The Bacillus thuringiensis (Bt) cry3Bb gene was successfully introduced into poplar plastid genome, leading to transplastomic poplar with high mortality to Plagiodera versicolora. Poplar (Populus L.) is one of the main resource of woody industry, but being damaged by insect pests. The feasibility and efficiency of plastid transformation technology for controlling two lepidopteran caterpillars have been demonstrated previously. Here, we introduced B. thuringiensis (Bt) cry3Bb into poplar plastid genome by biolistic bombardment for controlling P. versicolora, a widely distributed forest pest. Chimeric cry3Bb gene is controlled by the tobacco plastid rRNA operon promoter combined with the 5'UTR from gene10 of bacteriophage T7 (NtPrrn:T7g10) and the 3'UTR from the E. coli ribosomal RNA operon rrnB (TrrnB). The integration of transgene and homoplasmy of transplastomic poplar plants was confirmed by Southern blot analysis. Northern blot analysis indicated that cry3Bb was transcribed to both read through and shorter length transcripts in plastid. The transplastomic poplar expressing Cry3Bb insecticidal protein showed the highest accumulation level in young leaves, which reach up to 16.8 μg/g fresh weight, and comparatively low levels in mature and old leaves. Feeding the young leaves from Bt-Cry3Bb plastid lines to P. versicolora caused 100% mortality in the first-instar larvae after only 1 day, in the second-instar larvae after 2 days, and in the third-instar larvae for 3 days. Thus, we report a successful extension of plastid engineering poplar against the chrysomelid beetle.

中文翻译：

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质体表达的苏云金芽孢杆菌 (Bt) cry3Bb 使杨树中的食叶甲虫具有高死亡率。

将苏云金芽孢杆菌 (Bt) cry3Bb 基因成功导入杨树质体基因组，导致转质体杨树对花斑病菌的高死亡率。杨树(Populus L.)是木业的主要资源之一，但受到害虫的侵害。质体转化技术控制两种鳞翅目毛虫的可行性和有效性已被证明。在这里，我们通过基因枪轰击将 B. thuringiensis (Bt) cry3Bb 引入杨树质体基因组，以控制广泛分布的森林害虫 P. versicolora。嵌合 cry3Bb 基因由烟草质体 rRNA 操纵子启动子与来自噬菌体 T7 基因 10 (NtPrrn:T7g10) 的 5'UTR 和来自大肠杆菌核糖体 RNA 操纵子 rrnB (TrrnB) 的 3'UTR 控制。通过Southern印迹分析证实了转基因和转基因杨树同质性的整合。Northern印迹分析表明cry3Bb在质体中被转录为通读和较短长度的转录物。表达Cry3Bb杀虫蛋白的转基因杨树在幼叶中的积累水平最高，最高可达16.8 μg/g鲜重，而在成熟叶和老叶中积累水平相对较低。将来自 Bt-Cry3Bb 质体系的幼叶喂养到 P. versicolora 中，仅 1 天后一龄幼虫、2 天后二龄幼虫和三龄幼虫 3 天死亡率为 100%。因此，我们报告了质体工程杨树对菊科甲虫的成功扩展。