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Nitric Oxide is Involved in the Azospirillum brasilense-induced Lateral Root Formation in Tomato

Azospirillum spp. is a well known plant-growth-promoting rhizobacterium. Azospirillum-inoculated plants have shown to display enhanced lateral root and root hair development. These promoting effects have been attributed mainly to the production of hormone-like substances. Nitric oxide (NO) has recently been described to act as a signal molecule in the hormonal cascade leading to root formation. However, data on the possible role of NO in free-living diazotrophs associated to plant roots, is unavailable. In this work, NO production by Azospirillum brasilense Sp245 was detected by electron paramagnetic resonance (6.4 nmol. g⁻¹ of bacteria) and confirmed by the NO-specific fluorescent probe 4,5-diaminofluorescein diacetate (DAF-2 DA). The observed green fluorescence was significantly diminished by the addition of the specific NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO). Azospirillum-inoculated and noninoculated tomato (Lycopersicon esculentum L.) roots were incubated with DAF-2 DA and examined by epifluorescence microscopy. Azospirillum-inoculated roots displayed higher fluorescence intensity which was located mainly at the vascular tissues and subepidermal cells of roots. The Azospirillum-mediated induction of lateral root formation (LRF) appears to be NO-dependent since it was completely blocked by treatment with cPTIO, whereas the addition of the NO donor sodium nitroprusside partially reverted the inhibitory effect of cPTIO. Overall, the results strongly support the participation of NO in the Azospirillum-promoted LRF in tomato seedlings.

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ACR :

Agar Congo Red

ARF :

adventitious root formation

cPTIO :

2-(4-carboxyphenyl)-4,4,5,5,-tetramethylimidazoline-1-oxyl-3-oxide

DAF-2 DA :

4,5-diamino-fluorescein diacetate

EPR :

electron paramagnetic resonance

IAA :

indole acetic acid

L-NAME :

N^G -Nitro-L-arginine methyl ester hydrochloride

L-NIO :

L-N⁵ -(1-iminoethyl)-ornithine dihydrochloride

LR :

lateral root

LRF :

lateral root formation

NO :

nitric oxide

OAB :

Okon-Albrecht-Burris

PCIB :

α-(p-chlorophenoxy) isobutyric acid

SDW :

sterile distilled water

SNP :

sodium nitroprusside

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This work was supported by ANPCyT, CONICET, Fundación Antorchas and UNMdP, Argentina.

1. Unidad Integrada Facultad de Cs. Agrarias (UNMdP) Balcarce, Argentina

   Cecilia M. Creus, Elda M. Casanovas, María A. Pereyra & Carlos A. Barassi

2. Instituto de Investigaciones Biológicas, UNMdP, CC 1245, 7600, Mar del Plata, Argentina

   Magdalena Graziano & Lorenzo Lamattina

3. Fisicoquímica-PRALIB, Facultad de Farmacia y Bioquímica, UBA, Buenos Aires, Argentina

   Marcela Simontacchi & Susana Puntarulo

1. Cecilia M. Creus
2. Magdalena Graziano
3. Elda M. Casanovas
4. María A. Pereyra
5. Marcela Simontacchi
6. Susana Puntarulo
7. Carlos A. Barassi
8. Lorenzo Lamattina

Correspondence to Lorenzo Lamattina.

Creus, C.M., Graziano, M., Casanovas, E.M. et al. Nitric Oxide is Involved in the Azospirillum brasilense-induced Lateral Root Formation in Tomato. Planta 221, 297–303 (2005). https://doi.org/10.1007/s00425-005-1523-7

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• Received: 21 February 2005

• Accepted: 21 February 2005

• Published: 12 April 2005

• Issue Date: May 2005

• DOI: https://doi.org/10.1007/s00425-005-1523-7

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