Analysis Of Metabolic Differences And Mechanisms Of Fructus Aurantii Immaturus In Different Growth Stages

Fructus aurantii(FA)and Fructus aurantii immaturus(FAI)are fruits of citrus plants of rutaceae in different growth and development stages.It is a genuine medicinal material in Jiangxi province and also a traditional Chinese medicine for regulating qi in clinical practice.Due to the difference in chemical composition of Aurantii fruit,its efficacy is different.Flavonoids are the main active components.Fructus aurantii immarurus tends to break qi and dissipate accumulation,fructus aurantii tends to regulate qi width.FA and FAI not only has good effects on gastrointestinal diseases,but also improves immune function and antioxidant function.In this study,the dynamic changes of flavonoids in FAI and FA at different growth stages and the related genes of flavonoids synthesis were studied systematically.To elucidate the pharmacodynamic basis of FAI and FA,we studied the metabolic and transcription levels of FA at different growth stages,explored the core differential metabolites and genes regulating the biosynthesis of active components of FAI and FA.It provides scientific basis for further development of medicinal and edible value of FA.In this study,Citrus junos Sieb.ex Tanaka and Citrus aurantium‘Xiucheng’ from Zhangshu city,Jiangxi province were used as raw materials.UPLC-MS/MS technology was used to detect and analyze metabolites in different growth stages,which can identify and screen the differential metabolites.Illumina Nova Seq6000 high-throughput sequencing technology was used to conduct transcriptome sequencing for different developmental stages of Fructus aurantii immaturus.Differential genes were screened by DESeq and analyzed by GO enrichment and KEGG enrichment.The correlation between differential metabolites and differential expression genes was analyzed to explore the relationship between them.A total of293 different metabolites were detected,which could be divided into 17 classes.The first principal component of Fructus aurantii Immaturus was isolated in different growth stages.The reference genome of sweet orange was used in this study,a total of1023535000 clean reads containing 153.55 Gb of effective nucleotide sequences were obtained.GO enrichment analysis showed that the most term was enriched in molecular function,especially in tetrapyridine binding,heme binding and iron ion binding.KEGG enrichment analysis showed that the differentially expressed genes were mainly enriched in phenylpropanoid biosynthesis and flavonoid biosynthesis.The results showed that there were significant differences of Fructus aurantii immaturus in different growth stages.With the growth and development of Fructus aurantii immaturus,the differential metabolites and differential expression genes increased first and then decreased,and the differences of Fructus auranti I Immaturus30 days after flowering were the greatest,and the differences between Citrus junos Sieb.ex Tanaka and Citrus aurantium‘Xiucheng’ 60 days after flowering were the least.The core differential metabolites represented by nobiletin were obtained by intersection of different comparison combinations.Correlation network analysis showed that most of the differentially expressed genes had significant inhibitory effect on flavonoids.In the flavonoid biosynthesis pathway,most genes were highly expressed at 45 or 60 days after flowering,and the content of most flavonoid compounds in Fructus aurantii Immaturus was higher than that in Fructus aurantii,indicating that Fructus aurantii Immaturus has stronger medicinal effect than Fructus aurantii.