Cloning And Spatio-temporal Expression Pattern Analysis Of Flowering Genes Of Bupleurum Chinense DC.

Objective: To study the relationship between vegetative growth and reproductive growth of Bupleurum chinense DC.in the Umbelliferae family,to clone and sequence the c DNA related to flower formation of Bupleurum chinense DC.,and to analyze the differential expression of flowering genes in Bupleurum chinense DC..It is clear that the flower-forming genes are in the north.The distribution of time and space expression in Bupleurum.Methods: Firstly,by consulting related literature,the relationship between vegetative growth and reproductive growth in Umbelliferae medicinal plants was reviewed.Secondly,based on homologous cloning and whole-genome sequencing data,genes related to flower formation in Bupleurum chinense DC.were cloned.Then using SYBGreenⅠmethod realtime fluorescent quantitative PCR technology to obtain the dissolution curve and Ct value of the five housekeeping genes of Actin,α-tubulin,β-tubulin,Cyclophilinand EF-1α in the above Bupleurum chinense DC.samples,and the dissolution of internal reference genes The curve and amplification efficiency were tested,and the internal reference genes stably expressed in Bupleurum chinense DC.were screened out.Finally,using the q RT-PCR technology,the four floral genes were compared and analyzed in the form of double internal reference.Relative expression in different tissues such as roots,stems,leaves and flowers of Bupleurum chinense DC.and various stages of flower development.Results: Firstly,the relationship between vegetative growth and reproductive growth of Umbelliferae medicinal plants is analyzed for the first time.Based on the law of unity of opposites in materialist dialectics,the relationship between the two is analyzed and discussed,and the In actual production,the two prevention measures are balanced.Secondly,four genes related to flower formation were cloned from Bupleurum chinense DC.which were named Bc SVP,Bc PAF1,Bc CO and Bc FT.Bioinformatics shows that they are highly conservative,closely related to carrots that belong to the Umbelliferae family,and participate in the biological regulation of the flowering process of plants.Then the internal reference genes stably expressed in Bupleurum chinense DC.Actin and EF-1α,and the subsequent spatiotemporal expression analysis of the target gene is performed in the form of a double internal reference combination.Lastly,four genes related to flower formation in Bupleurum chinense DC.are expressed in both vegetative organs(roots,stems and leaves)and reproductive organs(flowers),but their expression levels are different.During the flowering process of Bupleurum chinense DC.,the total relative expression of Bc SVP was the lowest,and the expression of Bc FT was the highest.The latter was 13.48 times that of the former,and the total expression of Bc CO and Bc PAF1 were close to the same.Bc SVP is mainly expressed in vegetative organs,with the highest expression in roots,which is 4.75 times the expression in flowers.It shows a trend of rising first and then falling during the flowering process,and the expression reaches the maximum in the full bloom.Bc CO has the smallest expression in roots and the largest expression in leaves,which is 1.57 times the expression in roots,and the expression before flowering is small.The overall relative expression level during the flowering period was relatively stable,and the expression level was close to the same in the full flowering period,and the flowering period began to decrease,but it was always higher than before flowering.The expression abundance ratio of Bc PAF1 in the roots,stems,leaves and flowers of the organ tissues of Bupleurum chinense DC.was1.05:1:1:1.20:1.06.The expression levels in different tissues and organs are close to the same,with the progress of flowering,showing a trend of first rising and then falling.The expression level reached the maximum in the early stage of blooming,which was 2.69 times the expression level before flowering.Bc FT is mainly expressed in stems and leaves,and rarely in flowers.The expression level varies greatly among various tissues,and the expression level rises sharply as the flowering period progresses.The maximum expression level is in the full blooming stage,which is about 20 times the expression level before flowering.It began to decline during the flowering period,but the expression level was still at a high level,which was 5.68 times that before flowering.Conclusion: This study took Bupleurum chinense DC.,the main original plant of Bupleurum chinense DC.,which is commonly used in traditional Chinese medicine,as the research object.For the first time,4 genes related to flower formation were cloned from Bupleurum chinense DC..The differential expression levels of floral genes in roots,stems,leaves,flowers and other tissues at different developmental stages were analyzed by a double internal control method,and it was found that the four genes were all in the key parts of floral formation,It may bethe key regulation gene of Bupleurum to flower formation.This result is of far-reaching significance for the study of the flower-forming gene of Bupleurum chinense DC.and the clarification of its flower-forming mechanism.It provides important information to further clarify the evolution and function of genes related to flower formation,and also helps to formulate high-yield and high-quality measures in the production of medicinal materials.