First published online July 18, 2002; 10.1104/pp.001222
Plant Physiol, August 2002, Vol. 129, pp. 1627-1632
Reactive Oxygen Species in the Elongation Zone of Maize Leaves
Are Necessary for Leaf Extension1
Andrés A.
Rodríguez,
Karina A.
Grunberg, and
Edith L.
Taleisnik*
Instituto de Fitopatologia y Fisiologia Vegetal-Instituto Nacional
de Tecnología Agropecuaria, Camino a 60 Cuadras Km 5 1/2,
5119 Córdoba, Argentina
The production and role of reactive oxygen species
(ROS) in the expanding zone of maize (Zea mays) leaf
blades were investigated. ROS release along the leaf blade was
evaluated by embedding intact seedlings in
2',7'-dichlorofluorescein-containing agar and examining the
distribution of 2',7'-dichlorofluorescein fluorescence along leaf 4, which was exposed by removing the outer leaves before embedding the
seedling. Fluorescence was high in the expanding region, becoming
practically non-detectable beyond 65 mm from the ligule, indicating
high ROS production in the expansion zone. Segments obtained from the
elongation zone of leaf 4 were used to assess the role of ROS in leaf
elongation. The distribution of cerium perhydroxide deposits in
electron micrographs indicated hydrogen peroxide
(H2O2) presence in the apoplast.
2',7'-Dichlorofluorescein fluorescence and apoplastic
H2O2 accumulation were inhibited with diphenyleneiodonium (DPI), which also inhibited
O·2 generation, suggesting a
flavin-containing enzyme activity such as NADPH oxidase was involved in
ROS production. Segments from the elongation zone incubated in water
grew 8% in 2 h. KI treatments, which scavenged
H2O2 but did not inhibit
O·2 production, did not modify growth.
DPI significantly inhibited segment elongation, and the addition of
H2O2 (50 or 500 µM) to the
incubation medium partially reverted the inhibition caused by DPI.
These results indicate that a certain concentration of H2O2 is necessary for leaf elongation, but it
could not be distinguished whether H2O2, or
other ROS, are the actual active agents.
1
This work was supported by the Agencia de
Promoción Científica y Tecnológica of Argentina
(Fondo para la Investigación Cientifica y
Tecnológica grant no. 6869) and by the Fundación Antorchas (grant no. 13740/1-115). This work is part of A.A.R.'s doctorate.
*
Corresponding author; e-mail gertale{at}cordoba.com.ar; fax
0054-351-4974330.
© 2002 American Society of Plant Physiologists
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