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growth and photosynthetic pigments responses of two varieties of catharanthus roseus to triadimefon treatment.triadimefon, potential fungicide cum plant-growth retardant was used in this study to investigate its effect on the growth and the photosynthetic pigment contents of two varieties of catharanthus roseus (l.) g. don. the plants of both varieties were subjected to 15 mg l(-1) triadimefon treatment by soil drenching 30, 45, 60, and 75 days after planting (dap). plants were uprooted on 90 dap, and morphological parameters, like plant height, number of leaves, leaf area, root length and fresh and dry ...200818355749
inheritance of pollen-less anthers and "thrum" and "pin" flowers in periwinkle.two mutants, 1 with small, pollen-less anthers (or-ea) and another with "pin" flowers (ems 13-2), in contrast to "thrum" flowers found in normal periwinkle (catharanthus roseus) plants, were isolated after induced mutagenesis in strain or and cultivar, "dhawal," respectively. inheritance of these 2 traits, pollen-less anthers, and pin flowers was studied by crossing the mutants with their respective parental strains. segregation ratios observed in f(2) and testcross generations of the cross or-e ...200818339651
the leaf epidermome of catharanthus roseus reveals its biochemical specialization.catharanthus roseus is the sole commercial source of the monoterpenoid indole alkaloids (mias), vindoline and catharanthine, components of the commercially important anticancer dimers, vinblastine and vincristine. carborundum abrasion technique was used to extract leaf epidermis-enriched mrna, thus sampling the epidermome, or complement, of proteins expressed in the leaf epidermis. random sequencing of the derived cdna library established 3655 unique ests, composed of 1142 clusters and 2513 sing ...200818326827
construction of genetic linkage map of the medicinal and ornamental plant catharanthus roseus.an integrated genetic linkage map of the medicinal and ornamental plant catharanthus roseus, based on different types of molecular and morphological markers was constructed, using a f(2) population of 144 plants. the map defines 14 linkage groups (lgs) and consists of 131 marker loci, including 125 molecular dna markers (76 rapd, 3 rapd combinations; 7 issr; 2 est-ssr from medicago truncatula and 37 other pcr based dna markers), selected from a total of 472 primers or primer pairs, and six morph ...200718305345
differential responses in water use efficiency in two varieties of catharanthus roseus under drought stress.two varieties, rosea and alba, of catharanthus roseus (l.) g. don. were screened for their water use efficiency under two watering regimes, viz. 60 and 100% filed capacity in the present study. drought stress was imposed at 60% filed capacity from 30 to 70 days after sowing, while the control pots were maintained at 100% filed capacity throughout the entire growth period. leaf area duration, cumulative water transpired, water use efficiency, net assimilation rate, mean transpiration rate, harves ...200818187121
7-o-methylated anthocyanidin glycosides from catharanthus roseus.anthocyanins were isolated from orange-red flowers of catharanthus roseus cv 'equator deep apricot', and identified as rosinidin 3-o-[6-o-(alpha-rhamnopyranosyl)-beta-galactopyranoside] (1), and also 7-o-methylcyanidin 3-o-[6-o-(alpha-rhamnopyranosyl)-beta-galactopyranoside] (2) by chemical and spectroscopic methods. pigment 1 was found to be a major anthocyanin in the flowers of this cultivar. by contrast, the distribution of rosinidin glycosides is very limited in plants, and reported only in ...200818164044
know the medicinal herb: catharanthus roseus (vinca rosea). 200825606134
alterations in lipid peroxidation, electrolyte leakage, and proline metabolism in catharanthus roseus under treatment with triadimefon, a systemic fungicide.triadimefon (tdm), a systemic fungicide with non-traditional plant-growth regulator properties, was administered to catharanthus roseus (l.) g. don. plants in order to determine its effects on oxidative injury in terms of h2o2 content, lipid peroxidation (lpo), electrolyte leakage (el), protein and amino acid contents, as well as proline metabolism. the lpo, estimated as thiobarbituric acid-reactive substances (tbars), decreased under tdm treatment. it was found that h2o2 and el were reduced und ...200718068649
the suzuki-miyaura reaction in the chemical transformations of vindoline.vindoline and its analogues are important constituents of the madagascan periwinkle catharanthus roseus, and some of them are valuable chemotherapy drugs used in treatment for some types of cancer, including leukaemia, lymphoma, breast and lung cancer. the search for semi-synthetic congeners of natural substances is still an important task for organic chemistry. in this communication we report the synthesis of five new vindoline derivatives, 15-(2-methoxyphenyl)vindoline 11, 15-(3-methoxyphenyl) ...200718066403
molecular cloning and characterization of a vacuolar class iii peroxidase involved in the metabolism of anticancer alkaloids in catharanthus roseus.catharanthus roseus produces low levels of two dimeric terpenoid indole alkaloids, vinblastine and vincristine, which are widely used in cancer chemotherapy. the dimerization reaction leading to alpha-3',4'-anhydrovinblastine is a key regulatory step for the production of the anticancer alkaloids in planta and has potential application in the industrial production of two semisynthetic derivatives also used as anticancer drugs. in this work, we report the cloning, characterization, and subcellula ...200818065566
substrate specificity and diastereoselectivity of strictosidine glucosidase, a key enzyme in monoterpene indole alkaloid biosynthesis.strictosidine glucosidase (sgd) from catharanthus roseus catalyzes the deglycosylation of strictosidine, an intermediate from which thousands of monoterpene indole alkaloids are derived. the steady-state kinetics of sgd with a variety of strictosidine analogs revealed the substrate preferences of this enzyme at two key positions of the strictosidine substrate. additionally, sgd from c. roseus turns over both strictosidine and its stereoisomer vincoside, indicating that although this enzyme prefe ...200818061449
application of carborundum abrasion for investigating the leaf epidermis: molecular cloning of catharanthus roseus 16-hydroxytabersonine-16-o-methyltransferase.the madagascar periwinkle (catharanthus roseus) produces the well-known and remarkably complex anti-cancer dimeric alkaloids vinblastine and vincristine that are derived from the coupling of vindoline and catharanthine monomers. this study describes the novel application of a carborundum abrasion (ca) technique for large-scale isolation of leaf epidermis-enriched proteins in order to purify to apparent homogeneity 16-hydroxytabersonine-16-o-methyltransferase (16omt), which catalyses the second o ...200818053006
water deficit stress effects on reactive oxygen metabolism in catharanthus roseus; impacts on ajmalicine accumulation.in the present work, we have analysed the changes in the reactive oxygen metabolism of catharanthus roseus (l.) g. don. plants in terms of h(2)o(2) content, lipid peroxidation and the free radical quenching systems (non-enzymatic and enzymatic antioxidants) under drought stress. in addition to this, the root alkaloid ajmalicine was extracted and quantified from both control and drought stressed plants. the h(2)o(2) content was analysed from both stressed and unstressed control plants. lipid pero ...200817996429
[simultaneous determination of vindoline, catharanthine and anhydrovinblastine in catharanthus roseus by high performance liquid chromatography].a high performance liquid chromatographic (hplc) method with gradient elution was developed for the simultaneous determination of vindoline, catharanthine and anhydrovinblastine in catharanthus roseus. the analytes were separated on a waters (5)c18-ms-ii column (4.6 mm x 250 mm) and detected at 220 nm. the mobile phases were methanol-1% (v/v) diethylamine solution (adjusted to ph 7.3 with phosphate) for gradient elution at temperature of 25 degrees c. the calibration curves for vindoline, cathar ...200717970117
alterations in morphological parameters and photosynthetic pigment responses of catharanthus roseus under soil water deficits.an experiment was conducted in two varieties, rosea and alba, of catharanthus roseus plants with two watering treatments viz., 100 and 60% of field capacity, to understand the effects of water deficit on early growth, biomass allocation and photosynthetic pigment responses. we found that there were significant differences in early growth, dry matter accumulation and pigment variations between the two varieties. the root length, shoot length, total leaf area, fresh and dry weights were significan ...200817949951
purification and characterization of udp-glucose : curcumin glucoside 1,6-glucosyltransferase from catharanthus roseus cell suspension cultures.catharanthus roseus cell suspension cultures converted exogenously added curcumin to a series of curcumin glucosides that possessed drastically enhanced water solubility. a cdna clone encoding a glucosyltransferase responsible for glucosylation of curcumin to form curcumin 4'-o-glucoside was previously isolated, and in the present study a novel sugar-sugar glycosyltransferase, udp-glucose:curcumin glucoside glucosyltransferase (ucggt), was purified approximately 900-fold to apparent homogeneity ...200717940060
a simplified procedure for indole alkaloid extraction from catharanthus roseus combined with a semi-synthetic production process for vinblastine.dried leaves of catharanthus roseus were extracted with aqueous acidic 0.1 m solution of hcl. alkaloid-embonate complexes were obtained as precipitates by treating the extract with an alkaline (naoh) solution of embonic acid (4,4-methylene-bis-3-hydroxynaphtalenecarboxylic acid). the precipitate mainly consisted of catharanthine and vindoline embonates and it was directly used as the starting material for a semi-synthesis of the anti-cancer bisindole alkaloid vinblastine. the coupling reaction i ...200717909486
long-term maintenance of a transgenic catharanthus roseus hairy root line.stably transformed transgenic hairy root cultures have the potential to be a valuable production platform for a variety of secondary metabolites. this study reports that a transgenic hairy root culture of catharanthus roseus has been stably maintained for over 4.5 years. this culture carries a transgene that expresses the green fluorescent protein under the control of the glucocorticoid-inducible promoter. genomic pcr confirmed the presence of the gfp insert within the hairy roots, and induction ...200717900137
rapid identification of enzyme variants for reengineered alkaloid biosynthesis in periwinkle.monoterpene indole alkaloids from catharanthus roseus (madagascar periwinkle), such as the anticancer agents vinblastine and vincristine, have important pharmacological activities. metabolic engineering of alkaloid biosynthesis can provide an efficient and environmentally friendly route to analogs of these synthetically challenging and pharmaceutically valuable natural products. however, the narrow substrate scope of strictosidine synthase, the enzyme at the entry point of the pathway, limits a ...200717719488
allergy to periwinkle pollen (catharanthus roseus g. don.).catharanthus roseus g. don. (cr) or periwinkle plants are widely grown/ cultivated as garden plants in the tropics and subtropics. in spite of its predominantly entomophilous nature, cr pollen had been reported to be airborne and allergenic. the objective of this study was to discover the seasonal changes of cr pollen concentration in air, to determine its potential to cause respiratory allergy and to analyze its allergenic components. a 2-year aerobiological survey was conducted with a burkard ...200717655175
optimisation of supercritical fluid extraction of indole alkaloids from catharanthus roseus using experimental design methodology--comparison with other extraction techniques.response surface modelling, using modde 6 software for design of experiments and optimisation, was applied to optimise supercritical fluid extraction (sfe) conditions for the extraction of indole alkaloids from the dried leaves of catharanthus roseus. the effects of pressure (200-400 bar), temperature (40-80 degrees c), modifier concentration (2.2-6.6 vol%) and dynamic extraction time (20-60 min) on the yield of alkaloids were evaluated. the extracts were analysed by high-performance liquid chro ...201517654538
membrane-associated phosphoinositides-specific phospholipase c forms from catharanthus roseus transformed roots.we have previously reported that catharanthus roseus transformed roots contain at least two phosphatidylinositol 4,5-bisphosphate-phospholipase c (plc) activities, one soluble and the other membrane associated. detergent, divalent cations, and neomycin differentially regulate these activities and pure protein is required for a greater understanding of the function and regulation of this enzyme. in this article we report a partia purification of membrane-associated plc. we found that there are at ...200717652793
induction of drought stress tolerance by ketoconazole in catharanthus roseus is mediated by enhanced antioxidant potentials and secondary metabolite accumulation.a pot culture experiment was conducted to estimate the drought stress mitigating effect of ketoconazole (kcz), a fungicide cum plant growth regulator, in catharanthus roseus plants. the plants under pot culture were subjected to drought stress and drought stress with kcz from 30 days after sowing (das) and regular irrigation was kept as control. antioxidant contents and activities of antioxidant enzymes were estimated from root, stem and leaf of both control and treated plants. the alkaloid ajma ...200717643970
water deficit stress mitigation by calcium chloride in catharanthus roseus: effects on oxidative stress, proline metabolism and indole alkaloid accumulation.the present investigation was conducted to determine whether cacl(2) increases catharanthus roseus drought tolerance and if such tolerance is correlated with changes in oxidative stress, osmoregulation and indole alkaloid accumulation. c. roseus plants were grown under water deficit environments with or without cacl(2). drought induced oxidative stress was measured in terms of lipid peroxidation (lpo) and h(2)o(2) contents, osmolyte concentration, proline (pro) metabolizing enzymes and indole al ...200717643271
downregulation of terpenoid indole alkaloid biosynthetic pathway by low temperature and cloning of a ap2 type c-repeat binding factor (cbf) from catharanthus roseus (l). g. don.plants produce secondary metabolites in response to various external signals. coordinated transcriptional control of biosynthetic genes emerges as a major mechanism dictating the accumulation of secondary metabolites in plant cells. however, information about stress regulation of secondary metabolites and the molecular mechanisms regulating these specialized pathways are poorly understood. here, we show that terpenoid indole alkaloid (tia) biosynthetic pathway is differentially regulated in resp ...200717619193
a predictive nutritional model for plant cells and hairy roots.a structured nutritional model is proposed to describe growth and nutritional behavior of eschscholtzia californica suspension cells and catharanthus roseus and daucus carota hairy roots in in vitro culture. the model describes the cells specific growth rate from concentration of intracellular nutrients such as inorganic phosphate (pi), nitrogen sources (no(3) (-) and nh(4) (+)) and sugars. two-level michaelis-menten kinetics are used to describe pi and no(3) (-) uptake and simple michaelis-ment ...200817614323
spatial distribution and hormonal regulation of gene products from methyl erythritol phosphate and monoterpene-secoiridoid pathways in catharanthus roseus.the monoterpene indole alkaloids (mias) from madagascar periwinkle (catharanthus roseus) are secondary metabolites of high interest due to their therapeutical values. secologanin, the monoterpenoid moiety incorporated into mias, is derived from the plastidial methyl-d: -erythritol 4-phosphate (mep) pathway. here, we have cloned a cdna encoding hydroxymethylbutenyl diphosphate synthase (hds), a mep pathway enzyme, and generated antibodies to investigate the distribution of transcripts and protein ...200717611800
expression of deacetylvindoline-4-o-acetyltransferase in catharanthus roseus hairy roots.madagascar periwinkle [catharanthus roseus (l.) g don] is a pantropical plant of horticultural value that produces the powerful anticancer drugs vinblastine and vincristine that are derived from the dimerization of the monoterpenoid indole alkaloids (mias), vindoline and catharanthine. the present study describes the genetic engineering and expression of the terminal step of vindoline biosynthesis, deacetylvindoline-4-o-acetyltransferase (dat) in catharanthus roseus hairy root cultures. biochemi ...200717574634
alterations in osmoregulation, antioxidant enzymes and indole alkaloid levels in catharanthus roseus exposed to water deficit.catharanthus roseus (l.) g. don plants were grown in different water regimes in order to study the drought induced osmotic stress and proline (pro) metabolism, antioxidative enzyme activities and indole alkaloid accumulation. the plants under pot culture were subjected to 10, 15 and 20 days interval drought (did) stress from 30 days after sowing (das) and regular irrigation was kept as control. the plants were uprooted on 41das (10did), 46das (15did) and 51das (20did). the drought stressed plant ...200717560094
dynamic flux cartography of hairy roots primary metabolism.a dynamic model for plant cell and hairy root primary metabolism is presented. the model includes nutrient uptake (pi, sugars, nitrogen sources), the glycolysis and pentose phosphate pathways, the tca cycle, amino acid biosynthesis, respiratory chain, biosynthesis of cell building blocks (structural hexoses, organic acids, lipids, and organic phosphated molecules). the energy shuttles (atp, adp) and cofactors (nad/h, nadp/h) are also included. the model describes the kinetics of 44 biochemical r ...201317555780
rapid and non-invasive detection of plants senescence using a delayed fluorescence technique.senescence is a phase of leaf ontogeny marked by declining photosynthetic activity that is paralleled by a decline in chloroplast function. the photosystem ii in a plant is considered to be the primary site where delayed fluorescence (df) is produced. we report here a simple, rapid, and non-invasive technique for detecting plants senescence based on quantitative measurements of df. in the experimental study, various senescence symptoms induced by age or hormones were examined in the catharanthus ...200717549265
responses of transformed catharanthus roseus roots to femtomolar concentrations of salicylic acid.catharanthus roseus transformed roots were cultured in the presence of salicylic acid (sa) at concentrations between 0.1 fm and 100 pm and the effect on root growth was evaluated. significant morphological changes in the lateral roots were recorded on day two in the sa treatment. presence of sa increased root cap size and caused the appearance of lateral roots closer to the root tip. the bioassay was sensitive enough to allow testing of low concentrations of other growth regulators that may affe ...201617544287
flux quantification in central carbon metabolism of catharanthus roseus hairy roots by 13c labeling and comprehensive bondomer balancing.methods for accurate and efficient quantification of metabolic fluxes are desirable in plant metabolic engineering and systems biology. toward this objective, we introduce the application of "bondomers", a computationally efficient and intuitively appealing alternative to the commonly used isotopomer concept, to flux evaluation in plants, by using catharanthus roseus hairy roots as a model system. we cultured the hairy roots on (5% w/w u-(13)c, 95% w/w naturally abundant) sucrose, and acquired t ...200717532015
accumulation of monoterpenoid indole alkaloids in periwinkle seedlings (catharanthus roseus) as a model for the study of plant-environment interactions.alkaloids are part of the chemical arsenal designed to protect plants against an adverse environment. therefore, their synthesis and accumulation are frequently induced in response to certain environmental conditions and are mediated by chemical signals, which are formed as the first responses to the external stimulus. a set of experiments using catharanthus roseus seedlings is presented to illustrate this interaction. using the well characterized induction of monoterpenoid indole alkaloid synth ...200721591090
glycosylation of capsaicin and 8-nordihydrocapsaicin by cultured cells of catharanthus roseus.the glycosylation of capsaicin and 8-nordihydrocapsaicin was investigated using cultured cells of catharanthus roseus. in addition to capsaicin 4-o-beta-d-glucopyranoside (170 microg/g fr. wt of cells), the biotransformation products, capsaicin 4-o-(6-o-beta-d-xylopyranosyl)-beta-d-glucopyranoside (116 microg/g fr. wt of cells) and capsaicin 4-o-(6-o-alpha-l-arabinopyranosyl)-beta-d-glucopyranoside (83 microg/g fr. wt of cells), were isolated from the cell suspension after three days of incubati ...200717449077
transcript profiling of terpenoid indole alkaloid pathway genes and regulators reveals strong expression of repressors in catharanthus roseus cell cultures.the understanding of the complexities and molecular events regulating genes and the activators involved in terpenoid indole alkaloid (tia) metabolism is known to a certain extent in cell cultures of an important tia yielding plant, catharanthus roseus, though it is not yet complete. recently, the repressors of early tia pathway genes have also been identified. however, their roles in the regulation of tia pathway in c. roseus cell cultures remains yet unknown. we have made a comparative profilin ...200717333024
effect of the engineered indole pathway on accumulation of phenolic compounds in catharanthus roseus hairy roots.catharanthus roseus has been well-known to contain indole alkaloids effective for treatment of diverse cancers. we examined the intracellular accumulation profiles of phenolic compounds in response to ectopic overexpression of tryptophan feedback-resistant anthranilate synthase holoenzyme (asalphabeta) in c. roseus hairy roots. among 13 phenolic compounds measured, 6 phenolic compounds were detected in late exponential phase asalphabeta hairy roots. uninduced and induced asalphabeta hairy roots ...201317256967
rapid in vitro multiplication of plants from mature nodal explants of catharanthus roseus. 199817253257
involvement of rapid nucleotide synthesis in recovery from phosphate starvation of catharanthus roseus cells.growth of suspension-cultured catharanthus roseus cells ceased during phosphate starvation, but the cells grew again upon addition of pi even after long-term starvation. the metabolic fate of [(33)p]pi was studied in 1-week-old stationary phase cells in ordinary culture and in 1- or 2-week-old pi-starved cells. immediately after administration, the most heavily labelled organic compounds are nucleotides, followed by sugar phosphates. two weeks pi starvation slowed down the speed of incorporation ...200717185741
[determination of three endogenous hormones in catharanthus roseus (l.) g. don using solid-phase extraction and high performance liquid chromatography]. 200617165557
determination of biomass composition of catharanthus roseus hairy roots for metabolic flux analysis.metabolic flux analysis is a powerful diagnostic tool in metabolic engineering, and determination of biomass composition is indispensable to accurate flux evaluation. we report the elemental and biomolecular composition of catharanthus roseus hairy roots, a pharmaceutically significant plant system and an important metabolic engineering target. the molecular formula of the organic material in the hairy roots was c12.0h22.7n0.4o7.6 during mid-exponential growth. the abundances of lipids, lignin, ...200617137315
directed biosynthesis of alkaloid analogs in the medicinal plant catharanthus roseus.terpene indole alkaloids are plant natural products with diverse structures and biological activities. a highly branched biosynthetic pathway is responsible for the production of approximately 130 different alkaloids in madagascar periwinkle (c. roseus) from a common biosynthetic intermediate derived from tryptamine. although numerous biosynthetic pathways can incorporate unnatural starting materials to yield novel natural products, it was not clear how efficiently the complex, eukaryotic tia pa ...200617076499
transcriptome analysis in catharanthus roseus leaves and roots for comparative terpenoid indole alkaloid profiles.in catharanthus roseus (l.) g. don each tissue is known to produce a distinct spectrum of terpenoid indole alkaloids. since the invaluable antineoplastic bisindole alkaloids are restricted to the aerial parts of the plant and do not occur in its underground tissues, identification of the structural and regulatory factors operating distinctly in the shoot/leaf of the plant will be a necessity for modulation of bisindole alkaloid biosynthesis. this study aimed at elucidating the differential gene ...200617050644
inorganic phosphate uptake in intact vacuoles isolated from suspension-cultured cells of catharanthus roseus (l.) g. don under varying pi status.inorganic phosphate (pi) uptake across the vacuolar membrane of intact vacuoles isolated from catharanthus roseus suspension-cultured cells was measured. under low pi status, pi uptake into the vacuole was strongly activated compared to high pi status. since pi uptake across the vacuolar membrane is correlated with h+ pumping, we examined the dependency of h+ pumping on plant pi status. both h+ pumping and the activities of the vacuolar h+-pumps, the v-type h+-atpase and the h+-ppase were enhanc ...200716955272
predicting leaf-level fluxes of o3 and no2: the relative roles of diffusion and biochemical processes.pollutants like o(3) and no(2) enter leaves through the stomata and cause damage during reactions with components of biological cell membranes. the steady-state flux rates of these gases into the leaf are determined by a series of physical and biochemical resistances including stomatal aperture, reactions occurring within the cell wall and the ability of the leaf to remove the products of apoplastic reactions. in the present study, multiple regression models incorporating stomatal conductance, a ...200616913863
expressed sequence tags from madagascar periwinkle (catharanthus roseus).the madagascar periwinkle (catharanthus roseus) is well known to produce the chemotherapeutic anticancer agents, vinblastine and vincristine. in spite of its importance, no expressed sequence tag (est) analysis of this plant has been reported. two cdna libraries were generated from rna isolated from the base part of young leaves and from root tips to select 9,824 random clones for unidirectional sequencing, to yield 3,327 related sequences and 1,696 singletons by cluster analysis. putative funct ...200616870181
identification of a low vindoline accumulating cultivar of catharanthus roseus (l.) g. don by alkaloid and enzymatic profiling.the madagascar periwinkle [catharanthus roseus (l.) g. don] is a commercially important horticultural flower species and is the only source of the monoterpenoid indole alkaloids (mias), vinblastine and vincristine, key pharmaceutical compounds used to combat a number of different cancers. the present study uses high performance liquid chromatography for metabolic profiling of the mias extracted from seedlings and young leaves of 50 different flowering cultivars of c. roseus to show that, except ...200616806326
catharanthus roseus shoot cultures for the production of monoterpenoid indole alkaloids.a protocol for the establishment of in vitro shoot cultures of catharanthus roseus is described. shoots can be maintained for more than 1 yr without evidence of tissue vitrification, disaggregation, or callus formation. vindoline was the main alkaloid accumulated, reaching values similar to those found in leaves from field-grown plants, after a long period of culture. an induction methodology to reduce such waiting time is also presented.200616673929
methylation of sulfhydryl groups: a new function for a family of small molecule plant o-methyltransferases.in plants, type i and ii s-adenosyl-l-methionine-dependent o-methyltransferases (omts) catalyze most hydroxyl group methylations of small molecules. a homology-based rt-pcr strategy using catharanthus roseus (madagascar periwinkle) rna previously identified six new type i plant omt family members. we now describe the molecular and biochemical characterization of a seventh protein. it shares 56-58% identity with caffeic acid omts (comts), but it failed to methylate comt substrates, and had no act ...200616623883
gene-to-metabolite networks for terpenoid indole alkaloid biosynthesis in catharanthus roseus cells.rational engineering of complicated metabolic networks involved in the production of biologically active plant compounds has been greatly impeded by our poor understanding of the regulatory and metabolic pathways underlying the biosynthesis of these compounds. whereas comprehensive genome-wide functional genomics approaches can be successfully applied to analyze a select number of model plants, these holistic approaches are not yet available for the study of nonmodel plants that include most, if ...200616565214
micropropagation: a tool for the production of high quality plant-based medicines.medicinal plants are the most important source of life saving drugs for the majority of the world's population. the biotechnological tools are important to select, multiply and conserve the critical genotypes of medicinal plants. plant tissue culture techniques offer an integrated approach for the production of standardized quality phytopharmaceutical through mass-production of consistent plant material for physiological characterization and analysis of active ingredients. micropropagation proto ...200616472132
precursor limitations in methyl jasmonate-induced catharanthus roseus cell cultures.jasmonates enhance the expression of various genes involved in terpenoid indole alkaloid (tia) biosynthesis in catharanthus roseus. we applied precursor feeding to our c. roseus suspensions to determine how methyl jasmonate (mj) alters the precursor availability for tia biosynthesis. c. roseus suspensions were induced with mj (100 microm) on day 6 and fed loganin (0.30 mm), tryptamine (0.15 mm), loganin plus tryptamine, or geraniol (0.1-1.0 mm) on day 7. while mj increased ajmalicine production ...200616432630
isolation of a cdna encoding the alpha-subunit of caax-prenyltransferases from catharanthus roseus and the expression of the active recombinant protein farnesyltransferase.crfta/ggt_ia (af525030), a cdna encoding the ?-subunit of the two types of caax-prenyltransferase (caax-ptase), i.e. protein farnesyltransferase (pft) and type i protein geranylgeranyltransferase, was cloned from catharanthus roseus via a pcr strategy. crfta/ggt_ia is 1381-bp long and bears a 999-bp open reading frame encoding a protein of 332 residues (fta) that shares 66% identity with its lycopersicon esculentum orthologue. southern blot analysis revealed that fta is encoded by a single gene ...200516341273
epidermis is a pivotal site of at least four secondary metabolic pathways in catharanthus roseus aerial organs.catharanthus roseus produces a wide range of secondary metabolites, some of which present high therapeutic values such as antitumoral monoterpenoid indole alkaloids (mias), vinblastine and vincristine, and the hypotensive mia, ajmalicine. we have recently shown that a complex multicellular organisation of the mia biosynthetic pathway occurred in c. roseus aerial organs. in particular, the final steps of both the secoiridoid-monoterpene and indole pathways specifically occurred in the epidermis o ...200616322983
effect of long-term phosphate starvation on the levels and metabolism of purine nucleotides in suspension-cultured catharanthus roseus cells.the effect of long-term phosphate (pi) starvation of up to 3 weeks on the levels of purine nucleotides and related compounds was examined using suspension-cultured catharanthus roseus cells. levels of adenine and guanine nucleotides, especially atp and gtp, were markedly reduced during pi-starvation. there was an increase in the activity of rnase, dnase, 5'- and 3'-nucleotidases and acid phosphatase, which may participate in the hydrolysis of nucleic acids and nucleotides. accumulation of adenos ...200616321409
magnetic field exposure stiffens regenerating plant protoplast cell walls.single suspension-cultured plant cells (catharanthus roseus) and their protoplasts were anchored to a glass plate and exposed to a magnetic field of 302 +/- 8 mt for several hours. compression forces required to produce constant cell deformation were measured parallel to the magnetic field by means of a cantilever-type force sensor. exposure of intact cells to the magnetic field did not result in any changes within experimental error, while exposure of regenerating protoplasts significantly incr ...200616304695
localization of tabersonine 16-hydroxylase and 16-oh tabersonine-16-o-methyltransferase to leaf epidermal cells defines them as a major site of precursor biosynthesis in the vindoline pathway in catharanthus roseus.the madagascar periwinkle (catharanthus roseus) produces the well known and remarkably complex anticancer dimeric alkaloids vinblastine and vincristine, which are derived by the coupling of vindoline and catharanthine monomers. recent data from in situ rna hybridization and immunolocalization suggest that combinatorial cell factories within the leaf are involved in vindoline biosynthesis. in this study, the cell types responsible for vindoline biosynthesis were identified by laser-capture microd ...200516262708
effects of terpenoid precursor feeding on catharanthus roseus hairy roots over-expressing the alpha or the alpha and beta subunits of anthranilate synthase.among the pharmacologically important terpenoid indole alkaloids produced by catharanthus roseus are the anti-cancer drugs vinblastine and vincristine. these two drugs are produced in small yields within the plant, which makes them expensive to produce commercially. metabolic engineering has focused on increasing flux through this pathway by various means such as elicitation, precursor feeding, and introduction of genes encoding specific metabolic enzymes into the plant. recently in our lab, a f ...200616240438
development of a novel system for producing ajmalicine and serpentine using direct culture of leaves in catharanthus roseus intact plant.due to problems of production instability, the production of plant secondary metabolites using dedifferentiated cells (callus) is not always feasible on an industrial scale. to propose a new methodology, which does not use dedifferentiated cells, a novel system for producing useful secondary metabolites using the direct culture of intact plant leaves was developed. catharanthus roseus was used as a model medicinal plant to produce terpenoid indole alkaloids (tias) by suspension culture of the le ...200516233779
a novel hydroxylase from catharanthus roseus participating in the hydroxylation of 2-hydroxybenzoic acid.a novel 55-kda hydroxylase was isolated from cultured cells of catharanthus roseus by a three-step procedure: anion exchange chromatography, affinity chromatography and hydroxylapatite adsorption chromatography. the enzyme specifically catalyzed the hydroxylation of 2-hydroxybenzoic acid to give 2,5-dihydroxybenzoic acid. the enzyme activity was optimal at ph 7.8 and was completely inhibited by divalent cations, such as cu(2+) and hg(2+). the enzyme showed sequence similarity to certain plant fl ...200416233668
production of 5'-phosphodiesterase by catharanthus roseus cells promoted by heat-degraded products generated from uronic acid.polyalginate was autoclaved at 121 degrees c for 20 min and its molecular weight distribution was analyzed. the autoclaved alginate yielded alginate polymer, oligomer and heat degraded products (hdps). each of the separated substances promoted 5'-phosphodiesterase (5'-pdase) production in suspension culture of catharanthus roseus cells. hdps could also be generated from other uronic acids (galacturonic acid and glucuronic acid) by autoclave treatment. the most effective substance in the hdps was ...200216233285
development of an apparatus for monitoring protoplast isolation from plant tissues based on both dielectric and optical methods.in order to develop a method allowing objective determination of the optimal conditions for the isolation of protoplasts, the process of protoplast isolation from plant tissues was quantitatively evaluated. first, a specialized spectrophotometer cuvette (working volume = 2.0 ml) was designed for the continuous monitoring of protoplast isolation from plant tissues based on the optical method. homogeneous mixing of tissue sections and the protoplast suspension in the cuvette was accomplished by me ...199916232551
rapid identification of vinca alkaloids by direct-injection electrospray ionisation tandem mass spectrometry and confirmation by high-performance liquid chromatography-mass spectrometry.a simple and rapid method for the identification of vinca alkaloids from a crude extract of catharanthus roseus g. don (apocynaceae) by direct-injection electrospray ionisation (esi) and tandem mass spectrometry (ms/ms) has been developed. the alkaloids vindoline, vindolidine, vincristine and vinblastine were evaluated in a commercial extract of c. roseus using this method. catharanthine and its isomers 19s-vindolinine and vindolinine were detected in the commercial product by direct injection e ...200616223089
simultaneous determination of vincristine, vinblastine, catharanthine, and vindoline in leaves of catharanthus roseus by high-performance liquid chromatography.a simple reversed-phase liquid chromatographic method is developed for the simultaneous quantitation of the anticancerous drugs vincristine, vinblastine, and their precursors catharanthine and vindoline using a merck chromolith performance reversed-phase high-performance liquid chromatography column. a better resolution is obtained in comparison with available particulate-type c18 columns. the column provides good reproducibility and peak symmetry. chromatography is carried isocratically with a ...200516212789
transient effects of overexpressing anthranilate synthase alpha and beta subunits in catharanthus roseus hairy roots.catharanthus roseus produces two economically valuable anticancer drugs, vinblastine and vincristine. these drugs are members of the terpenoid indole alkaloids and accumulate in small quantities within the plant; thus these two drugs are expensive to produce. metabolic engineering efforts have focused on increasing the alkaloids in this pathway through various means such as elicitation, precursor feeding, and gene overexpression. recently we successfully expressed arabidopsis genes encoding a fe ...200616209565
expression of the arabidopsis feedback-insensitive anthranilate synthase holoenzyme and tryptophan decarboxylase genes in catharanthus roseus hairy roots.in plants, the indole pathway provides precursors for a variety of secondary metabolites. in catharanthus roseus, a decarboxylated derivative of tryptophan, tryptamine, is a building block for the biosynthesis of terpenoid indole alkaloids. previously, we manipulated the indole pathway by introducing an arabidopsis feedback-insensitive anthranilate synthase (as) alpha subunit (trp5) cdna and c. roseus tryptophan decarboxylase gene (tdc) under the control of a glucocorticoid-inducible promoter in ...200616188339
ajmalicine production in methyl jasmonate-induced catharanthus roseus cell cultures depends on ca2+ level.cytosolic ca(2+) and jasmonate mediate signals that induce defense responses in plants. in this study, the interaction between ca(2+) and methyl jasmonate (mj) in modulating defense responses was investigated by monitoring ajmalicine production in catharanthus roseus suspension cultures. c. roseus suspensions were treated with nine combinations of cacl(2) (3, 23, and 43 mm) and mj (0, 10, and 100 microm) on day 6 of growth. increased ca(2+) influx through the addition of extracellular cacl(2) su ...200516094527
biotransformation of alpha-santonin by cell suspension cultures of five plants.cell suspension cultures of five plants (catharanthus roseus, ginkgo biloba, platycodon grandiflorum, taxus cuspidata, phytolacca asinosa) were employed to bioconvert the eudesmanolide compound, alpha-santonin. reactions occurring were hydroxylation (c-1, c-11 and c-15), reduction of the double bond [1(2) or 3(4)], rearrangment of the eudesmanolide skeleton to a guaianolide skeleton and lactone-ring hydrolysis. four new compounds were identified.200516086262
fosmidomycin analogues as inhibitors of monoterpenoid indole alkaloid production in catharanthus roseus cells.substituted 3-[2-(diethoxyphosphoryl)propyl]oxazolo[4,5-b]pyridine-2(3h)-ones were obtained by functionalization at 6-position with various substituents (aryl, vinyl, carbonyl chains) via reactions catalysed with palladium. we found that these new fosmidomycin analogues inhibited the accumulation of ajmalicine, a marker of monoterpenoid indole alkaloids production in plant cells. some of them have greater inhibitory effect than fosmidomycin and fully inhibit alkaloid accumulation at the concentr ...200516054176
processing methods for differential analysis of lc/ms profile data.liquid chromatography coupled to mass spectrometry (lc/ms) has been widely used in proteomics and metabolomics research. in this context, the technology has been increasingly used for differential profiling, i.e. broad screening of biomolecular components across multiple samples in order to elucidate the observed phenotypes and discover biomarkers. one of the major challenges in this domain remains development of better solutions for processing of lc/ms data.200516026613
[role of elements and physiologically active compounds in the regulation of synthesis and accumulation of indole alkaloids in catharanthus roseus l].effects of various elements (co, ni, zn, w, mn, cr, b, mo, fe, and v), natural and synthetic auxins, cytokinins, and gibberellin on biosynthesis and accumulation of indole alkaloids was studied at increasing concentrations in the model system of madagascar periwinkle seedlings (catharanthus roseus l.). the main types of concentration dependences for the effect of physiologically active compounds under study were evaluated. a possible mechanism of the influence of zn and auxin on this process was ...200615977796
cytokinin and ethylene control indole alkaloid production at the level of the mep/terpenoid pathway in catharanthus roseus suspension cells.the madagascar periwinkle catharanthus roseus accumulates a number of terpenoid indole alkaloids, some of which have high therapeutic interest. the biotechnological approach with cells in vitro remains an alternative to the field culture of periwinkle for the production of such compounds. we previously reported that two phytohormones, cytokinin and ethylene, remarkably enhanced the accumulation of alkaloids in periwinkle cell suspensions. in this work, we investigated the effects of these hormon ...200515971133
phytic acid synthesis and vacuolar accumulation in suspension-cultured cells of catharanthus roseus induced by high concentration of inorganic phosphate and cations.we have established a new system for studying phytic acid, myo-inositol hexakisphosphate (insp(6)) synthesis in suspension-cultured cells of catharanthus. insp(6) and other intermediates of myo-inositol (ins) phosphate metabolism were measured using an ion chromatography method. the detection limit for insp(6) was less than 50 nm, which was sufficient to analyze ins phosphates in living cells. synthesis of ins phosphates was induced by incubation in high inorganic phosphate medium. insp(6) was m ...200515965017
caax-prenyltransferases are essential for expression of genes involvedin the early stages of monoterpenoid biosynthetic pathway in catharanthus roseus cells.caax-prenyltransferases (caax-ptases) catalyse the covalent attachment of isoprenyl groups to conserved cysteine residues located at the c-terminal caax motif of a protein substrate. this post-translational modification is required for the function and/or subcellular localization of some transcription factors and components of signal transduction and membrane trafficking machinery. caax-ptases, including protein farnesyltransferase (pft) and type-i protein geranylgeranyltransferase (pggt-i), are ...200515952070
cytochrome p450 2d6 (cyp2d6) inhibitory constituents of catharanthus roseus.the meoh-soluble fraction of the water extract of catharanthus roseus from indonesia, having shown potent inhibitory activity on the metabolism mediated by cyp2d6, was subjected to activity-guided isolation to yield two triterpenes, ursolic acid (1) and oleanolic acid (2), and three alkaloids, vindoline (3), ajmalicine (4), and serpentine (5). the isolated compounds were tested for their inhibitory activity on the metabolism mediated by cyp3a4 or cyp2d6 using [n-methyl-14c]erythromycin or [o-met ...200515930738
plant cell culture monitoring using an in situ multiwavelength fluorescence probe.a multiwavelength fluorescence probe is proposed for in situ monitoring of eschscholtzia californica and catharanthus roseus plant cell cultures. the potential of the probe as a tool for real-time estimation of biomass and production in secondary metabolites has been studied. the probe excitation range is 270-550 nm and the emission range is 310-590 nm, with a step of 20 nm for both excitation and emission filters. many endogenous fluorophores such as nad(p)h, riboflavins (riboflavin and derivat ...201315801802
effect of nitric oxide on catharanthine production and growth of catharanthus roseus suspension cells.sodium nitroprusside (snp) was used as the donor of nitric oxide (no) to investigate its effect on catharanthine synthesis and the growth of catharanthus roseus suspension cells. the results showed that snp at high concentrations (10.0 and 20.0 mmol/l) stimulated catharanthine formation of c. roseus cells, but inhibited growth of the cells. low concentrations of snp (0.1 and 0.5 mmol/l) enhanced the growth of c. roseus cells, but had no effect on catharanthine synthesis. the maximum total cathar ...200515744842
purification, molecular cloning, and cell-specific gene expression of the alkaloid-accumulation associated protein crps in catharanthus roseus.identification of molecular markers of monoterpenoid indole alkaloid (mia) accumulation in cell-suspension cultures of madagascar periwinkle (catharanthus roseus (l.) g. don) was performed by two-dimensional polyacrylamide gel electrophoresis. comparison of the protein patterns from alkaloid-producing and non-producing cells showed the specific occurrence of a 28 kda polypeptide restricted to cells accumulating mias. the polypeptide was purified by preparative two-dimensional gel electrophoresis ...200515737982
expression of terpenoid indole alkaloid biosynthetic pathway genes corresponds to accumulation of related alkaloids in catharanthus roseus (l.) g. don.madagascar periwinkle, catharanthus roseus (l.) g. don, a medicinally important plant, produces anticancer dimeric alkaloids, vinblastine and vincristine, in the leaves and accumulates antihypertensive alkaloids, ajmalicine and serpentine, in the roots. this plant grows wild in distant tropical and sub-tropical geographical locations with different agro-climates and shows wide variations in morphological and alkaloid yield-related traits. in order to understand the correlation between the expres ...200515714355
proteome analysis of the medicinal plant catharanthus roseus.a proteomic approach is undertaken aiming at the identification of novel proteins involved in the alkaloid biosynthesis of catharanthus roseus. the c. roseus cell suspension culture a11 accumulates the terpenoid indole alkaloids strictosidine, ajmalicine and vindolinine. cells were grown for 21 days, and alkaloid accumulation was monitored during this period. after a rapid increase between day 3 and day 6, the alkaloid content reached a maximum on day 16. systematic analysis of the proteome was ...200515682277
simultaneous quantification of some pharmaceutical catharanthus roseus leaf and root terpenoid indole alkaloids and their precursors in single runs by reversed-phase liquid chromatography.a new, rapid, sensitive, and reproducible reversed-phase liquid chromatographic (lc) method with photodiode array detection is described. it allows, in a single run of 30 min, simultaneous separation of 6 pharmaceutically and biologically important catharanthus roseus leaf and root terpenoid indole alkaloids (tias) and 3 of their precursors: tia precursors tryptophan, tryptamine, and loganine; and tias serpentine, catharanthine, ajmalicine, vincristine, vinblastine, and vindoline. the method inv ...200815675438
methyljasmonate accelerates catabolism of monoterpenoid indole alkaloids in catharanthus roseus during leaf processing.variations in alkaloid pattern during drying of leaves (leaf processing) showed that treatment with methyljasmonate can induce formation of bisindole alkaloids as a result of catabolism of the monomeric alkaloids catharanthine and vindoline. a two-fold increase in 3',4'-anhydrovinblastine was shown in treated leaves especially from day 8 until day 21. serpentine also increased in the same period under the treatment as a catabolic product of ajmalicine. basic peroxidases that are responsible for ...200515664467
[advances in the study of vincristine: an anticancer ingredient from catharanthus roseus].vincristine is a dimer-indo-alkaloid which is extracted from the leaves of catharanthus roseus. it is effective to treat acute lymphocytic cell leukemia, hodgkin disease and non-hodgkin disease clinically. but the severe side effects, such as neurotoxic and tissue damage, limit its application. in this paper, we summarize physical, chemical, pharmacological and pharmacokinetical properties of vcr and advances in decreasing its side effects. in clinic, association with other medication is adopted ...200315615402
enhancement of ajmalicine production in catharanthus roseus cell cultures with methyl jasmonate is dependent on timing and dosage of elicitation.the optimum growth stage for enhancing ajmalicine production in catharanthus roseus cultures with methyl jasmonate (mj) was after 6 d growth. mj added at 10 or 100 microm on day 6 gave a maximum ajmalicine production of 10.2 mg l(-1), a 300% increase over that of non-elicited cultures.200415604804
allelic differences at two loci govern different mechanisms of intraflower self-pollination in self-pollinating strains of periwinkle.periwinkle [catharanthus roseus (l.) g. don], an ornamental and medicinal plant, is a self-compatible, insect-pollinated plant species in which intraflower self-pollination does not occur because of spatial separation of the stigma and anthers. recently three self-pollinating strains-mj, vi, and or-were identified. self-pollination in these strains was found to be brought about by continuous increase in gynoecium length from anthesis to self-pollination, in contrast to non-self-pollinating strai ...200615598715
metabolic engineering of the indole pathway in catharanthus roseus hairy roots and increased accumulation of tryptamine and serpentine.transgenic hairy roots of catharanthus roseus were established with glucocorticoid inducible tryptophan decarboxylase (tdc) expression alone or in combination with inducible expression of a feedback-resistant anthranilate synthase alpha subunit (asalpha) from arabidopsis. northern blot analysis confirmed transcription of the anthranilate synthase gene upon induction in the double line (tdc+asalpha) and in vitro enzyme assays confirmed increased resistance to feedback inhibition by tryptophan. in ...200415491856
alkaloid metabolism in wounded catharanthus roseus seedlings.the effect of mechanical wounding on alkaloid metabolism was analyzed in catharanthus roseus seedlings. wounding induced an increase in ajmalicine accumulation, whereas catharanthine remained unaffected. a positive dual effect on vindoline was noticed. short and mid-term effects were detected between 12 and 24 h after mechanical damage was inflicted, and apparently resulted from the accelerated transformation of the tabersonine intermediaries to vindoline. long-term effects involved a generalize ...201615331091
salt-induced lipid changes in catharanthus roseus cultured cell suspensions.salt treatment strongly affected cell growth by decreasing dry weight. exposure of catharanthus roseus cell suspensions to increasing salinity significantly enhanced total lipid (tl) content. the observed increase is mainly due to high level of phospholipids (pl). hundred mm nacl treatment increased phospholipid species phosphatidylcholine (pc) and phosphatidylethanolamine (pe), whereas it reduced glycolipid ones monogalactosyldiacylglycerol (mgdg) and digalactosyldiacylglycerol (dgdg) but not s ...200415279997
alkaloid accumulation in catharanthus roseus cell suspension cultures fed with stemmadenine.feeding stemmadenine to catharanthus roseus cell suspension culture resulted in the accumulation of catharanthine, tabersonine and condylocarpine. condylocarpine is not an intermediate in the pathway to catharanthine or tabersonine when it is fed to the cultures. the results support the hypothesis that stemmadenine is an intermediate in the pathway to catharanthine and tabersonine.200415269549
novel bisindole derivatives of catharanthus alkaloids with potential cytotoxic properties.pantropical plant catharanthus roseus (l) g. don is known as a source of valuable bisindole alkaloids: vinblastine (vbl) and vincristine (vcr), oncolytics widely used as sulfates in therapy of malignant diseases. they are biosynthesized in the plant from monoindolic vindoline and catharanthine, both derived from l-tryptophan and loganine units. in the course of phytochemical screening of this plant cultivated in poland and considered as a home source of vbl and vcr we developed a new isolation m ...200315206784
vindoline synthesis in in vitro shoot cultures of catharanthus roseus.vindoline, the major alkaloid in cultures of catharanthus roseus shoots, reached 2 mg g(-1) dry wt after 27 d in culture. maximal vindoline accumulation coincided with maximum activities of deacetoxyvindoline 4-hydroxylase, deacetylvindoline acetyl-coa acetyl transferase and tryptophan decarboxylase. shoot exposure to jasmonate shortened the time required for the maximal vindoline accumulation to 14 d.200415200179
expression of a feedback-resistant anthranilate synthase in catharanthus roseus hairy roots provides evidence for tight regulation of terpenoid indole alkaloid levels.different plant species produce a variety of terpenoid indole alkaloids, which are of interest as plant defensive secondary metabolites and as valuable pharmaceuticals. although significant progress has been made, the mechanisms regulating the levels of this important class of compounds require continued elucidation. previous precursor feeding studies have indicated that alkaloid accumulation can be improved during the exponential growth phase of hairy root cultures through enhanced tryptophan a ...200415137084
flavonoid methylation: a novel 4'-o-methyltransferase from catharanthus roseus, and evidence that partially methylated flavanones are substrates of four different flavonoid dioxygenases.catharanthus roseus (madagascar periwinkle) flavonoids have a simple methylation pattern. characteristic are b-ring 5' and 3' methylations and a methylation in the position 7 of the a-ring. the first two can be explained by a previously identified unusual o-methyltransferase (cromt2) that performs two sequential methylations. we used a homology based rt-pcr strategy to search for cdnas encoding the enzyme for the a-ring 7 position. full-length cdnas for three proteins were characterized (cromt5, ...200415110688
map kinase-like activity in transformed catharanthus roseus hairy roots varies with culture conditions such as temperature and hypo-osmotic shock.mitogen activated protein (map) kinase-like activity was determined in extracts obtained from transformed catharanthus roseus hairy roots by the ability to phosphorylate myelin basic protein (mbp). both in solution and in gel kinase assays showed variation in activity, depending on root developmental stage. in gel kinase assays, using the extract soluble fraction, revealed a 56 kda polypeptide with phosphorylation activity on mbp. in addition, another 75 kda polypeptide was observed in the parti ...200415061086
co-expression of three mep pathway genes and geraniol 10-hydroxylase in internal phloem parenchyma of catharanthus roseus implicates multicellular translocation of intermediates during the biosynthesis of monoterpene indole alkaloids and isoprenoid-derived primary metabolites.in higher plants, isopentenyl diphosphate (ipp) is synthesised both from the plastidic 2-c-methyl-d-erythritol 4-phosphate (mep) and from the cytosolic mevalonate (mva) pathways. primary metabolites, such as phytol group of chlorophylls, carotenoids and the plant hormones abscisic acid (aba) and gibberellins (gas) are derived directly from the mep pathway. many secondary metabolites, such as monoterpene indole alkaloids (mias) in catharanthus roseus, are also synthesised from this source of ipp. ...200415053766
interaction of spermine with a signal transduction pathway involving phospholipase c, during the growth of catharanthus roseus transformed roots.in cantharanthus roseus transformed roots, the application of methylglyoxal bis(guanylhydrazone) (mgbg), an inhibitor of s-adenosylmethionine decarboxylase (samdc; ec 4.1.1.50), inhibited the root growth in a dose-dependent manner with a dl(50) of about 300 micro m. spermidine and spermine (spm) levels and samdc and phospholipase c (plc; ec 3.1.4.3) activities were reduced in the presence of the inhibitor. the inhibition was reversed by the addition of spm. radioactivity from [(14)c]spm was dete ...200415032886
biotransformation of triptolide and triptonide by cell suspension cultures of catharanthus roseus.catharanthus roseus cell suspension cultures were used to bioconvert both triptolide (1) and triptonide (2). the same reaction path was followed in both biotransformations. two biotransformed products were obtained and their structures identified as triptriolide (3) and 12beta,13alpha-dihydroxytriptonide (4), respectively, from 1 and 2. product 4 is a new compound.200415008455
histidine-containing phosphotransfer domain extinction by rna interference turns off a cytokinin signalling circuitry in catharanthus roseus suspension cells.we previously reported that cytokinins (ck) induce the fast and specific transcription of crrr1, a gene encoding a type a response regulator in catharanthus roseus cell cultures. here, we characterized the crhpt1 gene that encodes a histidine-containing phosphotransfer domain. crhpt1 was silenced through rna interference (rnai) to test its possible implication in the ck signalling pathway. in transgenic lines stably transformed with an intron-spliced construct, the degradation of crhpt1 transcri ...200414759521
production of unnatural glucosides of curcumin with drastically enhanced water solubility by cell suspension cultures of catharanthus roseus.catharanthus roseus cell suspension cultures converted exogenously supplied curcumin to a series of glucosides, none of which has been found in nature so far. the efficiency of glucosylation was dependent on culture stage of the cells and medium sucrose concentration. methyl jasmonate and salicylic acid enhanced the glucoside formation only when they were added to the cultures prior to the addition of curcumin. the glucoside yield was 2.5 micromol/g fresh weight of the cells at an optimal cultur ...200314644434
production of cell wall accumulative enzymes using immobilized protoplasts of catharanthus roseus in agarose gel.catharanthus roseus cells and protoplasts were used for production of peroxidase and alpha-galactosidase which are accumulated in the cell wall. only 4% (0.026 u ml(-1)) of the total peroxidase was secreted into the broth by cultured cells while in cultured protoplasts, 45% (0.12 u ml(-1)) was secreted. protoplasts were protected against the physical and osmotic stresses by immobilizing them in 3% agarose gel (high mass transfer, non-electric charge, low gelation temperature). in order to increa ...200314626409
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