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Induction of Defense-Related Responses in Cf9 Tomato Cells by the AVR9 Elicitor Peptide of Cladosporium fulvum Is Developmentally Regulated1

Guy Honée, Julia Buitink2, Thorsten Jabs3, José De Kloe, Fred Sijbolts, Marion Apotheker, Rob Weide, Titia Sijen4, Maarten Stuiver, and Pierre J.G.M. De Wit*

Department of Phytopathology, Wageningen Agricultural University, Binnenhaven 9, 6709 PD Wageningen, The Netherlands (G.H., J.B., T.J., R.W., T.S., P.J.G.M.D.W.); and Mogen International N.V., Einsteinweg 71, 2333 CB Leiden, The Netherlands (J.D.K., F.S., M.A., M.S.)

The AVR9 elicitor from the fungal pathogen Cladosporium fulvum induces defense-related responses, including cell death, specifically in tomato (Lycopersicon esculentum Mill.) plants that carry the Cf-9 resistance gene. To study biochemical mechanisms of resistance in detail, suspension cultures of tomato cells that carry the Cf-9 resistance gene were initiated. Treatment of cells with various elicitors, except AVR9, induced an oxidative burst, ion fluxes, and expression of defense-related genes. Agrobacterium tumefaciens-mediated transformation of Cf9 tomato leaf discs with Avr9-containing constructs resulted efficiently in transgenic callus formation. Although transgenic callus tissue showed normal regeneration capacity, transgenic plants expressing both the Cf-9 and the Avr9 genes were never obtained. Transgenic F1 seedlings that were generated from crosses between tomato plants expressing the Avr9 gene and wild-type Cf9 plants died within a few weeks. However, callus cultures that were initiated on cotyledons from these seedlings could be maintained for at least 3 months and developed similarly to callus cultures that contained only the Cf-9 or the Avr9 gene. It is concluded, therefore, that induction of defense responses in Cf9 tomato cells by the AVR9 elicitor is developmentally regulated and is absent in callus tissue and cell-suspension cultures, which consists of undifferentiated cells. These results are significant for the use of suspension-cultured cells to investigate signal transduction cascades.

1   This work was supported by grants to J.B. from the Life Science Foundation, which is subsidized by the Netherlands Organization for Scientific Research; and from the Ministry of Economic Affairs; the Ministry of Education, Culture, and Science; and the Ministry of Agriculture, Nature Management, and Fishery in the framework of the Industrial Relevant Research Program of The Netherlands Association of Biotechnology Centers in The Netherlands to G.H.
2   Present address: Department of Plant Physiology, Wageningen Agricultural University, Arboretumlaan 4, 6703 BD Wageningen, The Netherlands.
3   Present address: Institut für Biologie III, RWTH Aachen, Worringer Weg, 52074 Aachen, Germany.
4   Present address: Department of Genetics, Free University, de Boelelaan 1087, 1081 HV Amsterdam, The Netherlands.
*   Corresponding author; e-mail pierre.dewit{at}medew.fyto.wau.nl; fax 31-317-483412.

Plant Physiol. (1998) 117: 809-820
Copyright Clearance Center:   0032-0889/98/117/0809/12
© 1998 American Society of Plant Physiologists




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