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Rationale Tissue-specific metabolite profiling helps to find trace alkaloids masked during organ analysis, which contributes to understanding the alkaloid biosynthetic pathways in vivo and evaluating the quality of medical plants by morphology. As Macleaya cordata contains diverse types of benzylisoquinoline alkaloids (BIAs), the alkaloid metabolite profiling was carried out on various tissues of the root. Methods Laser microdissection with fluorescence detection was used to recognize and dissect different tissues from the root of M. cordata. Ultra-high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry was applied to analyze the trace alkaloids in tissues. These detected alkaloids were elucidated using their accurate molecular weights, MS/MS data, MS fragmentation patterns and the known biosynthetic pathways of BIAs. Finally, the distribution of alkaloids in dissected tissues and whole sections was mapped. Results Forty-nine alkaloids were identified from five microdissected tissues, and 24 of them were detected for the first time in M. cordata. Some types of alkaloids occurred specifically in dissected tissues. More alkaloids were detected in the cork and xylem vascular bundles which emit strong fluorescence under fluorescence microscopy. Some of the screened alkaloids were intermediates in sanguinarine and chelerythrine biosynthetic pathways, and others were speculated to be involved in the new branches of biosynthetic pathways. Conclusions The integrated method is sensitive, specific and reliable for determining trace alkaloids, which is also a powerful tool for metabolite profiling of tissue-specific BIAs in situ. The present findings should contribute to a better understanding of the biosynthesis of BIAs in M. cordata root and provide scientific evidence for its quality evaluation based on morphological characteristics. Copyright © 2016 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

Based on network pharmacological analysis and molecular docking techniques, the main components of M. cordata for the treatment of bovine relevant active compounds in M. cordata were searched for through previous research bases and literature databases, and then screened to identify candidate compounds based on physicochemical properties, pharmacokinetic parameters, bioavailability, and drug-like criteria. Target genes associated with hoof disease were obtained from the GeneCards database. Compound−target, compound−target−pathway−disease visualization networks, and protein−protein interaction (PPI) networks were constructed by Cytoscape. Gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed in R language. Molecular docking analysis was done using AutoDockTools. The visual network analysis showed that four active compounds, sanguinarine, chelerythrine, allocryptopine and protopine, were associated with the 10 target genes/proteins (SRC, MAPK3, MTOR, ESR1, PIK3CA, BCL2L1, JAK2, GSK3B, MAPK1, and AR) obtained from the screen. The enrichment analysis indicated that the cAMP, PI3K-Akt, and ErbB signaling pathways may be key signaling pathways in network pharmacology. The molecular docking results showed that sanguinarine, chelerythrine, allocryptopine, and protopine bound well to MAPK3 and JAK2. A comprehensive bioinformatics-based network topology strategy and molecular docking study has elucidated the multi-component synergistic mechanism of action of M. cordata in the treatment of bovine hoof disease, offering the possibility of developing M. cordata as a new source of drugs for hoof disease treatment. [ABSTRACT FROM AUTHOR]

Bopu powder® and Sangrovit® were developed from Macleayacordata and are widely used in agriculture and animal husbandry, but their impurities have been rarely reported in the literature. Impurity analysis is of great importance to the quality and safety of veterinary drugs. In this study, high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS) combined with a screening method was used to screen and characterize the impurities in Bopu powder® and Sangrovit®. A total of 58 impurities were screened from Bopu powder® and Sangrovit® using the screening strategies, of which 39 were identified by their accurate m/z value, characteristic MS/MS data, and fragmentation pathways of references. This established method was used for impurity analysis for the first time and proved to be a useful and rapid tool to screen and identify the impurities of Bopu powder® and Sangrovit®, especially for those at trace levels in a complex sample. In addition, this study marks the first comprehensive research into impurities in these two products and has great significance for the systematic detection of impurities in other plant-derived drugs. [ABSTRACT FROM AUTHOR]

Although Lilium brownii (L. brownii) bulbs are popular fresh vegetables, a series of quality problems still remain after harvest. In this study, fresh L. brownii bulbs were placed in the dark at 25, 4, and −20 °C and under light at 25 °C from 0 to 30 days; the chemical compositions were analyzed by ultraviolet spectrophotometry (UV) and high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS). During the 30-day storage period, the browning degree increased over the storage time and with increasing temperature, but the contents of proteins and free amino acids decreased and were aggravated by light. The total polyphenol content increased until the 6th day at 25 °C (dark or light), but it did not significantly accumulate at −20 or 4 °C. The reducing sugar content showed a dynamic balance, but the total polysaccharide content decreased constantly in the four storage conditions. The polyphenol oxidase (PPO) activity increased with storage time and increasing temperature, while it was inhibited by light. The increase rates of malondialdehyde (MDA) content at −20 °C and light (25 °C) were higher than those at 4 and 25 °C. In addition, 12 secondary metabolites were identified, most of which accumulated during the storage period, for example, 1-O-feruloyl-3-O-β-D-glucopyranosylglycerol; 1,3-O-di-p-coumaroylglycerol; 1-O-feruloyl-3-O-p-coumaroylglycerol; and 1,2-O-diferuloylglycerol. The variations in nutrient levels had a low correlation with browning, but the variations in MDA, PPO, and secondary metabolite (phenolic acids) levels had a high correlation with browning. In conclusion, fresh L. brownii bulbs should be stored at a low temperature (4 °C) and in dark condition, and browning bulbs are excellent materials for secondary metabolite utilization. [ABSTRACT FROM AUTHOR]

Simple Summary: Weaning stress is a serious problem in the goat production industry. This article demonstrates that weaning can induce intestinal oxidative damage, and adding Lactobacillus plantarum or Macleaya cordata can alleviate the oxidative damage of the lower gut in weaned young goats in an intestinal region-specific way. Weaning usually leads to stress in livestock, which has a negative impact on their growth and development. Research on oxidative stress and inflammation induced by weaning has not been reported in goats. Here, we focused on oxidative stress profile and inflammation status of the lower gut (jejunum, ileum, and colon) of goats. First, we illustrated the status of antioxidant activity and inflammation in the intestine of young goats on pre-(2 weeks postnatal, 2 wkpn) or post-(11 wkpn, weaning at day 45 postnatal)-weaned period of young goats. Malondialdehyde (MDA) was higher (p < 0.0001) in jejunum and ileum of the young goats in 11 wkpn than that in 2 wkpn, whereas superoxide dismutase (SOD) activity was lower (p = 0.012) in the lower gut of the young goats with 11 wkpn than that in 2 wkpn. Furthermore, we intended to explore the protective influence of a probiotic additive (Lactobacillus plantarum (LAC) P-8, 10 g/d) and a prebiotic additive (Sangrovit®, Macleaya cordata (MAC) extract 3.75% w/w premix, 0.3 g/d) on intestinal oxidative stress and inflammation status of early-weaned young goats (average weights of 5.63 ± 0.30 kg, weaned on d 45 postnatal). We observed that LAC reduced MDA in jejunum and ileum (p < 0.0001), increased SOD activity in ileum (p < 0.01), and increased glutathione peroxidase (GSH-Px) activity in jejunum (p < 0.05). Similarly, MAC reduced MDA contents (p < 0.0001), increased SOD activities (p < 0.01) in both of ileum and jejunum, and increased GSH-Px activity (p < 0.05) in jejunum. However, there were no differences in feed intake, average daily gain, inflammation parameters (interleukin 2 and interleukin 6), and colon oxidative stress profile (MDA, SOD, or GSH-Px) among treatments. These results provide evidence that weaning induces oxidative damage in the lower gut of young goats, and the oxidative damage in the small intestine can be reduced by adding the addition of LAC or MAC in diets depending on the region of the lower gut. [ABSTRACT FROM AUTHOR]

AbstractSanguinarine (SAN) is an alkaloid with multiple biological activities, mainly extracted from Sanguinaria canadensis or Macleaya cordata. The low bioavailability of SAN limits its utilization. At present, the nature and mechanism of SAN intestinal absorption are still unclear. The pharmacokinetics, single-pass intestinal perfusion test (SPIP), and equilibrium solubility test of SAN in rats were studied. The absorption of SAN at 20, 40, and 80 mg/L in different intestinal segments was investigated, and verapamil hydrochloride (P-gp inhibitor), celecoxib (MPR2 inhibitor), and ko143 (BCRP inhibitor) were further used to determine the effect of efflux transporter proteins on SAN absorption. The equilibrium solubility of SAN in three buffer solutions (pH 1.2, 4.5 and 6.8) was investigated. The oral pharmacokinetic results in rats showed that SAN was rapidly absorbed (Tmax=0.5 h), widely distributed (Vz/F = 134 L/kg), rapidly metabolized (CL = 30 L/h/kg), and had bimodal phenomena. SPIP experiments showed that P-gp protein could significantly affect the effective permeability coefficient (Peff) and apparent absorption rate constant (Ka) of SAN. Equilibrium solubility test results show that SAN has the best solubility at pH 4.5. In conclusion, SAN is a substrate of P-gp, and its transport modes include efflux protein transport, passive transport and active transport. [ABSTRACT FROM AUTHOR]

The protein phosphatase (PP2C) gene family, known to participate in cellular processes, is one of the momentous and conserved plant-specific gene families that regulate signal transduction in eukaryotic organisms. Recently, PP2Cs were identified in Arabidopsis and various other crop species, but analysis of PP2C in cotton is yet to be reported. In the current research, we found 87 (Gossypiumarboreum), 147 (Gossypiumbarbadense), 181 (Gossypiumhirsutum), and 99 (Gossypiumraimondii) PP2C-encoding genes in total from the cotton genome. Herein, we provide a comprehensive analysis of the PP2C gene family in cotton, such as gene structure organization, gene duplications, expression profiling, chromosomal mapping, protein motif organization, and phylogenetic relationships of each species. Phylogenetic analysis further categorized PP2C genes into 12 subgroups based on conserved domain composition analysis. Moreover, we observed a strong signature of purifying selection among duplicated pairs (i.e., segmental and dispersed) of Gossypiumhirsutum. We also observed the tissue-specific response of GhPP2C genes in organ and fiber development by comparing the RNA-sequence (RNA-seq) data reported on different organs. The qRT-PCR validation of 30 GhPP2C genes suggested their critical role in cotton by exposure to heat, cold, drought, and salt stress treatments. Hence, our findings provide an overview of the PP2C gene family in cotton based on various bioinformatic tools that demonstrated their critical role in organ and fiber development, and abiotic stress tolerance, thereby contributing to the genetic improvement of cotton for the resistant cultivar. [ABSTRACT FROM AUTHOR]

Inflammatory bowel disease, a disease featured by intestinal epithelial barrier destruction and dysfunction, has been a constant threat to animal health. The primary objective of this research was to assess the impact of the extract derived from lotus leaves (LLE) on lipopolysaccharide (LPS) induced damage to the intestines in mice, as well as to investigate the fundamental mechanism involved. The LLE was prepared using ultrasonic extraction in this experiment, and the LLE total flavonoid content was 117.02 ± 10.73 mg/g. The LLE had strong antioxidant activity in vitro, as assessed by 2, 2-diphenyl-1-picrylhydrazyl, ferric reducing antioxidant power, and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) methods. In the vivo experiment, different doses of LLE (50, 100, and 200 mg/kg) were administered for 2 weeks before LPS treatment in mice. The results revealed that LLE alleviates intestinal tissue damage in LPS-induced mice. In the jejunum tissue, LLE significantly upregulated mRNA and protein expression levels of tight junction proteins, such as ZO-1, occludin, and claudin-1, and decreased the contents of the inflammatory cytokines, interleukin (IL)-1β, IL-6, and tumor necrosis factor-α. Furthermore, the malondialdehyde and lactate dehydrogenase contents increased by LPS in the liver were significantly reduced after administration of LLE, and the total antioxidant capacity, superoxide dismutase, and reduced glutathione decreased by LPS were remarkably increased by LLE. It was found that LLE could relieve LPS-induced oxidative stress by upregulating mRNA and protein expression of Nrf2 and HO-1 in jejunum tissue. In conclusion, LLE alleviates LPS-induced intestinal damage through regulation of the Nrf2/HO-1 signal pathway to alleviate oxidative stress, reducing inflammatory factors and increasing the expression of tight junction proteins in mice. [ABSTRACT FROM AUTHOR]

This correction notice addresses inaccuracies in an article about two products used in the animal breeding industry in China. The names of the products and a picture were incorrectly used, and the term "impurity" should be replaced with "trace components." The correction provides the accurate names of the products and clarifies their use as safe and effective Chinese veterinary medicine. Additionally, errors in figures and sample preparation information are corrected. The scientific conclusions of the article remain unaffected by these changes. [Extracted from the article]

Acute colitis is a complex disease that can lead to dysregulation of the gut flora, inducing more complex parenteral diseases. Dandelion polysaccharides (DPSs) may have potential preventive and therapeutic effects on enteritis. In this study, LPS was used to induce enteritis and VC was used as a positive drug control to explore the preventive and therapeutic effects of DPS on enteritis. The results showed that DPS could repair the intestinal barrier, down-regulate the expression of TNF-α, IL-6, IL-1β, and other pro-inflammatory factors, up-regulate the expression of IL-22 anti-inflammatory factor, improve the antioxidant capacity of the body, and improve the structure of intestinal flora. It is proved that DPS can effectively prevent and treat LPS-induced acute enteritis and play a positive role in promoting intestinal health. [ABSTRACT FROM AUTHOR]

Sanguinarine (SAN), as the main active component of a traditional Chinese veterinary medicine, has been widely used in the animal husbandry and breeding industry. However, the metabolites of SA are still uncertain. Therefore, this research aimed to investigate the metabolites of SA based on rats in vivo. The blood, feces, and urine of rats were collected after the oral administration of 40 mg/kg SAN. Ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS) was employed to identify the metabolites of SAN. The elemental composition of sanguinarine metabolites was inferred by analyzing their exact molecular weight, and the structures of the metabolites were predicted based on their fragment ions and cleavage pathways. A total of 12 metabolites were identified, including three metabolites in the plasma, four in the urine, and nine in the feces. According to the possible metabolic pathways deduced in this study, SAN was mainly metabolized through reduction, oxidation, demethylation, hydroxylation, and glucuronidation. This present research has summarized the metabolism of SAN in rats, which is helpful for further studying the metabolic mechanism of SAN in vivo and in vitro. [ABSTRACT FROM AUTHOR]

Eggs are recognized for their rich nutrient profile, providing essential proteins and lipids with notable functional properties. This study examines the effects of incorporating Water Extract of Ampelopsis grossedentata (WEA) into poultry feed on egg quality, focusing on lipid content, choline, L-carnitine levels, and flavonoid compound deposition. Our results show significant increases in essential amino acids, flavonoids, and other bioactive compounds in eggs from WEA-treated hens, suggesting enhanced cardiovascular, antioxidant, and anti-inflammatory benefits. Additionally, we observed elevated levels of choline and betaine in egg yolks, alongside increased L-carnitine content, which may contribute to improved lipid metabolism and reduced cardiovascular disease risk. KEGG pathway analysis revealed upregulation of metabolites involved in critical metabolic pathways, enhancing the nutritional profile of eggs. Flavonoid compounds, traditionally associated with plant-based foods, were also significantly increased, with notable levels of 7, 4′-dihydroxyflavone, daidzein, and glycitein identified in WEA-treated eggs, indicating potential health benefits. These findings suggest that WEA supplementation can produce functional eggs with improved nutritional quality, offering a novel approach to enhancing egg production and meeting the growing demand for functional foods. Further research is needed to fully understand the bioavailability and health impacts of these enriched compounds. [ABSTRACT FROM AUTHOR]

Edible dock, also known as Rumex patientia L.× Rumex tianschanicus A. LOS., exhibiting the characteristics of high yield, high protein and high nutrition, is widely used in the functional food industry and Chinese herbal medicine while short of secondary metabolites research. This study used UPLC-qTOF MS with two ion polarity in conjunction with Molecular Networking from Global Natural Products Social (GNPS) platform. Then, these 16 flavonoid substances including quercetin derivatives and kaempferol derivatives, firstly were identified from leaves in edible dock and were systematically investigated by UPLC-qTOF MS/MS. MS/MS experiments found that quercetin derivatives have five common characteristic ions, including m/z 300, m/z 271, m/z 255, m/z 243, and m/z 151. In addition, kaempferol derivatives have three common characteristic ions, including m/z 284, m/z 255, and m/z 227. Subsequently, 7 representative constituents were quantified via UPLC-QQQ MS with chemometrics, and the total content of 7 major flavonoids in its dry leaves is 6.988 mg/g in the 21st day samples. The work provided abundant metabolites information of flavonoids in edible dock, and the common characteristic ions will facilitate the building blocks-based molecular network (BBMN) to predict novel framework compounds from quercetin derivatives and kaempferol derivatives, laying the basis for exploitation of edible dock in the fields of pharmaceuticals, health food, and livestock feed. [Display omitted] • GNPS with HR-MS enables rapid screening secondary metabolites in edible dock. • Flavonoids in edible dock were systematically investigated by LC-MS/MS. • 16 flavonoids identified for the first time in edible dock via Molecular Networking. • The total content of main flavonoids in its dry leaves was as high as 6.988 mg•g−1. [ABSTRACT FROM AUTHOR]

A novel home-made H2SO4-Nafion (HN) tube sampling system coupled to a line ion trap mass spectrometer (LTQ-MS) with a versatile ambient ionization source, hectowatt microwave plasma torch (HMPT), has manifested unique advantages for picking directly metal elements in aqueous samples and acquiring the fully characteristic MPT mass spectra of copper and zinc composite ions. Here, we report the development of a novel HN-HMPT-LTQ-MS for metal elements assay based on environmental water to analyze samples of Poyang Lake, China. Detailed multi-stage tandem mass spectra show that the general structural form of target ions is [M(NO3)x(H2O)y(OH)z]+ for the positive ion mode. Under the optimized conditions, the proposed method provided low limits of detection (LODs) of 0.23 μg.L−1 for 63Cu+ and 1.1 μg.L−1 for 66Zn+, with relative standard deviations (RSDs) of less than 12.7% by MPT-LTQ-MS. This new result has met the requirements of national standards (GB 5750.6–2006) and is only about one magnitude order larger than the LOD of ICP-MS method. A wide linear response range of about 4 orders of magnitude for the method with linear coefficients (R2) of 0.99709 – 0.99962 for copper and zinc tested was in accordance with that of ICP-MS. Except for the recovery of 79% for the third sample and 123.8% for the seventh sample, the present method also provided good recoveries (84 – 119.3%) in spiked 10 batches of drinking water samples. Furthermore, it is envisioned that the developed approach might build a powerful hectowatt-MPT-MS platform for food security detection, drug analysis, and origin traceability. [ABSTRACT FROM AUTHOR]

Sanguinarine and chelerytrine have antibacterial and anti-inflammatory effects and is the main active ingredients of growth promoters in animals. Currently, Sanguinarine and chelerytrine were extracted from the capsules of the medicinal plant Macleaya cordata. However, the biomass of M. cordata nonmedicinal parts (leaves) accounted for a large proportion and contained a rich presentation of protopine and allocryptopine which are the precursor compounds of sanguinarine and chelerytrine. The aim of this study was to develop a new method for producing sanguinarine and chelerytrine through yeast transformation of protopine and allocryptopine in M. cordata leaves. First, we isolated different genes from Papaver somniferum (PsP6H, PsCPR, PsDBOX), Eschscholtzia californica (EcP6H), Cucumis sativus (CuCPR), Arabidopsis thaliana (AtCPR) and M. cordata (Mc11229, Mc11218, Mc6408, Mc6407, Mc19967, Mc13802). Additionally, some of the gene sequences were codon optimized. Then, we transformed these genes into yeast cells to compare the catalytic efficiency. Second, we used the most efficient strains to biotransform the leaves of M. cordata. Finally, we obtained 85.415 ± 11.887 ng mL−1 sanguinarine and 4.288 ± 1.395 ng mL−1 chelerytrine, which was more than 2–3 times the content in leaves of M. cordata. Overall, we using the nonmedicinal parts of M. cordata and successfully obtained sanguinarine and chelerytrine by the plant–microbial hybrid synthesis method. [ABSTRACT FROM AUTHOR]

Dietary Macleaya cordata extract (MCE) can improve the meat quality of poultry. However, the specific mechanism by which MCE regulates the meat quality has not been clarified yet. Sanguinarine (SAN) is one of the important natural active components in MCE. Our study aims to explore the regulatory mechanism of dietary SAN supplementation on meat quality through transcriptomic and gut microbiome analysis, thereby providing a basis for regularing meat quality with MCE. 240 1-day-old broilers were divided into 4 groups according to different doses of SAN (0, 0.225, 0.75, and 2.25 mg/kg). The results indicated that SAN significantly improve the physicochemical quality indicators of breast and thigh muscle in broilers, improved the serum biochemical indexes. Through transcriptome sequencing analysis of the liver and ileum tissues of broilers, we found that the differentially expressed genes induced by SAN were mainly enriched in lipid metabolism, which were related to the peroxisome proliferator-activated receptor ( PPAR ) pathway. It reconfirmed that SAN can regulate lipid metabolism in the body by promoting the expression of genes related to cholesterol metabolism, fatty acid transport and oxidation by RT-PCR, this ultimately affects the physicochemical quality of muscle. Additionally, through 16S rRNA sequencing analysis, we found that dietary addition of SAN increased the relative abundance of Bacteroides, Lactobacillus and unclassified_f_Lachnospiraceae , while decreased the relative abundance of Alistipes in ceca. To further investigate the impact of gut microbiota on lipid metabolism, we conducted a correlation analysis of PPAR pathway factor expression in cecum tissue and microflora structure. The results showed that Bacteroides exhibited a positive correlation with the expression of most genes in the PPAR signaling pathway. Unclassified_f__Lachnospiraceae is positively correlated with PPARγ, Cytochrome P450 family 7 subfamily A member 1 ( CYP7A1 ) and Acyl-CoA synthetase long-chain family member 5 ( ACSL5 ). In conclusion, dietary addition of SAN can promote the genes expression of the PPAR pathway, target the regulation of intestinal microflora structure and abundance and regulate lipid metabolism, thereby improving meat quality of broilers. [ABSTRACT FROM AUTHOR]

Morphine, sanguinarine and chelerythrine are benzylisoquinoline alkaloids (BIAs), and these compounds possess strong biological activities. (S)-scoulerine is a commonly shared precursor of these compounds, and berberine bridge enzyme (BBE) is a key rate-limiting enzyme in the synthesis of (S)-scoulerine. We isolated the BBE gene from Macleaya cordata (McBBE) and used CEN.PK2-1C as a chassis strain. We compared the catalytic efficiency of five codon-optimized McBBE genes in Saccharomyces cerevisiae and finally obtained a yeast strain (YH03) that exhibited a 58-fold increase in yield (1.12 mg/L). Then, we truncated the N-terminus of McBBE by 8 and 22 amino acids and found that with the increase in the number of N-terminal truncated amino acids, the production of (S)-scoulerine gradually decreased. Additionally, we used CRISPR-Cas9 to integrate the McBBE gene at the delta site of the S. cerevisiae genome to achieve stable genetic inheritance and found that the yield of (S)-scoulerine was not significantly increased in the integrated strain. In conclusion, our work achieved high-efficiency expression of McBBE in S. cerevisiae, explored the influence of N-terminal truncation on the yield of (S)-scoulerine, and obtained a genetically stable S. cerevisiae strain with high McBBE expression. This study provides a reference for further complex metabolic engineering optimization and lays a foundation for the efficient biosynthesis of BIAs. [ABSTRACT FROM AUTHOR]

Sanguinarine (SAN) and chelerythrine (CHE) have been widely used as substitutes for antibiotics for decades. For a long time, SAN and CHE have been extracted from mainly Macleaya cordata, a plant species that is a traditional herb in China and belongs to the Papaveraceae family. However, with the sharp increase in demand for SAN and CHE, it is necessary to develop a new method to enhance the supply of raw materials. Here, we used methyl jasmonate (MJ), salicylic acid (SA) and wounding alone and in combination to stimulate aseptic seedlings of M. cordata at 0 h, 24 h, 72 h and 120 h and then compared the differences in metabolic profiles and gene expression. Ultimately, we found that the effect of using MJ alone was the best treatment, with the contents of SAN and CHE increasing by 10- and 14-fold, respectively. However, the increased SAN and CHE contents in response to combined wounding and MJ were less than those for induced by the treatment with MJ alone. Additionally, after MJ treatment, SAN and CHE biosynthetic pathway genes, such as those encoding the protopine 6-hydroxylase and dihydrobenzophenanthridine oxidase enzymes, were highly expressed, which is consistent with the accumulation of SAN and CHE. At the same time, we have also studied the changes in the content of synthetic intermediates of SAN and CHE after elicitor induction. This study is the first systematic research report about using elicitors to increase the SAN and CHE in Macleaya cordata. [ABSTRACT FROM AUTHOR]