Transgenic aequorin monitors cytosolic calcium transients in soybean cells challenged with β-glucan or chitin elicitors

Mithöfer, Axel; Ebel, Jürgen; Bhagwat, Arvind A.; Boller, Thomas; Neuhaus-Url, Gabriele

doi:10.1007/s004250050519

Cited by 105 publications

(108 citation statements)

References 45 publications

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“…None of these compounds was capable to alter the cytosolic Ca 2 + level. In a positive control, the β-glucan elicitor caused an elevation of the cytosolic Ca 2 + concentration, demonstrating the responsiveness of the cell culture as reported earlier (Mithöfer et al, 1999) (data not shown).…”

Section: Activation Of Early Signaling Elementssupporting

confidence: 85%

“…For instance, a transient increase of the cytosolic Ca 2 + concentration has been proven to be a necessary signaling element in the β-glucan elicitor-dependent stimulation of phytoalexin biosynthesis in soybean (Mithöfer et al, 1999). Using the transgenic, aequorin-expressing soybean cell line 6.6.12 (Mithöfer et al, 1999), we tested the effects of JA, JAMe, OPDA, coronatine and the synthetic mimics IndEt-IleMe and IndAz-IleMe, respectively, on the cytosolic Ca 2 + concentration at 10 µM each. None of these compounds was capable to alter the cytosolic Ca 2 + level.…”

Section: Activation Of Early Signaling Elementsmentioning

confidence: 99%

“…Harosoy 63) were grown as described previously by Hille et al (1982). The cultures were incubated in the dark at 26°C on a rotary shaker at 110 rpm and were subcultured in fresh medium every 7 d. The transgenic soybean suspension cell line 6.6.12, carrying the stably integrated plasmid pGNAequ/neo2 and expressing apoaequorin, was grown as described by Mithöfer et al (1999). Soybean seeds were obtained from Pioneer, Buxtehude, Germany (G. max L. cv.…”

Section: Plant Materialsmentioning

confidence: 99%

“…Transgenic 6.6.12 cell lines were used to reconstitute aequorin in vivo with 10 µM synthetic coelenterazine on a shaker (125 rpm) in the dark for 24 to 48 h. Ca 2 + -specific luminescence (470 nm) was measured and the cytosolic Ca 2 + concentration was calculated as described (Mithöfer et al, 1999). None of the compounds used interfered with the aequorin-dependent luminescence which was tested by the addition of ethanol and CaCl 2 to final concentrations of 10% (v/v) and 1 M, respectively.…”

Section: Aequorin Dependent Luminescence Measurementsmentioning

confidence: 99%

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The Role of Octadecanoids and Functional Mimics in Soybean Defense Responses

Fliegmann

Schüler

Boland

et al. 2003

Biological Chemistry

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Oxylipins of the jasmonate pathway and synthetic functional analogs have been analyzed for their elicitor-like activities in an assay based on the induced accumulation of glyceollins, the phytoalexins of soybean (Glycine max L.), in cell suspension cultures of this plant. Jasmonic acid (JA) and its methyl ester showed weak phytoalexin-inducing activity when compared to an early jasmonate biosynthetic precursor, 12-oxo-phytodienoic acid (OPDA), as well as to the bacterial phytotoxin coronatine and certain 6-substituted indanoyl-L-isoleucine methyl esters, which all were highly active. Interestingly, different octadecanoids and indanoyl conjugates induced the accumulation of transcripts of various defense-related genes to different degrees, indicating distinct induction competencies. Therefore, these signaling compounds and mimics were further analyzed for their effects on signal transduction elements, such as the transient enhancement of the cytosolic Ca 2 + concentration and MAP kinase activation, which are known to be initiated by a soybean pathogen-derived ␤-glucan elicitor. In contrast to the ␤-glucan elicitor, none of the other compounds tested triggered these early signaling elements. Moreover, endogenous levels of OPDA and JA in soybean cells were shown to be unaffected after treatment with ␤-glucans. Thus, OPDA and JA, which are functionally mimicked by coronatine and a variety of 6-substituted derivatives of indanoyl-L-isoleucine methyl ester, represent highly efficient signaling compounds of a lipid-based pathway not deployed in the ␤-glucan elicitor-initiated signal transduction.

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Section: Activation Of Early Signaling Elementssupporting

confidence: 85%

Section: Activation Of Early Signaling Elementsmentioning

confidence: 99%

Section: Plant Materialsmentioning

confidence: 99%

Section: Aequorin Dependent Luminescence Measurementsmentioning

confidence: 99%

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The Role of Octadecanoids and Functional Mimics in Soybean Defense Responses

Fliegmann

Schüler

Boland

et al. 2003

Biological Chemistry

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“…Transgenic soybean (Glycine max) 6.6.12 cell lines carrying the stably integrated plasmid pGNAequ/neo2 and expressing apoaequorin (Mithö fer et al, 1999) were used to reconstitute aequorin in vivo with 10 mM synthetic coelenterazine on a shaker (125 rpm) in the dark for 24 h. The Ca 21 -specific luminescence (470 nm) was measured in a final volume of 200 mL using a digital luminometer (Bio-Orbit 1250) as described (Mithö fer et al, 1999;Mithö fer and Mazars, 2002). Treatments with various compounds were performed by adding 1 to 10 mL of different concentrations of aqueous stock solutions to the cell suspension culture.…”

Section: Aequorin-dependent Luminescence Measurementsmentioning

confidence: 99%

Effects of Feeding Spodoptera littoralis on Lima Bean Leaves. III. Membrane Depolarization and Involvement of Hydrogen Peroxide

et al. 2006

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In response to herbivore (Spodoptera littoralis) attack, lima bean (Phaseolus lunatus) leaves produced hydrogen peroxide (H 2 O 2 ) in concentrations that were higher when compared to mechanically damaged (MD) leaves. Cellular and subcellular localization analyses revealed that H 2 O 2 was mainly localized in MD and herbivore-wounded (HW) zones and spread throughout the veins and tissues. Preferentially, H 2 O 2 was found in cell walls of spongy and mesophyll cells facing intercellular spaces, even though confocal laser scanning microscopy analyses also revealed the presence of H 2 O 2 in mitochondria/peroxisomes. Increased gene and enzyme activations of superoxide dismutase after HW were in agreement with confocal laser scanning microscopy data. After MD, additional application of H 2 O 2 prompted a transient transmembrane potential (V m ) depolarization, with a V m depolarization rate that was higher when compared to HW leaves. In transgenic soybean (Glycine max) suspension cells expressing the Ca 21 -sensing aequorin system, increasing amounts of added H 2 O 2 correlated with a higher cytosolic calcium ([Ca 21 ] cyt ) concentration. In MD and HW leaves, H 2 O 2 also triggered the increase of [Ca 21 ] cyt , but MD-elicited [Ca 21 ] cyt increase was more pronounced when compared to HW leaves after addition of exogenous H 2 O 2 . The results clearly indicate that V m depolarization caused by HW makes the membrane potential more positive and reduces the ability of lima bean leaves to react to signaling molecules.

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Oligoglucoside elicitor-mediated activation of plant defense

Ebel

1998

Bioessays

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Transgenic aequorin monitors cytosolic calcium transients in soybean cells challenged with β-glucan or chitin elicitors

Cited by 105 publications

References 45 publications

The Role of Octadecanoids and Functional Mimics in Soybean Defense Responses

The Role of Octadecanoids and Functional Mimics in Soybean Defense Responses

Effects of Feeding Spodoptera littoralis on Lima Bean Leaves. III. Membrane Depolarization and Involvement of Hydrogen Peroxide

Oligoglucoside elicitor-mediated activation of plant defense

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