Bt棉抗虫性表达的化学调控
摘要
本论文以转Bt基因抗虫棉新棉33~B为试材，室内盆栽与大田试验相结合，从不同发育期叶片氮素代谢生理的角度研究了Bt棉抗虫性不稳定表达的机理与促进抗虫性稳定表达的化学调控技术及其机理。结果表明：
1转Bt基因抗虫棉(新棉33~B)抗虫性不稳定，由项部幼叶(倒1叶)至基部衰老叶片(倒8叶)Bt毒蛋白含量呈降低趋势，其中以功能叶(倒3叶)毒蛋白含量最低；由苗蕾期至吐絮期叶片毒蛋白含量呈下降趋势，其中盛花期幼叶和功能叶中毒蛋白含量下降幅度较大，出现一抗虫性“低谷”。
2盛花期、盛铃期，由顶部幼叶至基部衰老叶片，叶片硝态氮含量、硝酸还原酶活力呈下降趋势，叶片可溶性蛋白和游离氨基酸含量呈上升趋势，其中功能叶含量最高。
3幼叶和功能叶中硝态氮含量、硝酸还原酶活力、可溶性蛋白含量和游离氨基酸含量以盛铃期最高，盛花期最低；衰老叶片中则以吐絮期最低。
4由苗蕾期至吐絮期，幼叶和衰老叶片中单宁的含量呈递增趋势，功能叶片中则以盛铃期最高。幼叶和功能叶片中黄酮类化合物的含量呈递减趋势，衰老叶片中以絮期最高。
5盛花期，Bt棉幼叶(倒1叶)中CTKs中的ZR+Z和IAA含量较低，CTKs(iPA+iP、DHZR+DHZ)、GAs(GA4、GA3)和ABA含量以盛花期最高，硝酸还原酶活性较低，氨基酸和蛋白质含量低，毒蛋白合成量低。
6盛花期，功能叶片中CTKs中的ZR+Z、IAA和ABA含量较低，CTKs中的iPA+iP和DHZR+DHZ、GAs含量较高，硝酸还原酶活性较低，氨基酸和蛋白质含量低，毒蛋白合成量低。
7衰老叶片中ABA、IAA、GAs和CTKs(ZR+Z、DHZR+DHZ)含量以盛铃期最低，iPA+iP含量以盛铃期最高。
8 B1P1处理提高了幼叶中IAA含量，协调了不同生育期幼叶中CTKs三组分(iPA+iP、ZR+Z、DHZR+DHZ)的活性表达。促进了幼叶氮素同化，提高了硝酸还原酶活力，提高了可溶性蛋白含量，促进了毒蛋白的合成；提高了盛花期功能叶中CTKs(iPA+iP、ZR+Z、DHZR+DHZ)、GAs和IAA的含量，促进了盛花期功能叶的氮素同化，可溶性蛋白含量和硝酸还原酶活力增高，毒蛋白含量增加；提高了衰老叶片中iPA+iP和IAA含量，降低了ABA含量，延缓叶片衰老，促进毒蛋白的合成。
9苗蕾期至絮期，B2P1S2处理提高了幼叶(苗蕾期除外)、功能叶和衰老叶片中CTKs(iPA+iP、ZR+Z、DHZR+DHZ)、GAs和IAA的含量，提高了硝酸还原酶活力，促进了游离氨基酸和可溶性蛋白的合成，促进了毒蛋白的合成。
10盛花期，B1P1S1、B1P2和B2P1处理提高了功能叶中CTKs(iPA+iP、ZR+Z、DHZR+DHZ)的含量，提高了硝酸还原酶活力，促进了可溶性蛋白和毒蛋白的合成；苗蕾期、盛铃期和絮期幼叶、衰老叶片中可溶性蛋白和毒蛋白含量、硝酸还原酶活力等变化不稳定。
11 B1P1、B1P2、B2P1、B1P1S1和B2P1s2五个处理中，B1P1和B2P1S2处理提高了除苗蕾期幼叶之外的所有生育期中幼叶、功能叶和衰老叶片中毒蛋白的含量，提高了Bt棉的抗虫性。
Taking Bt transgenic cotton NuCOTN33~B as trial material, this paper studied on the insect resistance instability mechanism of Bt cotton from the aspect of the leaf nitrogen metabolism physiology at different developing stage. Results were showed as follows.
1 The insect resistance of Bt cotton (NuCOTN33~B) was instability. Bt protein content of leaves showed fall trend from young leaves at the apex to old leaves at the bottom, of which the lowest one was at the functional leaves. From seedling stages to wadding stage, Bt protein content of all leaves was reduces. Bt protein content of the young leaves and the functional leaves at the flowering stage dropped deeply. The insect resistance showed a lower valley.
2 The content of nitrate and the NRA (nitrate reductase activity) showed fall trend from young leaves at the apex to old leaves at the bottom in the flowering-boiling stage. While contents of protein and dissociate amino acid showed up trend, of which the highest one was at the functional leaves.
3 Contents of nitrate, protein, amino acid and the NRA of young and old leaves were increased in boiling stage, but those were cut down in flowering stage. And the lowest one was at the old leaves in wadding stage.
4 From seedling-squaring stage to wading stage, the condensed tannins content of young and old leaves were enhanced, and the highest one was at functional leaves in boiling stage. The flavonoids content of young and functional leaves showed low trend, but the highest one was at old leaves in wadding stage.
5 In flowering stage, the content of ZR+Z and IAA in the young leaves is lower than the other developing stage, the content of iPA+iP, DHZR+DHZ, GAs (GA3, GA4) and ABA is higher than the other developing stage, the NRA is lower, the content of amino acid, protein and Bt protein are all lower than the other developing stage.
6 In flowering stage, the content of ZR+Z, IAA and ABA in the functional leaves is lower than the other developing stage, the content of iPA+iP, DHZR+DHZ, and GAs (GA3, GA4) is higher than the other developing stage, the NRA is lower, the content of amino acid, protein and Bt protein are all lower than the other developing stage.
7 In boiling stage, the content of CTKs (ZR+Z, DHZR+DHZ), IAA, GAs and ABA in the old leaves is the lowest, the content of iPA+iP is the highest.
8 By B1P1 treatment at the different developing stage, the IAA content in young leaves was increased, the activity of CTKs (iPA+iP, ZR+Z, DHZR+DHZ) were coordinated. The nitrogen assimilation and the NRA in the young leaves were improved, the content of protein was increased, and the synthetic process of the Bt toxin protein was improved too.
9 By B2P1S2 treatment from seedling-squaring stage to wadding stage, the content of CTKs (iPA+iP, ZR+Z, DHZR+DHZ), GAs and IAA in the young leaves were increased the NRA was improved, the content of protein and amino acid were heightened, the synthesize of Bt protein was enhanced.
10 By B1P1S1, B1P2 and B2P1 treatment, in flowering stage, the content of CTKs (iPA+iP, ZR+Z, DHZR+DHZ) in functional leaves was heightened, the NRA was improved, the content of leaf protein and Bt protein were increased, the content of protein, Bt protein and NRA in the young, the old leaves from seedling stage to wadding stage were changed within a narrow range.
11 The content of Bt protein in the young, the old and the functional leaves were increased from seedling stage to wadding stage, except the young leaves in seedling stage, the insect resistance was improved.
1转Bt基因抗虫棉(新棉33~B)抗虫性不稳定，由项部幼叶(倒1叶)至基部衰老叶片(倒8叶)Bt毒蛋白含量呈降低趋势，其中以功能叶(倒3叶)毒蛋白含量最低；由苗蕾期至吐絮期叶片毒蛋白含量呈下降趋势，其中盛花期幼叶和功能叶中毒蛋白含量下降幅度较大，出现一抗虫性“低谷”。
2盛花期、盛铃期，由顶部幼叶至基部衰老叶片，叶片硝态氮含量、硝酸还原酶活力呈下降趋势，叶片可溶性蛋白和游离氨基酸含量呈上升趋势，其中功能叶含量最高。
3幼叶和功能叶中硝态氮含量、硝酸还原酶活力、可溶性蛋白含量和游离氨基酸含量以盛铃期最高，盛花期最低；衰老叶片中则以吐絮期最低。
4由苗蕾期至吐絮期，幼叶和衰老叶片中单宁的含量呈递增趋势，功能叶片中则以盛铃期最高。幼叶和功能叶片中黄酮类化合物的含量呈递减趋势，衰老叶片中以絮期最高。
5盛花期，Bt棉幼叶(倒1叶)中CTKs中的ZR+Z和IAA含量较低，CTKs(iPA+iP、DHZR+DHZ)、GAs(GA4、GA3)和ABA含量以盛花期最高，硝酸还原酶活性较低，氨基酸和蛋白质含量低，毒蛋白合成量低。
6盛花期，功能叶片中CTKs中的ZR+Z、IAA和ABA含量较低，CTKs中的iPA+iP和DHZR+DHZ、GAs含量较高，硝酸还原酶活性较低，氨基酸和蛋白质含量低，毒蛋白合成量低。
7衰老叶片中ABA、IAA、GAs和CTKs(ZR+Z、DHZR+DHZ)含量以盛铃期最低，iPA+iP含量以盛铃期最高。
8 B1P1处理提高了幼叶中IAA含量，协调了不同生育期幼叶中CTKs三组分(iPA+iP、ZR+Z、DHZR+DHZ)的活性表达。促进了幼叶氮素同化，提高了硝酸还原酶活力，提高了可溶性蛋白含量，促进了毒蛋白的合成；提高了盛花期功能叶中CTKs(iPA+iP、ZR+Z、DHZR+DHZ)、GAs和IAA的含量，促进了盛花期功能叶的氮素同化，可溶性蛋白含量和硝酸还原酶活力增高，毒蛋白含量增加；提高了衰老叶片中iPA+iP和IAA含量，降低了ABA含量，延缓叶片衰老，促进毒蛋白的合成。
9苗蕾期至絮期，B2P1S2处理提高了幼叶(苗蕾期除外)、功能叶和衰老叶片中CTKs(iPA+iP、ZR+Z、DHZR+DHZ)、GAs和IAA的含量，提高了硝酸还原酶活力，促进了游离氨基酸和可溶性蛋白的合成，促进了毒蛋白的合成。
10盛花期，B1P1S1、B1P2和B2P1处理提高了功能叶中CTKs(iPA+iP、ZR+Z、DHZR+DHZ)的含量，提高了硝酸还原酶活力，促进了可溶性蛋白和毒蛋白的合成；苗蕾期、盛铃期和絮期幼叶、衰老叶片中可溶性蛋白和毒蛋白含量、硝酸还原酶活力等变化不稳定。
11 B1P1、B1P2、B2P1、B1P1S1和B2P1s2五个处理中，B1P1和B2P1S2处理提高了除苗蕾期幼叶之外的所有生育期中幼叶、功能叶和衰老叶片中毒蛋白的含量，提高了Bt棉的抗虫性。
Taking Bt transgenic cotton NuCOTN33~B as trial material, this paper studied on the insect resistance instability mechanism of Bt cotton from the aspect of the leaf nitrogen metabolism physiology at different developing stage. Results were showed as follows.
1 The insect resistance of Bt cotton (NuCOTN33~B) was instability. Bt protein content of leaves showed fall trend from young leaves at the apex to old leaves at the bottom, of which the lowest one was at the functional leaves. From seedling stages to wadding stage, Bt protein content of all leaves was reduces. Bt protein content of the young leaves and the functional leaves at the flowering stage dropped deeply. The insect resistance showed a lower valley.
2 The content of nitrate and the NRA (nitrate reductase activity) showed fall trend from young leaves at the apex to old leaves at the bottom in the flowering-boiling stage. While contents of protein and dissociate amino acid showed up trend, of which the highest one was at the functional leaves.
3 Contents of nitrate, protein, amino acid and the NRA of young and old leaves were increased in boiling stage, but those were cut down in flowering stage. And the lowest one was at the old leaves in wadding stage.
4 From seedling-squaring stage to wading stage, the condensed tannins content of young and old leaves were enhanced, and the highest one was at functional leaves in boiling stage. The flavonoids content of young and functional leaves showed low trend, but the highest one was at old leaves in wadding stage.
5 In flowering stage, the content of ZR+Z and IAA in the young leaves is lower than the other developing stage, the content of iPA+iP, DHZR+DHZ, GAs (GA3, GA4) and ABA is higher than the other developing stage, the NRA is lower, the content of amino acid, protein and Bt protein are all lower than the other developing stage.
6 In flowering stage, the content of ZR+Z, IAA and ABA in the functional leaves is lower than the other developing stage, the content of iPA+iP, DHZR+DHZ, and GAs (GA3, GA4) is higher than the other developing stage, the NRA is lower, the content of amino acid, protein and Bt protein are all lower than the other developing stage.
7 In boiling stage, the content of CTKs (ZR+Z, DHZR+DHZ), IAA, GAs and ABA in the old leaves is the lowest, the content of iPA+iP is the highest.
8 By B1P1 treatment at the different developing stage, the IAA content in young leaves was increased, the activity of CTKs (iPA+iP, ZR+Z, DHZR+DHZ) were coordinated. The nitrogen assimilation and the NRA in the young leaves were improved, the content of protein was increased, and the synthetic process of the Bt toxin protein was improved too.
9 By B2P1S2 treatment from seedling-squaring stage to wadding stage, the content of CTKs (iPA+iP, ZR+Z, DHZR+DHZ), GAs and IAA in the young leaves were increased the NRA was improved, the content of protein and amino acid were heightened, the synthesize of Bt protein was enhanced.
10 By B1P1S1, B1P2 and B2P1 treatment, in flowering stage, the content of CTKs (iPA+iP, ZR+Z, DHZR+DHZ) in functional leaves was heightened, the NRA was improved, the content of leaf protein and Bt protein were increased, the content of protein, Bt protein and NRA in the young, the old leaves from seedling stage to wadding stage were changed within a narrow range.
11 The content of Bt protein in the young, the old and the functional leaves were increased from seedling stage to wadding stage, except the young leaves in seedling stage, the insect resistance was improved.
引文
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