Burgess Elisabeth P J, Malone Louise A, Christeller John T, Lester Melissa T, Murray Colleen, Philip Bruce A, Phung Margaret M, Tregidga Emma L
The Horticulture and Food Research Institute of New Zealand Limited, Mt Albert Research Center, Auckland.
Transgenic Res. 2002 Apr;11(2):185-98. doi: 10.1023/a:1015297302990.
Fertile transgenic tobacco plants with leaves expressing avidin in the vacuole have been produced and shown to halt growth and cause mortality in larvae of two noctuid lepidopterans, Helicoverpa armigera and Spodoptera litura. Late first instar H. armigera larvae and neonate (< 12-h-old) S. litura larvae placed on leaves excised from T0 tobacco expressing avidin at 3.1-4.6 microM (micromoles/kg of fresh leaf tissue) had very poor growth over their first 8 days on the leaves, significant numbers had died by days 11 or 12 and all were dead by day 22 (H. armigera) or day 25 (S. litura). Similar results were obtained when late first instar H. armigera larvae were placed on leaves from T1 plants expressing avidin at six different average concentrations, ranging from 3.7 to 17.3 microM. Two larvae on the lowest expressing leaves survived to pupation, but there was total mortality among the other groups and no relationship between avidin concentration and the effects on the larvae. Synergistic effects between avidin-expressing tobacco plants and a purified Bt toxin, Cry1Ba, were demonstrated. Late instar H. armigera larvae fed with leaves from T2 plants expressing avidin at average concentrations of either <5.3 or > 12.9 microM, and painted with Cry1Ba protein at a rate equivalent to an expression level of 0.5% of total leaf protein, died significantly faster than larvae given either of the two treatments alone. Larvae fed with avidin-expressing leaves painted with the protease inhibitor, aprotinin, at a rate equivalent to 1% of total leaf protein had mortality similar to those given avidin-leaves alone. There was no evidence of antagonism between these two proteins.
已培育出叶片在液泡中表达抗生物素蛋白的可育转基因烟草植株，这些植株能使两种夜蛾科鳞翅目昆虫棉铃虫和斜纹夜蛾的幼虫停止生长并死亡。将棉铃虫一龄后期幼虫和斜纹夜蛾新生幼虫（<12小时龄）置于从表达抗生物素蛋白浓度为3.1 - 4.6微摩尔/千克鲜叶组织的T0烟草上切下的叶片上，它们在叶片上的前8天生长非常缓慢，到第11天或12天，大量幼虫死亡，到第22天（棉铃虫）或第25天（斜纹夜蛾）时全部死亡。当将棉铃虫一龄后期幼虫置于表达抗生物素蛋白的T1植株的叶片上时，也得到了类似结果，这些叶片的抗生物素蛋白平均浓度有6种不同水平，范围从3.7到17.3微摩尔。在表达量最低的叶片上有两条幼虫存活到化蛹，但其他组全部死亡，且抗生物素蛋白浓度与对幼虫的影响之间没有关系。研究还证明了表达抗生物素蛋白的烟草植株与纯化的Bt毒素Cry1Ba之间存在协同效应。用平均浓度<5.3或>12.9微摩尔且涂有相当于总叶蛋白表达水平0.5%的Cry1Ba蛋白的T2植株叶片喂养棉铃虫老龄幼虫，这些幼虫的死亡速度明显快于单独接受这两种处理之一的幼虫。用相当于总叶蛋白含量1%的蛋白酶抑制剂抑肽酶处理过的表达抗生物素蛋白的叶片喂养幼虫，其死亡率与单独用表达抗生物素蛋白的叶片喂养的幼虫相似。没有证据表明这两种蛋白之间存在拮抗作用。
