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Ion currents involved in early Nod-factor response in Medicago
sativa root hairs.
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Young root hairs and protoplasts of Medicago
sativa. The seeds were germinated according to the protocol described in Methods. The
plantlets were left for 6 h in a vertical position for root hairs to grow. A, root
hairs; B, protoplasts released from young root hair tips, after 3 min incubation
with enzyme cocktail. Bar scales = 10 µm. |
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Root hairs are a primary site for nutrient absorption and
for initiation of signaling processes linked to variations of the root environment:
plant-microbe interactions or abiotic changes. In many of these cases, the earliest
detectable response is the modification of plasma membrane transports, detected through
alteration of the electrical membrane potential. In spite of this, root hairs have not
been extensively used in electrophysiological research so far. Problems with cell shape
and current coupling are often prohibitive for microelectrode voltage clamp on intact root
hairs. In the present study, these difficulties have been overcome and the ion channel
currents are described for young root hairs from alfalfa seedlings (Medicago sativa
cv Sitel). Electrophysiological and pharmacological studies indicated an inward rectifying
K+ time-dependent current. This current was sensitive to tetraethylammonium and Cs+ (10 mM
each). Two others currents never shown in root hairs were described: an outward rectifying
time-dependent K+ current, inhibited by tetraethylammonium and Cs+ (10 mM each) allowing
K+ efflux under strong depolarizations and an instantaneous inward current identified as
an anion current, inhibited by 4-acetamido-4'-isothiocyanatostilbene-2,2', disulfonic acid
and anthracene-9-carboxylic acid (100 µM each). These results should contribute to the
understanding of root hair development and of signaling processes in M. sativa root
hairs.
Nod-factor [NodRm-IV(Ac,S)], isolated from the bacterium Rhizobium
meliloti, induces a well known depolarization in Medicago sativa (cv Sitel)
root hairs (Kurkdjian Plant Physiol. 107: 783-790, 1995). The analysis of this membrane
response using the discontinuous single electrode voltage clamp technique (dSEVC) shows
that anion channel, K+ channel and H+-ATPase pump currents are involved in young growing
root hairs. The early Nod-factor-induced depolarization is due to the increase of the
inward ion current and to the inhibition of the H+-pump. It involved an instantaneous
inward anion current (IIAC) and/or a time dependent inward K+ current (IRKC). These two
ion currents are then down-regulated while the H+-pump is stimulated allowing the long
term rectification of the membrane potential (Em). Our results support the idea that the
regulation of inward current plays a primary role in the Nod-factor induced electrical
response, the nature of ions carried by these currents depending on the activated anion
and/or K+ channels at the plasma membrane.
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