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1.
Peter Lundberg Hans J. Vogel Peter E. Brodelius 《In vitro cellular & developmental biology. Plant》1997,33(4):301-305
Summary The induction of metabolic changes in suspension cultured cells of Catharanthus roseus upon elicitation has been investigated. Addition of a yeast glucan preparation to the growth medium resulted in induction
of phenylalanine ammonia lyase. Phosphate uptake and metabolism of elicited cells was followed by 31P nuclear magnetic resonance. The uptake rate of Pi from the medium by oxygenated cells of C. roseus was reduced immediately after elicitation. Despite this reduced Pi uptake elicited cells had significantly increased amounts
of ATP (twofold increase within 6 h). Cytoplasmic levels of Pi, phosphomonoesters, and Uridine Diphasphate glucose (UDP-Glc)
were unaffected by eliciation. Furthermore, the cytoplasmic and vacuolar pH remained constant after addition of elicitor. 相似文献
2.
Effect of terpenoid precursor feeding and elicitation on formation of indole alkaloids in cell suspension cultures of Catharanthus roseus 总被引:2,自引:0,他引:2
The effects of terpenoid precursor feeding and elicitation by a biotic elicitor on alkaloid production of Catharanthus roseus suspension cultures were studied. After addition of secologanin, loganin or loganic acid an increase in the accumulation of ajmalicine and strictosidine and a decrease of tryptamine level was observed in non-elicited cells. Elicitation increased tryptamine accumulation in non-fed cells but it did not further increase alkaloid accumulation in precursor-fed cells. A decrease of tryptamine level was also observed, despite the induction of the tryptamine pathway after elicitation. Feeding mevalonic acid did not increase alkaloid accumulation in any studied case. 相似文献
3.
Phytoalexin Induction in French Bean : Intercellular Transmission of Elicitation in Cell Suspension Cultures and Hypocotyl Sections of Phaseolus vulgaris 总被引:2,自引:2,他引:0 下载免费PDF全文
Treatment of hypocotyl sections or cell suspension cultures of dwarf French bean (Phaseolus vulgaris L.) with an abiotic elicitor (denatured ribonuclease A) resulted in increased extractable activity of the enzyme l-phenylalanine ammonia-lyase. This induction could be transmitted from treated cells through a dialysis membrane to cells which were not in direct contact with the elicitor. In hypocotyl sections, induction of isoflavonoid phytoalexin accumulation was also transmitted across a dialysis membrane, although levels of insoluble, lignin-like phenolic material remained unchanged in elicitor-treated and control sections. In bean cell suspension cultures, the induction of phenylalanine ammonia-lyase in cells separated from ribonuclease-treated cells by a dialysis membrane was also accompanied by increases in the activities of chalcone synthase and chalcone isomerase, two enzymes previously implicated in the phytoalexin defense response. Such intercellular transmission of elicitation did not occur in experiments with cells treated with a biotic elicitor preparation heat-released from the cell walls of the bean pathogen Colletotrichum lindemuthianum. The results confirm and extend previous suggestions that a low molecular weight, diffusible factor of host plant origin is involved (in French bean) in the intercellular transmission of the elicitation response to abiotic elicitors. 相似文献
4.
H. U. Seitz U. Eilert V. De Luca W. G. W. Kurz 《Plant Cell, Tissue and Organ Culture》1989,18(1):71-78
Cell suspension cultures (cell line No 615) of Catharanthus roseus cv. Little Delicata responded to elicitor treatment by accumulating monoterpenoid indole alkaloids and phenolic compounds. The excretion of phenols into the culture medium resulted from the induction of the branch-point enzyme phenylalanine ammonia lyase. The accumulation of alkaloids, however, occurred several hours earlier than the elicitor-mediated induction of tryptophan decarboxylase through which shikimate pathway intermediates are channelled into tryptamine and related indole alkaloids. The results indicate that both pathways for phenol and indole alkaloid biosynthesis responded to elicitor treatment and that no obvious causal relationship between pathways could be deduced from this study.Abbreviations PAL
phenylalanine ammonia lyase
- TDC
tryptophan decarboxylase
Dedicated to Dr. Friedrich Constabel on the occasion of his 60th birthday 相似文献
5.
The effects of two synthetic elicitor indanoyl-isoleucine (In-Ile), N-linolenoyl-l-glutamine (Lin-Gln) and one biotic elicitor insect saliva (from Manduca sexta larvae) on plant cell cultures with respect to the induction of secondary metabolite production were investigated. Stimulated production of secondary metabolites, particularly anthocyanins in plant cells and phenolic acids in culture medium, was studied by using suspension culture of Vitis vinifera L. cv. Gamay Fréaux as a model system. In the treatments with In-Ile, the production of anthocyanins was enhanced 2.6-fold. In-Ile, Lin-Gln and saliva significantly elevated the accumulation of phenolic acids, particularly 3-O-glucosyl-resveratrol. The used elicitors did not suppress cell growth. Secondary metabolites were differently responsive to elicitation. 3-O-glucosyl-resveratrol was the predominant phenolic acid in V. vinifera cell culture, and its production was significantly stimulated by saliva, with 7.0-fold of the control level 24 h after treatment. The production of 4-(3,5-dihydroxy-phenyl)-phenol was significantly stimulated by In-Ile with 6.4-fold of the control level 24 h after treatment. 相似文献
6.
Summary More efficient bioreactors for the production and recovery of secondary metabolites from plant cell cultures are needed. Three factors that have the potential to increase productivity are adsorption in situ, elicitors, and cell immobilization. The effects of these factors on ajmalicine production from Catharanthus roseus are reported in this paper. Elicitation using autoclaved cultures of the mold, Phytophthora cactorum, stimulates a 60% increase in ajmalicine production. The response time to elicitor addition was under 11 h. Adsorption of ajmalicine from the extracellular medium with the neutral resin, Amberlite XAD-7, greatly enhanced the release of ajmalicine (less than 10% extracellular to 40%) with a 40% increase in total productivity. Immobilization in Caalginate beads resulted in a significant increase in the accumulation of ajmalicine in the medium. The effects of elicitation, adsorption and immobilization were synergistic. For a 23-day culture period the amount of ajmalicine in the medium for cells subjected to all three treatments was 90 mg/L compared to 2 mg/L for suspension cultures cultured under otherwise identical conditions. These results suggest that immobilized cell bioreactors may be feasible for continuous production of products normally stored intracellularly in vacuoles in plant cells. 相似文献
7.
《Plant Physiology and Biochemistry》2000,38(3):233-241
Cell suspension cultures of Eschscholtzia californica produce relatively large amounts of benzophenanthridine alkaloids upon elicitation. Sodium orthovanadate is used as an abiotic elicitor to induce alkaloid biosynthesis in cultures of E. californica. The response of the cell culture to this abiotic elicitor is very similar to that observed after elicitation with a biotic elicitor (a carbohydrate fraction from yeast extract). Treatment with orthovanadate leads to alkalinization of the growth medium, a 20-fold induction of the key enzyme tyrosine decarboxylase and increased alkaloid formation (up to 40 mg.L–1). Cells treated with the yeast elicitor excrete a large portion of alkaloids produced into the growth medium (up to 50 % of total alkaloids) while cells treated with orthovanadate release very small amounts of alkaloids into the medium (less than 10 % of total alkaloids). These results suggest that an active transport system, possibly specific for benzophenanthridine alkaloids, is present in the plasma membrane of E. californica cells. The nature of this putative vanadate-sensitive transporter is not known at present. 相似文献
8.
Elicitation of 2,3-Dihydroxybenzoic Acid and Ajmalicine in a Catharanthus roseus Suspension Culture*
Addition of an elicitor preparation from cell walls of Phytophthora megasperma f. sp. glycinea (Pmg elicitor) to a newly established cell suspension culture of Catharanthus roseus induced extracellular free 2,3-dihydroxybenzoic acid, suggesting its role in pathogen defense. The same substance also accumulated intracellularly in a bound form. Treatment of the crude Pmg elicitor preparation with trypsin abolished elicitor activity, suggesting that the active fraction is proteinaceous. The cells became more sensitive to low but not to elevated elicitor concentrations when they were pretreated with 2,6-dichloroisonicotinic (DCIA) or 5-chlorosalicylic (5CSA) acid for about 1 day before addition of the elicitor. This indicates that the elicitor reception/transduction system becomes improved by these compounds known to be related to systemic acquired resistance against plant pathogens. The newly established cell line initially accumulated also the indole alkaloid ajmalicine, a process enhanced by Pmg elicitor. This potency was lost during subculturing for about 1 year and was also not restored by preincubation with DCIA or 5CSA. In contrast, elicitation of 2,3-dihydroxybenzoic acid synthesis was undiminished, suggesting that the Pmg elicitor perception system was still functioning and not the cause for the decline in elicited indole alkaloid production. 相似文献
9.
Effects of elicitation with heavy metals such as copper, cadmium, chromium (abiotic elicitation) and supplementation of CaCl2 on production of dipyranocoumarins (inophyllums) in suspension cultures of leaf and stem callus of Calophyllum inophyllum were studied. The optimum timing for elicitor introduction was found to be the 10th day after initiating the suspension cultures.
Cadmium as abiotic elicitor in suspension cultures of stem callus was found best to elicit maximum production of inophyllums
A, C, and calophyllolide while cadmium in suspension cultures of leaf callus was found best for eliciting maximum production
of inophyllums B and P. Inophyllum D was the only dipyranocoumarin whose highest production was achieved when 1.0 mM chromium
was used as abiotic elicitor in suspension cultures of stem callus. Out of the three abiotic elicitors used, none could result
biomass growth. Only incorporation of CaCl2 in suspension cultures resulted biomass growth. A maximum of 35.26-fold biomass growth was achieved when suspension cultures
of stem callus were incorporated with 2.0 mM CaCl2. CaCl2 was noted to have no positive influence on production of most of the dipyranocoumarins under study. 相似文献
10.
Inhibition of elicitor-induced phytoalexin formation in cotton and soybean cells by citrate 总被引:3,自引:2,他引:1 下载免费PDF全文
Addition of an elicitor preparation from Verticillium dahliae to soybean or cotton cell suspension cultures induces the formation of the phytoalexins, glycelollin or sesquiterpene aldehydes, respectively. Recent work (PS Low, PF Heinstein 1986 Arch Biochem Biophys 249: 472-479) has shown that the induction of phytoalexin biosynthesis in these cells is preceded by rapid changes in the plant cell membrane which can be conveniently monitored by membrane associated fluorescent probes. Using this elicitation assay, we have found that citrate, a common metabolite of higher plants, acts as a potent inhibitor of elicitation when added prior to treatment with elicitor. The citrate concentration required to obtain a 50% inhibition of the elicitor-induced fluorescence transition in cultured cotton cells was found to be about 2 millimolar, while the concentration of citrate observed to inhibit elicitor-induced sesquiterpene aldehyde formation in the same cell suspensions was also 2 millimolar. Curiously, in the presence of elicitor, citrate at less than ID50 concentrations increased cell mass accumulation significantly above control incubations without elicitor. A similar inhibition of glyceollin formation with an increase in cell mass accumulation was also observed upon addition of 1 to 5 millimolar citrate to soybean cell suspension cultures. The physiological significance of the inhibition by citrate of phytoalexin formation in plant cell suspensions was supported by the observation that a similar inhibition of sesquiterpene aldehyde formation occurs in cotton plantlets elicited by cold shock or V. dahliae stress. The specificity of citrate as an inhibitor of phytoalexin formation was demonstrated by data showing that other di- and tricarboxylic-hydroxy acids did not inhibit, with the exception of malate which inhibited phytoalexin formation in soybean cells with roughly half the potency of citrate. These experiments not only demonstrate that citrate can act as a specific inhibitor of elicitation, but they further confirm the validity of monitoring elicitation and its modulation with fluorescent probes. 相似文献
11.
Qian Chen Zunwei Chen Li Lu Haihong Jin Lina Sun Qin Yu Hongke Xu Fengxia Yang Mengna Fu Shengchao Li Huizhong Wang Maojun Xu 《Biotechnology progress》2013,29(4):994-1001
Elicitations are considered to be an important strategy to improve production of secondary metabolites of plant cell cultures. However, mechanisms responsible for the elicitor‐induced production of secondary metabolites of plant cells have not yet been fully elucidated. Here, we report that treatment of Catharanthus roseus cell suspension cultures with PB90, a protein elicitor from Phytophthora boehmeriae, induced rapid increases of abscisic acid (ABA) and nitric oxide (NO), subsequently followed by the enhancement of catharanthine production and up‐regulation of Str and Tdc, two important genes in catharanthine biosynthesis. PB90‐induced catharanthine production and the gene expression were suppressed by the ABA inhibitor and NO scavenger respectively, showing that ABA and NO are essential for the elicitor‐induced catharanthine biosynthesis. The relationship between ABA and NO in mediating catharanthine biosynthesis was further investigated. Treatment of the cells with ABA triggered NO accumulation and induced catharanthine production and up‐regulation of Str and Tdc. ABA‐induced catharanthine production and gene expressions were suppressed by the NO scavenger. Conversely, exogenous application of NO did not stimulate ABA generation and treatment with ABA inhibitor did not suppress NO‐induced catharanthine production and gene expressions. Together, the results showed that both NO and ABA were involved in PB90‐induced catharanthine biosynthesis of C. roseus cells. Furthermore, our data demonstrated that ABA acted upstream of NO in the signaling cascade leading to PB90‐induced catharanthine biosynthesis of C. roseus cells. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:994–1001, 2013 相似文献
12.
When various autoclaved microbial cells suspensions (exogenous elicitors) were added to Catharanthus roseus cell cultures, its growth was inhibited but 5′-phosphodiesterase (PDase) production was stimulated. The greatest effect was with autoclaved Alteromonas macleodii: the dry cell concentration decreased from 13 to 10.9 mg/ml while PDase production increased from 0.022 to 0.235 U/ml. A combination of A. macleodii (as exogenous elicitor) and 0.1%(w/v) alginate oligomers (AO: acting as both endogenous elicitor and scavenger of active oxygen species) minimized the cell growth inhibition but enhanced PDase production (0.474 U/ml) about 20 times higher than the control (no addition). The method for the preparation of mixed alginate elicitors with high activities containing exogenous elicitor (autoclaved A. macleodii), endogenous elicitor (AO), and trans-4,5-dihydroxy-2-cyclopenten-1-one was developed. The mixed alginate elicitors significantly promoted PDase production (2.67 U/ml) by C. roseus, and the productivity was increased 120-fold compared to the control without cell growth inhibition. 相似文献
13.
Cell suspension cultures of Lycopersicon peruvianum (Solanaceae) were established as well-growing photoautotrophic, photomixotrophic and heterotrophic cultures and their growth parameters were characterized. Elicitor-induced responses of these cultures to the tomato pathogen Fusarium oxysporum f. sp. lycopersici were investigated after treatment of cells with autoclaved mycelium and culture filtrate of this fungus. The dominant reaction was an enhanced incorporation of phenolic constituents in the plant cell wall. Among the nine phenolics released by alkaline hydrolysis the most prominent compounds were p-hydroxybenzaldchyde, vanillin, p-coumaroyltryamine, feruloyltyramine, p-coumaric acid and ferulic acid. Phenolic incorporation in cell walls resulted in increased stability of cells against protoplasting with microbial enzymes. Chlorogenic acid, as the main soluble phenolic compound, showed differential accumulation in the three cell cultures lines as well as an elicitor-induced transient decrease. In heterotrophic cells decrease of chlorogenate occurred concomitant with accumulation of caffeoyl- and p-coumaroylshikimate as well as increased activities of p-coumaroyleoenzyme A: shikimic acid p-coumaroyltransferase. Upon elicitation increased activities of phenylalanine ammonia lyase and changes in peroxidase activities wore also detected. Sesquiterpenoid phytoalexines were not produced by either one of the cell culture lines and levels of tomatine were not significantly affected by elicitation. 相似文献
14.
Elicitor-induced nitric oxide burst is essential for triggering catharanthine synthesis in Catharanthus roseus suspension cells 总被引:11,自引:0,他引:11
Elicitor prepared from the cell walls of Penicillium citrinum induced multiple responses in Catharanthus roseus suspension cells, including rapid generation of nitric oxide (NO), sequentially followed by enhancement of catharanthine production by C. roseus cells. Elicitor-induced catharanthine biosynthesis was blocked by NO-specific scavenger 2-4-carboxyphenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide and nitric oxide synthase (NOS) inhibitor S,S-1,3-phenylene-bis(1,2-ethanediyl)-bis-isothiourea (PBITU). PBITU also strongly inhibited elicitor-induced NO generation by C. roseus suspension cells. The inhibiting effect of PBITU on elicitor-induced catharanthine production was reversed by external application of NO via the NO-donor sodium nitroprusside. The results strongly suggested that NO, generated by NOS or NOS-like enzymes in C. roseus suspension cells when treated with the fungal elicitor, was essential for triggering catharanthine synthesis. 相似文献
15.
Improvement of phenylethanoid glycosides biosynthesis in Cistanche deserticola cell suspension cultures by chitosan elicitor 总被引:2,自引:0,他引:2
Effect of chitosan elicitor on growth and phenylethanoid glycosides (PeGs) accumulation in Cistanche deserticola cell suspension cultures was investigated. PeGs accumulation was dramatically improved by addition of selected chitosan at optimal elicitation conditions. Furthermore, a strategy of repeated addition of the chitosan elicitor for enhancing PeGs accumulation was developed. The chitosan elicitor of 10 mg l(-1)-medium repeatedly added on days 15 and 17 improved PeGs accumulation further, and the final PeGs production in the treated cell cultures of C. deserticola reached 364.6 mg l(-1), which was 3.4-fold higher than that of the control without elicitation. The increase of PeGs accumulation in C. deserticola cell suspension cultures was related to the increase of phenylalanine ammonium lyase activity stimulated by the chitosan elicitor. 相似文献
16.
The effects of alginate on the physiological activities of plant cells were studied. Addition of alginate oligomer (AO) to
the suspension culture of Catharanthus roseus L. or Wasabia japonica cells promoted the production of antibiotic enzymes such as 5′-phosphodiesterase or chitinase respectively. Ajmalicine (a
secondary metabolite) production by C. roseus CP3 cells was also promoted when AO was added to the suspension culture. On the basis of these results, we assumed that alginate
is an elicitor-like substance. We therefore compared the effect of AO on C. roseus L. and W. japonica cells with those of chitosan oligomer (CO) and oligo-galacturonic acid (OGA), which are well known as an exogenous elicitor
and endogenous elicitor respectively. The effects of various concentrations of AO, OGA, and CO on the physiological activities,
membrane permeability and protoplast formation of C. roseus L. or W. japonica cells were investigated. AO and OGA showed similar physiological effects, which were quite different from those of CO. Since
alginate appeared to have similar effects to galacturonic acid, we concluded that alginate acts as an endogenous elicitor.
Both alginate and galacturonic acid are uronic acids, and we considered their structural similarity. The effects of esterification
of the carboxylic groups of alginate by propylene oxide were also studied. The greater the degree of esterification, the less
the secretion of 5′-phosphodiesterase. Hence we assumed that carboxylic groups have an important role in the initiation of
the elicitation reaction in plant cells, as shown in the case of galacturonic acid.
Received: 18 January 1999 / Received revision: 2 April 1999 / Accepted: 1 May 1999 相似文献
17.
Induction of sesquiterpene cyclase and suppression of squalene synthetase activities in plant cell cultures treated with fungal elicitor 总被引:4,自引:4,他引:0
Addition of elicitor, cell wall fragments of the fungus Phytophthora parasitica, to tobacco cell suspension cultures (Nicotiana tabacum) resulted in the rapid synthesis and secretion of large amounts of antibiotic sesquiterpenoids. Pulse-labeling experiments with [14C]acetate and [3H] mevalonate demonstrated that the induction of sesquiterpenoid biosynthesis, maximal by 6 to 9 hours after elicitor addition to the cell cultures, was paralleled by a rapid and large decline in the incorporation rate of radioactivity into sterols. Consequently, sterol accumulation was also inhibited upon addition of elicitor to the cell cultures. Sesquiterpene cyclase activity was absent from control cell cultures but induced to a maximum within 10 hours of elicitor addition to the cell cultures. The cyclase activity remained elevated for an additional 30 hours before declining. In contrast, squalene synthetase activity was suppressed to less than 15% of that found in control cells within 7 hours of elicitor addition. Our results suggest that the channeling of isoprenoid intermediates, and especially farnesyl diphosphate, into sesquiterpenoids occurred by a coordinated increase in the sesquiterpene cyclase and a decrease in the squalene synthetase enzyme activities. A reexamination of the data pertaining to the transient induction of 3-hydroxy-3-methylglutaryl coenzyme A reductase activity (EC 1.1.1.34) in elicitor-treated cells suggested that, while the reductase activity was necessary for sesquiterpenoid biosynthesis, it functioned more to maintain a sufficient level of intermediates between mevalonate and farnesyl diphosphate rather than as a rate limiting step controlling the synthesis rate of any one class of isoprenoids. 相似文献
18.
Changes in phenolic metabolism after elicitation with Colletotrichum gloeosporioides (CG) has been studied in Hypericum perforatum L. (HP) cell suspension cultures. Soluble phenolics were analysed by HPLC-DAD and HPLC-DAD-MS/MS. HP cultures elicited with the CG elicitor showed a significant increase in xanthone accumulation. Xanthone accumulation increased twelve fold when the cells were primed with methyl-jasmonate (MeJ) or salicylic acid (SA), before elicitation. HP cultures exposed only to MeJ produced a set of flavonoids, the flavones which represent a substantial part (approx. 40%) of the total flavonoids accumulated in these cells. The possible importance of xanthones as a component of defence mechanism of HP against biotic stress is discussed. 相似文献
19.
Summary Growth and alkaloid production of surface-immobilized C. roseus cells were studied in a 2-1 bioreactor. Media designed to maximize cell growth or alkaloid production were employed. Nitrate and carbohydrate consumption rates as well as growth rates and biomass yields of immobilized cultures were equal or somewhat lower than for cell suspension cultures. Respiration rate (O2 consumption and CO2 production rates) of immobilized C. roseus cell cultures was obtained by on-line analysis of inlet and outlet gas composition using a mass spectrometer. Respiration rate increased during the growth phase and decreased once the nitrogen or the carbon source was depleted from the medium. The respiration rate of immobilized C. roseus cells resembled rates reported in the literature for suspension cultures.
Offprint requests to: Denis Rho 相似文献
20.
Suspension cultures of Catharanthus roseus were used to evaluate ultraviolet-B (UV-B) treatment as an abiotic elicitor of secondary metabolites. A dispersed cell suspension culture from C. roseus leaves in late exponential phase and stationary phase were irradiated with UV-B for 5 min. The stationary phase cultures were more responsive to UV-B irradiation than late exponential phase cultures. Catharanthine and vindoline increased 3-fold and 12-fold, respectively, on treatment with a 5-min UV-B irradiation. 相似文献