关键词:Bt棉; 蕾; Bt杀虫蛋白; 氮代谢; 高温胁迫 Effect of High Temperature Stress on Bt Insecticidal Protein Content and Nitrogen Metabolism of Square in Bt Cotton HENG Li1, LI Ya-Bing2, HU Da-Peng1, WANG Gui-Xia1, LYU Chun-Hua1, ZHANG Xiang1, CHEN Yuan1, CHEN De-Hua1,* 1Key Laboratory of Crop Genetics and Physiology of Jiangsu Province / College of Agriculture, Yangzhou University, Yangzhou 225009, China
2Institute of Cotton Research, Chinese Academy of Agricultural Sciences, Anyang 455000, Henan
Fund:The study was supported by the National Natural Science Foundation of China (31171479, 31301263, 31471435), the Ministry of Education Research Fund for the Doctoral Program (20113250110001), Jiangsu “Three-innovation” Agricultural Project [SXGC(2014)317], and the Development Plan of Higher Education Advantaged Discipline in Jiangsu Province AbstractTo explore the reason for low insect resistance under high temperature climate in Bt cotton production, we selected two Bt cotton cultivars as experimental material with treatments of 38℃ for three, five and seven days in 2011, and 38℃/25℃ at day /night for four, seven and ten days, respectively at the artificial climate chamber in 2012. The results showed that the Bt insecticidal protein contents declined within seven days under 38℃, with the largest reduction occurred within three days. Compared with control, the square Bt insecticidal protein contents decreased by 18.71% and 26.54% for cultivar Sikang 3 (SK-3) and Sikang 1 (SK-1), respectively. Under 38℃/25℃ at day/night, the square Bt insecticidal protein contents had no obvious reduction within four days, and significant reduced after seven days with the reduction of 11.32% for SK-3 and 14.18% for SK-1. Under the treatment conditions the soluble protein contents, glutamate pyruvate transaminase (GPT) activities reduced, but the free amino acid contents, protease activities increased. There existed significant negative correlation of insecticidal protein content with free amino acid contents, and protease activities; and significantly positive correlation of insecticidal protein content with soluble protein contents, and GPT activities under high temperature stresses. Therefore, the reduced synthesis and the enhanced degradation for protein in the square under the high temperature condition resulted in the decrease of soluble protein content, including Bt insecticidal protein content. The larger reductions of the square Bt insecticidal protein content and nitrogen metabolic strength were detected in cultivar SK-1 as confound with cultivar SK-3 under the high temperature treatments.
Keyword:Bt cotton; Square; Bt insecticidal protein; Nitrogen metabolism; High temperature stress Show Figures Show Figures
表1 持续高温胁迫下棉蕾氮代谢主要物质含量和关键酶活性 Table 1 Main compound contents and key enzyme activities in nitrogen metabolism of square under high temperature stress
品种 Variety
胁迫时间 Stress time (d)
游离氨基酸含量 Free amino acid content (μ g g-1)
可溶蛋白含量 Soluble protein content (mg g-1)
GPT活性 GPT activity (μ mol g-1 h-1)
蛋白酶活性 Protease activity (mg g-1 h-1)
泗抗3号SK-3
0
105.097 E
36.066 A
15.403 A
1.788 CD
3
182.345 D
28.491 C
13.564 C
2.532 AB
5
206.401 BC
25.812 D
13.026 D
2.849 AB
7
222.253 A
20.772 E
12.650 E
3.101 A
泗抗1号SK-1
0
99.141 E
31.033 B
14.624 B
1.548 D
3
173.173 D
21.305 E
12.313 F
2.325 BC
5
195.440 C
19.851 E
11.864 G
2.622 AB
7
215.028 AB
16.786 F
11.581 G
2.944 AB
Values followed by different letters are significantly different among the treatments at P< 1% (Duncan’ s test). 标以不同大写字母的值在1%水平下差异显著(Duncan’ s新复极差测验)。
表1 持续高温胁迫下棉蕾氮代谢主要物质含量和关键酶活性 Table 1 Main compound contents and key enzyme activities in nitrogen metabolism of square under high temperature stress
表2 昼夜变温下高温对棉蕾氮代谢主要物质含量及关键酶活性的影响 Table 2 Main compound contents and key enzyme activities in nitrogen metabolism of cotton square under the variable high temperature at day/night
品种 Variety
胁迫持续时间 Stress time (d)
游离氨基酸含量 Free amino acid content (μ g g-1)
可溶蛋白含量 Soluble protein content (mg g-1)
GPT活性 GPT activity (μ mol g-1 h-1)
蛋白酶活性 Protease activity (mg g-1 h-1)
SK-3
0
564.74 C
4.13 A
18.32 A
1.93 C
4
559.07 C
4.09 A
17.62 A
1.92 C
7
633.46 A
3.64 B
16.09 AB
2.49 B
10
752.85 B
2.97 C
14.02 B
3.55 A
SK-1
0
494.21 C
3.43 A
15.98 A
1.56 C
4
498.63 C
3.56 A
15.42 A
1.61 C
7
606.29 B
3.06 A
13.86 B
2.51 B
10
740.88 A
2.45 B
11.11 C
3.31 A
Values followed by different letters are significantly different among the treatments at P< 1% (Duncan’ s test). 标以不同大写字母的值在1%水平下差异显著(Duncan’ s新复极差测验)。
表2 昼夜变温下高温对棉蕾氮代谢主要物质含量及关键酶活性的影响 Table 2 Main compound contents and key enzyme activities in nitrogen metabolism of cotton square under the variable high temperature at day/night
表3 高温胁迫下蕾中Bt杀虫蛋白含量与氮代谢主要物质含量及关键酶活性的相关系数 Table 3 Correlation coefficients between square Bt protein content and main compound contents and key enzyme activities under high temperature stresses
胁迫方式 Stress pattern
品种 Variety
游离氨基酸含量 Free amino acid content (μ g g-1)
可溶蛋白含量 Soluble protein content (mg g-1)
GPT活性 GPT activity (μ mol g-1 h-1)
蛋白酶活性 Protease activity (mg g-1 h-1)
高温持续胁迫 Duration stress under high temperature
SK-3
-0.994* *
0.992* *
0.993* *
-0.999* *
SK-1
-0.999* *
0.998* *
0.988* *
-0.997* *
昼夜变温胁迫 Variable temperature at the day and night
SK-3
-0.997* *
0.999* *
0.992* *
-0.996* *
SK-1
-0.999* *
0.996* *
0.993* *
-0.991* *
* * represents that the correlation is significant at the 1% possibility level. * * 表示相关性在1%水平上达极显著。
表3 高温胁迫下蕾中Bt杀虫蛋白含量与氮代谢主要物质含量及关键酶活性的相关系数 Table 3 Correlation coefficients between square Bt protein content and main compound contents and key enzyme activities under high temperature stresses
4 结论持续高温和昼夜变温下高温胁迫引起Bt棉蕾中杀虫蛋白表达量下降, 38℃高温持续胁迫3日内下降幅度最大; 38℃/25℃昼夜变温下高温胁迫持续7 d以上时, 棉蕾Bt杀虫蛋白含量显著下降; 高温胁迫下棉蕾Bt杀虫蛋白含量的变化与氮代谢密切相关。高温胁迫下引起游离氨基酸含量和蛋白酶活性升高, 可溶性蛋白含量和GPT活性下降。因此在棉花生长季节需及时注意高温气候对Bt棉蕾抗虫性的影响及程度, 防止棉铃虫等相关害虫对棉花的危害。 The authors have declared that no competing interests exist.
作者已声明无竞争性利益关系。The authors have declared that no competing interests exist.
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