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TRENDS in Plant Science Vol.6 No.4 April 2001

141

A Nod and a wave: calcium signals during nodulation

shares common signalling elements with
nodulation, including Ca2+ spiking. Thus,
Ca2+ spiking could be an integral component
of several signalling pathways.
The observation that several genes are
required for Ca2+ spiking and nodulation
establishes a strong correlation between
these two phenomena. Nevertheless, the
authors are careful to caveat that Ca2+ spiking
might be a correlative event activated by
steps in common with nodulation. A
comprehensive genetic study would enable
a causal dissection of the processes initiating
root nodulation, as well as providing an
insight into the roles for Ca2+ spiking in other
cellular processes. The characterization of
further mutants with contrasting Ca2+ spiking
phenotypes is eagerly anticipated, as is the
cloning of the DMI and SYM genes.

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root hair deformation occurs. This is

followed by the induction of early nodulin
(ENOD) genes. No mutant defective in the
earliest stages of nodulation was found to
exhibit Ca2+ spiking. Thus, the products of
the M. truncatula DMI1 and DMI2 genes,
and the pea SYM1, SYM8 and SYM10 genes,
are apparently involved in establishing Ca2+
spiking. Mutants in these genes do not
exhibit root deformation or the induction of
ENOD genes. By contrast, Ca2+ spiking was
observed in the mutants dmi3, sym9 and
sym30. These mutants do not exhibit root
deformation or the induction of ENOD
genes either, but the products of the DMI3,
SYM9 and SYM30 genes must act
subsequently to Ca2+ spiking on the
nodulation pathway. Ca2+ spiking was also
exhibited by the M. truncatula mutants nsp
and hcl and the pea mutants sym2A and
sym7. These mutants are compromised in
events subsequent to root hair deformation,
and their phenotype supports the
hypothesis that Ca2+ spiking is an early
event in the signalling pathway leading to
nodulation. Interestingly, the M. truncatula
mutants dim1, dim2 and dim3 and the pea
mutants sym8, sym9, sym19 and sym30 are
also unable to form associations with
mycorrhizal fungi. This suggests that the
development of mycorrhizal associations

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About 70% of the nitrogen available to plants

originates through nitrogen fixation by
symbiotic bacteria. A group of Gram-negative
bacteria, the rhizobia, are housed within the
root nodules of legumes. Nodulation is
initiated by the presence of appropriate
rhizobial endosymbionts in the rhizosphere.
To effect this, plant roots continuously release
elicitors of bacterial Nod gene expression,
and respond proactively to the presence of
bacterial Nod factors by initiating signalling
and developmental pathways that lead to
infection and nodule morphogenesis. Early
cellular responses to Nod factors include
membrane depolarization, Ca2+ influx and the
initiation of multiple, transient increases in
cytoplasmic Ca2+ concentration (termed Ca2+
waves or Ca2+ spikes). The phenotypic
characterization of non-nodulating mutants
of Medicago truncatula and pea by Rebecca
Wais and colleagues1 and Simon Walker and
colleagues2 has shows that Ca2+ spiking is an
early intracellular signal that leads to nodule
development,
The presence of Nod factors elicits Ca2+
spiking in root hairs of legumes after
~10 min. During each spike, the cytoplasmic
Ca2+ concentration ([Ca2+]cyt) increases
transiently by 200500 nM. The oscillations
in [Ca2+]cyt have a periodicity of 12 min and
persist for several hours. After ~6 hours,

1 Wais, R.J. et al. (2000) Genetic analysis of

calcium spiking responses in nodulation
mutants of Medicago truncatula. Proc. Natl.
Acad. Sci. U. S. A. 97, 1340713412
2 Walker, S.A. et al. (2000) Dissection of
nodulation signaling using pea mutants
defective for calcium spiking induced by Nod
factors and chitin oligomers. Proc. Natl. Acad.
Sci. U. S. A. 97, 1341313418

Philip J. White
philip-j.white@hri.ac.uk

Oxygen transport in the static plant cell system

TH

Haemoglobin is one of the best known and

more completely characterized proteins in
biochemistry. In mammals, its function is
the bi-directional transport of oxygen from
lungs to tissues and of carbon dioxide from
tissues to lungs. Thus, haemoglobin travels
in the circulatory system through veins and
capillaries inside the erythrocytes in the
blood stream. Haemoglobin was described
in plants a long time ago in the legume root
nodule, and its function was suggested to
be associated with the oxygen supply to the
symbiotic bacteroid. However, the question
remains as to how plant haemoglobin can
exert this function, being enclosed in the
static plant cell system.
In a recent article, Xxxxxx Kawashima and
colleagues1 describe the existence of
multiple symbiotic haemoglobins
(leghemoglobins) in pea. These are grouped
into two types, PsLbA and PsLbB, which

differ in oxygen affinity as well as in cellular

localization. PsLb5-10, the only protein
representative of the PsLbA type, has a
higher oxygen-binding capacity and the
corresponding transcripts are detected
throughout the central tissue of effective
nodules. The transcripts for PsLb120-1,
which is representative of the PsLbB type, are
localized in a region from infection zone II to
the distal part of nitrogen fixation zone.
The presence of two types of
leghemoglobins of different oxygen affinity
could serve to create and maintain an
oxygen gradient across the nodule tissue.
This primary gradient might be enriched by
the existence of multiple leghemoglobins of
each type, but also the oxygen affinities of
the leghemoglobins might be modulated in
response to the metabolic reactions that
occur inside the cells. For example, recently
two carbonic anhidrases were described of

different cellular expression across the

nodule section in alfalfa (Medicago sativa).
Carbonic anhidrases might modify the pH
status of the cells, affecting in turn the
oxygen-binding capacities of haemoglobins
(the Bohr effect).
Thus, the possibility exists that a
coordinate action between multiple
symbiotic haemoglobins and carbonic
anhidrases could be important in the
creation and maintenance of oxygen and
carbon dioxide gradients required for
nodule metabolism.
1 Kawashima K. et al. (2001) Two types of pea
leghemoglobin genes showing different O2binding affinities and distinct patterns of spatial
expression in nodules. Plant Physiol.
125, 641651

Emilio Cervantes
ecervant@gugu.usal.es

http://plants.trends.com 1360-1385/01/$ see front matter 2001 Elsevier Science Ltd. All rights reserved.