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High-frequency shoot multiplication in Curcuma longa L using thidiazuron.

The effects of plant growth regulators, explant types, and culture regimens were investigated on in vitro shoot proliferation from terminal bud explants of Curcuma longa. Each bud was longitudinally divided into four equal pieces, each 1 cm in length, and used as explants. These were then cultured on MS medium supplemented with 18.17 microM thidiazuron for 4 weeks prior to transfer to MS medium without growth regulator for 8 weeks. Under these conditions, a shoot induction rate of 18.22+/-0.62 shoots/explant was obtained after 12 weeks of cultures. Spontaneous rooting was achieved. The regenerated plants were transferred to soil under greenhouse conditions and subsequently grown successfully in the field.

Culture Techniques↗

An efficient protocol for genetic transformation and shoot regeneration of turmeric (Curcuma longa L.) via particle bombardment.

Turmeric (Curcuma longa L.) is an important spice crop plant that is sterile and cannot be improved by conventional breeding. An efficient method for stable transformation for turmeric, C. longa L., was developed using particle bombardment. Callus cultures initiated from shoots were bombarded with gold particles coated with plasmid pAHC25 containing the bar and gusA genes each driven by the maize ubiquitin promoter. Transformants were selected on medium containing glufosinate. Transgenic lines were established on selection medium from 50% of the bombarded calluses. Transgenic shoots regenerated from these were multiplied and stably transformed plantlets were produced. Polymerase chain reaction (PCR) and histochemical GUS assay confirmed the stable transformation. Transformed plantlets were resistant to glufosinate.

Aminobutyrates↗

Antifungal activity of turmeric oil extracted from Curcuma longa (Zingiberaceae).

Turmeric oil and curcumin, isolated from Curcuma longa L., were studied against fifteen isolates of dermatophytes, four isolates of pathogenic molds and six isolates of yeasts. The inhibitory activity of turmeric oil was tested in Trichophyton-induced dermatophytosis in guinea pigs. The results showed that all 15 isolates of dermatophytes could be inhibited by turmeric oil at dilutions of 1:40-1:320. None of the isolates of dermatophytes were inhibited by curcumin. The other four isolates of pathogenic fungi were inhibited by turmeric oil at dilutions of 1:40-1:80 but none were inhibited by curcumin. All six isolates of yeasts tested proved to be insensitive to both turmeric oil and curcumin. In the experimental animals, turmeric oil (dilution 1:80) was applied by dermal application on the 7th day following dermatophytosis induction with Trichophyton rubrum. An improvement in lesions was observed in 2-5 days and the lesions disappeared 6-7 days after the application of turmeric oil.

Animals↗

Xanthorrhizol, a natural sesquiterpenoid from Curcuma xanthorrhiza, has an anti-metastatic potential in experimental mouse lung metastasis model.

Xanthorrhizol is a sesquiterpenoid compound isolated from the rhizome of Curcuma xanthorrhiza. In this study, the anti-metastatic activity of xanthorrhizol was evaluated by using an in vivo mouse lung metastasis model and a tumor mass formation assay. Interestingly, xanthorrhizol dramatically inhibited the formation of tumor nodules in the lung tissue and the intra-abdominal tumor mass formation. Next, to examine the mechanism of the anti-metastatic action of xanthorrhizol in the mouse lung metastasis, expression patterns of the several intracellular signaling molecules were evaluated using the lung tissues with tumor nodules. Higher expression levels of cyclooxygenase-2 (COX-2), matrix metalloproteinase-9 (MMP-9), and phosphorylated extracellular signal-regulated kinase (ERK) were observed in the metastatic group compared with control, but these were attenuated by the treatment of xanthorrhizol. In conclusion, xanthorrhizol exerts anti-metastatic activity in vivo and this effect could be highly linked to the metastasis-related multiplex signal pathway including ERK, COX-2, and MMP-9.

Animals↗

Biological effects of indigenous medicinal plants Curcuma longa and Alpinia galanga.

The ethanolic extracts of Curcuma longa and Alpinia galanga exhibited excellent (100%) phytotoxic activity against Lemma minor. These extracts were also found to possess good antifungal activities against Trichophyton longifusus (65% and 60%, respectively), while in the brine shrimp lethality bioassay were found to be toxic with LD50 of 33 and 109 mincrog/ml, respectively. These extracts were found quite inert in antibacterial bioassay, while the extract from C. longa, tested for insecticidal activity, was also found to be devoid of any activity.

Alpinia↗

Isolation and characterization of antioxidant and antibacterial compound from mango ginger (Curcuma amada Roxb.) rhizome.

The chloroform extract of mango ginger (Curcuma amada Roxb.) rhizome was subjected to antioxidant activity-guided purification by repeated silica gel column chromatography to obtain a pure antioxidant compound. The structure was deduced by analyzing UV, IR, liquid chromatography-mass spectrometry (LC-MS) and two-dimensional heteronuclear multiple quantum coherence transfer spectroscopy (2D-HMQCT) NMR spectral data, and named it as "Amadannulen", a novel compound. It exhibited DPPH radical scavenging activity, super oxide radical scavenging activity, lipid peroxidation inhibitory activity and metal chelating activity. Amadannulen also showed antibacterial activity against both Gram-positive and Gram-negative bacteria tested. It also exhibited bactericidal activity against M. luteus, B. cereus and B. subtilis.

Anti-Bacterial Agents↗

Solid-phase microextraction-gas chromatographic-mass spectrometric analysis of volatile compounds from Curcuma wenyujin Y.H. Chen et C. Ling.

A solid-phase microextraction coupled with gas chromatography-mass spectrometry (SPME-GC-MS) for analysis of the volatile compounds from Curcuma wenyujin Y.H. Chen et C. Ling is described. SPME parameters (fiber type, extraction temperature and time, headspace volume and desorption time) and GC conditions were tested. The powdered sample of C. wenyujin Y.H. Chen et C. Ling was directly analyzed by SPME-GC-MS and 72 compounds were identified. The results from SPME-GC-MS were compared with those obtained from steam distillation gas chromatography-mass spectrometry (SD-GC-MS) with a good agreement. The results show that SPME-GC-MS method is a fast, simple and efficient way for the analysis of volatile components from traditional Chinese medicines (TCMs).

Curcuma↗

Optimization of GC-MS conditions based on resolution and stability of analytes for simultaneous determination of nine sesquiterpenoids in three species of Curcuma rhizomes.

GC-MS is a powerful tool for analysis of volatile oil, and resolutions of analytes were exclusively used as marker for optimization of the conditions. However, volatile oil usually contains heat labile components which may degrade and result in wrong results during GC analysis. In present study, based on both resolutions and stabilities of 11 sesquiterpenoids, GC-MS conditions were optimized for simultaneously quantitative determination of nine compounds including beta-elemene, curzerene, curcumol, isocurcumenol, germacrone, curdione, curcumenol, neocurdione and curcumenone in Ezhu. However, the other two compounds, i.e. furanodienone and furanodiene, were still thermal sensitive and not available for GC analysis. The results showed that both resolutions and stabilities of analytes should be considered for optimization of GC conditions because the properties of most components in volatile oil are unknown. Under optimum conditions, a capillary column (30 m x 0.25 mm i.d.) coated with 0.25 microm film 5% phenyl methyl siloxane was used for separation. Pulsed splitless inlet with temperature of 190 degrees C was selected for sample injection (0.2 microl). The calibration curves of nine sesquiterpenoids showed good linearity (r2>0.9989) within test ranges. The optimized method showed good repeatability for quantification of these nine components in Ezhu with intra- and inter-day variations of less than 1.42% and 2.79%, respectively. The validated method was successfully applied to quantify 9 sesquiterpenoids in 18 samples of 3 species of Curcuma used as Ezhu.

Calibration↗

Analysis of volatile components of Curcuma sichuanensis X. X. Chen by gas chromatography-mass spectrometry.

Volatile components of Curcuma sichuanensis X. X. Chen growing in Sichuan, China, were extracted by steam distillation and analyzed by using gas chromatography and gas chromatography-mass spectrometry. A total of 44 volatile essential oil components were identified in the extract of C. sichuanensis X. X. Chen, representing 87.1% of the total integrated chromatographic peaks. The major compounds were found to be epi-curzerenone (26.9%), germacrone (12.4%), isocurcumenol (9.7%), beta-elemene (6.4%) and curzerene (6.2%). The results of semi-quantitative analysis indicated that levels of total sesquiterpene fraction (85.4%) were more than 55 times higher than those of monoterpene components (1.5%).

China↗

A water extract of Curcuma longa L. (Zingiberaceae) rescues PC12 cell death caused by pyrogallol or hypoxia/reoxygenation and attenuates hydrogen peroxide induced injury in PC12 cells.

A number of studies indicate that free radicals are involved in the neurodegeneration in Alzheimer's disease (AD). The role of superoxide anion (O2*-) in neuronal cell injury induced by reactive oxygen species (ROS) was examined in PC12 cells using pyrogallol (1,2,3-benzenetrior), a donor to release O2*-. Pyrogallol induced PC12 cell death at concentrations, which evidently increased intracellular O2*-, as assessed by O2*- sensitive fluorescent precursor hydroethidine (HEt). A water extract of Curcuma longa L. (Zingiberaceae) (CLE), having O2*- scavenging activity rescued PC12 cells from pyrogallol-induced cell death. Hypoxia/reoxygenation injury of PC12 cells was also blocked by CLE. The present study was also conducted to examine the effect of CLE on H2O2 -induced toxicity in rat pheochromocytoma line PC12 by measuring cell lesion, level of lipid peroxidation and antioxidant enzyme activities. Following a 30 min exposure of the cells to H2O2 (150 microM), a marked decrease in cell survival, activities of glutathione peroxidase and catalase as well as increased production of malondialdehyde (MDA) were found. Pretreatment of the cells with CLE (0.5-10 microg/ml) prior to H2O2 exposure significantly elevated the cell survival, antioxidant enzyme activities and decreased the level of MDA. The above-mentioned neuroprotective effects are also observed with tacrine (THA, 1 microM), suggesting that the neuroprotective effects of cholinesterase inhibitor might partly contribute to the clinical efficacy in AD treatment. Further understanding of the underlying mechanism of the protective effects of these radical scavengers reducing intracellular O2*- on neuronal cell death may lead to development of new therapeutic treatments for hypoxic/ischemic brain injury.

Animals↗

Inhibitory effect of Curcuma comosa on NO production and cytokine expression in LPS-activated microglia.

Curcuma comosa is an indigenous plant of Thailand, which has been traditionally and widely used as an anti-inflammatory agent for the treatment of postpartum uterine bleeding and uterine inflammation. However, the scientific investigation on its anti-inflammatory activity has not been reported. In the present study, we investigated the anti-inflammatory effect of the extract from C. comosa on the responses in microglia stimulated with lipopolysaccharide (LPS). Pretreatment of highly aggressively proliferating immortalized (HAPI) cells, a rat microglial cell line, with the hexane extract of C. comosa rhizome at 10(-9) to 10(-5) g/ml significantly suppressed the levels of NO released from these cells. The attenuation in iNOS protein and mRNA expression was also observed suggesting an interference at transcriptional level. In addition, C. comosa extract inhibited interferon regulatory factor-1 expression which is an essential transcription factor governing the iNOS expression. Moreover, the levels of mRNA expressions of MCP-1 and IL-6 induced by LPS were also prominently decreased in the presence of C. comosa extract. These results suggest that C. comosa extract possesses a strong anti-inflammatory activity and has a potential to be developed as a therapeutic compound for diverse neurological disorders associated with inflammation.

Animals↗

Biosynthesis of curcuminoids and gingerols in turmeric (Curcuma longa) and ginger (Zingiber officinale): identification of curcuminoid synthase and hydroxycinnamoyl-CoA thioesterases.

Members of the Zingiberaceae such as turmeric (Curcuma longa L.) and ginger (Zingiber officinale Rosc.) accumulate at high levels in their rhizomes important pharmacologically active metabolites that appear to be derived from the phenylpropanoid pathway. In ginger, these compounds are the gingerols; in turmeric these are the curcuminoids. Despite their importance, little is known about the biosynthesis of these compounds. This investigation describes the identification of enzymes in the biosynthetic pathway leading to the production of these bioactive natural products. Assays for enzymes in the phenylpropanoid pathway identified the corresponding enzyme activities in protein crude extracts from leaf, shoot and rhizome tissues from ginger and turmeric. These enzymes included phenylalanine ammonia lyase, polyketide synthases, p-coumaroyl shikimate transferase, p-coumaroyl quinate transferase, caffeic acid O-methyltransferase, and caffeoyl-CoA O-methyltransferase, which were evaluated because of their potential roles in controlling production of certain classes of gingerols and curcuminoids. All crude extracts possessed activity for all of these enzymes, with the exception of polyketide synthases. The results of polyketide synthase assays showed detectable curcuminoid synthase activity in the extracts from turmeric with the highest activity found in extracts from leaves. However, no gingerol synthase activity could be identified. This result was explained by the identification of thioesterase activities that cleaved phenylpropanoid pathway CoA esters, and which were found to be present at high levels in all tissues, especially in ginger tissues. These activities may shunt phenylpropanoid pathway intermediates away from the production of curcuminoids and gingerols, thereby potentially playing a regulatory role in the biosynthesis of these compounds.

Catechols↗

Antidepressant activity of aqueous extracts of Curcuma longa in mice.

Curcuma longa (turmeric) is a well-known indigenous herbal medicine. The aqueous extracts, when administered orally to the mice from 140 to 560 mg/kg for 14 days, were able to elicit dose-dependent relation of immobility reduction in the tail suspension test and the forced swimming test in mice. The effects of the extracts at the dose of 560 mg/kg were more potent than that of reference antidepressant fluoxetine. The extracts, at the dose of 140 mg/kg or above for 14 days, significantly inhibited the monoamine oxidize A (MAO) activity in mouse whole brain at a dose-dependent manner, however, oral administration of the extract only at a dose of 560 mg/kg produced observable MAO B inhibitory activity in animal brain. Fluoxetine showed only a tendency to inhibit MAO A and B activity in animal brain in the study. Neither the extracts of C. longa nor fluoxetine, at the doses tested, produced significant effects on locomotor activity. These results demonstrated that C. longa had specifically antidepressant effects in vivo. The activity of C. longa in antidepression may mediated in part through MAO A inhibition in mouse brain.

Administration, Oral↗

Fungicidal property of Curcuma longa L. rhizome-derived curcumin against phytopathogenic fungi in a greenhouse.

Fungicidal activity of Curcuma longa rhizome-derived materials against Botrytis cineria, Erysiphe graminis, Phytophthora infestans, Puccinia recondita, Pyricularia oryzae, and Rhizoctonia solani was tested using a whole plant method in vivo. It was compared with synthetic fungicides and four commercially available compounds derived from C. longa. The response varied with the tested plant pathogen. At 1000 mg/L, the hexane extract of C. longa showed fungicidal activities against E.graminis, P. infestans, and R. solani, and the ethyl acetate extract of C. longa showed fungicidal activities against B. cineria, P. infestans, Pu. recondita, and R. solani. Curcumin was isolated from the ethyl acetate fraction using chromatographic techniques and showed fungicidal activities against P. infestans, Pu. recondita, and R. solani with 100, 100, and 63% control values at 500 mg/L and 85, 76, and 45% control values at 250 mg/L, respectively. In the test with components derived from C. longa, turmerone exhibited weak activity against E. graminis, but no activity was observed from treatments with borneol, 1,8-cineole, sabinene, and turmerone. In comparison, potent fungicidal activity with chlorothalonil against P. infestans at 50 mg/L and dichlofluanid against B. cinerea at 50 mg/L was exhibited. These results may be an indication of at least one of the fungicidal actions of curcumin derived from C. longa.

Curcuma↗

Extraction of essential oil and pigments from Curcuma longa [L] by steam distillation and extraction with volatile solvents.

Curcuma longa [Linn] (turmeric), of the Zingiberaceae family, has a great importance in the food, textile, and pharmaceutical industries. The aim of this work was to identify the best processing conditions to maximize the yields of essential oil and pigments, as well as their content of ar-turmerone, (alpha and beta)-turmerone, and the curcuminoids, respectively. Autoclave pressure and distillation time were the variables studied for the steam distillation process. The highest yields of essential oil (0.46 wt %) and pigment (0.16 wt %)-expressed as curcumin, demethoxycurcumin, and bisdemethoxycurcumin-were obtained at a pressure of 1.0 x 10(5) Pa and a time of 2 h. On the other hand, with extraction by volatile solvents, the best yield of essential oil (5.49 wt %) was obtained when using 0.175, 0.124, 0.088 mm particles (Foust, A. S.; Wenzel, L. A.; Clump, C. W.; Maus, L.; Andersen, L. B. Princípios das Operações Unitárias; Editora Guanabara Dois S.A.: Rio de Janeiro, Brazil, 1982), at 40 degrees C, and 6 h of extraction. However, the best yield of pigment (7.98 wt %) was obtained under the same conditions, except for the temperature (30 degrees C).

Curcuma↗

Curcuminoids and sesquiterpenoids in turmeric (Curcuma longa L.) suppress an increase in blood glucose level in type 2 diabetic KK-Ay mice.

Turmeric, the rhizome of Curcuma longa L., has a wide range of effects on human health. The chemistry includes curcuminoids and sesquiterpenoids as components, which are known to have antioxidative, anticarcinogenic, and antiinflammatory activities. In this study, we investigated the effects of three turmeric extracts on blood glucose levels in type 2 diabetic KK-A(y) mice (6 weeks old, n = 5/group). These turmeric extracts were obtained by ethanol extraction (E-ext) to yield both curcuminoids and sesquiterpenoids, hexane extraction (H-ext) to yield sesquiterpenoids, and ethanol extraction from hexane-extraction residue (HE-ext) to yield curcuminoids. The control group was fed a basal diet, while the other groups were fed a diet containing 0.1 or 0.5 g of H-ext or HE-ext/100 g of diet or 0.2 or 1.0 g of E-ext/100 g of diet for 4 weeks. Although blood glucose levels in the control group significantly increased (P < 0.01) after 4 weeks, feeding of 0.2 or 1.0 g of E-ext, 0.5 g of H-ext, and 0.5 g of HE-ext/100 g of diet suppressed the significant increase in blood glucose levels. Furthermore, E-ext stimulated human adipocyte differentiation, and these turmeric extracts had human peroxisome proliferator-activated receptor-gamma (PPAR-gamma) ligand-binding activity in a GAL4-PPAR-gamma chimera assay. Also, curcumin, demethoxycurcumin, bisdemethoxycurcumin, and ar-turmerone had PPAR-gamma ligand-binding activity. These results indicate that both curcuminoids and sesquiterpenoids in turmeric exhibit hypoglycemic effects via PPAR-gamma activation as one of the mechanisms, and suggest that E-ext including curcuminoids and sesquiterpenoids has the additive or synergistic effects of both components.

Adipocytes↗

Curcuma longa L. constituents inhibit sortase A and Staphylococcus aureus cell adhesion to fibronectin.

The inhibitory activity of Curcuma longa L. (turmeric) rhizome constituents against sortase A, a bacterial surface protein anchoring transpeptidase, from Staphylococcus aureus ATCC 6538p was evaluated. The activity of the isolated compounds (1-4) was compared to that of the positive control,p-hydroxymecuribenzoic acid (pHMB). The biologically active components of C. longa rhizome were characterized by spectroscopic analysis as the curcuminoids curcumin (1), demethoxycurcumin (2), and bisdemethoxycurcumin (3). Curcumin was a potent inhibitor of sortase A, with an IC50 value of 13.8 +/- 0.7 microg/mL. Bisdemethoxycurcumin (IC50 = 31.9 +/- 1.2 microg/mL) and demethoxycurcumin (IC50 = 23.8 +/- 0.6 microg/mL) were more effective than pHMB (IC50 = 40.6 +/- 1.2 microg/mL). The three isolated compounds (1-3) showed no growth inhibitory activity against S. aureus strain Newman, with minimum inhibitory concentrations (MICs) greater than 200 microg/mL. Curcumin also exhibited potent inhibitory activity against S. aureus cell adhesion to fibronectin. The suppression of fibronectin-binding activity by curcumin highlights its potential for the treatment of S. aureus infections via inhibition of sortase activity. These results indicate that curcumin is a possible candidate in the development of a bacterial sortase A inhibitor.

Aminoacyltransferases↗

Metabolic profiling of turmeric (Curcuma longa L.) plants derived from in vitro micropropagation and conventional greenhouse cultivation.

Turmeric (Curcuma longa) was considered only a culinary spice in many parts of the world until the notable anti-inflammation curcuminoids were discovered from this herb. Because it is a sterile triploid and is propagated vegetatively by rhizome division, turmeric is susceptible to pathogens that accumulate and are transmitted from generation to generation, and amplification of particularly useful stocks is a slow process. An in vitro propagation method has been developed to alleviate these problems. Metabolic profiling, using GC-MS and LC-ESI-MS, was used to determine if chemical differences existed between greenhouse-grown and in vitro micropropagation derived plants. The major chemical constituent curcuminoids, a group of diarylheptanoid compounds, as well as major mono- and sesquiterpenoids were identified and quantified. Principal component analysis and hierarchical cluster analysis revealed chemical differences between lines (T3C turmeric vs Hawaiian red turmeric) and tissues (rhizome, root, leaf, and shoot). However, this analysis indicated that no significant differences existed between growth treatments (conventional greenhouse-grown vs in vitro propagation derived plants).

Agriculture↗