Evaluation of the chemical profile from four germplasms sources of Pruni Semen using UHPLC-LTQ-Orbitrap-MS and multivariate analyses

Pruni Semen,the seed of several unique Prunus plants,is a traditional purgative herbal material.To determine the authentic sources of Pruni Semen,46 samples from four species were collected and analyzed.Ten compounds including multiflorin A(Mul A),a notable purative compound,were isolated and identified by chemical separation and nuclear magnetic resonance spectroscopy.Seventy-six communal components were identified by ultra-high performance liquid chromatography with linear ion trap-quadrupole Orbitrap mass spectrometry,and acetyl flavonoid glycosides were recognized as characteristic constituents.The flavonoids were distributed in the seed coat and cyanogenic glycosides in the kernel.Based on this,methods for identifying Pruni Semen from different sources were established using chemical fingerprinting,quantitative analysis of the eight principal compounds,hierarchical cluster analysis,principal component analysis,and orthogonal partial least squares discriminant analysis.The results showed that the samples were divided into two categories:one is the small seeds from Prunus humilis(Ph)and Prunus japonica(Pj),and the other is the big seeds from Prunus pedunculata(Pp)and Prunus triloba(Pt).The average content of Mul A was 3.02,6.93,0.40,and 0.29 mg/g,while the average content of amygdalin was 18.5,17.7,31.5,and 30.9 mg/g in Ph,Pj,Pp,and Pt,respectively.All the above information suggests that small seeds might be superior sources of Pruni Semen.This is the first comprehensive report on the identification of chemical components in Pruni Semen from different species.

ERF5.1 modulates carotenoid accumulation by interacting with CCD4.1 in Lycium

Carotenoids are important natural pigments and have medical and health functions for humans.Carotenoid cleavage dioxygenase 4(CCD4)and ethylene responsive factor(ERF)participate in carotenoid metabolism,but their roles in Lycium have not been discovered.Here,we annotated LbCCDs from the Lycium reference genome and found that LbCCD4.1 expression was significantly correlated with the carotenoid metabolites during Lycium five fruit developmental stages.Over-expression of LbCCD4.1 in NQ’s leaves resulted in a series of significantly lower contents of carotenoid metabolites,includingβ-carotene andβ-cryptoxanthin.Moreover,LbERF5.1,a transcription factor belonging to the ERF family that was located in the nucleus,was isolated.Significant reductions in the carotenoids,especially lutein,violaxanthin and their derivatives,were observed in over-expressing ERF5.1 transgenic NQ’s leaves.Over-expression or virus-induced gene silencing of LbERF5.1 in NQ’s leaves induced a consistent up-or down-expression,respectively,of LbCCD4.1.Furthermore,yeast one-hybrid and dual-luciferase reporter assays showed that ERF5.1 interacted with the promoter of CCD4.1 to increase its expression,and LbERF5.1 could bind to any one of the three predicted binding sites in the promoter of LbCCD4.1.A transcriptome analysis of LbERF5.1 and LbCCD4.1 over-expressed lines showed similar global transcript expression,and geranylgeranyl diphosphate synthase,phytoene synthase,lycopeneδ-cyclase cytochrome,cytochrome P450-type monooxygenase 97A,cytochrome P450-type monooxygenase 97C,and zeaxanthin epoxidase in the carotenoid biosynthesis pathway were differentially expressed.In summary,we uncovered a novel molecular mechanism of carotenoid accumulation that involved an interaction between ERF5.1 and CCD4.1,which may be used to enhance carotenoid in Lycium.