Preliminary Study On Ramie Flower Development Related AP2/ERF Gene

Ramie is perennial plant.The Ramie breeding objectives mainly focus on the yield,fiber quality,processing quality,stress resistance,forage quality and flowering which largely affect the growth after August.According to the growth cycle of the ramie,the ramie conducive to the three period of vegetative growth.Flowering and fruiting will lead to a decline in the yield and quality of ramie fiber obviously at three period.The purpose of this study is that according to the different varieties of ramie in different parts of the expression of transcription factors in different periods,to find transcription factors associated with flowering which contained the AP2/ERF domain.Flowering related genes will be cloned.All the goals are to lays the foundation for further study on the regulation of flowering in ramie.The main results of this paper are as follows:1?The transcriptome data of prophase,seven transcription factors have the higher credibility.At the same time,bioinformatics sites ExPASy,NCBI and Smart were used to nucleotide sequence analysis,amino acid translation and amino acid sequences of conserved domain search.Seven transcription factors 15247 n,6896,18197,17478,15426 N,9156,16687 were selected.After analysis,the analysis results that 18197,17478,15247,6896,9156 contains AP2 domain.At the same time,the seven selected transcription factors are far from relatives and have certain representativeness which provide theoretical direction for further studies.2?In this study,using 2 ramie gene,3 ramie varieties as materials,actin as the reference gene to establish real-time quantitative PCR system.The results show that the extraction method of reagent box of RNA can meet the requirements of RT-qPCR.The amplification curve and melting curve are consistent with the experimental requirements.Actin gene can be used as reference for research on relative quantitative analysis of ramie.The relative quantitative analysis showed that the RT-qPCR method established in this study can apply to the study of gene expression level.3?By using fluorescence quantitative PCR technology to study non flowering period of 7 possible genes of “Hei pi dou MS5”,“Hei pi dou MS9”,“Da ye lu MS1”,“Huang xiao ye”,“Wan zi zhu ma”,Yuan qing No.5”,“Lu xian qing pi”,“Liang ping qing ma”,“1313”,“1394” a total of 10 species.It found that most of ramie varieties,transcription factors 15247 n,9156,6896,17478,15426 N and 16687 are expressed obviously.9156,6896 and 17478 which all contain the AP2 domain.From the plant parts,9156,15247 n,15426N,9156,17478 in 08 varieties of Ramie Male flower and 9156 in 09 parts of female flower are expresed higher.Especially the 9156 is very significant.The 08 varieties(GBN08)is a monoecious ramie and ramie 09 is female(GBN09),are selected from the sterile offspring.High expression of 9156 gene may confirmed a possible that flowering related to the 9156.4?By gene clone technology,CDS sequence of 9156 ramie have cloned.The full-length encoding region is 1302 bp,encoding 433 amino acids.Through bioinformatics analysis,molecular mass is 47.05359 Kda of 9156 protein.9156 protein is little which is helpful to the study of prokaryotic expression or pattern plants.The theory PI of protein(isoelectric point)was 9.04 which belong to the basic protein.