<sup>31</sup>P NMR Study of Elicitor Treated <i>Phaseolus vulgaris</i> Cell Suspension Cultures

Ojalvo, Israel; Rokem, J. Stefan; Navon, Gil; Goldberg, Israel

doi:10.1104/pp.85.3.716

Cited by 53 publications

(33 citation statements)

References 16 publications

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“…The same applies to the phaseollin-producing P. vulgaris cells of Ojalvo et al (1987). In L. flavum cells, in which production depends solely on the cell line under observation and no extemal stimulus is needed, a lower cytoplasmic pH was also observed for the producing cells (Table 111).…”

Section: Cytoplasmic Acidification and Secondary Metabolite Productionmentioning

confidence: 75%

“…Cytoplasmic acidification upon activation cQ a defense response by a glucan elicitor has also been observed in Phaseolus vulgaris cells (Ojalvo et al, 1987) and in parsley cells by a Phyfophthora elicitor (Kneusel et al, 1989), whereas pH decreases in the total cell sap of tobacco cells were detected after treatment with Pseudomonas sp. (Atkinson et al, 1985).…”

Section: Cytoplasmic Acidification and Secondary Metabolite Productionmentioning

confidence: 99%

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Cytoplasmic Acidification and Secondary Metabolite Production in Different Plant Cell Suspensions (A Comparative Study)

Hagendoorn¹,

Wagner²,

Segers³

et al. 1994

Plant Physiol.

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In this study, a correlation is described between low cytoplasmic pH, measured with the fluorescent probes 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein (acetoxymethyl ester) and bis-[3-propyl-5-oxoisoxazol-4-yl]pentamethine oxonol, and the production of secondary metabolites for several plant cell-suspension systems. Anthraquinone production in Morinda citrifolia suspensions is negligible in the presence of 2,4-dichlorophenoxyacetic acid (2,4-D), whereas with naphthalene acetic acid (NAA) a significant accumulation is realized. NAA-grown cells showed a lower cytoplasmic pH than did 2,4-D-grown cells. Addition of 2,4-D or parachlorophenoxy acetic acid to NAA-grown cells resulted in an inhibition of anthraquinone production and an increase of the cytoplasmic pH, whereas addition of parachlorophenyl acetic acid had no effect on either parameter. lignin produdion in Petunia hybrida cells could be induced by subculturing them in a medium without iron. These cells showed a lower cytoplasmic pH than control cells. Addition of Fe3+ led to a decreased lignin content and an increased cytoplasmic pH. Two cell lines of Linum flavum showed a different level of coniferin and lignin concentration in their cells. Cells that accumulated coniferin and lignin had a lower cytoplasmic pH than cells that did not accumulate these secondary metabolites. Apparently, in different species and after different kinds of treatment there is a correlation between acidification of the cytoplasm and the production of different secondary metabolites. The possible role of this acidification in secondary metabolite production i s discussed.Some plant tissues are able to produce substantial amounts of secondary metabolites. However, the production of these compounds in plant cell and tissue cultures is generally low. A number of treatments have been described that lead to an increased production; these treatments often stimulate the defense response in the cell (Scheel and Parker, 1990).In a previous paper (Hagendoom et al., 1991a), we ana- 723lyzed the effect of elicitors on the plasma membrane of Petunia hybrida cell suspensions. It appeared that changes in the activity of the plasma membrane ATPase, and subsequently of ApH, were sufficient to increase PAL activity and product formation in these cells. This conclusion was based on two different experimental approaches. First, we tested a number of plasma membrane ATPase inhibitors and ionophores that specifically influence ApH, A*, or both for their ability to activate secondary metabolism. Second, we directly showed changes in cytoplasmic pH upon activation of a secondary metabolic pathway with the aid of the fluorescent probe oxonol VI.In this paper we show that this relation between low cytoplasmic pH and secondary metabolite production is not limited to P. hybrida and the products of the PPP but can also be observed in different model systems. We investigated the production of secondary metabolites by cell suspensions derived from three plant species belonging to three different fa...

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Section: Cytoplasmic Acidification and Secondary Metabolite Productionmentioning

confidence: 75%

Section: Cytoplasmic Acidification and Secondary Metabolite Productionmentioning

confidence: 99%

Cytoplasmic Acidification and Secondary Metabolite Production in Different Plant Cell Suspensions (A Comparative Study)

Hagendoorn¹,

Wagner²,

Segers³

et al. 1994

Plant Physiol.

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“…It has been reported that these early events (ion leakage and extracellular alkalinization) are correlated with a decrease in intracellular pH (22) and a depolarization of the plasma membrane (23,24). These responses could result from either …”

Section: Discussionmentioning

confidence: 99%

Responses of Cultured Tobacco Cells to Cryptogein, a Proteinaceous Elicitor from Phytophthora cryptogea

Blein¹,

Milat²,

Ricci³

1991

Plant Physiol.

126

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In culture, the phytopathogenic fungus Phytophthora cryptogea secretes a protein which elicits hypersensitive-like necroses and protects tobacco plants against invasion by the pathogen Phytophthora parasitca var. nicotianae. This protein, named cryptogein, has been purified and its amino acid sequence determined. In this work, we studied the effect of cryptogein on tobacco cell suspension cultures. Cryptogein was lethal at about 0.10 micromolar. When added at sublethal doses, it elicited the production of ethylene and phytoalexins. It also induced a rapid increase in pH and conductivity of the extracellular medium without affecting the integrity of the plasma membrane. Cryptogein reduced the fusicoccin-induced acidification of the extracellular medium. The concentration which inhibited the fusicoccin response by 50% was 0.8 nanomolar, while 1 micromolar erythrosine B, an ATPase inhibitor, was needed to produce the same inhibition. However, cryptogein did not inhibit the activity of a purified plasma membrane ATPase. Results of binding studies with whole cells suggested the presence of elicitor-binding sites with a high affinity for cryptogein. The involvement of the plasma membrane during the initial interaction between elicitor and cells is discussed.

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“…A transient decrease of intracellular pH has been observed repeatedly (Ojalvo et al, 1987;Kneusel et al, 1989;Guern et al, 1992;Mathieu et al, 1996a) among the changes in the ionic composition after elicitor contact, with only a few exceptions (Horn et al, 1992). This raises the question of whether changes in the intracellular pH could be involved in the signal path toward the elicitation of phytoalexin biosynthesis.…”

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confidence: 99%

Shifts of Intracellular pH Distribution as a Part of the Signal Mechanism Leading to the Elicitation of Benzophenanthridine Alkaloids

et al. 1998

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Cultured cells of Eschscholtzia californica (Californian poppy)respond to a yeast elicitor preparation or Penicillium cyclopium spores with the production of benzophenanthridine alkaloids, which are potent phytoalexins. Confocal pH mapping with the probe carboxy-seminaphthorhodafluor-1-acetoxymethylester revealed characteristic shifts of the pH distribution in challenged cells: within a few minutes after elicitor contact a transient acidification of cytoplasmic and nuclear areas occurred in parallel with an increase of the vacuolar pH. The change of proton concentration in the vacuole and in the extravacuolar area showed a nearly constant relation, indicating an efflux of vacuolar protons into the cytosol. A 10-min treatment with 2 mM butyric or pivalic acid caused a transient acidification of the cytoplasm comparable to that observed after elicitor contact and also induced alkaloid biosynthesis. Experimental depletion of the vacuolar proton pool reversibly prevented both the elicitor-triggered pH shifts and the induction of alkaloid biosynthesis. pH shifts and induction of alkaloid biosynthesis showed a similar dependence on the elicitor concentration. Net efflux of K ؉ , alkalinization of the outer medium, and browning of the cells were evoked only at higher elicitor concentrations. We suggest that transient acidification of the cytoplasm via efflux of vacuolar protons is both a necessary and sufficient step in the signal path toward biosynthesis of benzophenanthridine alkaloids in Californian poppy cells.The induction of the biosynthesis of phytoalexins by external elicitors is one component of the multifactorial response of plants to pathogens. In plant tissues the production of plant-specific phytoalexins is usually in concert with the widely distributed reactions of the hypersensitive response complex: cross-linking of cell wall proteins, accumulation of phenolics, lignification, and production of antimicrobial exoenzymes (e.g. chitinases) and of various other pathogenesis-related proteins. An integrative signal system obviously coordinates these activities among the invaded cells and their neighbors and at the systemic level (for recent reviews, see Ricci et al., 1993;Ebel and Cosio, 1994;Kombrink and Somssich, 1995). In cultured cells the primary responses evoked by the contact with exogenous elicitors can be grouped into (a) an oxidative burst, i.e. the generation of reactive oxygen species (Apostol et al., 1989; Sutherland, 1991;Levine et al., 1994;Lamb and Dixon, 1997), and (b) perturbations of the cellular ionic balance, i.e. efflux of K ϩ and Cl Ϫ , influx of Ca 2ϩ , external alkalinization, and cellular acidification (Mathieu et al., 1991;Bach et al., 1993).Whereas in distinct species both groups of responses appear to be causally related (Nü rnberger et al., 1997), other findings argue that at least in some plants both responses are not necessarily coupled but rather belong to different signal paths that selectively convert signals from elicitor-binding sites at the plasma membrane into the activ...

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³¹P NMR Study of Elicitor Treated Phaseolus vulgaris Cell Suspension Cultures

Cited by 53 publications

References 16 publications

Cytoplasmic Acidification and Secondary Metabolite Production in Different Plant Cell Suspensions (A Comparative Study)

Cytoplasmic Acidification and Secondary Metabolite Production in Different Plant Cell Suspensions (A Comparative Study)

Responses of Cultured Tobacco Cells to Cryptogein, a Proteinaceous Elicitor from Phytophthora cryptogea

Shifts of Intracellular pH Distribution as a Part of the Signal Mechanism Leading to the Elicitation of Benzophenanthridine Alkaloids

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31P NMR Study of Elicitor Treated Phaseolus vulgaris Cell Suspension Cultures

Cited by 53 publications

References 16 publications

Cytoplasmic Acidification and Secondary Metabolite Production in Different Plant Cell Suspensions (A Comparative Study)

Cytoplasmic Acidification and Secondary Metabolite Production in Different Plant Cell Suspensions (A Comparative Study)

Responses of Cultured Tobacco Cells to Cryptogein, a Proteinaceous Elicitor from Phytophthora cryptogea

Shifts of Intracellular pH Distribution as a Part of the Signal Mechanism Leading to the Elicitation of Benzophenanthridine Alkaloids

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³¹P NMR Study of Elicitor Treated Phaseolus vulgaris Cell Suspension Cultures