Identification Analysis And Functional Verification Of TCP Gene Family In Chenopodium Quinoa

TEOSINTE BRANCHED 1/CYCLOIDEA/PROLIFERATING CELL FACTORS(TCP)transcription factor is a kind of plant-specific protein,which participates in the regulation of seed germination,branch development,mosaic development,day and night law,hormone response and defense response.In this study,bioinformatics methods were used to identify and analyze the members of the quinoa TCP transcription factor family at the genome-wide level.At the same time,some quinoa TCP genes were selected to construct an overexpression vector,which was ectopically expressed in Arabidopsis thaliana.Perform analysis.The main results of this research are as follows:1.There are 31 members in the quinoa genome that encode the TCP protein.The analysis of physical and chemical properties showed that the sequence length,the number of encoded amino acids,the molecular weight of the protein and the protein isoelectric point of different quinoa TCP genes differ greatly.The phylogenetic tree analysis of the TCP genes of quinoa and Arabidopsis showed that the quinoa TCP family was clearly divided into two subfamilies,Class I and Class II,of which Class II was divided into two clades,namely CIN clades And CYC/TB1 clade.Conserved domain analysis showed that TCP proteins of the same subfamily had similar conserved domain composition.Analysis of the quinoa TCP gene structure revealed that 25 members have no introns,and the other 6 genes have only 1 intron.2.Tissue expression analysis of some quinoa TCP genes by fluorescence quantitative PCR.The results show that,CqTCP2.1,CqTCP11,CqTCP14.1,CqTCP14.2 are mainly expressed in leaves and flower buds.CqTCP14.2 is not expressed in the developing seeds,but also expressed in other tissues.CqTCP15.1 and CqTCP15.2 have been detected to express in all tissues,but they are mainly highly expressed in vegetative organs(roots,stems,leaves).CqTCP18.1 is mainly expressed in flower buds,followed by expression in stems,leaves,and inflorescences,and low in roots and developing seeds.3.In order to explore the function of CqTCP gene,a part of CqTCP gene overexpression vector was constructed and expressed in Arabidopsis,and its phenotype was observed.The results showed that the CqTCP11 transgenic Arabidopsis plants were smaller overall,the leaf area became smaller and curled downward,and the flowering of CqTCP11 overexpression plants was delayed,and the number of rosette leaves during flowering was also significantly higher than that of the wild type,indicating that the CqTCP11 gene is regulating It plays an important role in leaf development and flowering time.The CqTCP18.1 gene is involved in the regulation of branch development.Branch statistics show that the number of primary branches of CqTCP18.1 is not significantly different from that of the wild type,but the number of secondary branches is significantly less than that of the wild type.The Arabidopsis lines overexpressing CqTCP14.1 and CqTCP14.2 genes had smaller leaf area and curled downwards,and also showed a dwarf plant phenotype.The number of primary and secondary branches of CqTCP14.1 was significantly less than that of the wild type.In addition,after drought treatment,the fruit pods and leaves of wild-type Arabidopsis thaliana quickly turned yellow,while the CqTCP14 transgenic line was still partially green.Therefore,it is speculated that CqTCP14 plays a role in regulating leaf development and drought stress.The Arabidopsis lines overexpressing CqTCP15.1 and CqTCP15.2 have a faster germination rate than wild-type seeds.Therefore,CqTCP15 may play a role in the process of seed germination.CqTCP2.1 gene may regulate leaf development.Arabidopsis lines overexpressing CqTCP2.1 showed a phenotype of dwarf plants,smaller leaf area,and reduced number of rosette leaves.4.Exploring the application of TRV virus vector-mediated gene silencing in quinoa.First,the sprouting seeds of quinoa were infected by the vacuum infection method,and the results were consistent with the phenotype of the control group.Then,4-6 pairs of true were selected.Quinoa at the leaf stage was infected with leaves,and green fluorescence appeared in the leaves,indicating that it might be feasible to establish a TRV-VIGS system for quinoa by infecting the leaves.