| Medicinal plants are fundamental to the sustainable development of Traditional Chinese medicine(TCM).The excavation and ultilization of medicinal plant taxa are important parts of the TCM modernization.Pharmacophylogeny as a theory to study the phylogenetic relationship,chemical composition,and activities,provided an innovative model of scientific research for the sustainable utilization of Chinese medicinal plant resources and the discovery of natural plant-derived new drugs.In recent years,the emerging of new technologies and methods from systems biology has provided new impetus for the further development of Pharmacophylogeny.Transcriptomics and metabolomics have unique advantages in the study of plant genetic relationship and phytochemistry.In this paper,these two technologies were applied for the Pharmacophylogenetic study(phylogenetic relationships,phytochemistry,and therapeutic effects)of two medicinal plant taxa(genera Dichocarpum and Paeonia),which could promote the disciplinary innovation of Pharmacophylogeny and facilitate the sustainable development of Chinese medicinal plant resources.Part I:A preliminary exploration of the Pharmacophylogeny of Chinese genus DichocarpumThe genus Dichocarpum(Ranunculaceae)is endemic to East Asia,with about 19 species worldwide,11 of which are distributed in China.There are 7 species have traditional medicinal records with the effect of clearing heat and detoxifying,relieving swelling and pain,educating and tonic,treating epilepsy and brightening eyes,etc.,showing their vital medicinal potential.However,research on the phytochemistry and modern pharmacological activity of this genus is extremely lacking.In order to explore the medicinal plant value of this genus,chemical composition and pharmacological activity studies were performed in this study via multi-omics and other techniques.Traditional medicinal records of plants are an important basis for modern drug development.In the current study,in vitro biological screening revealed that 8 Dichocarpum plant extracts had inhibitory activity of AchE(acetylcholinesterase)based on the traditional efficacy of treating neurological disorders(epilepsy)from D.auriculatum and D.trifoliolatum,among which the D.auriculatum extract showed the best activity.It is also found that 10 Dichocarpum plant extracts had agonistic effects on cannabinoid receptors(CB1 and CB2),among which the D.trifoliolatum extracts showed the best agonistic activity of CB2.A total of 11 compounds were isolated and identified from D.auriculatum for the first time based on multiple chromatographic and spectroscopic techniques.Six compounds showed significant inhibitory activity against AchE,with columbamine(IC50=0.24 ?M)and palmatine(IC50=0.34 ?M)was comparable to that of the positive drug(Tacrine,IC50=0.45 ?M).Corydaline(53.86%),columbamine(39.13%),and palmatine(39.13%)showed some intensity of agonistic effects on CB2.The results of content determination of active compounds based on LC-MS showed that these active compounds were widely distributed in the Chinese Dichocarpum,with relatively high contents in the roots(medicinal parts)of D.auriculatum.This study provided a scientific basis for the traditional efficacy of the genus Dichocarpum,especially for the D.auriculatum,and suggested that the plants from this genus had potential active components.Natural products are the material basis for the activity of medicinal plants.In order to further reveal the chemical composition of the genus Dichocarpum,the phytochemical analytical study of this genus was performed using UPLC-Q-TOF-MS/MS metabolomics technology.A total of 128 metabolites,including alkaloids,flavonoids,triterpenoid saponins,steroids,phenolic acids,and lactones,were identified from the genus Dichcocarpum.Alkaloids and flavonoids were widely distributed in 10 species,and magnoflorine could be considered as a characteristic compound.A molecular network of plant metabolites of 10 Dichocarpum species was constructed based on the GNPS network platform to reveal the existence of three chemical families(isoquinoline alkaloids,flavonol glycosides,and triterpenoid saponins),and the results showed that D.fargesii contained a unique cluster of triterpenoid saponins.Further comparative transcriptome analysis revealed that triterpene saponin biosynthesis-related genes were the main differential genes that distinguished the D.fargesii from other species.The results of this study elucidated the chemical characteristics of the Chinese Dichcocarpum and reveal the unique genetic clues and metabolic phenotype of D.fargesii,which can be further investigated,such as the targeted isolation of triterpenoid saponins.Based on the available literature,ITS sequence analysis,and transcriptome sequencing analysis,the correlation between the phylogenetic relationship,chemical components,and activities of Chinese Dichcocarpum plans was explored.The clustering analysis of chemotaxonomy based on metabolomics and the phylogenetic analysis based on ITS sequences were consistent,and both classified 10 Chinese Dichcocarpum spp.into three groups,which was different from the existing morphology-based classification system(Flora of China).The pharmacophylogenetic analysis showed that six species from the 6 species of Sect.Dichocarpum were clustered into one group,mainly containing protoberberine,aporphine,and flavonoids.The D.fargesii plants from different origins were clustered into one group,morphologically with specialized seeds,which were effective in strengthening the spleen and stomach and have abundant triterpenoid saponins.The remaining 3 species were clustered into one group,mainly containing bisbenzylisoquinoline alkaloids and triterpenoid saponins.The results of the above study preliminarily elucidated the medicinal pharmacophylogenetic correlation of the Chinese Dichcocarpum,and provided scientific basis for the improvement of the taxonomic system and the utilization of medicinal resources of this genus.Part II:A study on the Pharmacophylogeny of the genus PaeoniaThe family Paeoniaceae is a single family and genus with about 33 species and 26 subspecies worldwide,most of which are used medicinally and several of which are edible.Cortex Moutan,Radix paeoniae alba,and Radix paeoniae rubra are commonly used as herbal medicines.More than 400 compounds have been reported from the genus Paeonia,and studies have shown that Paeonia plant extracts and compounds have a variety of biological activities such as antioxidant,anti-inflammatory,cardiovascular protection,and neuroprotection.In order to further explore the medicinal potential of the genus Paeonia,which is an important medicinal plant group with abundant resources,and to comprehensively exploit the plant resources,a comprehensive study of phylogenetic relationships,phytochemistry,and therapeutic effects of the genus Paeonia was conducted in this study using multi-omics and other techniques.Many wild species of Paeonia spp.have shown medicinal value in some parts of China as substitutes for the species specified in the Pharmacopoeia of the People's Republic of China(CP),but their chemical composition has been less studied.To provide evidence for the correct origin of the specified species in the CP and to explore the potential medicinal value of the resources of this genus,metabolomics techniques were used to metabolize the medicinal parts of 15 Paeonia species.The results showed that the pharmacopoeiaspecified species(Paeonia lactiflora and Paeonia vetchii)were significantly different from other wild species in the Sect.Paeonia,and the chemical profiles of the two oil peonies were similar.Further results of content determination showed that paeoniflorin compounds were higher in the roots of most species,and the content of paeoniflorin in the roots of all species met the criterion of CP except for P.delavayi var.angustiloba and P.mairei.The roots of Sect.Moutan were significant enrichment plant parts for paeonol.The results indicated the inadequacy of the quality control indexes of the TCM Cortex Moutan,Radix paeoniae alba,and Radix paeoniae rubra,and suggested that most Paeonia wild species are potential medicinal resources that deserve further research and exploitation.The genus Paeonia is an isolated taxon whose phylogenetic position has not been determined.In this study,for the first time,we explored the phylogenetic position and subgeneric relationships from the perspective of the biosynthetic regulatory pathways of the unique chemical constituent,paeoniflorin.The root transcriptomes of 15 species of Paeonia spp.were determined and a total of 34738-59114 unigene were obtained,58.65%of which were annotated.Most of the terpene biosynthesis pathway-related gene genes were highly expressed in the roots of P.mairei,and 53 genes were strongly correlated with the content of paeoniflorin and its derivatives.A phylogenetic tree of 15 species of the genus Paeonia was constructed from the full-length sequence of pinene synthase for the first time,and the results were consistent with the existing Paeonia systematic classification.The results of this study indicated that the phylogenetic relationships within the genus Paeonia are closely related to the key enzymes of the synthesis pathway of paeoniflorin and its derivatives,providing a new insight to further reveal the phylogenetic position of the genus Paeonia from the perspective of key enzymes and gene evolution,and could be used as a basis of the resolution of the paeoniflorin biosynthetic pathway.Based on metabolomics techniques,this paper analyzed and compared for the first time the chemical composition of ten plant parts(root bark,wood core,branches,leaves,petals,stamens,ovary,seed coat,seed kernels,and fruit shells)of the oil peony P.suffruticosa,and identified 55 differential metabolites.The biological activity assays showed that ten plant parts extracts exhibited good antioxidant activities,with gallic acid,PGG,resveratrol,kaempferol,and rutin being the main antioxidant active compounds.In addition,a major by-product of peony seed oil production showed the agonistic activity on cannabinoid receptors for the first time.The results of this study had important reference values for the comprehensive development and utilization and the sustainable development of the oil peony industry.In summary,this study employed metabolomics and transcriptomics to investigate the Pharmacophylogeny of two medicinal plant taxa(genera Dichocarpum and Paeonia).Compared with traditional phytochemical studies,mass spectrometry-based metabolomics techniques can identify metabolite information in plants more rapidly and comprehensively,and molecular networks analysis are more advantageous in plants with few study on phytochemistry.Transcriptomics correlates metabolite biosynthesis with plant phylogeny,contributing to the discovery of pharmacophylogenetic patterns in medicinal plants.In the future,the combination of systems biology theory and multi-omics technology with the theory of pharmacophylogeny will strongly guide the development and utilization of medicinal plant resources and promote the disciplinary innovation of pharmacophylogeny. |
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