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Achieving High Expression of Cry in Green Tissues and Negligible Expression in Endosperm Simultaneously via rbcS Gene Fusion Strategy in Rice

文献类型： 外文期刊

作者： Chen, Hao 1 ; Huang, Yuqing 1 ; Ye, Mengnan 1 ; Wang, Ya 1 ; He, Xiuying 2 ; Tu, Jumin 1 ;

作者机构： 1.Zhejiang Univ, Inst Crop Sci, Coll Agr & Biotechnol, Hangzhou 310058, Peoples R China

2.Guangdong Acad Agr Sci, Rice Res Inst, Guangzhou 510640, Peoples R China

3.Guangdong Key Lab New Technol Rice Breeding, Guangzhou 510640, Peoples R China

4.Minist Agr & Rural Affairs, Coconstruct Minist & Prov, Key Lab Genet & Breeding High Qual Rice Southern C, Guangzhou 510640, Peoples R China

5.Henan Acad Agr Sci, Cereal Crops Res Inst, Zhengzhou 450002, Peoples R China

关键词： OsrbcS; Cry1Ab/Cry1Ac; fuse; insect resistant; transgenic rice

期刊名称：INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES （ 影响因子：5.6； 五年影响因子：6.2 ）

ISSN： 1661-6596

年卷期： 2023 年 24 卷 10 期

页码：

收录情况： SCI

摘要： 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 mu g g(-1) in the leaf, higher than the Actin I promoter-driven control, T51-1, about 1.78 mu g g(-1) in the leaf, but negligible (only 0.00012-0.00117 mu g 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.

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