| Soybean is a typical short-day(SD)plant,it delays to flowering in long-day(LD).The flowering time of soybean is influenced by planting area,growing season,and sunshine length,so the area of suitable planting is generally relatively narrow.Moreover,photoperiodic response is an essential factor affecting the regional adaptability of soybean varieties.In recent years,researchers have been discovered many genes loci that regulate flowering,and some of the important genes that control flowering time have been successfully cloned,and their functions have been discovered.However,there are still many loci which regulate flowering time in soybean that have not been validated.A candidate gene GmRNPC3 related to soybean flowering time was identified by real-time fluorescence quantitative experiment,and the gene function of Arabidopsis thaliana was further studied.1.A new quantitative trait loci(QTL)related to soybean flowering was initially identified.There were four candidate genes on this locus.Through tissue expression pattern analysis of the candidate genes in two parents,GmRNPC3 was finally predicted to be a candidate gene related to soybean flowering time.2.Bioinformatics analysis showed that GmRNPC3 is a small spliceosomal protein located on chromosome 16 of soybean,which belongs to the RNPC gene family,whose RNPC domain is highly conserved.According to evolutionary tree analysis,GmRNPC3 is closely related to beans and has low homology with plants such as rice and corn.Structure prediction of GmRNPC3 protein showed that GmRNPC3 protein had no transmembrane domain.GmRNPC3 protein was hydrophobic,no signal peptide sequence was found,and the protein subcellular localization prediction result was nucleus.3.GmRNPC3 was connected to the p1302-GFP vector to construct the fusion expression vector,and it was injected into the tobacco epidermal cells to observe the subcellular localization of the protein.The results showed that GmRNPC3 protein was mainly located in the nucleus.The expression pattern of GmRNPC3 was analyzed by qRTPCR,and it was found that GmRNPC3 was expressed in all organ of soybean.Moreover,the expression level of flower was higher than that of B13 in Guizao 1.4.GmRNPC3 gene was inserted into the p TF101 overexpression vector and heterogeneously transformed into wild Arabidopsis thaliana to select overexpressed strains.The phenotype of the flowering period of transgenic plants were analyzed,and the results showed that the transgenic plants showed significantly early flowering under LD compared with wild Arabidopsis thaliana.5.T-DNA insertion mutants of Arabidopsis homologous gene has significantly delayed flowering with Col-0 phenotypic analysis under LD.After the GmRNPC3 gene remediation mutant,transgenic remediation plants were obtained.The phenotype of the transgenic remediation plants were analyzed in this study.The results showed that the late flowering phenomenon of the mutants were remediation,and the transgenic remediation plants had significantly earlier flowering than Col-0 plants and mutant plants.Through overexpression of the target gene,this study preliminarily identified the function of the GmRNPC3 gene,which may be involved in the regulation of soybean flowering time.This study provides reference for further research on the flowering time regulation mechanism of soybean. |
