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The key role of peltate glandular trichomes in symbiota comprising clavicipitaceous fungi of the genus periglandula and their host plants.

Steiner U, Kucht SH, Ahimsa-Müller MA, Grundmann N, Li SM, Drewke C, Leistner E - Toxins (Basel) (2015)

Bottom Line: Clavicipitaceous fungi producing ergot alkaloids were recently discovered to be epibiotically associated with peltate glandular trichomes of Ipomoea asarifolia and Turbina corymbosa, dicotyledonous plants of the family Convolvulaceae.Molecular biological studies and microscopic investigations led to the observation that the trichomes do not only secrete sesquiterpenes and palmitic acid but also seem to absorb ergot alkaloids from the epibiotic fungal species of the genus Periglandula.Thus, the trichomes are likely to have a dual and key function in a metabolic dialogue between fungus and host plant.

View Article: PubMed Central - PubMed

Affiliation: Institut für Nutzpflanzenwissenschaften und Ressourcenschutz, Universität Bonn, Nußallee 9, D-53115 Bonn, Germany. u-steiner@uni-bonn.de.

ABSTRACT
Clavicipitaceous fungi producing ergot alkaloids were recently discovered to be epibiotically associated with peltate glandular trichomes of Ipomoea asarifolia and Turbina corymbosa, dicotyledonous plants of the family Convolvulaceae. Mediators of the close association between fungi and trichomes may be sesquiterpenes, main components in the volatile oil of different convolvulaceous plants. Molecular biological studies and microscopic investigations led to the observation that the trichomes do not only secrete sesquiterpenes and palmitic acid but also seem to absorb ergot alkaloids from the epibiotic fungal species of the genus Periglandula. Thus, the trichomes are likely to have a dual and key function in a metabolic dialogue between fungus and host plant.

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Specificity of a polyclonal antibody directed against the His-tagged 4-(γ,γ-dimethylallyl)tryptophan synthase (DmaW) from P. ipomoeae. The specificity against corresponding tryptophan prenylating enzymes (FgaPT2 and 7-DMATS) from A. fumigatus is significantly lower. The amount of DmaW enzyme from P. ipomoeae was doubled when compared to FgaPT2 and 7-DMATS.
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toxins-07-01355-f007: Specificity of a polyclonal antibody directed against the His-tagged 4-(γ,γ-dimethylallyl)tryptophan synthase (DmaW) from P. ipomoeae. The specificity against corresponding tryptophan prenylating enzymes (FgaPT2 and 7-DMATS) from A. fumigatus is significantly lower. The amount of DmaW enzyme from P. ipomoeae was doubled when compared to FgaPT2 and 7-DMATS.

Mentions: Production and purification of the recombinant His6-tagged DmaW enzyme was previously described [17]. Seqlab Sequence Laboratories Göttingen GmbH, Germany, was commissioned to produce rabbit immuno serum containing antibodies against His6-DmaW by using three injections of 200 µg enzyme in 50 mM Tris-HCl pH 7, and complete five in a two month protocol. Preimmune serum was also taken and used as a control. The specificity of the resulting antibody was tested against His-tagged DmaW from P. ipomoeae and two related His-tagged tryptophan prenylating enzymes from Aspergillus fumigatus, such as 4-(γ,γ-dimethylallyl)tryptophan synthase (FgaPT2) and 7-(γ,γ-dimethylallyl)tryptophan synthase (7-DMATS). The two latter enzymes were prepared as described for FgaPT2 [51] and for 7-DMATS [52,53]. The prenylation reaction of all three tryptophan prenylating enzymes was now tested (Figure 7) in the presence of anti-DmaW serum using enzyme concentrations of 0.01 mg/mL 7-DMATS, 0.01 mg/mL FgaPT2 but 0.02 mg/mL DmaW, each calculated as monomer and solved in 90 µL 50 mM Tris-HCl pH 7,5 and of anti-DmaW serum with increasing concentrations. Incubation was carried out for one hour at 30 °C. The samples were then centrifuged for 30 min at 13.000× g and 4 °C. The supernatant was transferred into a new tube. After addition of 1 mM L-tryptophan, 1 mM DMAPP (dimethylallyl diphosphate), and 5 mM CaCl2, the assays were incubated at 30 °C for one hour. The enzymatic reaction was then stopped with 12.5 µL TCA (trichloroacetic acid, 1.5 M). The samples were analysed by HPLC [17]. As expected, the highest specificity of the antibody was observed for the DmaW enzyme from P. ipomoeae when compared to those from A. fumigatus (Figure 7).


The key role of peltate glandular trichomes in symbiota comprising clavicipitaceous fungi of the genus periglandula and their host plants.

Steiner U, Kucht SH, Ahimsa-Müller MA, Grundmann N, Li SM, Drewke C, Leistner E - Toxins (Basel) (2015)

Specificity of a polyclonal antibody directed against the His-tagged 4-(γ,γ-dimethylallyl)tryptophan synthase (DmaW) from P. ipomoeae. The specificity against corresponding tryptophan prenylating enzymes (FgaPT2 and 7-DMATS) from A. fumigatus is significantly lower. The amount of DmaW enzyme from P. ipomoeae was doubled when compared to FgaPT2 and 7-DMATS.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4417971&req=5

toxins-07-01355-f007: Specificity of a polyclonal antibody directed against the His-tagged 4-(γ,γ-dimethylallyl)tryptophan synthase (DmaW) from P. ipomoeae. The specificity against corresponding tryptophan prenylating enzymes (FgaPT2 and 7-DMATS) from A. fumigatus is significantly lower. The amount of DmaW enzyme from P. ipomoeae was doubled when compared to FgaPT2 and 7-DMATS.
Mentions: Production and purification of the recombinant His6-tagged DmaW enzyme was previously described [17]. Seqlab Sequence Laboratories Göttingen GmbH, Germany, was commissioned to produce rabbit immuno serum containing antibodies against His6-DmaW by using three injections of 200 µg enzyme in 50 mM Tris-HCl pH 7, and complete five in a two month protocol. Preimmune serum was also taken and used as a control. The specificity of the resulting antibody was tested against His-tagged DmaW from P. ipomoeae and two related His-tagged tryptophan prenylating enzymes from Aspergillus fumigatus, such as 4-(γ,γ-dimethylallyl)tryptophan synthase (FgaPT2) and 7-(γ,γ-dimethylallyl)tryptophan synthase (7-DMATS). The two latter enzymes were prepared as described for FgaPT2 [51] and for 7-DMATS [52,53]. The prenylation reaction of all three tryptophan prenylating enzymes was now tested (Figure 7) in the presence of anti-DmaW serum using enzyme concentrations of 0.01 mg/mL 7-DMATS, 0.01 mg/mL FgaPT2 but 0.02 mg/mL DmaW, each calculated as monomer and solved in 90 µL 50 mM Tris-HCl pH 7,5 and of anti-DmaW serum with increasing concentrations. Incubation was carried out for one hour at 30 °C. The samples were then centrifuged for 30 min at 13.000× g and 4 °C. The supernatant was transferred into a new tube. After addition of 1 mM L-tryptophan, 1 mM DMAPP (dimethylallyl diphosphate), and 5 mM CaCl2, the assays were incubated at 30 °C for one hour. The enzymatic reaction was then stopped with 12.5 µL TCA (trichloroacetic acid, 1.5 M). The samples were analysed by HPLC [17]. As expected, the highest specificity of the antibody was observed for the DmaW enzyme from P. ipomoeae when compared to those from A. fumigatus (Figure 7).

Bottom Line: Clavicipitaceous fungi producing ergot alkaloids were recently discovered to be epibiotically associated with peltate glandular trichomes of Ipomoea asarifolia and Turbina corymbosa, dicotyledonous plants of the family Convolvulaceae.Molecular biological studies and microscopic investigations led to the observation that the trichomes do not only secrete sesquiterpenes and palmitic acid but also seem to absorb ergot alkaloids from the epibiotic fungal species of the genus Periglandula.Thus, the trichomes are likely to have a dual and key function in a metabolic dialogue between fungus and host plant.

View Article: PubMed Central - PubMed

Affiliation: Institut für Nutzpflanzenwissenschaften und Ressourcenschutz, Universität Bonn, Nußallee 9, D-53115 Bonn, Germany. u-steiner@uni-bonn.de.

ABSTRACT
Clavicipitaceous fungi producing ergot alkaloids were recently discovered to be epibiotically associated with peltate glandular trichomes of Ipomoea asarifolia and Turbina corymbosa, dicotyledonous plants of the family Convolvulaceae. Mediators of the close association between fungi and trichomes may be sesquiterpenes, main components in the volatile oil of different convolvulaceous plants. Molecular biological studies and microscopic investigations led to the observation that the trichomes do not only secrete sesquiterpenes and palmitic acid but also seem to absorb ergot alkaloids from the epibiotic fungal species of the genus Periglandula. Thus, the trichomes are likely to have a dual and key function in a metabolic dialogue between fungus and host plant.

Show MeSH