Abstract

The chemical ecology of plant-insect interactions has been driving our understanding of ecosystem evolution into a more comprehensive context. Chlosyne lacinia (Lepidoptera: Nymphalidae) is an olygophagous insect herbivore, which mainly uses host plants of Heliantheae tribe (Asteraceae). Herein, plant-insect interaction between Tithonia diversifolia (Heliantheae) and Chlosyne lacinia was investigated by means of untargeted LC-MS/MS based metabolomics and molecular networking, which aims to explore its inherent chemical diversity. C. lacinia larvae that were fed with T. diversifolia leaves developed until fifth instar and completed metamorphosis to the adult phase. Sesquiterpene lactones (STL), flavonoids, and lipid derivatives were putatively annotated in T. diversifolia (leaves and non-consumed abaxial surface) and C. lacinia (feces, larvae, pupae, butterflies, and eggs) samples. We found that several furanoheliangolide-type STL that were detected in T. diversifolia were ingested and excreted in their intact form by C. lacinia larvae. Hence, C. lacinia caterpillars may have, over the years, developed tolerance mechanisms for STL throughout effective barriers in their digestive canal. Flavonoid aglycones were mainly found in T. diversifolia samples, while their glycosides were mostly detected in C. lacinia feces, which indicated that the main mechanism for excreting the consumed flavonoids was through their glycosylation. Moreover, lysophospholipids were predominately found in C. lacinia samples, which suggested that they were essential metabolites during pupal and adult stages. These findings provide insights into the natural products diversity of this plant-insect interaction and contribute to uncovering its ecological roles.

Highlights

  • The integration of ecological and chemical researches has enlightened how plant-insect interactions shape community dynamics and ecosystem evolution [1,2]

  • Plants that were collected at the interface between two forest ecosystems exhibited an amplification of defensive compounds, which was related to sesquiterpene lactones (STL)

  • C. lacinia larvae that were fed with T. diversifolia leaves were reared in the laboratory (16/8 h of light/dark; 25 ± 2 ◦ C) and exhibited a survival rate of 100%

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Summary

Introduction

The integration of ecological and chemical researches has enlightened how plant-insect interactions shape community dynamics and ecosystem evolution [1,2]. Plant phytochemical diversity has been directly and positively correlated with the diversity of plant-associated insects [3]. In this context, the metabolomics approach is a valuable tool for unraveling biological interfaces, including the ecological bases for interactions among metabolites and the interplay between plants and insects [4,5,6,7]. Seasonal and spatial effects on plant secondary metabolite contents were associated with variation in the concentrations of chlorogenic acids and flavonoids [9]. We explored the plant-insect interaction between T. diversifolia and C. lacinia by means of insect performance bioassay, untargeted LC-MSn , and molecular networking analyses. We thereby unraveled the natural products diversity in this plant-insect interaction and investigated the influence of the plant metabolites on the insect metabolism

Results and Discussion
Natural Products Diversity
Experimental Design
Sample Preparation
LC-MSn Analysis
Data Processing and Analysis
Chemical Analysis and Dereplication
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