Cloning And Sequence Analysis Of Full-length CDNA Of Ginseng BBM Gene

Ginseng is a perennial herb that has been used in China for hundreds of years.The excellent varieties of ginseng are rare,the growth cycle is long,and the breeding process is difficult.With the increasing demand for ginseng,ginseng breeding has become the focus of scientists.Cultivation is the primary problem of ginseng' s reproduction.Somatic Embryogenesis is an important means of inducing plant' s regeneration in vitro.Somatic embryogenesis can induce the formation of new individuals under specific conditions without the fusion of cells,which has become an important way to study the development of plants' embryo.Ginseng can be cultivated more excellent varieties in a shorter time by this way,and the research on somatic embryogenesis of ginseng can provide a good solution for the problems of the difficulties of ginseng cultivation.However,the differentiation of embryogenic cells cultured in vitro requires the action of inducing factors and the expression of related genes.According to the related references,heterologous expression of Arabidopsis thaliana or Brassica napus BBM gene in tobacco enhances its regeneration ability and induces spontaneous healing.The formation of injured tissues,roots and buds suggests that the BBM gene may be involved in the somatic embryogenesis of ginseng.In this thesis,ginseng seeds were used as material to design the degenerate primers of this gene,and the BBM gene in ginseng seeds was successfully cloned by RACE method.In recent years,a large number of transcription factors have been isolated from plants.The AP2/EREBP family is one of them,and its transcription factors are closely related to plant growth and development and stress response.Searching for the BBM gene in Arabidopsis thaliana on the NCBI revealed that the BBM gene belongs to the transcription factor of the AP2(APETALA2)subfamily.The BBM gene plays a key regulatory role in somatic embryogenesis.In this study,the full-length cDNA sequence of ginseng BBM gene was cloned by RACE technology,and the gene was analyzed by bioinformatics.The research results are as follows:1.The full length cDN A of the BBM gene cloned from ginseng is 2256 bp in length and contains an open reading frame(ORF)consisting of 2010 bases encoding a protein consisting of 669 amino acids.2.The relative molecular weight of the protein encoded by the BBM gene is 73.57 KDa,and the isoelectric point(PI value)is 6.02;the protein contains euANT2-euANT6.bbm-1 and other motifs and two AP2 domains(AP2-R1 and AP2-)R2).a transcription factor belonging to the AP2 subfamily.The ginseng BBM gene is closely related to the BBM genes of tobacco,soybean and alfalf-a.In this study,the ginseng BBM genc(PgBBM)was successfully cloned by RACE method,and sequence analysis and protein function prediction were carried out.This lays a theoretical foundation for the induction of human ginseng somatic embryos by BBM gene and related research.