Abstract

BackgroundStick and leaf insects (Phasmatodea) are an exclusively leaf-feeding order of insects with no record of omnivory, unlike other “herbivorous” Polyneoptera. They represent an ideal system for investigating the adaptations necessary for obligate folivory, including plant cell wall degrading enzymes (PCWDEs). However, their physiology and internal anatomy is poorly understood, with limited genomic resources available.ResultsWe de novo assembled transcriptomes for the anterior and posterior midguts of six diverse Phasmatodea species, with RNA-Seq on one exemplar species, Peruphasma schultei. The latter’s assembly yielded >100,000 transcripts, with over 4000 transcripts uniquely or more highly expressed in specific midgut sections. Two to three dozen PCWDE encoding gene families, including cellulases and pectinases, were differentially expressed in the anterior midgut. These genes were also found in genomic DNA from phasmid brain tissue, suggesting endogenous production. Sequence alignments revealed catalytic sites on most PCWDE transcripts. While most phasmid PCWDE genes showed homology with those of other insects, the pectinases were homologous to bacterial genes.ConclusionsWe identified a large and diverse PCWDE repertoire endogenous to the phasmids. If these expressed genes are translated into active enzymes, then phasmids can theoretically break plant cell walls into their monomer components independently of microbial symbionts. The differential gene expression between the two midgut sections provides the first molecular hints as to their function in living phasmids. Our work expands the resources available for industrial applications of animal-derived PCWDEs, and facilitates evolutionary analysis of lower Polyneopteran digestive enzymes, including the pectinases whose origin in Phasmatodea may have been a horizontal transfer event from bacteria.Electronic supplementary materialThe online version of this article (doi:10.1186/1471-2164-15-917) contains supplementary material, which is available to authorized users.

Highlights

  • Stick and leaf insects (Phasmatodea) are an exclusively leaf-feeding order of insects with no record of omnivory, unlike other “herbivorous” Polyneoptera

  • Previous research has confirmed that the endogenous cellulase genes we demonstrated are most highly expressed in the anterior midgut are most highly active in the anterior midgut [27], making it the site of both cellulase translation and action

  • If we extend the results of cellulases to those of the other Plant cell wall degrading enzyme (PCWDE), we hypothesize that phasmid digestive enzymes are active in the same region of the gut where they are expressed, making the pleated anterior midgut (AMG) the site of primary plant cell wall and polymer digestion and the posterior midgut (PMG) the site of secondary digestion of smaller oligomers at most

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Summary

Introduction

Stick and leaf insects (Phasmatodea) are an exclusively leaf-feeding order of insects with no record of omnivory, unlike other “herbivorous” Polyneoptera. They represent an ideal system for investigating the adaptations necessary for obligate folivory, including plant cell wall degrading enzymes (PCWDEs). Their physiology and internal anatomy is poorly understood, with limited genomic resources available. Its high coverage is desirable when profiling transcripts in Such a combination of low genome resource availability and enigmatic physiology exists in the stick and leaf insects (order Phasmatodea), or phasmids. Phasmids are an ideal system for studying the evolution of herbivory in the lower Polyneoptera

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