A fast breeding strategy creates fragrance- and anthocyanin-enriched rice lines by marker-free gene-editing and hybridization

文献类型： 外文期刊

第一作者： Shi, Tiantian

作者： Shi, Tiantian;Gao, Ying;Xu, Andi;Wang, Rui;Sun, Yinglu;Chen, Luoying;Liu, Yuanhang;Luo, Rong;Liu, Jun;Lyu, Mingjie;Chen, Luoying;Wang, Huan;Wang, Huan

作者机构：

关键词： F25 inbred line; Fragrance; Anthocyanin; CRISPR; Cas9 edit system; Hybridization

期刊名称：MOLECULAR BREEDING （ 影响因子：3.1； 五年影响因子：3.1 ）

ISSN： 1380-3743

年卷期： 2023 年 43 卷 4 期

页码：

收录情况： SCI

摘要： As rice is a staple food for nearly half of the world's population, rice varieties with excellent agronomic traits as well as high flavor and nutritional quality such as fragrant rice and purple rice are naturally favored by the market. In the current study, we adopt a fast breeding strategy to improve the aroma and anthocyanin content in the excellent rice inbred line, F25. The strategy skillfully used the advantages of obtaining editing pure lines in T0 generation of CRISPR/Cas9 editing system and easy observation of purple character and grain shape, integrated the subsequent screening of non-transgenic lines, and the elimination of undesirable edited variants from gene-editing and cross-breeding at the same time as the separation of the progeny from the purple cross, thus expediting the breeding process. Compared with conventional breeding strategies, this strategy saves about 6-8 generations and reduces breeding costs. Firstly, we edited the OsBADH2 gene associated with rice flavor using an Agrobacterium-mediated CRISPR/Cas9 system to improve the aroma of F25. In the T0 generation, a homozygous OsBADH2-edited F25 line (F25B) containing more of the scented substance 2-AP was obtained. Then, we crossed F25B (female) with a purple rice inbred line, P351 (male), with high anthocyanin enrichment to improve the anthocyanin content of F25. After nearly 2.5 years of screening and identification over five generations, the undesirable variation characteristics caused by gene-editing and hybridization and the transgenic components were screened out. Finally, the improved F25 line with highly stable aroma component, 2-AP, increased anthocyanin content and no exogenous transgenic components were obtained. This study not only provides high-quality aromatic anthocyanin rice lines that meet the market demand, but also offers a reference for the comprehensive use of CRISPR/Cas9 editing technology, hybridization, and marker-assisted selection to accelerate multi-trait improvement and breeding process.

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