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Molecular, Biochemical, and Organismal Analyses of Tomato Plants Simultaneously Attacked by Herbivores from Two Feeding Guilds

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Abstract

Previous work identified aphids and caterpillars as having distinct effects on plant responses to herbivory. We sought to decipher these interactions across different levels of biological organization, i.e., molecular, biochemical, and organismal, with tomato plants either damaged by one 3rd-instar beet armyworm caterpillar (Spodoptera exigua), damaged by 40 adult potato aphids (Macrosiphum euphorbiae), simultaneous damaged by both herbivores, or left undamaged (controls). After placing insects on plants, plants were transferred to a growth chamber for 5 d to induce a systemic response. Subsequently, individual leaflets from non-damaged parts of plants were excised and used for gene expression analysis (microarrays and quantitative real-time PCR), C/N analysis, total protein analysis, proteinase inhibitor (PI) analysis, and for performance assays. At the molecular level, caterpillars up-regulated 56 and down-regulated 29 genes systemically, while aphids up-regulated 93 and down-regulated 146 genes, compared to controls. Although aphids induced more genes than caterpillars, the magnitude of caterpillar-induced gene accumulation, particularly for those associated with plant defenses, was often greater. In dual-damaged plants, aphids suppressed 27% of the genes regulated by caterpillars, while caterpillars suppressed 66% of the genes regulated by aphids. At the biochemical level, caterpillars induced three-fold higher PI activity compared to controls, while aphids had no effects on PIs either alone or when paired with caterpillars. Aphid feeding alone reduced the foliar C/N ratio, but not when caterpillars also fed on the plants. Aphid and caterpillar feeding alone had no effect on the amount of protein in systemic leaves; however, both herbivores feeding on the plant reduced the amount of protein compared to aphid-damaged plants. At the organismal level, S. exigua neonate performance was negatively affected by prior caterpillar feeding, regardless of whether aphids were present or absent. This study highlights areas of concordance and disjunction between molecular, biochemical, and organismal measures of induced plant resistance when plants are attacked by multiple herbivores. In general, our data produced consistent results when considering each herbivore separately but not when considering them together.

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Acknowledgements

We thank Deborah Tam, Joseph Sherman, Joshua Lighton, and Spencer Williams for laboratory assistance, and Robert Holdcraft for help with figures and tables. We appreciate comments from Anurag Agrawal, Stuart Campbell, Jin-ho Kang, Dan Kliebenstein, Scott McArt, Martin de Vos, Caroline von Dahl, and anonymous reviewers. Funding was provided by the National Science Foundation Plant Genome Research Initiative (No. 0820367 to ROM and SMH), the National Research Initiative of the USDA Cooperative State Research, Education and Extension Service (No. 2004-01540 to ROM and SMH, and 2006-35302-17431 to JST), the USDA Cooperative State Research, Education and Extension Service Special Grant (No. 2006-34155-17118 to CRS), and the Western Illinois University research council for partial funding of this project.

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Authors and Affiliations

  1. Department of Entomology, Philip E. Marucci Center for Blueberry and Cranberry Research & Extension, Rutgers University, Chatsworth, NJ, 08019, USA

    Cesar R. Rodriguez-Saona

  2. Department of Biological Sciences, Western Illinois University, Macomb, IL, 61455, USA

    Richard O. Musser & Sue M. Hum-Musser

  3. Department of Entomology, Max Planck Institute for Chemical Ecology, Beutenberg Campus, Jena, 07745, Germany

    Heiko Vogel

  4. Departments of Entomology and Ecology & Evolutionary Biology, Cornell University, Ithaca, NY, 14853, USA

    Jennifer S. Thaler

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  1. Cesar R. Rodriguez-Saona
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Correspondence to Cesar R. Rodriguez-Saona.

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Cesar R. Rodriguez-Saona and Richard O. Musser contributed equally to this work

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Supplemental Material Table S1

shows mean expression for all treatments and probabilities for one-way ANOVA and t-test comparison for each treatment. This list includes only those genes where herbivory significantly up- or down-regulated gene expression (P ≤ 0.05) and changed gene expression by at least 2-fold compared to controls. Treatments are A=non-wounded control (NW); B=aphid feeding (aphid), C=caterpillar feeding (caterpillar), and D=caterpillar and aphid feeding (caterpillar and aphid). (XLS 112 kb)

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Rodriguez-Saona, C.R., Musser, R.O., Vogel, H. et al. Molecular, Biochemical, and Organismal Analyses of Tomato Plants Simultaneously Attacked by Herbivores from Two Feeding Guilds. J Chem Ecol 36, 1043–1057 (2010). https://doi.org/10.1007/s10886-010-9854-7

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