湖北大学张江、武玉永团队利用质体转基因技术培育了新的抗虫杨树

发布时间：2019-12-06浏览量：10696

【字体大小：大中小】

杨树是重要的木材资源，然而却受到严重的虫害破坏。利用基因工程技术是解决杨树虫害问题的重要手段。我国是世界上杨树人工林面积最大的国家，也是唯一批准转基因杨树商业化种植的国家。最近，湖北大学会员张江、武玉永团队利用质体转基因技术培育了新的转基因抗虫杨树。

质体转基因具有表达量高、无位置效应、母性遗传等优势，然而尚未应用于杨树中。研究人员将具有鞘翅目杀虫活性的Bt基因cry3Bb通过基因枪导入杨树质体基因组，以防治杨树中的主要害虫柳蓝叶甲（Plagiodera versicolora）。cry3Bb基因表达盒还含有烟草质体rRNA操纵子启动子、T7噬菌体g10（NtPrrn:T7g10）的5’-UTR和大肠杆菌核糖体RNA操纵子rrnB（TrrnB）。研究者利用Southern和Northern技术证实了外源基因的整合和表达情况，并检测出外源Cry3Bb蛋白在转基因杨树嫩叶中的含量最高可达到16.8 μg/g鲜重，成熟叶片和老叶中含量有所下降。使用转化体嫩叶喂食柳蓝叶甲幼虫生测的结果显示，1龄幼虫喂食1天后即全部死亡，2龄和3龄幼虫也分别在喂食2天和3天后全部死亡。

该质体转化技术开发的杨树转化体具有很好的害虫防治效果。

Plant Cell Reports, 03 December 2019

Plastid-expressed Bacillus thuringiensis (Bt) cry3Bb confers high mortality to a leaf eating beetle in poplar

Author

Shijing Xu, Yiqiu Zhang, Shengchun Li, Ling Chang, Yuyong Wu, Jiang Zhang*

*: State Key Laboratory of Biocatalysis and Enzyme Engineering, School of Life Sciences, Hubei University, China

Abstract

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 geneis 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.

本文经授权转载自 莱肯生物公众号