Plant domestication often reduces plant defenses by selection on chemical and physical defense traits. Thus, it is expected that herbivorous insects perform better on crop varieties than on their wild relatives. However, recent studies show that this pattern is not ubiquitous. We examined how varieties of squash (Cucurbita spp.) domesticated for different purposes (for consumption or as ornamentals), differ in plant defense traits and in their interactions with a leaf and a root herbivore. Two types of defenses were measured: cucurbitacins, which are toxic phytochemicals, and trichomes that are physical barriers for most herbivores. We addressed the following questions: (i) what is the variation in cucurbitacin content and leaf trichome density among varieties? (ii) does purpose of domestication explain differences in defense traits among varieties? and (iii) are herbivore feeding preferences and performance altered by the defense traits of squash varieties? We found great variation in cucurbitacin content among varieties, but not according to their purpose of domestication. Cucurbitacins were found mostly in cotyledons and roots and in very small quantities in the leaves. In contrast, trichome density was greater on the varieties selected for consumption than on the ornamental varieties. The performance of a leaf herbivore (Spodoptera latifascia) and a root herbivore (Diabrotica balteata), was not different among squash varieties. Moreover, in a choice experiment, larvae of the root herbivore preferred to feed on squash varieties with high cucurbitacin content. Whereas, in the field, native leaf herbivores preferred to feed on varieties selected for consumption. Our results contribute to a better understanding on how varietal selection may differentially affect plant defenses. This knowledge could help in the development of crop varieties with enhanced insect resistance.

The domestication of plants has commonly resulted in the loss of plant defense metabolites, with important consequences for the plants' interactions with herbivores and their natural enemies. Squash domestication started 10′000 years ago and has led to the loss of cucurbitacins, which are highly toxic triterpenes. The banded cucumber beetle (Diabrotica balteata), a generalist herbivore, is adapted to feed on plants from the Cucurbitaceae and is known to sequester cucurbitacins, supposedly for its own defense. However, the evidence for this is inconclusive. In this study we tested the impact of squash domestication on the chemical protection of D. balteata larvae against a predatory rove beetle (Dalotia coriaria). We found that cucurbitacins do not defend the larvae against this common soil dwelling predator. In fact, D. balteata larvae were less attacked when they fed on cucurbitacin‐free roots of domesticated varieties compared to high‐cucurbitacin roots of wild plants. This study appears to be the first to look at the consequences of plant domestication on belowground tritrophic interactions. Our results challenge the generalized assumption that sequestered cucurbitacins protect this herbivore against natural enemies, and instead reveals an opposite effect that may be due to a tradeoff between coping with cucurbitacins and avoiding predation.

Cucurbitaceae plants produce cucurbitacins, bitter triterpenoids, to protect themselves against various insects and pathogens. Adult banded cucumber beetles (Diabrotica balteata), a common pest of maize and cucurbits, sequester cucurbitacins, presumably as a defensive mechanism against their natural enemies, which might reduce the efficacy of biological control agents. Whether the larvae also sequester and are protected by cucurbitacins is unclear. We profiled cucurbitacin levels in four varieties of cucumber, Cucumis sativus, and in larvae fed on these varieties. Then, we evaluated larval growth and resistance against common biocontrol organisms including insect predators, entomopathogenic nematodes, fungi and bacteria. We found considerable qualitative and quantitative differences in the cucurbitacin levels of the four cucumber varieties. While two varieties were fully impaired in their production, the other two accumulated high levels of cucurbitacins. We also observed that D. balteata larvae sequester and metabolize cucurbitacins, and although the larvae fed extensively on both belowground and aboveground tissues, the sequestered cucurbitacins were mainly derived from belowground tissues. Cucurbitacins had no detrimental effects on larval performance and, surprisingly, did not provide protection against any of the natural enemies evaluated. Our results show that D. balteata larvae can indeed sequester and transform cucurbitacins, but sequestered cucurbitacins do not impact the biocontrol potential of common natural enemies used in biocontrol. Hence, this plant trait should be conserved in plant breeding programs, as it has been demonstrated in previous studies that it can provide protection against plant pathogens and generalist insects.

Main conclusion Cucurbita argyrosperma domestication affected plant defence by downregulating the cucurbitacin synthesis-associated genes. However, tissue-specific suppression of defences made the cultivars less attractive to co-evolved herbivores Diabrotica balteata and Acalymma spp. Abstract Plant domestication reduces the levels of defensive compounds, increasing susceptibility to insects. In squash, the reduction of cucurbitacins has independently occurred several times during domestication. The mechanisms underlying these changes and their consequences for insect herbivores remain unknown. We investigated how Cucurbita argyrosperma domestication has affected plant chemical defence and the interactions with two herbivores, the generalist Diabrotica balteata and the specialist Acalymma spp. Cucurbitacin levels and associated genes in roots and cotyledons in three wild and four domesticated varieties were analysed. Domesticated varieties contained virtually no cucurbitacins in roots and very low amounts in cotyledons. Contrastingly, cucurbitacin synthesis-associated genes were highly expressed in the roots of wild populations. Larvae of both insects strongly preferred to feed on the roots of wild squash, negatively affecting the generalist’s performance but not that of the specialist. Our findings illustrate that domestication results in tissue-specific suppression of chemical defence, making cultivars less attractive to co-evolved herbivores. In the case of squash, this may be driven by the unique role of cucurbitacins in stimulating feeding in chrysomelid beetles.