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We investigated the effect of methyl jasmonate (MeJA) on the content of asperosaponin VI and the expression of genes involved in its synthesis. Dipsacus aspero seedlings were treated with MeJA at different concentrations of 50, 100, 150, 200 and 300 μmol·L-1, and leaves and roots were sampled following treatment for 1, 3 and 5 days. The content of asperosaponin VI and superoxide anion in the roots, malondialdehyde (MDA) content in leaves and superoxide dismutase were determined. The results show that 150 μmol·L-1 MeJA significantly increased the accumulation of asperosaponin VI in roots. The content of asperosaponin VI was greatest after treatment for 3 days, and was 2.16 times higher than the control. After MeJA treatment, SOD activity decreased and MDA content increased in leaves. Moreover, superoxide anion content in roots increased. The expression of squalene epoxidase (DaSE1) and geranyl diphosphate synthase (DaGPS), key enzymes in the synthesis of asperosaponin VI, were up-regulated compared with the control group. These results indicate that an optimal concentration of 150 μmol·L-1 MeJA increases the accumulation of asperosaponin VI by up-regulating the expression of key enzymes involved in the synthesis of asperosaponin VI, which facilitates resistance to adversity stress stimulated by MeJA.
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Objective: To obtain the transcriptome sequence database induced by methyl jasmonate (MeJA) and identify the genes related to the biosynthesis of triterpenoid saponin in Polygala tenuifolia. Methods: The seedlings grown for 30 d were respectively treated with sterile water, 50 μmol/L MeJA and 100 μmol/L MeJA for 24 h. The transcriptome data of seedlings of P. tenuifolia were obtained by Illunima HiseqTM 2000 150PE sequencing and de novo splicing of Unigene was realized by Trinity software. The GO classification, KOG functional annotation, metabolism of KEGG metabolic pathway, protein function annotation analysis, differential gene analysis and screening were completed based on BLAST. Results: A total of 52.19 Gb clean data were obtained after the transcriptome of P. tenuifolia being assembled by Trinity software, and 54 426 Unigenes were assembled with an average length of 1 604 bp. All Unigenes were annotated in the public databases NR, NT, KEGG, Swissprot, GO, and Pfam. Through differential analysis of genes responding to MeJA, a total of 3 390 differentially expressed genes (DEGs) were found, of which 1 287 were up-regulated and 2 103 were down-regulated. The response of DEGs showed that the total number and up-regulated number of P. tenuifolia seedlings treated by 100 μmol/L MeJA was the highest. KEGG enrichment analysis showed that differentially expressed genes were significantly enriched in metabolic pathways including phenylpropanoid biosynthesis, cysteine and methionine metabolism, starch and sucrose metabolism, carbon fixation in photosynthetic organisms and terpenoid backbone biosynthesis. Furthermore, a total of 59 Unigenes involved in anthraquinones biosynthesis were found according to the assignment of KEGG pathway. Expression analysis showed that AACT, HMGS, HMGR, MK, PMK, MPD, DXS, IDI, FPPS, SQS, SE and β-AS were up-regulated after being induced by MeJA. Conclusion: In this study, the transcriptome of P. tenuifolia seedlings treated with methyl jasmonate was analyzed, and candidate genes related to triterpenoid skeleton biosynthesis of P. tenuifolia were obtained. MeJA can induce the expression of genes related to triterpenoid skeleton synthesis, which provided a wealth of data resources for the molecular biology research and also laid the foundation for the analysis of the secondary metabolic pathways of triterpenoid saponins in P. tenuifolia.
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To explore the mechanism hydroxysafflor yellow A (HSYA) biosynthesis and regulation, the effect of methyl jasmonate (MeJA) treatment on gene expression related to the biosynthesis of hydroxysafflor yellow A (HSYA) was analyzed, and expression differences in genes involved in HSYA biosynthesis in safflower of different colors was quantified. MeJA at concentrations of 0, 50, 100, and 200 μmol·L-1 was sprayed onto safflower florets to determine the optimal concentration of MeJA. Safflower was treated with 100 μmol·L-1 MeJA and florets were harvested 0, 3, 6, 12 and 24 h after treatment. The content of MeJA was determined by high performance liquid chromatography (HPLC). RNA was extracted from safflower florets treated with 100 μmol·L-1 MeJA for 6 h. The transcription of key genes involved in the biosynthesis of HSYA was quantified by qRT-PCR and differentially expressed genes were identified. The content of HSYA increased after treatment with MeJA, with 100 μmol·L-1 MeJA treatment for 6 h having the greatest effect on HSYA accumulation. qRT-PCR results showed that MeJA could significantly increase the transcription of HSYA biosynthesis genes including PAL2, PAL4, 4CL2, 4CL4, 4CL5, CHS3, CHS4 and CHI2. The content of HSYA differed between safflowers of different colors with a trend of red>orange-yellow>yellow>white. The results of qRT-PCR showed that the expression of CHS1 and CHI2 in red, orange and yellow safflower was significantly higher than that in white safflower. These results indicate that MeJA promotes the accumulation of HSYA by up-regulating the expression of genes involved in the biosynthesis of HSYA such as PAL2, PAL4, 4CL2, 4CL4, 4CL5, CHS3, CHS4 and CHI2, and the variation of HSYA content in safflower of different colors was related to a difference in the level of expression of CHS1 and CHI2.
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In order to analyze the expression of genes involved in steroidal saponin biosynthesis pathway in Polygonatum cyrtonema tubers, it is very important to select internal reference genes that are stably expressed at different development stages and in response to abiotic stress. According to the previously established P. cyrtonema transcriptome database and reported internal reference genes in plant, this study systematically analyzed eight candidate internal reference genes including histone H2 A, glyceraldehyde-3-phosphate dehydrogenase, ACTIN, β-tubulin, ubiquitin-conjugating enzyme-E2-10, elongation factor 1-alpha isoform, 18 S rRNA and α-tubulin 4 for expression stability in P. cyrtonema tubers at different development stages and in response to methyl jasmonate(MeJA) stress by using Real time fluorescence quantitative PCR(qPCR). Based on the statistical analysis of qPCR results by using GeNorm, NormFinder and BestKeeper softwares, the expression of ubiquitin-conjugating enzyme-E2-10 and elongation factor 1-alpha isoform are the most stable in P. cyrtonema tubes at different development stages and in response to MeJA stress. The two internal reference genes were further validated by analyzing the expression of 4 genes involved in steroidal saponin biosynthesis pathways. In conclusion, ubiquitin-conjugating enzyme-E2-10 and elongation factor 1-alpha isoform can be used as the most appropriate internal reference genes for qPCR analysis in P. cyrtonema. This study also provide a foundation for future investigate the molecular mechanism of steroidal saponin biosynthesis pathways in P. cyrtonema.
Subject(s)
Gene Expression Profiling , Polygonatum , Real-Time Polymerase Chain Reaction , Stress, Physiological , TranscriptomeABSTRACT
Centella asiatica is an important medicinal plant which contains various phytocompounds. Asiatic acid and asiaticosideare two major compounds which are responsible for its various pharmaceutical activities. The present study analyzesthe effect of elicitor, i.e., methyl jasmonate on the synthesis of asiaticoside and asiatic acid (ATA) in shoot, callus, andcell suspension cultures of C. asiatica. A high-performance liquid chromatography analysis showed that the elicitationwith 100 µM concentration of methyl jasmonate enhanced asiaticoside content by 69-fold in callus culture, 39-fold inshoot cultures, and ATA by 1.9-fold in cell suspension culture. Thus, elicitation with methyl jasmonate is an effectivemethod of increasing the rate of biosynthesis of asiaticoside and ATA in plant cell cultures of C. asiatica
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Objective To study the regulation among the content of L-borneol and four endogenous hormones and the activity of three anti-oxidant enzymes in Blumea balsamifera leaves located at different leaf positions and the concentration of inducer and the sampling time. Methods Methyle Jasmonate (MeJA) of 0.01 mmol/L, 0.10 mmol/L, 1.00 mmol/L, and 10.00 mmol/L was chosen for exogenous inducer in this experiment. The leaves of B. balsamifera located at different leaf positions (tender leaves, mature leaves, aged leaves) were experimental materials. The active content of L-borneol, the content of four endogenous hormones of auxin (IAA), abscisic acid (ABA), gibberellic acid (GA3), and zeatin (ZT), and the activity of three antioxidant enzymes of peroxidase (POD), catalase (CAT), and superoxide dismutase (SOD) were detection indexes. Results The results showed that the effect of 1.00 mmol/L MeJA on the accumulation of L-borneol was good. The changes of anti-oxidant enzymes induced by different concentrations of MeJA were complex. For the content of POD, except that the B. balsamifera leaves treated with 10.00 mmol/L MeJA were lower than that in the control, POD were significantly higher than those of the control (P < 0.05) at 72 h in other conditions. Under the induction of 10.00 mmol/L MeJA, the CAT content of the B. balsamifera leaves at three leaf positions was highest at 24 h, and the activity of CAT was decreased rapidly over time. Under the MeJA treatments of other concentrations, the activities of CAT in young leaves and old leaves were significantly lower than those in the control at 72 h, but higher than those in the control at 72 h except for the treatment of 0.1 mmol/L MeJA in mature leaves. The content of SOD in the three leaf positions was lower than the control except that the B. balsamifera leaves treated with 1 mmol/L MeJA was significantly higher than that of the control after 48 h. The rest of the concentration of superoxide dismutase were lower than the control. Low concentration of MeJA (≤ 0.10 mmol/L) could promote the accumulation of IAA, GA3, and ZT in leaves of B. balsamifera, whereas the high concentration of MeJA (≥ 10.00 mmol/L) could promote the accumulation of ABA. Conclusion Under the induction of exogenous MeJA (1.00 mmol/L), B. balsamifera can promote the accumulation of active ingredients, providing a theoretical basis for its cultivation and production.
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Objective: To study the effects of salicylic acid (SA) and methyl jasmonate (MeJA) on growth, activity of related enzymes and chemical components in Polygala callus. Methods: The callus of Polygala was used as material. After 30 d of dark culture at different concentrations of SA (0, 4, 8, 12, 16, 20, 24, 28, 32 mg/L) and MeJA (0, 100, 200, 400, 600, 800, 1 000 μmol/L), the growth of callus, anti-oxidant enzyme activity, the content of MDA, total phenolic, total flavonoid, polygalaxanthone III, and 3,6’-disinapoyl sucrose were determined. Results: MeJA inhibited the growth of Polygala callus, and 12 mg/L SA promoted the growth of Polygala callus. SA and MeJA promoted the activity of SOD, CAT, POD, and MDA in the callus of Polygala. With the increase of SA and MeJA concentration, the activities of SOD, CAT and POD increased first and then decreased, and the content of MDA continued to rise. When the concentration of SA was 20 mg/L, the activities of CAT and SOD reached the maximum, which were 248.45 U/mg and 4451.06 U/mg, respectively. When the concentration of SA was 16 mg/L, the activity of POD reached the maximum, which was 7.22 U/mg. When the concentration of SA was 32 mg/L, the MDA content reached the maximum value of 25.09 nmol/mg. When the concentration of MeJA was 600 μmol/L, the activities of CAT, SOD, and POD reached the maximum, which were 273.30, 1451.06 and 15.27 U/mg, respectively. When the concentration of MeJA was 1000 μmol/L, the MDA content reached the maximum value of 27.10 nmol/mg. SA inhibited the accumulation of total flavonoids in Polygala callus, and had no significant effect on total phenols. MeJA promoted the accumulation of total phenols and total flavonoids in Polygala callus. When MeJA concentration was 600 μmol/L, the content of total flavonoids was the highest. When the concentration of MeJA was 400 μmol/L, the total phenolic content was the highest. Both SA and MeJA promoted the accumulation of polygalaxanthone III and 3,6’-disinapoyl sucrose in Polygala callus. When the concentration of SA was 32 mg/L, the concentration of polygalaxanthone III, 3,6’-disinapoyl sucrose was the highest. When the concentration of MeJA was 1 000 μmol/L, the content of polygalaxanthone III, 3,6’-disinapoyl sucrose was the highest. Conclusion: MeJA had an inhibitory effect on the growth of Polygala callus, and 12 mg/L SA can promote this effect. SA and MeJA promoted the activity of SOD, POD, CAT, the content of MDA and the accumulation of polygalaxanthone III and 3,6’-disinapoyl sucrose in Polygala callus. SA inhibited the accumulation of total flavonoids in Polygala callus, and had no significant effect on total phenolics. MeJA promoted the accumulation of total phenolics and total flavonoids in Polygala callus.
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Objective: To clone the full-length cDNA of the ATP/ADP transporter protein (AATP) genes in Panax ginseng to provide resources and some knowledge necessary for the further gene function study. Methods: The mRNA sequence of the AATP genes in other plants were downloaded on the website of NCBI and used to perform local Blast alignment in the transcriptome of Jilin ginseng from 14 tissues. The AATP gene in Panax ginseng was cloned by PCR, and analyzed using bioinformatics software and online resources. The expression pattern of PgAATP1 gene in 14 tissues of Panax ginseng was analyzed using the expression profile of transcriptome and its expression level under methyl jasmonate was deceted by quantitative real-time PCR. Results: A full-length cDNA sequence was successfully cloned from Panax ginseng and named as PgAATP1, which was 1866 bp in length and encoded 621 amino acids. The relative molecular mass of PgAATP1 protein calculated was 67 897.23, and the isoelecric point calculated was 9.58. It was found that the protein was similar to the plastid AATP in other species. The expression of this gene was high in all tissues but higherin fruit flesh and leaf blade, and the expression of PgAATP1 gene was up-regulated by methyl jasmonate. Conclusion: We have obtained the full-length of PgAATP1 gene. This gene expressed higher in tissues of vigorous starch synthesis and responsing to methyl jasmonate.
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OBJECTIVE： To study the effects of exogenous methyl jasmonate on the contents of 4 kinds of secoiridoid substance （gentiopicrin， swertiamain， sweroside and amarogentin） and biochemical indexes in the leaves of Gentiana rigescens， and to provide scientific evidence for the application of methyl jasmonate in standardized planting of G. rigescens. METHODS： 2， 4， 6， 8， 10 d after spraying 10， 50， 100， 200 mg/L methyl jasmonate （spraying amount was 500 mL） for G. rigescens strain， the leaves of medicinal material were collected as sample， and the other leaves without spraying methyl jasmonate were collected as control. HPLC method was used to determine the contents of 4 kinds of secoiridoid substance in the leaves of G. rigescens after spraying 4 kinds of concentrations of methyl jasmonate for 10 d. The concentration of methyl jasmonate was optimized （the content of effective component was the highest）. HPLC method was adopted to determine the contents of 4 kinds of secoiridoid substance in the leaves of G. rigescens after spraying the optimal concentration of methyl jasmonate for different time. The levels of relevant biochemical indexes （SOD， POD， CAT， MDA） were determined. RESULTS： 10 d after spraying 10， 50， 100， 200 mg/L methyl jasmonate for G. rigescens strains， the contents of 4 kinds of secoiridoid substance in the leaves were increased to different extent. Compared with untreated leaves， 100 mg/L methyl jasmonate had the best effect after spraying， and the contents of gentiopicrin， swertiamain and sweroside in treated leaves were 1.88， 2.36 and 1.87 times of those in untreated leaves， respectively （P＜0.05）. 2， 4， 6， 8， 10 d after spraying 100 mg/L methyl jasmonate， the contents of 4 kinds of secoiridoid substance in treated leaves were higher than untreated leaves at corresponding stage； the content of secoiridoid had significant difference after spraying for 4 d （P＜0.05）. The contents of active components in general were relatively high after spraying for 6 d， and the contents of gentiopicrin， swertiamain， sweroside and amarogentin in treated leaves were 1.88， 1.88， 1.47， 1.82 times of those in untreated leaves， respectively （P＜0.05）. The levels of relevant biochemical indexes in treated leaves were increased significantly since 4 d of spraying， compared with untreated leaves （P＜0.05）. CONCLUSIONS： After spraying 100 mg/L methyl jasmonate for 6 d， the contents increase of 4 kinds of secoiridoid substance in the leaves of G. rigescens are most obvious， which may be associated with improving the levels of related biochemical indexes.
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Flavonoids, especially chalcones such as hydroxysafflor yellow A and carthamin are the main active ingredients of safflower. To study the biosynthesis pathway of safflower flavonoids is of great significance for the quality control of safflower. Chalcone synthase (CHS) is an enzyme that plays an important role in regulation of the synthesis of flavonoids. However, for the time being, the role of CHS is not yet clear in the biosynthesis of safflower flavonoids. As a plant signaling regulator, JA/MeJA can activate CHS gene expression in plants. CtCHS1, one of the CHS genes in safflower, was elucidated in our previous work. In our continuous search for CtCHSs functions from this plant, other CHS genes CtCHS2 and CtCHS4 in safflower were examined. The floret was stimulated with methyl jasmonate (MeJA) and the transcriptome expression of CtCHS2 and CtCHS4 was analyzed by qRT-PCR at different time points of 0, 3, 6, and 12 h after stimulation. Further metabolites under stimulation by MeJA were analyzed by UHPLC/Q-TOF-MS. The results showed that the expression of CtCHS4 in response to MeJA significantly increased at 3 and 6 h, while the expression of CtCHS2 showed a trend of decrease after induction. Meanwhile, the accumulation of rutin, hydroxysafflor yellow A, D-phenylalanine, kaempferol-3-O-β-rutinoside and carthamin increased obviously. Especially, accumulation of hydroxysafflor yellow A was increased significantly at 3, 6 and 12 h after induction (P ≥ 0.05 or 0.01), but the change in kaempferol, kaempferol-3-O-β-D-glucoside, luteolin, quercetin-3-β-D-glucoside was not significant. The accumulation of hydroxysafflor yellow A and carthamin was positively correlated with the expression abundance of CtCHS4 with Pearson correlation analysis method (r ≥ 0.8). The data suggest that CtCHS4 may be a key gene for forming hydroxysafflor yellow A and carthamin and plays an important role in the accumulation of safflower chalcones. The CtCHS4-pMAL-C5X recombinant vector was successfully expressed in BL21 (DE3) Plys to express the product naringenin in vitro under the catalytic substrates p-coumaryol-COA and malonyl-CoA. The results of this study provide a new insight into synthetic genes involved in flavonoids biosynthetic pathway to elucidate the biosynthesis pathway of safflower chalcones.
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The usage of wild ginseng (Panax ginseng C.A. Meyer) has been limited due to short supply and high price. Therefore, sufficient production as well as efficient extraction of mountain ginseng are required for the development as products. In this study, wild ginseng adventitious root cultures were prepared for efficient production with advantages of fast growth and stable production. Treatment of methyl jasmonate (MJ) to wild ginseng adventitious root cultures increased the extraction yield and antioxidative activity. Further investigation on effect of extraction conditions suggested the importance of ethanol concentration on antioxidative activity and extraction yield of MJ-treated wild ginseng adventitious root cultures. Optimized extraction condition of MJ-treated wild ginseng adventitious root cultures for maximum extraction yield and antioxidative activity was determined using response surface methodology with three-level-three-factor Box-Behnken design (BBD). Extraction of 1 g MJ-treated wild ginseng adventitious root culture with 30 ml of 9% ethanol at 30 ℃ produced 310.2 mg extract with 71.0% antioxidative activity at 100 µg/ml. Taken together, MJ-treated wild ginseng adventitious root culture is valuable source for wild ginseng usage and optimized extraction condition can be used for the development of functional products or folk remedies.
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Ethanol , Medicine, Traditional , PanaxABSTRACT
Hyoscyamine and scopolamine are important secondary metabolites of tropane alkaloid in Atropa belladonna with pharmacological values in many aspects.In this study, the seedlings of A.belladonna were planted by soil culture and treated with different concentrations of methyl jasmonate (MeJA). The contents of hyoscyamine and scopolamine,the upstream products in alkaloid synthesis,and the expression levels of key enzyme genes PMT, TR Ⅰ and H6H in secondary metabolites of A. belladonna seedlings were measured to clarify the mechanism of MeJA regulating alkaloids synthesis.The results showed that MeJA(200 μmol·L⁻¹) treatment was more favorable for the accumulation of alkaloids.The content of putrescine was almost consistent with the change of key enzymes activities in the synthesis of putrescine,the both increased first and then decreased with the increased MeJA concentration and the content of putrescine reached the highest at 200 μmol·L⁻¹ MeJA.Further detection of gene expression of PMT, TR Ⅰ and H6H in TAs synthesis pathway showed that no significant trend in PMT gene expression levels.The expression levels of TR Ⅰ and H6H in leaves and roots under 200 μmol·L⁻¹ MeJA were the highest.It can be speculated that the regulation of the formation of hyoscyamine and scopolamine by MeJA mainly through affecting the expression of key enzyme genes.Appropriate concentration of MeJA increased the gene expression of TR Ⅰ in both leaves and roots as well as H6H in roots,promoting the accumulation of alkaloids and the conversion of hyoscyamine to scopolamine.
Subject(s)
Acetates , Pharmacology , Atropa belladonna , Genetics , Metabolism , Cyclopentanes , Pharmacology , Gene Expression Regulation, Plant , Hyoscyamine , Metabolism , Oxylipins , Pharmacology , Plant Leaves , Metabolism , Plant Roots , Metabolism , Scopolamine , MetabolismABSTRACT
Objective To improve the yield of the effective medicinal ingredients of Atropa belladonna, to provide economic and efficient method for the actual production of A. belladonna, therefore to provide a basic reference for the related research on the mechanism of secondary metabolism of medicinal plants. Methods In this study, the influences of four kinds of elicitors, including methyl jasmonate (MJ), AgNO3, salicylic acid (SA), yeast extract (YE), on the accumulation of active components and the expression level of key metabolic enzyme genes, including putrescine N-methyl transferase (pmt), tropinone reductase-I (trI), and hyoscyamine-6-β-hydroxylase (h6h), were studied in hairy roots of A. belladonna. 0.5 g fresh hairy roots of A. belladonna were cultivated in B5 liquid medium. 12 d later, these mediums were replaced with the new medium containing one kind of four elicitors. Hairy roots were taken samples after 2 d to mensurate the fresh weight, dry weight, the content of tropane alkaloids, some physiological indexes, and three key genes expression level. Results MJ inhibited the growth and tropane alkaloids accumulation of hairy roots. The gene expression level of pmt and trI also decreased compared with control group (CK). The contents of tropane alkaloids and the expression level of pmt, trI and h6h were all increased compared with CK by the treatment of AgNO3, while the growth of hairy rootswas inhibited; SA contributed to the increased content of hyoscyamine, but with no obvious influence on growth of hairy roots. As to YE, the content of tropane alkaloids and the expression level of pmt, trI, h6h were all increased correspondingly. Further more, YE was benefit for the growth of hairy roots. Conclusion Elicitors had selective influence on growth and tropane alkaloids accumulation in hairy roots of A. belladonna. The best elicitor on accumulation of tropane alkaloids was AgNO3. YE could effectively improve of the growth of hairy and contents of tropane alkaloids. This study concluded that these elicitors influence the secondary metabolism of A. belladonna by regulating and controlling the expression level of some genes of key metabolic Enzyme, which could provide an effective method to enhance the medicinal composition in the culture of hairy roots of A. belladonna.
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Objective To study the effects of abiotic elicitors methyl jasmonate (MeJA) and salicylic acid (SA) on the alkaloids accumulation and related enzymes metabolism inPinellia ternata suspension cell cultures. Methods Using the leaf petioles-derived suspension cell cultures as the study object, the culture duration, concentrations of MeJA and SA were determined to get the optimal alkaloids accumulation, and the activities of metabolic enzymes IMP dehydrogenase and sAMP synthase were also measured.Results A 9-fold of dried biomass and a 3-fold of alkaloids accumulation were observed inP. ternata suspension cell cultures after culture for 21 d. Both MeJA and SA could significantly promote the accumulation of alkaloids inP. ternata suspension cells. 150 μmol/L MeJA enhanced alkaloids content (4.7 mg/gDW) by 3.6 folds in comparison with control group, whereas 50 μmol/L SA showed a 2.5-fold increase. Meanwhile, 100 μmol/L MeJA and 50 μmol/L SA promoted the increase in IMP dehydrogenase activity by 3.0 and 3.7 fold respectively, and 150 μmol/L MeJA and 100 μmol/L SA showed the increase by 2.6 and 4.4 fold respectively.Conclusion Proper adding exogenous MeJA and SA can promote the accumulation of alkaloids inPinellia ternata suspension cell cultures.
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The present investigation was carried out to evaluate anti-inflammatory and membrane stabilizing properties of methyl jasmonate (MJ) in experimental rat models of acute and chronic inflammation. The effects of MJ on acute inflammation were assessed using carrageenan-induced rat's paw edema model. The granuloma air pouch model was employed to evaluate the effects of MJ on chronic inflammation produced by carrageenan in rats. The number of white blood cells (WBC) in pouch exudates was estimated using light microscopy. The levels of biomarkers of oxidative stress, such as malondialdehyde (MDA), glutathione (GSH) and activity of antioxidant enzymes in the exudates, were determined using spectrophotometry. The membrane stabilizing property of MJ was assessed based on inhibition of hemolysis of rat red blood cells (RBC) exposed to hypotonic medium. Our results indicated that MJ (25-100 mg·kg, i.p.) produced significant anti-inflammatory activity in carrageenan-induced paw edema in rats (P < 0.05). MJ reduced the volume of pouch exudates and the number of WBC in carrageenan-induced granulomatous inflammation. It also exhibited potent antioxidant and membrane stabilizing activities. In conclusion, these findings suggest the therapeutic potentials of methyl jasmonate in disease conditions associated with inflammation and its anti-inflammatory activity may be related to its antioxidant and membrane stabilizing activities.
Subject(s)
Animals , Humans , Male , Rats , Acetates , Anti-Inflammatory Agents , Cell Membrane , Chemistry , Allergy and Immunology , Cyclopentanes , Disease Models, Animal , Edema , Drug Therapy , Allergy and Immunology , Erythrocytes , Chemistry , Glutathione , Allergy and Immunology , Malondialdehyde , Allergy and Immunology , Oxylipins , Plant Extracts , Rats, WistarABSTRACT
Changium smyrnioides is an endangered and endemic medicinal herb in China which contains rich furanocoumarins. Bergaptol, bergapten and xanthotoxin are natural furanocoumarins in Ch. smyrnioides, among which bergaptol is mainly contained in in vitro cultures while the latter ones distribute in all organs and cultures of the plant. In this study, methyl jasmonate was used to elicit furanocoumarins in both cultivated plant and suspension cells. The accumulations of biomass and 3 furanocoumarins as well as the activity of cell, phenylalanine ammonia-lyase and antioxidase were detected. The results showed that methyl jasmonate induced the biosynthesis of furanocoumarins markedly and suspension cells from petiole produced more furanocoumarins than those from leaf. In the case of suspension cells, the concentration at 100 μmol•L⁻¹ triggered the highest yield of furanocoumarins and the 10th day of the culture period was the proper time for treatment. After 4 days the yields of bergaptol, bergapten and xanthotoxin in suspension cells from petiole were enhanced to 2.83,14.04,0.62 mg•L⁻¹ respectively. The biomass and viability of treated suspension cells decreased. At the same time, the activity of antioxidase increased, which indicated that methyl jasmonate induced cell defense. In both in vivo and in vitro conditions, cells from petiole seemed to be more sensitive to methyl jasmonate treatment compared to those from leaf. Bergaptol and xanthotoxin mainly accumulated in medium and cell respectively. Bergapten was detected in both cell and medium. The elicitation treatment only enormously affected the yields but did not significantly involve the distributions of 3 furanocoumarins. This is the first systematic study focusing on the elicitation effects of methyl jasmonate and a series of changes which lead to the increase of furanocoumarins in Ch. smyrnioides cell suspension cultures. Methyl jasmonate appears to be an effective elicitor in the research and further efforts should be made to reveal the mechanism in detail.
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Methyl jasmonate (MeJA) is a lipid-derived plant hormone that mediates diverse biological phenomena. Application of MeJA onto rice spikelet could exhibit abnormal floral organ development. Although jasmonic acid (JA) has been proved to be involved in maize tassel sex determination process, the roles of JA and its precursor MeJA in maize tassel development still remain obscure. In this study, we found that tassel development was decelerated by application of 2 mM MeJA. Exogenous MeJA also influenced the number of palea and stamens of tassel spikelets. Exogenous MeJA increased the expression level of some key regulator genes, which may responsible for the phenotypic change in MeJA-treated tassel, and may mediate the crosstalk between MeJA and other hormones.
jasmonate (MeJA) é um derivado lipídico vegetal hormônio que medeia Diversos fenómenos biológicos.Aplicação de MeJA para arroz spikelet poderá apresentar Desenvolvimento anormal DOS órgãos florais.Apesar de Ácido jasmónico (Ja), precursor de MeJA, mostrou ser envolvido no processo de determinação do sexo de milho tassel, OS papéis DOS dois compostos de milho tassel Desenvolvimento ainda permanecem obscuros.No presente estudo, descobrimos que o Desenvolvimento FOI desacelerada pelo pedido do tassel de 2 mm MeJA.MeJA exógeno também influenciou o número de palea e estames de tassel spikelets.MeJA aumentou o nível de expressão exógena, um regulador chave genes, o que Pode o responsável PELA alteração fenotípica EM MeJA tratados tassel, e podem mediar a interferência entre MeJA e outras hormonas.
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Genes, Regulator , Zea mays , MorphogenesisABSTRACT
ABSTRACT One-year-old Glycyrrhiza uralensis Fisch. ex DC, Fabaceae, was treated with three exogenous phytohormones in June and July, namely gibberellin, auxin (indole-3-acetic acid), methyl jasmonate at different concentrations. Control plants were treated with water. Roots of controls and hormones-treated G. uralensis plants were harvested at different times, and the contents of seven main chemical components were determined. Root glycyrrhizic acid content of plants treated in June increased significantly compared with controls, and the difference was significant. As for plants treated in July, root glycyrrhizic acid content increased in which sprayed with appropriate concentrations of hormones, but the effects of hormones were more evident in plants treated in June coincided with the vigorous growth period than those treated in July. Gibberellin at 40 mg/l and auxin at 40 mg/l applied in the two treatment periods significantly promoted the accumulation of glycyrrhizic acid in G. uralensis root. Treatment with methyl jasmonate at 100 and 25 mg/l in June and July, respectively, also increased glycyrrhizic acid content significantly. The determination of major active compositions indicated that liquiritin, isoliquiritin, isoliquiritin apioside and liquiritin apioside contents were positively related to glycyrrhizic acid content. The study preliminarily found phytohormones and the main chemical components associated with glycyrrhizic acid content, and these discoveries could provide a basis for establishing a chemical control network with glycyrrhizic acid as the core, confirming the secondary product metabolic pathways in the network and completely uncovering synthesis mechanism underlying glycyrrhizic acid-combined functional gene polymorphism.
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Objective: In plant, squalene epoxidase (SE) catalyzes the first oxygenation step in the biosynthetic pathway of triterpenoid and phytosterol, representing one of the rate-limiting enzymes in this pathway. Bupleurum chinense is an important medicinal herb with its major active constituents such as triterpenoid saponins and saikosaponins. In order to obtain the series of enzymatic genes involved in saikosaponin biosynthesis, a cDNA of SE, designated BcSE1, was cloned from B. chinense. Methods: The BcSE1 gene was cloned by homology-based PCR and 5'/3' RACE methods from the adventitious roots of B. chinense. The physical and chemical parameters of BcSE1 protein were predicted by protparam. In order to discover hints in amino acid sequences on the dominant functions in the biosynthesis of saponin or phytosterol, sequences of SE from other plants were downloaded from NCBI for sequences alignment and phylogenetic analysis. BcSE1 was cloned into a yeast mutant KLN1 (MATa, erg1::URA3, leu2, ura3, and trp1) to verify the enzyme activity of BcSE1. Additionally, the tissue-specific expression and methyl jasmonate (MeJA) inducibility of BcSE1 were investigated using quantitative real-time PCR. Results: The predicted protein of BcSE1 is highly similar to SEs from other plants sharing amino acid sequence identities of up to 88%. The BcSE1 can functionally complement with yeast SE gene (ERG1) when expressed in the KLN1 mutant (MATa, erg1::URA3, leu2, ura3, and trp1). Using as controls with β-amyrin synthase (β-AS) which is presumed to catalyze the first committed step in saikosaponin biosynthesis and a cycloartenol synthase (CAS) relating to the phytosterol biosynthesis, the transcript of BcSE1 was significantly elevated by MeJA in adventitious roots of B. chinense and the transcript of BcSE1 was most abundant in the fruits and flowers of plants, followed by that in the leaves and roots, and least in stems. Conclusion: It is the first time to illustrate the molecular information of SE in B. chinense and to clone the full-length SE gene in plants of genus Bupleurum L.
ABSTRACT
Objective: To clone and characterize a late embryogenesis abundant protein SmLEA2 with its promoter region from Salvia miltiorrhiza, and to predict its probable function. Methods: SmLEA2 was cloned by PCR and RT-PCR from genomic DNA and cDNA. Protein structure and phylogenetic relationships were carried out by bioinformatic analysis. Gene expression in different organs and different development periods was detected by qPCR. Gene expression was also detected under different treatments. Results: By analyzing the cDNA library for S. miltiorrhiza with BLAST program, one of these sequences showed a high homology with late embryogenesis abundant (LEA) protein and was named as SmLEA2 (GenBank: HQ676610). We obtained 1 961 bp gene sequences of SmLEA2, which contained an intron and a single opening reading frame (ORF) of 960 bp encoding 319 amino acid peptides. Bioinformatic analysis showed that the putative SmLEA2 protein was a hydrophilic protein without signal peptides and transmembrane domains. The SmLEA2 protein was predicted with a molecular weight of 35 340 and a theoretical isoelectric point of 4.77. SmLEA2 was expressed in the roots, stems, and leaves of S. miltiorrhiza, and the most abundant in the stems. With the development of the flowers and seeds, its expression increased gradually. Expression of SmLEA2 could be induced by methyl jasmonate (MeJA) and abscisic acid (ABA). Conclusion: Thus, we speculate that SmLEA2 may be involved in seed development and plant defenses.