Research Article

Analysis of specific sequences in mutant rice generated by introduction of exogenous corn DNA

Published: December 29, 2015
Genet. Mol. Res. 14 (4) : 18980-18989 DOI: 10.4238/2015.December.29.5

Abstract

Rice variation induced by the introduction of exogenous DNA has become an important method of improving rice varieties and creating new germplasms. In this study, we transferred maize genomic DNA fragments to the receptor of Nipponbare rice using a modified “pollen-tube pathway” method. Material from mutant rice B1 and B2 were acquired and 14 specific bands were obtained from the material using amplified fragment length polymorphism analysis. From the 14 specific sequences obtained, there were 3791 bp, including 144 base mutations with a base mutation rate of 3.80%. Specific bands resulted from base mutation of selective bases or restriction endonuclease recognition sequences, or insertion or deletion of DNA fragments. The frequency of single-base mutations was significantly higher than that of double-base mutations, three-sequential base mutations, and multiple-sequential base mutations. The site frequency of base substitution (87.04%) was significantly higher than that of base insertion (3.70%) or deletion (9.26%). In all cases of base substitution, the frequency of transition (76.47%) was significantly higher than transversion (23.53%). The above results indicate that transferring foreign-species DNA into rice cells can induce base mutations in the receptor, with base substitutions occurring at the highest frequency, and the dominant type of base substitutions being transition. Preliminary analysis reveals that the molecular mechanism of transferring exogenous DNA into rice causes mutations, which provides theoretical data on biological mutagenesis for further research.

Rice variation induced by the introduction of exogenous DNA has become an important method of improving rice varieties and creating new germplasms. In this study, we transferred maize genomic DNA fragments to the receptor of Nipponbare rice using a modified “pollen-tube pathway” method. Material from mutant rice B1 and B2 were acquired and 14 specific bands were obtained from the material using amplified fragment length polymorphism analysis. From the 14 specific sequences obtained, there were 3791 bp, including 144 base mutations with a base mutation rate of 3.80%. Specific bands resulted from base mutation of selective bases or restriction endonuclease recognition sequences, or insertion or deletion of DNA fragments. The frequency of single-base mutations was significantly higher than that of double-base mutations, three-sequential base mutations, and multiple-sequential base mutations. The site frequency of base substitution (87.04%) was significantly higher than that of base insertion (3.70%) or deletion (9.26%). In all cases of base substitution, the frequency of transition (76.47%) was significantly higher than transversion (23.53%). The above results indicate that transferring foreign-species DNA into rice cells can induce base mutations in the receptor, with base substitutions occurring at the highest frequency, and the dominant type of base substitutions being transition. Preliminary analysis reveals that the molecular mechanism of transferring exogenous DNA into rice causes mutations, which provides theoretical data on biological mutagenesis for further research.