Genome-wide association study of callus induction variation to explore the callus formation mechanism of rice

Rice (Oryza sativa) is one of the most widely cultivated food crops, worldwide. Tissue culture is extensively used in rice breeding and functional genome research. The ability to induce callus determines whether a particular rice variety can be subjected to tissue culture and Agrobacterium-mediated transformation. Over the past two decades, many quantitative trait loci (QTLs) related to callus induction traits have been identified; however, individual genes associated with rice callus induction have not been reported. In this study, we characterized three callus-induction traits in a global collection of 510 rice accessions. A genome-wide association study of the rice population in its entirety as well as subpopulations revealed 21 significant loci located in rice callus induction QTLs. We identified three candidate callus induction genes, namely CRL1, OsBMM1, and OsSET1, which are orthologs of Arabidopsis LBD17/LBD29, BBM, and SWN, respectively, which are known to affect callus formation. Furthermore, we predicted that 14 candidate genes might be involved in rice callus induction and showed that RNA interference (RNAi)-mediated disruption of OsIAA10 inhibited callus formation on tissue culture medium. Embryo growth in the OsIAA10 RNAi line was not inhibited by synthetic auxin (2,4-D) treatment, suggesting that OsIAA10 may perceive auxin and activate the expression of downstream genes, such as CRL1, to induce callus formation. The significant loci and candidate genes identified here may provide insight into the mechanism underlying callus formation in rice.