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- Title
Achieving High Expression of Cry in Green Tissues and Negligible Expression in Endosperm Simultaneously via rbcS Gene Fusion Strategy in Rice.
- Authors
Chen, Hao; Huang, Yuqing; Ye, Mengnan; Wang, Ya; He, Xiuying; Tu, Jumin
- Abstract
To allay excessive public concern about the safety of transgenic foods, and to optimize insect-resistant genes expression to delay the evolution of resistance in pests, we developed a promising strategy to fuse the GOI (gene of interest) with OsrbcS (rice small subunit of ribulose bisphosphate carboxylase/oxygenase) in transgenic rice, which acted as a carrier, driven by the OsrbcS native promoter to sequester its expression in green tissues. Using eYFP as a trial, we reported a high-level accumulation of eYFP in green tissue and almost none in the seed and root of the fused construct compared to the non-fused construct. After applying this fusion strategy in insect-resistant rice breeding, recombinant OsrbcS-Cry1Ab/Cry1Ac expressed rice plants conferred high resistance to leaffolders and striped stem borers, among which two single-copy lines possessed normal agronomic performance in the field. Specifically, Cry1Ab/Cry1Ac protein levels in single-copy construct transgenic lines ranged from 1.8 to 11.5 µg g−1 in the leaf, higher than the Actin I promoter-driven control, T51-1, about 1.78 µg g−1 in the leaf, but negligible (only 0.00012–0.00117 µg g−1) in endosperm by ELISA analysis. Our study provided a novel approach to creating Cry1Ab/Cry1Ac-free endosperm rice with a high level of insect-resistant protein in green tissues through the simultaneous usage of the OsrbcS promoter and OsrbcS as a fusion partner.
- Subjects
ENDOSPERM; GENE fusion; GENE expression; RIBULOSE bisphosphate carboxylase; RICE; TRANSGENIC rice
- Publication
International Journal of Molecular Sciences, 2023, Vol 24, Issue 10, p9045
- ISSN
1661-6596
- Publication type
Article
- DOI
10.3390/ijms24109045