Comparative Transcriptome Analysis of Cytoplasmic Male Sterile Line and Its Restorer Line in Soybean
[Objective]The"three line"method is the main way to breed hybrid soybeans.At present,only a few soybean restoration genes have been cloned.Thus it will be helpful in the breeding strong restorer lines with multi genes aggregation,thereby improving the fertility stability of hybrid F1,if more new Rf genes can be discovered.[Method]Using soybean cytoplasmic male sterile line JLCMS5A and its restorer line JLR2 as materials,transcriptome sequencing technology was used to analyze the transcriptional level changes of JLCMS5A and JLR2 flower buds at different stages,and to explore the relevant genes and pathways regulating restorer of fertility and flower bud development processes.Further gene annotation,sequence difference analysis,RT-qPCR validation,phylogenetic analysis,and protein structure prediction were performed on differentially expressed genes(DEGs)with Rf gene features encoding PPR(pentatrioptide repeat)proteins to explore Rf related genes.[Result]Transcriptome sequencing identified 17 181 DEGs,of which 3 856 were related to developmental stage and 2 808 were related to fertility.GO(gene ontology)functional annotation indicated that fertility-related DEGs were mainly enriched in functional categories such as ADP binding,nucleic acid binding transcription factor activity,and protein kinase activity,while developmental stage-related DEGs were mainly enriched in functional categories such as protein heterodimerization activity,DNA replication,and DNA binding.KEGG(kyoto encyclopedia of genes and genes)enrichment analysis showed that fertility-related DEGs mainly participate in metabolic pathways closely related to protein,carbohydrates,and signal transduction,such as endoplasmic reticulum protein processing,glucoside biosynthesis,and plant hormone signal transduction.Development-related DEGs mainly participated in metabolic pathways closely related to DNA replication,mismatch repair,starch and sucrose metabolism,and cell division and energy degradation.Analysis of candidate Rf genes revealed that the Glyma.09G176400 in JLR2 may play a certain role in regulating the restorer-of-fertility process of soybean cytoplasmic male sterility.[Conclusion]3 856 DEGs and 2 808 DEGs which respectively related to developmental stage and fertility were identified by transcriptomic techniques and molecular biological methods.Whereafter,a total of 15 DEGs encoding PPR protein were identified,which had the characteristics of restorer-of-fertility gene.Finally,a restorer-of-fertility related gene Glyma.09G176400 was excavated.
soybeancytoplasmic male sterilityrestorer-of-fertility genetranscriptome sequencingdifferentially expressed gene
国家科技期刊平台
NSTL
万方数据
