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ObjectiveTo compare the effects of total alkaloids， matrine， and sophoridine extracted from Sophora alopecuroides on the activity of pheochromocytoma cells （PC12 cells）. MethodThe effect of S. alopecuroides total alkaloids， matrine， and sophoridine at concentrations of 2， 1， 0.5， 0.25， 0.125， and 0.062 5 g·L-1 on the proliferation of PC12 cells was evaluated using the 3-（4，5-dimethylthiazol-2-yl）-2，5-diphenyl tetrazolium bromide （MTT） assay. The lactate dehydrogenase （LDH） leakage rate was measured by enzyme-linked immunosorbent assay （ELISA）. Flow cytometry was used to assess cell apoptosis rate， cell cycle distribution， and intracellular Ca2+ levels. Real-time quantitative polymerase chain reaction （Real-time PCR） was performed to determine the mRNA transcription levels of B-cell lymphoma-2 （Bcl-2） and Bcl-2-associated X protein （Bax）. Protein expression levels of apoptosis-related proteins Caspase-3， Caspase-8， Bcl-2， and Bax were detected by Western blot. ResultCompared to the control group， S. alopecuroides total alkaloids， matrine， and sophoridine inhibited the proliferation of PC12 cells， increased LDH leakage rate， enhanced intracellular Ca2+ fluorescence intensity， and induced cell apoptosis in concentration-dependent manner （P<0.05， P<0.01）. Among them， S. alopecuroides total alkaloids had the strongest inhibitory effect on cell proliferation and induction of apoptosis in PC12 cells （P<0.01）. After treatment with S. alopecuroides total alkaloids， matrine， and sophoridine， the cell cycle progression of PC12 cells was arrested at G1/G0 in the S. alopecuroides total alkaloids group， and at G1/S in the matrine and sophoridine groups. The expression levels of Bax mRNA were significantly increased （P<0.05， P<0.01）， while the expression levels of Bcl-2 mRNA were significantly decreased （P<0.05， P<0.01）. All treatments significantly downregulated the expression of the anti-apoptotic protein Bcl-2 （P<0.05， P<0.01） and upregulated the expression of the pro-apoptotic proteins Bax， Caspase-3， and Caspase-8 （P<0.05， P<0.01）， with the most significant protein expression changes observed in the S. alopecuroides total alkaloids group. ConclusionAmong the S. alopecuroides total alkaloids， matrine， and sophoridine， S. alopecuroides total alkaloids exhibit the strongest inhibitory effect on the activity of PC12 cells， and its mechanism of action may be related to the inhibition of anti-apoptotic protein expression， upregulation of pro-apoptotic protein expression， and activation of the mitochondrial Caspase-8 apoptotic pathway.
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Sophora alopecuroides, a plant of the family Leguminosae, is one of the Daodi herbs in Ningxia. The active constituents of Sophora alopecuroides are abundant and complex, including alkaloids, flavonoids, volatile oils, steroids, polysaccharides, fatty acids and so on. In recent decades, a great number of domestic and overseas studies have been carried out on Sophora alopecuroides alkaloids, which have anti-hepatitis, anti-liver fibrosis, anti-cirrhosis, anti-liver failure and anti-liver cancer and other pharmacological effects. Clinically, Matrine-related drugs are used to treat hepatitis B virus infection and other diseases. This review aims to summarize the main active ingredients of Sophora alopecuroides, mainly focusing on the research progress in their treatment activities for liver diseases.
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Introduction@#<i>Sophora alopecuroides L</i> has broadly been utilized in traditional medicine and all crude drugs including root, herb, and seed are used to treat numerous diseases. This herb is included in 181 Tibetan-Mongolian medicinal prescriptions and ranks 8<sup>th</sup> among Mongolian medicinal plants in terms of frequency of administration. The <i>S.alopecuroidesL . </i> root standard was developed by the Institute of Traditional Medicine and Technology in 2017 and approved by “ҮФӨ-0307-2017”. Herb and seed are still used in medicine. Therefore, their standard parameters need to be determined and verified.@*Materials and methods@#The quantitative pharmacognosy analysis of herb and seed was carried out in accordance with the methodology specified in the “General requirements for medicinal plant raw materials” of the National Pharmacopoeia of Mongolia. To determine the total alkaloid in standard matrine, a bromcresol green complex was formed, which was measured by spectrophotometer.@*Conclusion@#By developing, standards for the crude drugs of herb and seed of <i>S.alopecuroides L. </i> which are included in numerous medicinal prescriptions, will confirm the rationale for the use of medicinal raw materials and to expand the utilization’s possibilities.
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Sophora alopecuroides belongs to the genus Sophora of the family Papilionoideae. It is mainly distributed in the desert and semidesert areas of northern China, and has high medicinal value and ecological function. Previous studies have reported the chemical composition and ecological functions of S. alopecuroides. However, only a few reports are available on the genomic information of S. alopecuroides, especially the chloroplast genome, which greatly limits the study of the evolutionary relationship between other species of Papilionoideae. Here, we report the complete chloroplast genome of S. alopecuroides. The size of the chloroplast genome is 155,207 bp, and the GC content is 36.44%. The S. alopecuroides chloroplast genome consists of 132 genes, including 83 protein-coding genes, 41 transfer RNA (tRNA) genes, and eight ribosomal RNA (rRNA) genes. Phylogenetic analysis revealed the taxonomic position of S. alopecuroidesin Papilionoideae, and the genus Sophora and the genus Ammopiptanthus were highly related. Comparative genomics analysis revealed the gene rearrangement in the evolution of S. alopecuroides. The comparison between S. alopecuroides and the species of the Papilionoideae identified a novel 23 kb inversion between the trnC-GCA and trnF-GAA which occurred before the divergence of Sophora and Ammopiptanthus of Thermopsideae. This study provided an essential data for the understanding of phylogenetic status of S. alopecuroides.
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To further study and fully exploit the medicinal plant Sophora alopecuroides, the molecular markers related with the phenotypic traits of alkaloid content in S. alopecuroides should be detected. In this study, SSR molecular markers were used to analyze the genetic diversity and genetic structure of 23 S. alopecuroides populations, in combination with the association analysis between molecular markers and the alkaloid contents. The results showed that P, H, I, G_(st) and N_m values were 40.10%, 0.335 3, 0.504 5, 0.433 7 and 0.625 9 respectively, in 23 S. alopecuroides populations. This indicated that there was less gene exchange and higher genetic differentiation among different S. alopecuroides populations. The results of SSR unweighted pair-group method with arithmetic means(UPGMA) cluster showed that the S. alopecuroides populations relationship from Xinjiang was far from the populations of other regions, but the populations of S. alopecuroides from Gansu, Inner Mongolia and Qinghai were closely relevant to those from Ningxia. The 23 populations were further divided into 2 genetic subpopulations by the population structure analysis. Through association analysis, a total of 26 loci in 13 SSR markers were found to be significantly associated(P<0.005)with the content of MA, OMA, SC and OSC, and the rate of explanation on the phenotype variance of related markers ranged from 36.45% to 77.93%. Among the locus, 1 each were related with MA and OSC content at interpretation rate reached as high as 50% with high threshold(P<0.000 1). These results could provide support for the discovery of important genes in the alkaloid biosynthetic and metabolic pathway of S. alopecuroides.
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Alkaloids , China , Genetic Variation , Microsatellite Repeats , Phenotype , Plants, Medicinal , Sophora , GeneticsABSTRACT
Objective: To investigate the mechanism by which total alkaloids of Sophora alopecuroides (TASA) and matrine (MT) impair biofilm to increase the susceptibility of Staphylococcus epidermidis (S. epidermidis) to ciprofloxacin. Methods: The minimum biofilm inhibitory concentration (mBIC) was determined using a 2-fold dilution method. Structure of biofilm of S. epidermidis was examined by Confocal Laser Scanning Microscope (CLSM). The cellular reactive oxygen species (ROS) was determined using a DCFH-DA assay. The key factors related to the regulation of ROS were accessed using respective kits. Results: TASA and MT were more beneficial to impair biofilm of S. epidermidis than ciprofloxacin (CIP) (P < 0.05). TASA and MT were not easily developed resistance to biofilm-producing S. epidermidis. The mBIC of CIP decreased by 2–6-fold following the treatment of sub-biofilm inhibitory concentration (sub-BIC) TASA and MT, whereas the mBIC of CIP increased by 2-fold following a treatment of sub-BIC CIP from the first to sixth generations. TASA and MT can improve the production of ROS in biofilm-producing S. epidermidis. The ROS content was decreased 23%−33% following the treatment of sub-mBIC CIP, whereas ROS content increased 7%−24% following treatment with TASA + CIP and MT + CIP combination from the first to sixth generations. Nitric oxide (NO) as a ROS, which was consistent with the previously confirmed relationship between ROS and drug resistance. Related regulatory factors-superoxide dismutase (SOD) and glutathione peroxidase (GSH) could synergistically maintain the redox balance in vivo. Conclusion: TASA and MT enhanced reactive oxygen species to restore the susceptibility of S. epidermidis to ciprofloxacin.
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OBJECTIVE: To optimize extraction process for active ingredients in seeds of Sophora alopecuroides, to provide a reference for scale production. METHODS: Active ingredients from Sophora alopecuroides were extracted by ethanol, with average yield of oxysophocarpine and oxymatrine as index, some factors affecting index were firstly evaluated by Plackett-Burman design, then taking oxysophocarpine and oxymatrine as indexes respectively, extraction conditions were optimized by Box-Behnken design, experimental data was fitted by multiple linear regression and binomial formula fitting, extraction process was optimized by response surface method, and prediction was carried out through comparing the observed and predicted value. RESULTS: Extracting times, crushing degree and solvent times had significant effects on yields of oxysophocarpine and oxymatrine; binomial equation fitted well with good predictability. optimum extraction technology of Sophora alopecuroides was as following:crushed through 65 mesh sieve, extracted 4 times with 12-fold the amount of 60% ethanol for 2 h each time; yield of oxysophocarpine and oxymatrine was 92.3%, 78.6% respectively, both deviations were small by comparing with the predicted value. CONCLUSION: This extraction process is reasonable and feasible by Plackett-Burman design and response surface analysis with good predictability. This study can provide experimental basis for further scale production of Sophora alopecuroides.
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Objective: To establish the chemical fingerprint of Sophora alopecuroides extracts based on high performance liquid chromatography (HPLC), and determine the LD50 of different extracts of S. alopecuroides to analyze its “spectrum toxicity” relationship. Methods: A series of extracts were prepared by 75% ethanol reflux (ER), water decoction (WD), 75% ethanol ultrasound (EU) and water ultrasound (WU), and their fingerprints were established to determine the acute toxicity LD50 of different extracts. The relationship between chemical composition and acute toxicity LD50 of S. alopecuroides extracts were studied by means of fingerprint similarity evaluation system. Results: The LD50 of ER, WD, EU, and WU extracts were 38.397, 24.994, 18.536, and 19.957 g/kg, respectively. The ocular lesions of mice viscera were mainly manifested in liver and kidney, and the toxicity of ER extracts was the greatest. The 10 common peaks of S. alopecuroides extracts can be divided into two categories; Peaks 4 and 10, oxymatrine and sophocarpidine were negatively correlated with acute toxicity LD50. Conclusion: The spectral toxicity relationship analysis method of S. alopecuroides was constructed. The unidentified peaks 4, 10 and oxymatrine and sophocarpidine were the main chemical components of the toxicity reaction, which laid a good foundation for clinical application and scientific and rational development of S. alopecuroides.
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Objective: In this study, flow cytometry and K-mer analysis were used to estimate genome size and hybridity percentage of Sophora alopecuroides, so as to provide reference for its genome sequencing. Methods: Glycine max served as an internal reference, the multiple relationship between DNA content in Glycine max and S. alopecuroides cells was detected by flow cytometry, and the genome size of S. alopecuroides was calculated. Genome sequencing of S. alopecuroides was carried out using NGS technology, genome size and hybridity percentage of S. alopec uroides were estimated by K-mer analysis. Results: The genome size of S. alopecuroides was about 1 749 Mb measured by flow cytometry. K-mer analysis showed that the genome size of S. alopecuroides was about 1 648 Mb, the hybridity percentage was 1.12%, and the percentage of repetitive sequence was high. Conclusion: The genome size of S. alopecuroides was about 1.7 Gb with high hybridity percentage. The technology of combining Illumina and PacBio is recommended for future genome sequencing.
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The Chinese herbal Sophora alopecuroides is widely used to clean intestine and eliminate dampness, and it has good therapeutic effects on treating bacillary dysentery and inflammatory bowel disease, etc. in clinics. However, the mechanism of treatment is not yet well understood. The present study was aimed to explore the mechanism of Sophora alopecuroides treatment of large intestine dampness-heat syndrome (LIDHS). The LIDHS model was performed by the comprehensive factors, including high temperature and humidity environment, high-sugar and high-fat diet, and intraperitoneal injection of Escherichia coli. The blood routine, serum proinflammatory cytokine levels and histopathological changes of intestine were detected and observed. Meanwhile, the serum metabolomic approach was conducted using the method of ultra performance liquid chromatography coupled to quadrupole time-of-flight mass/mass spectrometry (UHPLC-Q/TOF-MS/MS). The results showed that Sophora alopecuroides has good therapeutic effects on the LIDHS rat models. After treatment with Sophora alopecuroides, the abnormality of blood routine indexes as well as proinflammatory cytokines, including IL-1β, IL-2, IL-6 and TNF-α in vivo, tended to be normal, and the histopathological changes of intestine were improved. Through metabolic profiling and protocol analysis, 9 potential metabolic markers may be closely related with the treatment mechanism of Sophora alopecuroides on this disease, including taurine, L-tryptophan, LysoPE, LysoPC, LPA, DG, chenodeoxycholic acid disulfate, traumatic acid and 7-ketodeoxycholic acid, which were involved in taurine and hypotaurine metabolism, glycerophospholipid metabolism, glycerolipid metabolism, tryptophan metabolism and primary bile acid biosynthesis etc. The serum metabolomic approach can be applied to clarify the therapeutic mechanism of Sophora alopecuroides on LIDHS, and provide the theoretical basis for Sophora alopecuroides in clinical practice.
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It was aimed at exploring the potential pharmacological effects of alkaloids in Sophora alopecuroides by means of network pharmacology in this study. The main alkaloids in S. alopecuroides were collected for analysis of drug properties, prediction of potential targets and screening of signaling pathways. DAVID analysis tool combined with KEGG database was used to annotate and analyze the signaling pathway. The alkaloids-targets-signaling pathways network was built through Cytoscape software. Results showed that 17 alkaloids in S. alopecuroides involved 49 targets (170 times in all) and 22 important signaling pathways. Three nodes in model of network pharmacology were cross-linked, and the metabolic pathways were coordinated and regulated by each other. It indicated that alkaloids in S. alopecuroides may have therapeutic effect on diseases of cancer, metabolic disorder, endocrine system, digestive system, nervous system and so on.
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Alkaloids , Pharmacology , Phytochemicals , Pharmacology , Signal Transduction , Sophora , ChemistryABSTRACT
The aim of this paper was to investigate the potential pharmacological effect of flavonoids in Sophora alopecuroides by network pharmacology. This study predicted the potential targets of 11 flavonoids of S. alopecuroides with help of reversed pharmacophore matching target recognition service platform (PharmMapper). The pathway information was acquired from DAVID and KEGG databases. Cytoscape software was used to construct the "ingredient-target-pathway" network of flavonoids active components of S. alopecuroides. The flavonoids active components of S. alopecuroides play anti-inflammatory, blood sugar regulating and other pharmacological effects by regulating 62 targets (such as INSR，KDR，MET) and intervening 44 pathways, such as B cell receptor signaling pathway, insulin signaling pathway, neurotrophin signaling pathway, and T cell receptor signaling pathway. In this study, the mechanism of "muti components-multitargets-multiple pathway" of flavonoids was studied. It reflects the multi-components, multi-targets and multiple pathway features of traditional Chinese medicine. Meanwhile, it provides a scientific basis for the elucidation the mechanism of S. alopecuroides as a medicine, and the development and utilization resources of S. alopecuroides.
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Objective To establish a method of detecting the expression of Lysine decarboxylase (LDC) -a key enzyme for the synthesis of alkaloid in the host promoted by the endophytic fungal elicitor of Sophora alopecuroides by using real-time fluorescence quantitative PCR (qRT-PCR). Methods Target gene primers QLDC-F/QLDC-R and reference gene primers Lectin-F/Lectin-R were designed according to LDC and Lectin gene sequences of S. alopecuroids; Five-fold gradient dilution of cDNA was used as the standard sample for the construction of the standard curve of target gene and the reference gene. Reaction system and reaction conditions of qRT-PCR were optimized, and the sensitivity of semi-quantitative PCR and qRT-PCR were analyzed and compared. Under different eliciting time of endophytic fungal elicitors NDZKDF13 of S. alopecuroides, the content of oxymatrine in the host was determined by HPLC, the expression of LDC gene was detected by qRT-PCR, and the relationship between LDC gene expression and the accumulation of OMA was analyzed. Results The results of qRT-PCR were better when the cDNA content in the system was 200 ng/μL and the annealing temperature was 61 ℃. The standard curve of the target gene and the reference gene was constructed, in which the cycle threshold and template concentration showed a good linear relationship, the amplification efficiency was above 99%, and the sensitivity was 25 times that of semi-quantitative PCR. Under the induction effect of endophytic fungal elicitor NDZKDF13, expression of host LDC gene reached the peak on the 6th day, which was 25.58 times that of the control. The increase of OMA content lagged the change of the LDC gene expression and reached the highest amount on the 9th day after the induction. Conclusion The qRT-PCR technique was successfully applied to the functional gene research of S. alopecuroides. Through the optimization of various conditions, a platform for accurate and simple detection of functional gene expression in S. alopecuroides was established.
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Objective To evaluate the effect of total alkaloids of Sophora alopecuroides L on the nervous behavior and the expression of neurotransmitters in rats. Methods 48 male SD rats were randomly divided into 4 groups:blank group,total alkaloids of Sophora alopecuroides L treatment with 4 mg·kg-1 ·d-1 ( low dose group) ,8 mg·kg-1 ·d-1( medium dose group) and 16 mg·kg-1 ·d-1( high dose group) groups. After successive intragastric administration for 30 days,the locomotor activity was applied to test the nervous behavior and emotional state of rats in each group. After behavioral tests were finished,the contents of trypto-phan (Trp),5-hydroxytryptophan (5-HTP),5-serotonin (5-HT),5-hydroxyindoleacetic acid (5-HIAA), norepinephrine (NE),epinephrine (E) and dopamine (DA) were detected by ELISA in serum and brain. Results In the experiment of locomotor activity,compared with blank group ((95.33±12.75) times),the numbers of horizontal movement of Sophora alopecuroides L in medium and high dose group ( ( 61. 64 ± 5.91),(64.62±5.79)times both P0.05). Correlation analysis indicated that the degree of au-tonomic activity in rats with the content of 5-HT,5-HIAA and DA in serum was negatively correlated (P<0.05, P<0.01) ,the degree of emotional stress and the content of 5-HT,5-HIAA in brain was negatively cor-related (P<0.05, P<0.01) . Conclusion The total alkaloids of Sophora alopecuroides L can reduce the ac-tivity of rats and increase the degree of emotional stress. And the mechanism may be correlated with the in-creasing level of 5-HT and 5-HIAA in serum and brain.
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Objective: To study the structure of Sophora alopecuroides polysaccarides (SAP) and its antitumor activity on CT26 transplanted tumor in mice. Methods: The structure was characterized by high-performance gel permeation chromatography (HPGPC), gas chromatography-mass spectrometer (GC-MS), infrared spectroscopy (IR), and nuclear magnetic resonance spectroscopy (NMR). And then the anticancer activity of CT26-bearing mice was investigated in vivo. Results: The results showed that SAP was consisted of D-mannopyranose and D-galactopyranose residues. The main chain of the galactomannan comprises β-(1,4)-linked D-mannopyranose residues, with branches of galactose, linked to the carbohydrate core through α-D-Gal (1,6) linkage. SAP could inhibit the tumor proliferation of CT26-bearing mice in a dose-dependent manner with inhibitory rate of 44.03% at high dose. Conclusion: Structure determination of SAP provides the theoretical basis for structure-activity relationship study. And it is expected to be further developed as a new antitumor drug with high efficiency and low toxicity.
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OBJECTIVE:To select the optimum conditions for germination test of Sophora alopecuroides seed,and to provide reference for formulating seed test rule and standardization of S. alopecuroides. METHODS:S. alopecuroides seeds were soaked in 98% H2SO4 for 30 min and 35 ℃ warm water for 29 h,and then treated under different temperature conditions (15 ℃,20 ℃, 25℃and 30℃),different germination beds(on paper,between paper,on sand,in sand)and different light conditions(2 000 lx lighting 16 h,dark). Optimal germination condition was screened by using germination rate,germinative potential and germinative index as indicators. S. alopecuroides seeds were cultured under this condition for 7 days,and then germination rate was determined. RESULTS:Different temperatures had no significant effect on germination rate,but influenced germinative potential and germina-tive index;those indicators reached maximal value at 20 ℃. Different germination beds affected each indicator,and under condi-tion of on sand,those indicators were the highest. Light treatment had no significant effect on germination indicators. Under suit-able condition,the seed sprouted since first day of germination bed treatment;germination rate was more than 90% on second day,and reached maximal value on fifth day and didn’t increase any longer. CONCLUSIONS:The suitable condition of seed ger-mination was soaking in 98% H2SO4 30 min+35 ℃ warm water for 29 h,on sand under light at 20 ℃. Initial count on second day of germination bed treatment and final count on forth day were analyzed statistically as well as germination rate. This method can be used as standard quality test of the seed of S. alopecuroides.
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According to the research strategy of resource chemistry of Chinese medicinal materials and Chinese medicinal resources recycling utilization, this study intends to explore the potential resource-oriented utilization value of the seed of Sophora flavescens by contrasting with its kindred plant S. alopecuroides. This study established a rapid UPLC-Q-TOF-MS/MS and UPLC-TQ-MS/MS method to determine the alkaloids in the seed of S. flavescens. Results of UPLC-Q-TOF-MS/MS analysis showed that the alkaloids in the seed of S. flavescens were highly similar with S. alopecuroides.In the determination of 7 kinds of alkaloids, the total content was 11.203 and 15.506 mg•g⁻¹ in the seed of S. flavescens and S. alopecuroides, respectively. The content of oxymatrine, oxysophocarpine and sophoridine is high in the seed of S. flavescens. The results indicated that the seeds of S. flavescens. could be an important material resource to obtain alkaloids.
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Objective: To deliver multiple component drugs to colon site and sustain a synchronous release for better therapeutic effect. For achieving this purpose, colon specific pellet containing total alkaloids of Sophora alopecuroides (TASA) was prepared. Methods: The pellet was prepared by extrasion-spheronizing and subsequently coated with three layers of two polymers. Results: The pellet core consisted of 40% TASA, 1:2 in ratio of Bletilla striata polysaccharide (BSP), an enzyme-degradable material, to microcrystalline cellulose (MCC), filler, and 1% CMC-Na solution as binder by optimization. Concerning of the three coated layers, the outer layer was coated with Eudragit RS30D for controlling drug release in colon, the intermediate layer and the inner layer were coated with same polymer, Eudragit S100, for preventing drug release in upper gastrointestinal tract, which required 23.2%, 21.7%, and 9.3% weigh gain, respectively. The coated pellets released 1.20% of sophoridine and 1.98% of matrine in media mimicking the stomach condition for 2 h, and 23.88% of sophoridine and 22.91% of matrine in media mimicking the intestine for 3 h and finally 90.25% of sophoridine and 89.94% of matrine in colonic conditions within 24 h. And the similarity factor f of sophoridine and matrine of release curve for investigated formulation was internal in (50-100) and > 80, demonstrating that sophoridine and matrine in formulation achieved a synchronous release. Conclusion: The coated pellets achieve a certain colon-specific release and synchronous release.
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Objective:To study the quality standard for Fujie lotion. Methods:The qualitative identification of Sophora alopecu-roides L, Cnidium monnieri ( L. ) Cuss and borneol was detected by TLC. The quantitative determination of matrine was detected by HPLC. Results:The identification by TLC was highly specific, and the content determination method was accurate and repeatable. The linear range of matrine was 0. 013 0-1. 30 mg·ml-1, and the average recovery was 97. 1% with RSD of 1. 7%(n=6). Conclu-sion:The standard can effectively control the quality of Fujie lotion.
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OBJECTIVE: To study the effect of lysine decarboxylase (LDC) gene on the accumulation of matrine (MA) and oxymatrine (OMA) in cotyledon of Sophora alopecuroides L germinating seeds. METHODS: The S. alopecuroides germinating seeds were stressed by different mass fractions of PEG 6000, and the contents of MA and OMA were determined by high performance liquid chromatography (HPLC) and the expression level of LDC was analyzed by real-time fluorescence quantitative PCR (qPCR) after 72 h treatment. RESULTS: The contents of MA and OMA decreased in the cotyledon under light stress (PEG mass fraction 20%). The analysis of qPCR revealed that the LDC expression level was decreased first, and then increased with the stress rising. The changes of the contents of MA and OMA were parallel with the expression level of LDC especially under light and severe stress. CONCLUSION: There is certain association between the accumulation of MA and OMA and the gene expression quantity of LDC. The results is of significance for illustrating the role of LDC in the biosynthetic pathways of MA and OMA.