Development and Application of Functional Markers of Rice Lipoxygenase Mutants
Rice was seriously aged during long-term storage,mainly due to the lipid degradation of rice itself.There were three LOX isozymes in mature seed embryos,and 80％～90％of the total activity is LOX-3.The ab-sence of this enzyme could delay the aging and deterioration of rice.Its active deletion body(Daw Dam)could re-duce the peroxidation level of unsaturated fatty acids in rice during storage and reduced the accumulation of un-pleasant aging odors including hexanal,glutaraldehyde and pentanol.Genetic analysis studies confirmed that the deletion of LOX-3was controlled by a single recessive gene.Sequencing confirmed that compared with the wild-type(WT)lox3 gene,the mutant(MT)lox3 gene encoded by the gene had a substitution mutation from G to A in nucleotide 1 497 of the mRNA recoding region,resulting in the early termination of translation.Using this molecular marker,lox3 genotypes of some important rice germplasm resources were identified,and three new rice lines with lox3 deletion were selected.A primer combination was designed in this study,in which the outer primers lox3-o-F and lox3-o-R were located on both sides of the mutation site,and the inner primers lox3-I-F and lox3-I-R were located on the mutation site.Using this primer combination,the genomic DNA of rice varieties was amplified by PCR.The lox3 gene was wild-type(WT),mutant deletion type and heterozygous type.The gene marker was lo-cated in the lox3 coding region,and the detection was more accurate than other marker methods.The design of four primers could complete the reaction through one ordinary PCR,and the band type difference was large.Cheap agarose gel electrophoresis could be used to distinguish the band type.The primer combination of the invention could effectively identify the lox3 genotype of rice storage tolerance gene,and genotypes lox3 by using one-time ordinary PCR and agarose gel electrophoresis.Compared with other detection methods,the detection operation was simpler,more accurate and lower cost.The development of this marker could provide technical reference for the efficient utilization of lox3 genotype.The storage tolerance of rice was evaluated through storage experiments,which showed that LOX-3 of rice embryo was closely related to the storage characteristics of rice.
RiceLipoxygenaseLOX-3Marking development and applicationStorage resistance
NETL
NSTL
万方数据
维普
