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作物学报 ›› 2022, Vol. 48 ›› Issue (10): 2567-2574.doi: 10.3724/SP.J.1006.2022.14205

• 耕作栽培·生理生化 • 上一篇    下一篇

土壤增施氮肥对棉蕾Bt杀虫蛋白表达量影响及氮代谢机制

李涵佳(), 李远, 刘震宇, 张晨霞, 徐泽, 吴天凡, 陈媛, 张祥, 陈源, 陈德华()   

  1. 扬州大学 / 江苏省作物遗传生理国家重点实验室培育点 / 粮食作物现代产业技术协同创新中心, 江苏扬州 225009
  • 收稿日期:2021-10-30 接受日期:2022-02-25 出版日期:2022-10-12 网络出版日期:2022-03-22
  • 通讯作者: 陈德华
  • 作者简介:第一作者联系方式: E-mail: 1311190875@qq.com
  • 基金资助:
    国家自然科学基金项目(31471435);国家自然科学基金项目(31671613);江苏省自然科学基金项目(BK20191439);江苏省高等教育优势学科发展计划项目(PAPD)

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 Published:2022-10-12 Published online:2022-03-22
  • Contact: CHEN De-Hua
  • 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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摘要：

为明确土壤增施氮肥对Bt棉生殖器官棉蕾杀虫蛋白表达量影响, 2017—2018年于扬州大学遗传生理重点实验室以Bt棉常规品种泗抗1号(SK-1)、杂交品种泗抗3号(SK-3)为材料, 在常规施氮量300 kg hm-2基础上, 设计施氮量分别增加25%、50%、75%、100%的处理, 探讨土壤增施氮肥对Bt棉棉蕾中杀虫蛋白表达量的影响及其氮代谢生理机制。结果表明, 2个类型品种棉蕾中Bt杀虫蛋白含量均随增施氮量提高呈先增加后降低的趋势, 与对照相比, 施氮量增加25%~100%, 棉蕾Bt蛋白增加4.5%~132.7%, Bt蛋白的最大含量基本出现在常规施氮1.50~1.75倍(450~525 kg hm-2)。氮代谢生理机制则表明, 棉蕾中可溶性蛋白(soluble protein, SP)含量、游离氨基酸(free amino acid, AA)含量、谷氨酸草酰乙酸转氨酶(glutamic oxalacetic transaminase, GOT)活性和谷氨酰胺合成酶(glutamine synthetase, GS)活性的变化趋势与Bt蛋白含量表现一致, 而蛋白质分解关键酶(蛋白酶、肽酶)活性则随施氮量的增加呈下降趋势。因此, 在常规施氮基础上适量增施氮肥有利于棉蕾Bt蛋白的合成, 进而提高抗虫性。

关键词: Bt棉, 增施氮肥, 杀虫蛋白, 氮代谢

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

表1

增施氮肥对棉蕾Bt杀虫蛋白含量的影响"

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

表2

增施氮肥对棉蕾可溶性蛋白含量的影响"

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

表3

增施氮肥对棉蕾游离氨基酸含量的影响"

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

表4

增施氮肥对棉蕾GOT活性的影响"

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

表5

增施氮肥对棉蕾GS活性的影响"

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

表6

增施氮肥对棉蕾蛋白酶活性的影响"

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

表7

增施氮肥对棉蕾肽酶活性的影响"

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

图1

棉蕾氮代谢主要化合物及关键酶活性与Bt杀虫蛋白含量的关系 SP: 可溶性蛋白; AA: 游离氨基酸; GOT: 谷氨酸草酰乙酸转氨酶; GS: 谷氨酰胺合成酶。相关系数的正负及大小表示对Bt蛋白含量贡献的正负效应和大小。**表示极显著相关(P < 0.01)。"

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