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作物学报 ›› 2020, Vol. 46 ›› Issue (3): 440-447.doi: 10.3724/SP.J.1006.2020.94080

• 耕作栽培·生理生化 • 上一篇    下一篇

高温胁迫终止后Bt棉蕾杀虫蛋白的恢复特征及相关生理机制

刘震宇,王桂霞,李丽楠,蔡泽洲,梁潘潘,吴莘玲,张祥,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:2019-05-27 接受日期:2019-09-26 出版日期:2020-03-12 网络出版日期:2019-10-14
  • 通讯作者: 陈德华
  • 作者简介:E-mail: 1127317278@qq.com
  • 基金资助:
    本研究由国家自然科学基金项目(31671613);国家现代农业产业技术体系棉花岗位专家项目(CARS-18-18);江苏省高等教育优势学科发展计划(PAPD)资助

Recovery characteristics of Bt insecticidal protein and relative physiological mechanisms after high temperature stress termination in square of Bt cotton

Zhen-Yu LIU,Gui-Xia WANG,Li-Nan LI,Ze-Zhou CAI,Pan-Pan LIANG,Xin-Ling WU,Xiang ZHANG,De-Hua CHEN()   

  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:2019-05-27 Accepted:2019-09-26 Published:2020-03-12 Published online:2019-10-14
  • Contact: De-Hua CHEN
  • Supported by:
    This study was supported by the National Natural Science Foundation of China(31671613);the China Agriculture Research System (Cotton Post Expert Project)(CARS-18-18);the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD)

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摘要：

以大面积生产应用的Bt棉常规种‘泗抗1号’和杂交种‘泗抗3号’为材料, 于扬州大学遗传生理重点实验室人工气候室, 以盆栽方式设计高温胁迫试验。2017年设蕾期38℃高温持续胁迫72 h终止、2018年设38℃高温持续胁迫96 h终止处理后不同时间(0、12、24、48、72和96 h)研究棉蕾Bt杀虫蛋白表达量变化及相关氮代谢生理。结果表明, 盛蕾期38℃持续胁迫72 h、96 h终止后棉蕾Bt杀虫蛋白表达量均能恢复到相应对照水平, 并且恢复所需时间随胁迫持续时间的延长而增加。胁迫72 h终止后, 常规种泗抗1号经72 h、杂交种泗抗3号经48 h蕾的Bt杀虫蛋白表达量与对照已无显著差异; 胁迫96 h终止后, 泗抗1号经96 h、泗抗3号经72 h, 棉蕾Bt杀虫蛋白表达量与对照已无显著差异。相关分析表明, 高温胁迫终止后, 蕾中可溶性蛋白表达量、谷氨酸丙酮酸转氨酶(GPT)活性、谷氨酸草酞乙酸转氨酶(GOT)活性与Bt杀虫蛋白表达量呈极显著正相关 (相关系数分别为0.964 **、0.981 **、0.971 **); 而游离氨基酸含量、蛋白酶和肽酶的活性与Bt杀虫蛋白表达量呈极显著负相关(相关系数分别为-0.894 **、-0.912 **、-0.834 **)。因此, 生产上可根据高温持续时间预测棉蕾Bt杀虫蛋白表达量的恢复程度, 合理防治棉铃虫等相关害虫。

关键词: Bt棉, 棉蕾, 高温胁迫, Bt杀虫蛋白, 氮代谢

Abstract:

The experiments were conducted at Key Laboratory of Genetics and Physiology of Yangzhou University. The conventional cultivar Sikang-1 and hybrid cultivar Sikang-3 were used as the experimental materials. The potted cotton at squaring stage was moved to artificial climatic chamber with 38℃ for 72 h , and 38℃ for 96 h respectively in 2017 and 2018 cotton growth season, and moved to natural conditions, then the squares were collected to measure the Bt protein concentration and nitrogen metabolic physiology at 0, 12, 24, 48, 72, 96 hours respectively after the high temperature termination. The square Bt insecticidal protein contents were able to recover to the corresponding control level, the time for the recovery was in need of longer as the stressed high temperature period increased. In 72 h high temperature treatment, the square Bt insecticidal protein content could recover to the corresponding control level at 72 h for Sikang-1 and at 48 h for Sikang-3. In 96 h high temperature treatment, the square Bt insecticidal protein content could recover to the corresponding control level at 96 h for Sikang-1 and at 72 h for Sikang-3. There were significantly positive correlations of Bt toxin content with soluble protein content, glutamate pyruvate transaminase activity and glutamate oxaloacetate transaminase activity (the correlation coefficients were 0.964 **, 0.981 **, and 0.971 **, respectively), and significantly negative correlations of Bt toxin content with free amino acid contents, and activitied of protease and peptidase (the correlation coefficients were -0.894 **, -0.912 **, and -0.834 **, respectively). Therefore, the recovery degree of Bt insecticidal protein content of cotton square can be predicted according to the duration of high temperature stress, reasonably controlling Helicoverpa armigera and related pests in production.

Key words: Bt cotton, square, high temperature stress, Bt protein, nitrogen metabolism

图1

高温持续胁迫72 h终止后棉蕾Bt蛋白表达量变化 不同小写字母表示0.05显著水平下相同品种、相同处理间与对照间的差异。T: 高温胁迫72 h; CK: 对照。"

图2

高温持续胁迫96 h终止后棉蕾Bt蛋白表达量变化 不同小写字母表示0.05显著水平下相同品种、相同处理间与对照间的差异。T: 高温胁迫96 h; CK: 对照。"

表1

高温持续胁迫终止后蕾中可溶性蛋白与游离氨基酸含量变化"

测定指标
Tested index		 品种
Variety		 处理
Treatment		 恢复时间 Recovery time
0 h 12 h 24 h 48 h 72 h 96 h
可溶性蛋白含量
Soluble protein content
(mg g-1 FM)		 SK-1 CK 10.19 a 10.25 a 10.25 a 10.19 a 10.16 a 10.26 a
38℃-72 h 4.77 d 5.12 d 6.45 c 8.56 b 10.36 a 10.36 a
38℃-96 h 4.15 e 4.44 e 5.43 d 6.86 c 8.47 b 10.09 a
SK-3 CK 12.10 a 12.20 a 12.27 a 12.25 a 12.18 a 12.24 a
38℃-72 h 6.26 d 7.42 c 9.17 b 12.05 a 12.23 a 12.27 a
38℃-96 h 5.93 e 6.80 d 7.88 c 10.89 a 12.02 a 12.23 a
游离氨基酸含量
Free amino acid content
(μmol g-1 FM)		 SK-1 CK 255.21 a 250.16 a 249.88 a 251.92 a 253.55 a 249.60 a
38℃-72 h 747.74 a 693.73 b 626.64 c 476.67 d 240.14 d 243.28 d
38℃-96 h 974.31 a 923.16 b 854.25 c 689.28 d 500.86 e 252.38 e
SK-3 CK 277.06 a 273.20 a 281.32 a 277.44 a 275.88 a 272.99 a
38℃-72 h 681.54 a 607.52 b 520.30 c 286.08 d 279.63 d 272.75 d
38℃-96 h 905.24 a 829.97 b 742.43 c 346.71 d 273.32 e 274.59 e

表2

高温持续胁迫终止后蕾中GOT与GPT活性变化"

测定指标
Tested index		 品种
Variety		 处理
Treatment		 恢复时间 Recovery time
0 h 12 h 24 h 48 h 72 h 96 h
GOT
(μmol g-1 h-1)		 SK-1 CK 12.71 a 12.79 a 12.69 a 12.95 a 12.40 a 12.49 a
38℃-72 h 3.79 e 4.62 d 6.05 c 8.59 b 12.77 a 12.61 a
38℃-96 h 2.20 f 2.84 e 3.99 d 6.52 c 9.40 b 13.59 a
SK-3 CK 15.37 a 15.25 a 15.34 a 15.24 a 15.34 a 15.19 a
38℃-72 h 6.36 d 8.51 c 11.76 b 15.84 a 15.62 a 15.72 a
38℃-96 h 4.31 e 5.77 d 7.71 c 13.53 a 15.93 a 15.82 a
GPT
(μmol g-1 h-1)		 SK-1 CK 14.19 a 14.24 a 14.35 a 14.19 a 14.25 a 14.30 a
38℃-72 h 4.73 e 5.78 d 7.14 c 9.72 b 14.35 a 14.61 a
38℃-96 h 3.30 f 4.09 e 5.08 d 7.42 c 10.54 b 14.21 a
SK-3 CK 17.05 a 16.63 a 17.57 a 17.07 a 17.60 a 17.34 a
38℃-72 h 6.45 d 8.88 c 11.59 b 17.57 a 17.30 a 17.25 a
38℃-96 h 4.81 e 5.86 d 8.17 c 14.64 a 17.20 a 17.49 a

表3

高温持续胁迫终止后蕾中蛋白酶与肽酶活性变化"

测定指标
Tested index		 品种
Variety		 处理
Treatment		 恢复时间 Recovery time
0 h 12 h 24 h 48 h 72 h 96 h
蛋白酶活性
Protease activity
(μg g-1 h-1)		 SK-1 CK 34.62 a 34.08 a 34.88 a 33.86 a 35.07 a 34.58 a
38℃-72 h 125.79 a 114.46 b 100.05 c 73.23 d 35.43 d 34.88 d
38℃-96 h 153.95 a 145.35 b 131.47 c 106.43 d 76.00 e 35.35 e
SK-3 CK 39.84 a 40.76 a 40.98 a 40.67 a 39.74 a 40.19 a
38℃-72 h 114.88 a 99.46 b 83.75 c 39.08 d 41.16 d 39.94 d
38℃-96 h 141.91 a 122.30 b 105.78 c 62.09 d 43.01 e 41.49 e
肽酶活性
Peptidase activity
(μmol g-1 h-1)		 SK-1 CK 1.28 a 1.28 a 1.29 a 1.27 a 1.28 a 1.27 a
38℃-72 h 3.43 a 3.24 a 3.06 ab 2.46 b 1.26 d 1.27 d
38℃-96 h 4.07 a 3.91 b 3.72 c 3.15 d 2.50 e 1.30 f
SK-3 CK 1.64 a 1.61 a 1.62 a 1.62 a 1.61 a 1.60 a
38℃-72 h 3.20 a 2.77 b 2.32 c 1.56 d 1.62 d 1.59 d
38℃-96 h 3.79 a 3.48 b 3.11 c 2.07 d 1.59 e 1.59 e
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