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Benzylisoquinoline alkaloids (BIAs) represent a significant class of secondary metabolites with crucial roles in plant physiology and substantial potential for clinical applications. CYP82 genes are involved in the formation and modification of various BIA skeletons, contributing to the structural diversity of compounds. In this study, Corydalis yanhusuo, a traditional Chinese medicine rich in BIAs, was investigated to identify the catalytic function of CYP82s during BIA formation. Specifically, 20 CyCYP82-encoding genes were cloned, and their functions were identified in vitro. Ten of these CyCYP82s were observed to catalyze hydroxylation, leading to the formation of protopine and benzophenanthridine scaffolds. Furthermore, the correlation between BIA accumulation and the expression of CyCYP82s in different tissues of C. yanhusuo was assessed their. The identification and characterization of CyCYP82s provide novel genetic elements that can advance the synthetic biology of BIA compounds such as protopine and benzophenanthridine, and offer insights into the biosynthesis of BIAs with diverse structures in C. yanhusuo.Key messageCYP82s identified from Corydalis yanhusuo can catalyze the hydroxylation of different substrates. These newly identified CYP82s provide genetic elements for the production of BIAs by synthetic biology.

Climate warming accelerates the timing of flowering and insect pollinator emergence, especially in spring. If these phenological shifts progress independently between species, features of plant-pollinator mutualisms may be modified. However, evidence of phenological mismatch in pollination systems is limited. We investigated the phenologies of a spring ephemeral, Corydalis ambigua , and its pollinators (bumble bees), and seed-set success over 10-14 years in three populations. Although both flowering onset and first detection of overwintered queen bees in the C. ambigua populations were closely related to snowmelt time and/or spring temperature, flowering tended to be ahead of first pollinator detection when spring came early, resulting in lower seed production owing to low pollination service. Relationships between flowering onset time, phenological mismatch, and seed-set success strongly suggest that phenological mismatch is a major limiting factor for reproduction of spring ephemerals. This report demonstrates the mechanism of phenological mismatch and its ecological impact on plant-pollinator interactions based on long-term monitoring. Frequent occurrence of mismatch can decrease seed production and may affect the population dynamics of spring ephemerals.

The flowering phenology of early-blooming plants is largely determined by snowmelt timing in high-latitude and high-altitude ecosystems. When the synchrony of flowering and pollinator emergence is disturbed by climate change, seed production may be restricted due to insufficient pollination success. We revealed the mechanism of phenological mismatch between a spring ephemeral (Corydalis ambigua) and its pollinator (overwintered bumblebees), and its impact on plant reproduction, based on 19 years of monitoring and a snow removal experiment in a cool-temperate forest in northern Japan. Early snowmelt increased the risk of phenological mismatch under natural conditions. Seed production was limited by pollination success over the 3 years of the pollination experiment and decreased when flowering occurred prior to bee emergence. Similar trends were detected on modification of flowering phenology through snow removal. Following snowmelt, the length of the pre-flowering period strongly depended on the ambient surface temperature, ranging from 4 days (at greater than 7°C) to 26 days (at 2.5°C). Flowering onset was explained with an accumulated surface degree-day model. Bumblebees emerged when soil temperature reached 6°C, which was predictable by an accumulated soil degree-day model, although foraging activity after emergence might depend on air temperature. These results indicate that phenological mismatch tends to occur when snow melts early but subsequent soil warming progresses slowly. Thus, modification of the snowmelt regime could be a major driver disturbing spring phenology in northern ecosystems.

Corydalis yanhusuo W.T. Wang is a traditional herb. Benzylisoquinoline alkaloids (BIAs) are the main pharmacological active ingredients that play an important role in sedation, relieving pain, promoting blood circulation, and inhibiting cancer cells. However, there are few studies on the biosynthetic pathway of benzylisoquinoline alkaloids in Corydalis yanhusuo , especially on some specific components, such as tetrahydropalmatine. We carried out widely targeted metabolome and transcriptomic analyses to construct the biosynthetic pathway of benzylisoquinoline alkaloids and identified candidate genes. In this study, 702 metabolites were detected, including 216 alkaloids. Protoberberine-type and aporphine-type alkaloids are the main chemical components in C . yanhusuo bulbs. Key genes for benzylisoquinoline alkaloids biosynthesis, including 6-OMT , CNMT , NMCH , BBE , SOMT1 , CFS , SPS , STOX , MSH , TNMT and P6H , were successfully identified. There was no significant difference in the content of benzylisoquinoline alkaloids and the expression level of genes between the two suborgans (mother-bulb and son-bulb). The expression levels of BIA genes in the expansion stage (MB-A and SB-A) were significantly higher than those in the maturity stage (MB-C and SB-C), and the content of benzylisoquinoline alkaloids was consistent with the pattern of gene regulation. Five complete single genes were likely to encode the functional enzyme of CoOMT, which participated in tetrahydropalmatine biosynthesis in C . yanhusuo bulbs. These studies provide a strong theoretical basis for the subsequent development of metabolic engineering of benzylisoquinoline alkaloids (especially tetrahydropalmatine) of C . yanhusuo .

The aim of the present study was to investigate the alkaloids of Tibetan medicine Corydalis conspersa Maxim. and their hepatoprotective effect against carbon tetrachloride (CCl4)-induced acute liver injury (ALI). The ethanol extract of this herbal medicine was subjected to a phytochemical study. Network pharmacology (NP) and molecular docking were used to predict the active constituents and mechanism of action against ALI. Seven alkaloid components were isolated and identified from this herb medicine, including acetylcorynoline (1, ACE), corynoline (2), scoulerine (3), protopine (4), bulbocapnine (5, BBC), palmatine (6), and isocorydine (7, ISO), among which compounds 1, 3, and 5 were isolated from this plant for the first time. Pharmacological experiments have shown that compounds 1, 5, 7, and the total alkaloids (TTA) of the plant exhibit good improvement effects on CCl4-induced ALI in mice. NP and molecular docking predicted that their mechanism of action may be related to targets such as STAT3, SRC, EGFR, PIK3CA, and HSP90AA1. These research findings provide a theoretical basis for the development of the medicinal value of Tibetan medicine Corydalis conspersa Maxim.

Background The medicinal value of the herb Corydalis yanhusuo W. T. Wang has driven its cultivation expansion. However, long-term monoculture of C. yanhusuo causes severe soil degradation and increases the risk of soil-borne diseases. Crop rotation improves soil sustainability and reduces pathogen pressure. Further research is necessary to establish successful crop rotation systems for C. yanhusuo and to comprehend their effects on soil microbial populations and physicochemical qualities. Methods In this study, we established three crop cultivation systems, including CR ( C. yanhusuo –rice rotation), CS ( C. yanhusuo –soybean rotation), and CN (continuous cropping of C. yanhusuo ), to examine sustainable models suitable for C. yanhusuo cultivation and their regulatory effects on the soil. To assess alterations in the physicochemical characteristics of the soil, we employed chemical analysis. The microbial makeup and roles of bacterial and fungal communities were compared using high-throughput amplicon sequencing. We investigated the effects of these rotations on soil ecosystem functions through the use of structural equation modeling. Results The results showed that CR and CS increased C. yanhusuo tuber yield and tetrahydropalmatine and protopine contents compared to CN. The concentrations of soil organic matter, total/hydrolysable nitrogen, total/available phosphorus, and soil enzyme activity were significantly higher in CR and CS compared to CN. Firmicutes grew in rhizosphere soil in CS, while Actinobacteria abundance increased in CR. The abundance of Bacillus, Sphingomonas, Rhodanobacter, and Trichoderma increased. Meanwhile, that of Flavobacterium, Fusarium, and Plectosphaerella decreased in CR and CS. CR and CS had increased bacterial and fungal community network linkages and modularity. Structural equation modelling revealed that crop rotation affects soil nutrients through microbial community dynamics. These soil nutrients, in turn, influence the yield and quality of C. yanhusuo. Conclusion In this study, we have deepened our understanding of the effects of rice and soybeans on the microbial community and physicochemical properties of C. yanhusuo cultivation soil. The findings can inform the development of ecological restoration. methods for the sustained production of C. yanhusuo, increasing green agricultural development.

The antimicrobial properties of herbs from Papaveraceae have been used in medicine for centuries. Nevertheless, mutual relationships between the individual bioactive substances contained in these plants remain poorly elucidated. In this work, phytochemical composition of extracts from the aerial and underground parts of five Papaveraceae species (Chelidonium majus L., Corydalis cava (L.) Schweigg. and Körte, C. cheilanthifolia Hemsl., C. pumila (Host) Rchb., and Fumaria vaillantii Loisel.) were examined using LC-ESI-MS/MS with a triple quadrupole analyzer. Large differences in the quality and quantity of all analyzed compounds were observed between species of different genera and also within one genus. Two groups of metabolites predominated in the phytochemical profiles. These were isoquinoline alkaloids and, in smaller amounts, non-phenolic carboxylic acids and phenolic compounds. In aerial and underground parts, 22 and 20 compounds were detected, respectively. These included: seven isoquinoline alkaloids: protopine, allocryptopine, coptisine, berberine, chelidonine, sanguinarine, and chelerythrine; five of their derivatives as well as non-alkaloids: malic acid, trans-aconitic acid, quinic acid, salicylic acid, trans-caffeic acid, p-coumaric acid, chlorogenic acid, quercetin, and kaempferol; and vanillin. The aerial parts were much richer in phenolic compounds regardless of the plant species. Characterized extracts were studied for their antimicrobial potential against planktonic and biofilm-producing cells of S. aureus, P. aeruginosa, and C. albicans. The impact of the extracts on cellular metabolic activity and biofilm biomass production was evaluated. Moreover, the antimicrobial activity of the extracts introduced to the polymeric carrier made of bacterial cellulose was assessed. Extracts of C. cheilanthifolia were found to be the most effective against all tested human pathogens. Multiple regression tests indicated a high antimicrobial impact of quercetin in extracts of aerial parts against planktonic cells of S. aureus, P. aeruginosa, and C. albicans, and no direct correlation between the composition of other bioactive substances and the results of antimicrobial activity were found. Conclusively, further investigations are required to identify the relations between recognized and unrecognized compounds within extracts and their biological properties.

Many species of Corydalis (Papaveraceae) have been used as medicinal plants in East Asia, and the most well-known species are Corydalis yanhusuo and C. decumbens in the Pharmacopoeia of China. However, authentication of these species remains problematic because of their high morphological variation. Here, we selected 14 closely related species and five genomic regions (chloroplast: matK, trnG, rbcL, psbA-trnH; nuclear: ITS) to explore the utility of DNA barcoding for authenticating these herbs. In addition, the Poisson tree process (PTP) and automatic barcode gap discovery (ABGD) were also used and compared with DNA barcoding. Our results showed that the ITS region was not suitable for molecular analysis because of its heterogeneous nature in Corydalis. In contrast, matK was an ideal region for species identification because all species could be resolved when matK was used along with the other three chloroplast regions. We found that at least five traditional identified species were lumped into one genetic species by ABGD and PTP methods; thus, highlighting the overestimation of species diversity using the morphological approach. In conclusion, our first attempt of molecular analysis of Corydalis herbs presented here successfully resolved the identification of medicinal species and encouraged their taxonomic re-assessment.

Chloroplast genomes are valuable for inferring evolutionary relationships. We report the complete chloroplast genomes of 36 Corydalis spp. and one Fumaria species. We compared these genomes with 22 other taxa and investigated the genome structure, gene content, and evolutionary dynamics of the chloroplast genomes of 58 species, explored the structure, size, repeat sequences, and divergent hotspots of these genomes, conducted phylogenetic analysis, and identified nine types of chloroplast genome structures among Corydalis spp. The ndh gene family suffered inversion and rearrangement or was lost or pseudogenized throughout the chloroplast genomes of various Corydalis species. Analysis of five protein-coding genes revealed simple sequence repeats and repetitive sequences that can be potential molecular markers for species identification. Phylogenetic analysis revealed three subgenera in Corydalis . Subgenera Cremnocapnos and Sophorocapnos represented the Type 2 and 3 genome structures, respectively. Subgenus Corydalis included all types except type 3, suggesting that chloroplast genome structural diversity increased during its differentiation. Despite the explosive diversification of this subgenus, most endemic species collected from the Korean Peninsula shared only one type of genome structure, suggesting recent divergence. These findings will greatly improve our understanding of the chloroplast genome of Corydalis and may help develop effective molecular markers.

The increasingly wide application of chloroplast (cp) genome super‐barcode in taxonomy and the recent breakthrough in cp genetic engineering make the development of new cp gene resources urgent and significant. Corydalis is recognized as the most genotypes complicated and taxonomically challenging plant taxa in Papaveraceae. However, there currently are few reports about cp genomes of the genus Corydalis. In this study, we sequenced four complete cp genomes of two endangered lithophytes Corydalis saxicola and Corydalis tomentella in Corydalis, conducted a comparison of these cp genomes among each other as well as with others of Papaveraceae. The cp genomes have a large genome size of 189,029–190,247 bp, possessing a quadripartite structure and with two highly expanded inverted repeat (IR) regions (length: 41,955–42,350 bp). Comparison between the cp genomes of C. tomentella, C. saxicola, and Papaveraceae species, five NADH dehydrogenase‐like genes (ndhF, ndhD, ndhL, ndhG, and ndhE) with psaC, rpl32, ccsA, and trnL‐UAG normally located in the SSC region have migrated to IRs, resulting in IR expansion and gene duplication. An up to 9 kb inversion involving five genes (rpl23, ycf2, ycf15, trnI‐CAU, and trnL‐CAA) was found within IR regions. The accD gene was found to be absent and the ycf1 gene has shifted from the IR/SSC border to the SSC region as a single copy. Phylogenetic analysis based on the sequences of common CDS showed that the genus Corydalis is quite distantly related to the other genera of Papaveraceae, it provided a new clue for recent advocacy to establish a separate Fumariaceae family. Our results revealed one special cp genome structure in Papaveraceae, provided a useful resources for classification of the genus Corydalis, and will be valuable for understanding Papaveraceae evolutionary relationships. Our results revealed one special cp genome structure in Papaveraceae, provided a useful resources for classification of the genus Corydalis, and will be valuable for understanding Papaveraceae evolutionary relationships.