Abstract
Sulfur (S) and phosphorus (P) are essential elements for plant growth and physiological functioning. Their deficiency can limit N2 fixation and nodule development in nodulated legumes. The location of S within nodule tissues could provide insights into S metabolism and its little-known relationship with N2 fixation. Determinate and indeterminate nodules were inoculated with specific rhizobia and grown hydroaeroponically under sufficient versus deficient P supplies. Cryogenic and freeze-dried thin sections of nodules at the flowering stage were mapped using synchrotron micro-X-ray fluorescence to determine the S distribution within the nodule tissues with a spatial resolution of 2 or 3 μm. A large accumulation of S was found in the middle cortex for both types of nodules. S was also found in all of the other tissues but with a significantly lower signal. In the middle cortex, P deficiency decreased the S maximum fluorescence intensity by 20% and 25% for the determinate and indeterminate nodules, respectively. In addition, Mg and Cl maps were also collected showing that Mg was mostly localized in the middle and inner cortex, forming a Mg-rich ring consisting of several cell layers for the determinate nodules compared with only one cell layer for the indeterminate nodules. Cl was mainly accumulated in the outer cortex. It is concluded that the accumulation of S in the middle cortex is consistent with its involvement in the ionic equilibrium of the nodule, and in the osmotic variation of the inner cortex cell-size, which would regulate nodule permeability to oxygen.
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Acknowledgments
The authors would like to thank the ID21 and LUCIA staff and Nicolas Leclercq for FlyScan development and support during the experiments.
Funding
This work was supported by the Great Federative Project FABATROPIMED, financed by the Agropolis Fondation in Montpellier and the PHC Tassili project under the references ID 1001-009,16M and DU973, respectively, and by synchrotrons ESRF and SOLEIL through in-house beamtime allocation and proposal 20151042, respectively.
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Rivard, C., Amenc, L., Benlahrech, S. et al. Synchrotron micro-X-ray fluorescence shows sulfur accumulation in the middle cortex of N2-fixing legume nodules. Protoplasma 256, 1025–1035 (2019). https://doi.org/10.1007/s00709-019-01360-8
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DOI: https://doi.org/10.1007/s00709-019-01360-8