Nitrogen transfer and assimilation between the arbuscular mycorrhizal fungus Glomus intraradices Schenck & Smith and Ri T-DNA roots of Daucus carota L. in an in vitro compartmented system

2004 ◽  
Vol 50 (4) ◽  
pp. 251-260 ◽  
Author(s):  
Jean-Patrick Toussaint ◽  
Marc St-Arnaud ◽  
Christiane Charest

Nitrogen metabolism was examined in monoxenic cultures of carrot roots (Daucus carota L.) colonized with the arbuscular mycorrhizal (AM) fungus Glomus intraradices Schenck & Smith. Glutamine synthetase and glutamate dehydrogenase activities were significantly increased in mycorrhizal roots for which only the extraradical mycelium had exclusive access to NH4NO3 in a distinct hyphal compartment inaccessible to the roots. This was in comparison with the water controls but was similar to the enzyme activities of non-arbuscular-mycorrhizal (non-AM) roots that had direct access to NH4NO3. In addition, glutamate dehydrogenase activity was significantly enhanced in AM roots compared with non-AM roots. Carrot roots took up 15NH4+ more efficiently than 15NO3–, and the extraradical hyphae transfered 15NH4+ to host roots from the hyphal compartment but did not transfer 15NO3–. The extraradical mycelium was shown, for the first time, to have a different glutamine synthetase monomer than roots. Our overall results highlight the active role of AM fungi in nitrogen uptake, transfer, and assimilation in their symbiotic root association.Key words: arbuscular mycorrhizal fungus, Ri T-DNA carrot roots, in vitro root-organ culture, nitrogen metabolism.


Chemosphere ◽  
2008 ◽  
Vol 73 (3) ◽  
pp. 344-352 ◽  
Author(s):  
Derek G. Hillis ◽  
Pedro Antunes ◽  
Paul K. Sibley ◽  
John N. Klironomos ◽  
Keith R. Solomon


2008 ◽  
Vol 54 (2) ◽  
pp. 103-110 ◽  
Author(s):  
Manuel González-Guerrero ◽  
Lewis H. Melville ◽  
Nuria Ferrol ◽  
John N.A. Lott ◽  
Concepción Azcón-Aguilar ◽  
...  

Arbuscular mycorrhizal fungi, obligate symbionts of most plant species, are able to accumulate heavy metals, thereby, protecting plants from metal toxicity. In this study, the ultrastructural localization of Zn, Cu, and Cd in the extraradical mycelium and spores of the arbuscular mycorrhizal fungus Glomus intraradices grown in monoxenic cultures was investigated. Zinc, Cu, or Cd was applied to the extraradical mycelium to final concentrations of 7.5, 5.0, or 0.45 mmol/L, respectively. Samples were collected at time 0, 8 h, and 7 days after metal application and were prepared for rapid freezing and freeze substitution. Metal content in different subcellular locations (wall, cytoplasm, and vacuoles), both in hyphae and spores, was determined by energy-dispersive X-ray spectroscopy. In all treatments and fungal structures analysed, heavy metals accumulated mainly in the fungal cell wall and in the vacuoles, while minor changes in metal concentrations were detected in the cytoplasm. Incorporation of Zn into the fungus occurred during the first 8 h after metal addition with no subsequent accumulation. On the other hand, Cu steadily accumulated in the spore vacuoles over time, whereas Cd steadily accumulated in the hyphal vacuoles. These results suggest that binding of metals to the cell walls and compartmentalization in vacuoles may be essential mechanisms for metal detoxification.





1999 ◽  
Vol 77 (9) ◽  
pp. 1391-1397
Author(s):  
Genevieve Louise Mark ◽  
John E Hooker ◽  
Alexander Hahn ◽  
Chris T Wheeler

Micropropagated, rooted, and calli explants of Casuarina equisetifolia L. were inoculated with Frankia UGL 020605S and the arbuscular mycorrhizal fungus (AMF) Glomus mosseae, in single and dual co-culture, in vitro. Different micropropagation media formulations were evaluated for their capacity to stimulate germination of G. mosseae spores and growth of Frankia. Murashige and Skoog basal nutrient (half strength) medium, supplemented with 6-benzylaminopurine (BAP), 2,4-dichlorophenoxyacetic acid (2,4-D), and pyruvate was selected for the in vitro co-culture of C. equisetifolia callus explants, G. mosseae, and Frankia. This medium (M4) supported 70% AMF spore germination with 44 and 34% of the germinating spores producing single and branched hyphal strands, respectively. Hoaglands (quarter strength, modified by Hoaglands and Arnon (1950)) nutrient medium (M5) with no supplements was selected for the in vitro co-culture of rooted C. equisetifolia explants, G. mosseae, and Frankia and supported 57% AMF spore germination with 29 and 40% of the germinating spores producing single and branched hyphal strands, respectively. Both media supported significant growth of Frankia. In both cases agar was substituted with Terragreen(r). AMF appressoria and intercellular hyphae were observed in rooted C. equisetifolia at 28 days; arbuscule formation occurred at 56 days postinoculation. Frankia infection was evident after 28 days. This was observed in both dual and single in vitro co-cultures. No specific immunofluorescent or immunogold reactions to monoclonal antibodies (mABs) anti-Frankia < 8C5 > and anti-G. mosseae < F5G5 > were evident in C. equisetifolia callus explants.Key words: arbuscular mycorrhizal fungi (AMF), Frankia, Casuarina, micropropagation, immunofluorescent labelling.



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