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A defensin from tomato with dual function in defense and development

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Abstract

Defensins are antimicrobial peptides that are part of the innate immune system, contributing to the first line of defense against invading pathogens. Defensins and defensin-like peptides are functionally diverse, disrupting microbial membranes and acting as ligands for cellular recognition and signaling. Here we show that the tomato defensin DEF2 is expressed during early flower development. Defensin mRNA abundance, peptide expression and processing are differentially regulated in developing flowers. Antisense suppression or constitutive overexpression of DEF2 reduces pollen viability and seed production. Furthermore, overexpression of DEF2 pleiotropically alters the growth of various organs and enhances foliar resistance to the fungal pathogen Botrytis cinerea. Partially purified extracts from leaves of a DEF2-overexpressing line inhibited tip growth of B. cinerea. Besides providing insights into regulation of defensin expression, these data demonstrate that plant defensins, like their animal counterparts, can assume multiple functions related to defense and development.

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Acknowledgments

We thank M. P. Junkin and C. T. Batson for assistance with genotyping and antimicrobial assays, respectively; L. Meninghelli for vegetative propagation of transgenic tomato plants during the T0 generation; K. Cook for generating cross-sections of flowers; J. Fowler for use of the epifluorescent microscope; R. Cole and M. Asahina for helpful suggestions with microscopy and real-time PCR, respectively; J. Beckman for insights into production of anti-peptide antibody; J. Myers, T. Chen, V. Zarsky, and K. VandenBosch for critical reading of the manuscript. This study was supported by the current research information system (CRIS) project ORE00374.

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  1. Henrik U. Stotz

    Present address: Julius-von-Sachs-Institut fuer Biowissenschaften, Pharmazeutische Biologie, Biozentrum, Universitaet Wuerzburg, 97082, Wuerzburg, Germany

Authors and Affiliations

  1. Horticulture Department, Oregon State University, Corvallis, OR, 97331, USA

    Henrik U. Stotz

  2. Loma Linda University School of Medicine, Loma Linda, CA, 92354, USA

    Brandi Spence

  3. Biology Department, Macon State College, Macon, GA, 31206, USA

    Yueju Wang

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  1. Henrik U. Stotz
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Correspondence to Yueju Wang.

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Supplemental Fig. S1.

Predicted primary and tertiary structures of DEF2. (A) Predicted structure of DEF2. SWISS-MODEL was used to compare DEF2 to the solved structure of the defensin 1GPT from barley (Hordeum vulgare). Disulfide bonds are labeled pink. (B) ClustalW alignment of predicted amino acid sequences of selected genes from tomato, Arabidopsis, and radish (Raphanus sativus). Identical residues are shaded in black, similar residues are shaded in gray. The predicted cleavage site of the signal peptide is marked with an arrow. Note that floral defensins from tomato have a C-terminal domain in addition to the mature defensin domain. Identifiers for three SGN unigenes are shown. (JPG 1062 kb)

Supplemental Fig. S2.

Effect of altered DEF2 expression on pollen viability. Staining of alive and dead pollen grains from untransformed (UT) or transgenic sense (S8) or antisense (A2) tomato cv. Zhongshu 5 with fluorescein diacetate (green) and propidium iodide (red), respectively. (JPG 974 kb)

Supplemental Fig. S3.

No apparent effect of DEF2 on ovary development. Cross sections through ovaries from untransformed (UT) and transgenic (S8 and A2) plants; bar, 1 mm. Arrow indicates an individual ovule. (TIF 4483 kb)

Supplemental Fig. S4.

Expression of defensin peptides and DEF2 mRNA in primary antisense transformants. (TIF 910 kb)

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Stotz, H.U., Spence, B. & Wang, Y. A defensin from tomato with dual function in defense and development. Plant Mol Biol 71, 131–143 (2009). https://doi.org/10.1007/s11103-009-9512-z

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