First published online March 22, 2002; 10.1104/pp.010690
Plant Physiol, May 2002, Vol. 129, pp. 211-224
Structure-Function Analysis of Nod Factor-Induced Root Hair
Calcium Spiking in Rhizobium-Legume Symbiosis1
Rebecca J.
Wais,2
David H.
Keating,3 and
Sharon R.
Long*
Department of Biological Sciences, 371 Serra Mall (R.J.W., S.R.L.)
and Howard Hughes Medical Institute (D.H.K., S.R.L.), Stanford
University, Stanford, California 94305-5020
In the Rhizobium-legume symbiosis, compatible bacteria and
host plants interact through an exchange of signals: Host compounds promote the expression of bacterial biosynthetic nod
(nodulation) genes leading to the production of a
lipochito-oligosaccharide signal, the Nod factor (NF). The particular
array of nod genes carried by a given species of
Rhizobium determines the NF structure synthesized and
defines the range of legume hosts by which the bacterium is recognized.
Purified NF can induce early host responses even in the absence of live
Rhizobium One of the earliest known host responses to NF is an
oscillatory behavior of cytoplasmic calcium, or calcium spiking, in
root hair cells, initially observed in Medicago spp. and
subsequently characterized in four other genera (D.W. Ehrhardt, R. Wais, S.R. Long [1996] Cell 85: 673-681; S.A. Walker, V. Viprey, J.A. Downie [2000] Proc Natl Acad Sci USA 97: 13413-13418;
D.W. Ehrhardt, J.A. Downie, J. Harris, R.J. Wais, and S.R. Long,
unpublished data). We sought to determine whether live Rhizobium
trigger a rapid calcium spiking response and whether this response is
NF dependent. We show that, in the Sinorhizobium
meliloti-Medicago truncatula interaction,
bacteria elicit a calcium spiking response that is indistinguishable
from the response to purified NF. We determine that calcium spiking is
a nod gene-dependent host response. Studies of calcium
spiking in M. truncatula and alfalfa (Medicago
sativa) also uncovered the possibility of differences in early
NF signal transduction. We further demonstrate the sufficiency of the
nod genes for inducing calcium spiking by using
Escherichia coli BL21 (DE3) engineered to express 11 S. meliloti nod genes.
1
This work was supported in part by the
Department of Energy (grant no. DE-FG03-90ER200210). S.R.L. is an
investigator of the Howard Hughes Medical Institute.
2
Present address: Incyte Genomics, 3160 Porter Drive,
Palo Alto, CA 94304.
3
Present address: Department of Microbiology and
Immunology, Stritch School of Medicine, Loyola University Chicago, 2160 South First Avenue, Maywood, IL 60153.
*
Corresponding author; e-mail sharon.long{at}stanford.edu; fax
650-725-8309.
© 2002 American Society of Plant Physiologists
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