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How Agrobacterium rhizogenes triggers de novo root formation in a recalcitrant woody plant: an integrated histological, ultrastructural and molecular analysis

Published online by Cambridge University Press:  01 January 2000

GIUSEPPINA FALASCA
Affiliation:
Dipartimento di Biologia Vegetale, Università di Roma ‘La Sapienza’, P. le Aldo Moro 5, I-00185 Rome, Italy
MASSIMO REVERBERI
Affiliation:
Dipartimento di Biologia Vegetale, Università di Roma ‘La Sapienza’, P. le Aldo Moro 5, I-00185 Rome, Italy
PAOLA LAURI
Affiliation:
Istituto Sperimentale per la Frutticoltura, 00040 Ciampino Aeroporto, Rome, Italy
EMILIA CABONI
Affiliation:
Istituto Sperimentale per la Frutticoltura, 00040 Ciampino Aeroporto, Rome, Italy
ANGELO DE STRADIS
Affiliation:
Centro di Servizi Interdipartimentale di Microscopia, Università degli Studi della Basilicata, 85100 Potenza, Italy
MARIA MADDALENA ALTAMURA
Affiliation:
Dipartimento di Biologia Vegetale, Università di Roma ‘La Sapienza’, P. le Aldo Moro 5, I-00185 Rome, Italy
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Abstract

Adventitious rooting might be induced in recalcitrant woody genotypes by infection with Agrobacterium rhizogenes, and, in some cases, might also require exogenous auxin. The objective of the present study was to determine how agrobacteria trigger root formation in the stem of a recalcitrant woody microcutting, which cytological events result from the combined presence of infection and exogenous auxin, and which types of roots are induced by infection. Microcuttings of a recalcitrant walnut (Juglans regia), infected or not with A. rhizogenes strain 1855, were cultured with either indolebutyric acid (IBA), IAA, or without exogenous hormones, to induce rhizogenesis. They were cytohistologically and ultrastructurally investigated at various times in culture. Southern blot and PCR analyses were performed to verify the frequency of transgenic, chimeric and bacterium-containing roots. The infection was sufficient per se to stimulate rhizogenesis. Rooting on the infected cuttings was enhanced by exogenous IBA, which accelerated and increased root meristemoid formation, in comparison with without-hormone treatment. Meristemoids were organized both directly by the cambial cells and indirectly by the callus, and showed a pluricellular origin. Inter and intracellular bacteria were observed in the stem throughout the culture period (30 d). They were preferentially present in the vessels, and mainly in those showing polyphenol deposition. In the infected IAA-treated cultures, a high level of secondary xylem formation occurred instead of rhizogenesis. Nontransformed roots were preferentially produced by the infected cuttings treated with the auxins. Bacterium-containing and chimeric roots were produced by infected cuttings independently of the treatment. Thus, in a recalcitrant walnut, nontransformed root meristemoids are stimulated by combining infection and exogenous indolebutyric acid. Furthermore, the persistence of bacteria in the stem during the culture and the pluricellular origin of the meristemoids explain the presence of the bacterium-containing and chimeric roots.

Type
Research Article
Copyright
© Trustees of the New Phytologist 2000

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