Transcriptome Analysis And Cloning Of Key Genes For Flavonoid Synthesis In Potentilla Fruticosa L.

Potentilla fruticosa L.,a deciduous shrub,belongs to the genus Potentilla in the family Rosaceae.According to the literature and modern researches,it was sweet in flavor and cold in property.It also offers a number of health benefits,including the abilities to relieve summer heat,support the heart and brain,regulate menstruation,invigorate the spleen and stomach,and work as an anti-inflammatory,hypoglycemic,and antibacterial.Although flavonoids and tannins are the main active components,there is no relevant report on Potentilla fruticosa L.flavonoid biosynthesis.Therefore,in this study,the transcriptome was sequenced using second-generation high-throughput sequencing technology,and the upstream genes PAL,CHI,and F3 H,which are involved in the synthesis of flavonoids,were discovered,cloned,and bioinformatics analysis.To assist future research into the specific roles played by genes in the anabolic process of flavonoids and to establish the groundwork for uncovering the molecular regulatory mechanism of flavonoid production in Potentilla fruticosa L.The specific findings of the study are as follows:1.For the first time,the transcriptomes of Potentilla fruticosa L.at different altitudes was sequenced by Illumina platform,and 160,691 unigenes were obtained,with an average sequence length of 704.09 bp,and a functional annotation rate of100%.A total of 2,062 unigenes,or 1.3% of the total,were collaboratively annotated by the five major databases Nr,Swiss-Prot,Pfam,GO,and KEGG.These databases each annotated 99,736,49,121,44,284,11,836 and 31,582 unigenes,respectively.A total of 100,879 unigenes were successfully annotated in at least one database,representing a total of 62.8%.2.Using Pf L as a point of reference,analysis the difference of unigene expression in Pf H.Potentilla fruticosa L.samples collected at high and low altitudes were screened for a total of 8,618 DEGs;5,199 of these genes showed up-regulation,while 3,419 showed down-regulation.According to the findings of the KEGG enrichment analysis,the pathways for the biosynthesis of phenylpropanoid and flavonoids,respectively,contain annotations for 58 and 21 DEGs,respectively.The biosynthesis of flavonoids in Potentilla fruticosa L.is mainly mediated by the four enzymes PAL,CHS,CHI,and F3 H.3.The sequence information of Pf PAL1,PfCHI1 and PfF3H1 genes was originally cloned,and they each had coding sections containing 2,175 bp,1,325 bp,and 1,020 bp,as well as 724,410,and 339 amino acids,respectively.According to the results of the phylogenetic analysis,Pf PAL1 was closely related to PAL1 of Fragaria Vesca and PAL2 of Fragaria × ananassa,PfCHI1 was closely related to CHI of Fragaria Vesca,and PfF3H1 was closely related to F3 H of Fragaria Vesca.4.The results of the bioinformatics analysis are shown below: the physical and chemical properties of Pf PAL1,PfCHI1,and PfF3H1 are hydrophilic.Pf PAL1 and PfF3H1 are stable proteins,and PfCHI1 is unstable.None of them has a signal peptide,which indicate that they are non-secretory proteins.Pf PAL1 and PfF3H1 do not have a transmembrane domain structure,indicating that they are non-transmembrane proteins,but PfCHI1 is a transmembrane protein because it contains one transmembrane helix.Secondary structure analysis revealed that the main component of Pf PAL1 is alpha-helix,and the main components of PfF3H1 and PfCHI1 are random coils.