Creation of Rice Grain Size Mutants by Editing OsOFP30 via CRISPR/Cas9 System
[Objective]To study the role of OsOFP30,a transcription factor of the rice OFP family,in regulating grain size and to create new mutant materials of grain size,thereby providing a new reference for the improvement of rice grain size.[Method]The japonica rice line Zhonghua 11 was utilized as a transgenic recipient to generate OsOFP30 mutants using the CRISPR/Cas9 system.Through continuous screening of the T0,Ti,and T2 generations,three types of homozygous mutants without foreign fragment insertions were obtained:the long segment deletion mutation OsOFP30-89,the single base insertion mutation OsOFP30+1,and OsOFP30+1A.Grain size was measured at maturity stages,and the potential reasons for the different grain sizes were analyzed through bioinformatics and sequencing.[Results]Compared with the wild-type,the grain width and 1000-grain weight of all three types of mutants were significantly reduced.For OsOFP30-89 and OsOFP30+1,only the grain length and grain thickness are significantly reduced,with no obvious differences in panicle shape indicators from the wild type.The grain length and grain thickness of OsOFP30+1A show no obvious differences from the wild type,and only the number of primary branches is significantly lower than that of the wild type.Bioinformatics analysis revealed that the three types of mutants introduced premature translation termination due to a frameshift mutation.The OsOFP30 89 mutant protein consists of 252 amino acids,while the OsOFP30+1G and OsOFP30+1A mutant proteins consist of 282 amino acids.Sequence analyses showed that OsOFP30+1G and OsOFP30+1A differ by a single base,with G and A at the 323rd position,respectively.This variation resulted in a change between serine and asparagine at the 108th position of the mutant protein.[Conclusion]The OsOFP30 gene plays a crucial role in regulating rice grain size.This study successfully created new mutant materials affecting grain size,which will provide valuable references for the improvement of rice grain size.
万方数据
维普
