Abstract
Most fungi are known to synthesize siderophores under iron limitation. However, arbuscular mycorrhizal fungi (AM fungi) have so far not been reported to produce siderophores, although their metabolism is iron-dependent. In an approach to isolate siderophores from AM fungi, we have grown plants of Tagetes patula nana in the presence of spores from AM fungi of the genus Glomus (G. etunicatum, G. mossae & unidentified Glomus sp.) symbiotically under iron limitation and sterile conditions. A siderophore was isolated from infected roots after 2–3 weeks of growth in pots containing low-iron sand with Hoagland solution. HPLC analysis of the root cell lysate revealed a peak at a retention time of 6.7 min which showed iron-binding properties in a chrome azurol S test. The compound was isolated by preparative HPLC and the structure was determined by high resolution electrospray FTICR-MS and GC/MS analysis of the hydrolysis products. From an observed absolute mass to charge ratio (m/z) of 401.11925 [M+H]+ with a relative mass error of ∆ = 0.47 ppm an elemental composition of C16H21N2O10 [M+H]+ was derived, suggesting a molecular weight of 400 Da for glomuferrin. Corresponnding ion masses of m/z 423.10 and m/z 439.06 were asigned to the Na-adduct and K-adduct respectively. A mass of 455.03836 confirmed an Fe- complex with an elemental composition of C16H19N2O10Fe (∆ = 0.15 ppm). GC/MS analysis of the HCl lysate (6 N HCL, 12 h) revealed 1,4 butanediamine. Thus the proposed structure of the isolated siderophore from Glomus species consisted of 1,4 butanediamine amidically linked to two dehydrated citrate residues, similar to the previously identified bis-amidorhizoferrin. Thus, the isolated siderophore (glomuferrin) is a member of the rhizoferrin family previously isolated from fungi of the Mucorales (Zygomycetes).
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Acknowledgements
We thank T. Wubet for provideing the Glomus spores, I Kottke for help in the Glomus biology, G. Nicholson and A. Kulik for GC/MS-analysis and MS –analysis and Prof. Rolf Reuter for assistence in performing the FM CLSM analysis. Usha Kalidindi from from the Division of Fruits and Horticulture Technology, Indian Agricultural Research Institute, New Delhi-110012 helped in the isolation procedures during a Grant for 3 months within the International Scientific exchange programme of Indian National Science Academy (INSA), New Delhi, and the DFG (Bonn, Germany).
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Winkelmann, G. A search for glomuferrin: a potential siderophore of arbuscular mycorrhizal fungi of the genus Glomus . Biometals 30, 559–564 (2017). https://doi.org/10.1007/s10534-017-0026-x
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DOI: https://doi.org/10.1007/s10534-017-0026-x