Cry2Ab及Cry1Ac杀虫蛋白对棉铃虫中肠蛋白酶活性的影响
The effects of Cry2Ab and Cry1Ac insecticidal proteins on proteaseactivity in the midgut of Helicoverpa armigera
魏纪珍 梁革梅 高希武 高 珍 张万娜 等
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作者单位：1. 中国农业大学农学与生物技术学院昆虫学系北京100193;2. 中国农业科学院植物保护研究所植物病虫害生物学国家重点实验室北京100193
中文关键词：棉铃虫，Cry1Ac，Cry2Ab，总酶，类胰凝乳蛋白酶，类胰蛋白酶
英文关键词：Helicoverpa armigera，Cry1Ac，Cry2Ab，midgut total protease，trypsin-like enzyme，chymotrypsin-likeenzyme
中文摘要： 为明确Cry2Ab 和Cry1Ac 2 种Bt 杀虫蛋白单用与混用对棉铃虫Helicoverpa armigera (Hübner)中肠主要
蛋白酶活性的影响，本文测定了取食含不同Bt 蛋白人工饲料后棉铃虫中肠总蛋白酶、类胰蛋白酶和类胰凝乳蛋
白酶活性的差异。结果发现:Cry2Ab 处理12 h 后对棉铃虫中肠总蛋白酶影响不大;对类胰蛋白酶的影响最大，除
最高浓度处理外，其他浓度处理后棉铃虫类胰蛋白酶的活性明显高于对照;但对类胰凝乳蛋白酶活性的影响呈倒
“V”字型，只有6. 67 μg / g Cry2Ab 处理后的棉铃虫酶活力显著高于对照，其他浓度处理与对照差异不显著或略低
于对照;随着取食含Cry2Ab 饲料时间的增加，棉铃虫中肠类胰蛋白酶和类胰凝乳蛋白酶的活性比对照显著增加;
与对照相比，处理36 h 后类胰蛋白酶活性最高可增加到6. 43 倍。Cry1Ac 处理棉铃虫12 h 后总蛋白酶、类胰蛋白
酶和类胰凝乳蛋白酶活性都明显增加，而且与处理浓度呈正相关;但是24 h 后，处理后棉铃虫的总蛋白酶和类胰
凝乳蛋白酶活性明显降低，只有类胰蛋白酶活性仍高于对照，但活性增长倍数低于12 h 时的处理。Cry2Ab 和
Cry1Ac 2 种蛋白混用处理棉铃虫后，2 种酶的酶活力基本低于Cry1Ac 和Cry2Ab 单用的酶活力之和;只有2 种蛋
白浓度均为2. 22 μg / g 混用时，处理12 h 后类胰蛋白酶和类胰凝乳蛋白酶的活性高于2 种蛋白单用时酶活力之
和，且都显著的高于对照。
英文摘要：Differences in the midgut activity of total protease，trypsin-like enzyme and chymotrypsin-like enzyme in cotton
bollworm Helicoverpa armigera (Hübner) fed on artificial diets containing different proteins was investigated in order to
clarify the effects of Cry2Ab and Cry1Ac insecticidal proteins on protease activity in the midgut of H． armigera and provide
basic information for the rational utilization of double Bt toxin cotton． The results indicate that although Cry2Ab had little
effect on total protease activity after 12 h it had a major effect on trypsin-like enzyme． The trend in chymotrypsin-like
enzyme activity was like the reversed letter V． Compared to the control，the activity of the chymotrypsin-like enzyme was
significantly higher than the control only when the concentration of Cry2Ab was 6． 67 μg / g，and other concentrations were
either not significantly different，or slightly lower，than the control． Together with feeding time，the activities of trypsinlike
and chymotrypsin-like enzymes increased dramatically compared to the control． After 36 h the activity of the
chymotrypsin-like enzyme had increased by 6. 43 times relative to the control． The activities of total protease，trypsin-likeenzyme and chymotrypsin-like enzyme all increased significantly compared to the control after H． armigera had fed on a
Cry1Ac diet for 12 h and the trends were correlated with the treatment concentration． However，the activities of the
trypsin-like and chymotrypsin-like enzymes obviously declined to below the control after 24 h． Only the trypsin-like
enzyme was still higher than in the control． The activities of these three enzymes in H． armigera treated with Cry1Ac +
Cry2Ab were lower than the sum of treatment using Cry2Ab and Cry1Ac in isolation． But activities of these enzymes in
bollworms fed on a diet containing 2． 22 μg / g Cry2Ab and 2． 22 μg / g Cry1Ac was significantly higher than the sum of
their activities in insects fed on either Cry2Ab or Cry1Ac．