Abstract
Key message
The Bacillus thuringiensis (Bt) cry3Bb gene was successfully introduced into poplar plastid genome, leading to transplastomic poplar with high mortality to Plagiodera versicolora.
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 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.
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This work was supported by Grants from the National Key Research and Development Program of China (2017YFD0600101).
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JZ conceived and designed the project. SX, YZ, and YW conducted experiments. SL, LC, YW and JZ analyzed the data. JZ wrote the article with contributions of all the authors. All the authors read and approved the manuscript.
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The authors have filed two patent applications (201910077185.9, 201910077220.7) that related to the production of insect-resistant poplar by plastid transformation.
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Communicated by Amit Dhingra.
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Xu, S., Zhang, Y., Li, S. et al. Plastid-expressed Bacillus thuringiensis (Bt) cry3Bb confers high mortality to a leaf eating beetle in poplar. Plant Cell Rep 39, 317–323 (2020). https://doi.org/10.1007/s00299-019-02492-0
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DOI: https://doi.org/10.1007/s00299-019-02492-0