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Elevation of the Cytosolic Free [Ca2+] Is
Indispensable for the Transduction of the Nod Factor Signal in
Alfalfa1
Hubert H. Felle*,
Éva Kondorosi,
Ádám Kondorosi, and
Michael Schultze
Botanisches Institut I, Justus-Liebig-Universität,
Senckenbergstrasse 17, D-35390 Giessen, Germany (H.H.F.); Institut des
Sciences Végétales, Centre National de la Recherche
Scientifique, Avenue de la Terrasse, F-91198 Gif-sur-Yvette, France
(E.K., A.K., M.S.); and Institute of Genetics, Biological Research
Center, Hungarian Academy of Sciences, P.O. Box 521, H-6701 Szeged,
Hungary (A.K.)
In root hairs of alfalfa
(Medicago sativa), the requirement of Ca2+
for Nod factor signaling has been investigated by means of
ion-selective microelectrodes. Measured 50 to 100 µm behind the
growing tip, 0.1 µM NodRm-IV(C16:2,S) increased the
cytosolic free [Ca2+] by about 0.2 pCa, while the same
concentration of chitotetraose, the nonactive glucosamine backbone, had
no effect. We demonstrate that NodRm-IV(C16:2,S) still depolarized the
plasma membrane at external Ca2+ concentrations below
cytosolic values if the free EGTA concentration remained low ( 0.01
mM). Externally added Sr2+ was able to replace
Ca2+, and to some extent even enhanced the
Nod-factor-induced depolarization, whereas with Mg2+ it was
decreased. This suggests that the Nod factor response is triggered by
Ca2+ from external stores. The addition of the endomembrane
Ca2+-ATPase inhibitor
2,5-di(t-butyl)-1,4-benzohydroquinone, which presumably mobilizes
Ca2+ from Ins(1,4,5)P3-sensitive stores,
mimicked the Nod factor response, i.e. increased the cytosolic free
[Ca2+], triggered Cl -efflux, depolarized
the plasma membrane, and alkalized the root hair space. In all cases a
refractory state toward Nod factor perception was produced, indicating
a shortcut of Nod factor signal transduction by releasing
Ca2+ from internal stores. These latter results strongly
support the idea that an elevation of cytosolic free
[Ca2+] is indispensable for the transduction of the Nod
factor signal, which is consistent with the role of Ca2+ as
a second messenger.
1
This work was supported by the Commission of the
European Community (TMR contract no. ERBFMRX-CT98-0243).
*
Corresponding author; e-mail hubert.felle{at}bio.uni-giessen.de; fax
49-641-99-35119.
Plant Physiol. (1999) 121: 273-280
Copyright Clearance Center: 0032-0889/99/121//08
© 1999 American Society of Plant Physiologists
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