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Antisense Expression of a NBS-LRR Sequence in Sunflower (Helianthus annuus L.) and Tobacco (Nicotiana tabacum L.): Evidence for a Dual Role in Plant Development and Fungal Resistance

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Abstract

A partial sunflower cDNA clone, PLFOR48, segregating with a resistance marker to Plasmopara halstedii, the causal agent of downy mildew, has been cloned from the mildew resistant sunflower line, RHA 266. PLFOR48 encodes a putative protein with a nucleotide-binding site and a leucine-rich repeat domain, showing significant homology with previously cloned resistance genes belonging to the TIR-NBS-LRR family. Southern blot analysis of non-transgenic sunflower suggests that PLFOR48 is part of a multigenic family. The potential role of PLFOR48 sequence in sunflower resistance to mildew was studied, by assessing loss of function, using expression of the antisense cDNA in RHA 266 sunflower line. Quite unexpectedly, transgenic sunflower lines displayed severe developmental abnormalities, and in particular, on the main meristems of homozygote T2 progeny, thus hampering any further challenge inoculation with Plasmopara halstedii. The presence of homologous sequences to PLFOR48 in Nicotiana tabacum var Samsun NN, as demonstrated by Southern blotting, drove us to consider tobacco as an additional model to investigate the potential role of this sequence in fungal resistance. Expression of the same antisense cDNA in transgenic tobacco lines gave rise to higher degree of susceptibility to Phytophthora parasitica, as well as to severe alterations in seed development. These results suggest that PLFOR48 and homologous sequences could be involved in both regulating developmental pathways and controlling resistance to fungal pathogens.

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Abbreviations

LRR:

Leucine-rich repeat

NB-ARC:

signalling motif shared by plant resistance proteins and regulators of cell death in animals

NBS:

nucleotide binding site

PCD:

programmed cell death

Pl :

Plasmopara

Ppn :

Phytophthora parasitica var nicotianae

TIR:

Toll-interleukin-1 receptor

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Authors and Affiliations

  1. Laboratoire de Biotechnologies et Amélioration des Plantes (BAP), INP-ENSAT, Pôle de Biotechnologies Végétales, IFR 40, 18 Chemin de Borde Rouge, BP107, Auzeville, Castanet Tolosan, 31326, France

    Tarek Hewezi, Laurence Thion, Gilbert Alibert & Jean Kallerhoff

  2. Genetics and Cytology Department, National Research Center, Dokki-Cairo, Egypt

    Tarek Hewezi

  3. UMR INRA-UBP 1095 “Amélioration et Santé des Plantes” Bâtiment Biologie Végétale Recherche, Université Blaise Pascal, Campus des Cézeaux, 24 Avenue des Landais, 63177, Aubière Cédex

    Saïd Mouzeyar & Paul Nicolas

  4. Pôle de Biotechnologies Végétales, UMR 5546 Université Paul Sabatier-CNRS, France 24, Chemin de Borde-Rouge, BP 17, Auzeville, Castanet Tolosan, 31326, France

    Martina Rickauer

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  1. Tarek Hewezi
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  2. Saïd Mouzeyar
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  3. Laurence Thion
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  4. Martina Rickauer
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  5. Gilbert Alibert
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  6. Paul Nicolas
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  7. Jean Kallerhoff
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Correspondence to Jean Kallerhoff.

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Hewezi, T., Mouzeyar, S., Thion, L. et al. Antisense Expression of a NBS-LRR Sequence in Sunflower (Helianthus annuus L.) and Tobacco (Nicotiana tabacum L.): Evidence for a Dual Role in Plant Development and Fungal Resistance. Transgenic Res 15, 165–180 (2006). https://doi.org/10.1007/s11248-005-3518-3

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  • DOI: https://doi.org/10.1007/s11248-005-3518-3

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