Improved Transformation and Regeneration of Indica Rice: Disruption of SUB1A as a Test Case via CRISPR-Cas9

International Journal of Molecular Sciences

Julia Bailey-Serres¹  Yuya Liang²  Backki Kim²  Sudip Biswas²  Endang M. Septiningsih² 

[1] Center for Plant Cell Biology, Botany and Plant Sciences, University of California, Riverside, CA 92521, USA;Department of Soil and Crop Sciences, Texas A&M University, College Station, TX 77843, USA;

关键词: gene editing;    CRISPR-Cas9;    rice;    indica;    SUB1A;    tissue culture;   

DOI  :  10.3390/ijms22136989

来源: DOAJ

【 摘 要 】

Gene editing by use of clustered regularly interspaced short palindromic repeats (CRISPR) has become a powerful tool for crop improvement. However, a common bottleneck in the application of this approach to grain crops, including rice (Oryza sativa), is efficient vector delivery and calli regeneration, which can be hampered by genotype-dependent requirements for plant regeneration. Here, methods for Agrobacterium-mediated and biolistic transformation and regeneration of indica rice were optimized using CRISPR-Cas9 gene-editing of the submergence tolerance regulator SUBMERGENCE 1A-1 gene of the cultivar Ciherang-Sub1. Callus induction and plantlet regeneration methods were optimized for embryogenic calli derived from immature embryos and mature seed-derived calli. Optimized regeneration (95%) and maximal editing efficiency (100%) were obtained from the immature embryo-derived calli. Phenotyping of T₁ seeds derived from the edited T₀ plants under submergence stress demonstrated inferior phenotype compared to their controls, which phenotypically validates the disruption of SUB1A-1 function. The methods pave the way for rapid CRISPR-Cas9 gene editing of recalcitrant indica rice cultivars.

【 授权许可】

Unknown