Abstract

For nearly 400 million years, insects and plants have been embattled in an evolutionary arms race. Insects have developed diverse feeding strategies and behaviors in an effort to circumvent and overcome an extensive collection of plant defense tactics. Sap-sucking insects often inject saliva into hosts plants, which contains a suite of effector proteins and even microbial communities that can alter the plant’s defenses. Lacking salivary glands, leaf-feeding beetles represent an interesting group of phytophagous insects. Feeding beetles regurgitate onto leaf surfaces and it is thought that these oral secretions influence insect-plant interactions and even play a role in virus-vector specificity. Since the molecular and biological makeup of the regurgitant is virtually unknown, we carried out RNA sequencing and 16S rDNA analysis on a major soybean pest, Epilachna varivestis, to generate the first ever beetle “regurgitome” and characterize its microbiome. Interestingly, the regurgitant is comprised of a rich molecular assortment of genes encoding putative extracellular proteins involved in digestion, molting, immune defense, and detoxification. By carrying out plant inoculation assays, we reinforced the fundamental role of the regurgitant in beetle-borne virus specificity. Ultimately, these studies begin to characterize the importance of regurgitant in virus transmission and beetle-plant interactions.

Highlights

  • The interactions between host plants and their phytophagous insects are intrinsically complex and subject to remarkable evolution, where both have adapted strategies to avoid each other’s defense systems

  • A cDNA library derived from the regurgitant of roughly 200 E. varivestis individuals was sequenced, which produced 56,289,018 paired end reads of 300 nt

  • The unprocessed reads have been deposited in the NCBI Sequence Read Archive (SRA) under the accession SRR4934 939

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Summary

Introduction

The interactions between host plants and their phytophagous insects are intrinsically complex and subject to remarkable evolution, where both have adapted strategies to avoid each other’s defense systems. Plants have developed an extraordinary array of physical barriers, constitutive chemical mechanisms, and direct and indirect inducible defenses intended to counter/offset the effects of insect attack [1,2,3,4,5]. Insects have adapted tactics to combat the diverse arsenal of plant defenses, allowing them to feed, grow, and reproduce on their host plants [6]. A wide range of phytophagous insects possess highly modified piercing-sucking mouthparts, enabling them to use phloem sap as their exclusive food source. Saliva is injected into plant tissues to aid in penetration, ingestion of nutrients, and modulate.

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