.26±0.07 a 1.37±0.26 a 1.46±0.07 a 1.82±0.05 a
SK-3 CK 0.54±0.03 f 0.66±0.03 d 0.72±0.01 c 0.63±0.01 de 0.59±0.07 d 0.90±0.07 d
N1 0.90±0.09 e 0.90±0.01 cd 0.90±0.07 b 0.73±0.06 cde 1.03±0.1 bc 1.35±0.08 bc
N2 1.02±0.02 de 0.90±0.07 cd 0.96±0.07 b 0.76±0.08 cde 1.05±0.07 bc 1.36±0.04 bc
N3 1.20±0.01 abc 1.20±0.18 ab 1.26±0.03 a 1.03±0.05 abc 1.16±0.07 b 1.41±0.16 ab
N4 1.32±0.00 a 1.26±0.13 a 1.32±0.13 a 1.26±0.02 ab 1.21±0.06 b 1.64±0.05 ab
方差分析ANOVA
品种 Variety NS NS NS NS NS NS
处理 Treatment ** ** ** ** ** **
品种×处理 Variety×treatment NS NS NS NS NS NS

Table 6

Effects of increased nitrogen application on protease activities in the leaf subtending to cotton boll (μg g-1 min-1)"

品种
Cultivar		 处理
Treatment		 2017年花后天数 Days after flowering in 2017 2018年花后天数Days after flowering in 2018
15 d 20 d 25 d 15 d 20 d 25 d
SK-1 CK 15.04±0.71 ab 15.74±1.35 a 15.13±1.86 a 16.08±0.31 bc 19.22±1.37 a 16.53±1.27 b
N1 13.49±1.27 bc 14.20±0.65 bc 11.87±1.55 b 13.84±0.71 cd 16.11±0.43 bc 13.05±1.41 c
N2 12.32±1.11 c 12.89±1.21 cd 8.84±1.00 de 13.15±0.17 d 13.52±0.83 de 11.92±0.09 cd
N3 10.07±0.38 d 11.99±0.23 d 9.47±1.01 cd 8.27±0.54 ef 12.97±0.97 ef 10.84±0.16 cde
N4 7.88±0.58 e 8.34±0.36 e 7.51±0.36 e 6.31±0.22 f 8.46±0.39 g 10.19±0.51 de
SK-3 CK 15.66±1.29 a 15.10±1.23 ab 13.73±0.99 a 21.58±0.26 a 17.60±0.15 ab 19.94±0.90 a
N1 14.17±0.99 ab 12.09±0.91 d 12.16±0.97 b 17.42±0.77 b 15.76±0.30 bcd 16.06±0.85 b
N2 11.99±1.55 c 11.67±1.37 d 10.98±1.37 bc 16.54±0.22 b 14.03±0.59 cde 13.22±0.57 c
N3 9.84±0.78 d 9.64±0.75 e 9.86±1.93 cd 15.71±1.67 bc 11.69±1.08 ef 10.32±0.30 de
N4 7.98±1.05 e 8.99±0.12 e 8.65±0.37 de 8.84±0.28 e 10.87±0.37 f 8.72±0.28 e
方差分析ANOVA
品种 Variety NS * ** ** NS NS
处理 Treatment ** ** ** ** ** **
品种×处理 Variety×treatment NS NS NS * NS *

Table 7

Effects of increased nitrogen application on peptidase activities in the leaf subtending to cotton boll (U g-1 FW h-1)"

品种
Cultivar		 处理
Treatment		 2017年花后天数 Days after flowering in 2017 2018年花后天数Days after flowering in 2018
15 d 20 d 25 d 15 d 20 d 25 d
SK-1 CK 7.22±0.24 abc 7.04±0.12 ab 8.14±0.65 b 7.88±0.22 ab 10.89±0.93 a 5.19±0.47 a
N1 6.53±0.52 abcd 5.43±0.57 bc 8.09±1.51 b 7.31±0.98 ab 8.51±0.86 ab 4.48±0.72 abc
N2 5.37±0.76 cde 4.80±0.56 cde 6.49±0.52 bc 6.71±0.33 ab 7.83±0.02 b 3.99±0.26 abc
N3 5.03±0.22 de 3.48±0.15 de 3.99±0.38 d 6.58±0.48 ab 7.12±0.36 b 3.73±0.38 abc
N4 3.37±0.29 e 3.07±0.32 e 3.20±0.32 d 5.46±0.53 b 4.13±0.37 c 3.02±0.09 c
SK-3 CK 8.41±1.40 a 7.24±0.58 a 7.47±0.64 bc 8.61±1.28 a 9.39±0.61 ab 4.76±0.47 ab
N1 7.58±1.11 ab 5.59±0.86 abc 10.51±0.38 a 7.66±0.50 ab 9.03±0.44 ab 4.49±0.41 abc
N2 7.65±0.82 ab 5.09±1.31 cd 3.77±0.15 d 6.72±0.15 ab 8.45±0.89 ab 4.17±0.48 abc
N3 6.02±0.23 bcd 4.60±0.70 cde 3.50±0.15 d 6.58±1.11 ab 7.41±0.47 b 3.55±0.02 bc
N4 5.28±0.76 cde 3.37±0.06 de 5.97±0.34 c 5.81±0.32 ab 4.11±0.49 c 3.14±0.34 c
方差分析ANOVA
品种 Variety NS NS NS NS NS NS
处理 Treatment ** ** ** ** ** **
品种×处理 Variety×treatment NS NS NS NS NS NS

Table 8

Effects of increased nitrogen application on LAI in cotton"

品种
Cultivar		 处理
Treatment		 2017年花后天数 Days after flowering in 2017 2018年花后天数Days after flowering in 2018
15 d 20 d 25 d 15 d 20 d 25 d
SK-1 CK 3.4±0.1 e 3.7±0.1 e 3.9±0.1 f 3.6±0 g 3.8±0.1 f 4.1±0.1 e
N1 3.7±0 d 4.0±0.3 cde 4.2±0 e 3.9±0.1 f 4.2±0 def 4.4±0.1 d
N2 4.2±0.1 c 4.3±0.1 bc 4.6±0.1 d 4.3±0.1 d 4.5±0.1 bcd 4.7±0 c
N3 4.4±0.1 bc 4.5±0.1 ab 4.9±0 c 4.5±0.1 bc 4.7±0.1 abc 4.9±0.1 c
N4 4.5±0 ab 4.7±0.3 a 5.1±0 bc 4.7±0 a 4.8±0 ab 5.2±0.1 b
SK-3 CK 3.6±0.2 de 3.8±0.1 de 4.1±0.2 ef 3.7±0 g 3.9±0 ef 4.2±0 e
N1 3.8±0 d 4.1±0 cd 4.5±0 d 4.1±0.1 e 4.3±0 cde 4.5±0 d
N2 4.5±0 ab 4.6±0 ab 4.9±0 c 4.4±0 cd 4.7±0 abc 4.8±0 c
N3 4.6±0 ab 4.8±0 a 5.2±0 ab 4.6±0.1 ab 5.0±0.4 a 5.3±0.1 b
N4 4.7±0 a 4.9±0 a 5.3±0.1 a 4.7±0 a 5.1±0.1 a 5.5±0 a
方差分析ANOVA
品种 Variety ** ** ** * NS *
处理 Treatment ** ** ** ** ** **
品种×处理 Variety×treatment NS NS NS NS NS **
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