Preliminary Study On The Regulation Of Quality Variation Of Introduced Lonicerae Japonicae Flos By MicroRNA

OBJECTIVE:This study was based on the microRNA-mediated variation mechanism of honeysuckle quality,and studied from the active components of honeysuckle and microRNA.In order to provide a scientific basis for revealing the mechanism of microRNA regulation of the active constituents of honeysuckle,and to select excellent germplasm of honeysuckle.In the same time,the results laye the foundation for quality evaluationMethods:(1)UPLC-MS/MS was used to detect the contents of 16 main active constituents in the three major categories of flavonoids,chlorogenic acid and cycline in the origin and introduction of the seeds,and statistical analysis was carried out;(2)Using high-throughput sequencing technology to construct microRNA libraries of origin and introduction of honeysuckle,and use bioinformatics methods to analyze information about five microRNA libraries(3)The cleavage site of the differential miR6300 and its predicted target gene Unigene 19822 in 5 microRNA libraries was verified by 5 RLM-RACE method.The expression levels of honeysuckle in different flowering stages were detected by qRT-PCRusing miR6300 and Unigene19822;(4)The high-throughput sequencing technology was used to construct the decoction microRNA library of fresh and dried honeysuckle,and then compared with the honeysuckle flower bud microRNA library.The bioinformatics method was used to predict the human target gene of the selected miRNA and analyze it.Results:(1)Compared with the honeysuckle of origin,the content of main active components in the introduced honeysuckle was significantly reduced,indicating that the introduction may have a certain impact on the quality of honeysuckle,which led to changes in its clinical efficacy;(2)Statistically obtained the unique sequence and consensus sequence between the origin and the origin,and introduced microRNA,possible new microRNA and its target gene from the place of origin and origin;(3)5'RLM-RACE confirmed that the cleavage site of miR6300 may be located 1 bp downstream of the target gene binding site.qRT-PCR results showed that miR6300 was significantly negatively correlated with the target gene Unigene 19822 at different flowering stages;(4)miR162a-3p with significant difference in expression of honeysuckle from different producing areas and present in decoction,and four human target genes were predicted by bioinformatics analysis.Their functions were nuclear receptor co-repressors(NcoR),plexin-A3(PLXNA3),axis filament dynein heavy chain(DNAH3).Among them,nuclear receptor co-repressor(NcoR)may be related to the clinical efficacy of honeysuckle in lowering blood sugar and lowering blood fatConclusion:MicroRNA may be involved in the regulation of the quality variation of introduced honeysuckle to some extent,affecting the accumulation of active components in vivo.This provides a scientific basis for revealing the mechanism of microRNA regulation of the active constituents of honeysuckle,and lays a foundation for the breeding and quality evaluation of honeysuckle.