MALDI‐TOF MS identification of microbiota associated with pest insect Diabrotica speciosa

Abstract

1 Resistance development in pest insects has guided the advance of cleaner and more effective strategies for pest control. An interesting and promising strategy is the manipulation of insects via their gut microbiota. 2 To evaluate the feasibility of this strategy, Diabrotica speciosa, a highly polyphagous pest insect from South America that causes substantial damage to several important crops, was reared under controlled conditions. Aerobic culturable bacteria were isolated from the gut of D. speciosa and identified using proteomic fingerprints obtained by matrix-assisted laser desorption ionization coupled to time-of-flight mass spectrometry (MALDI-TOF MS), as well as by genomic methods via partial sequencing of the 16S rRNA gene. 3 Seventy-three strains belonging to 17 genera and up to 29 different species were isolated. -Proteobacteria of the orders Pseudomonadales and Enterobacteriales were the predominant. A core gut microbiome for the genus Diabrotica could be inferred when microbiotas from different species from the genus were compared. Molecular and spectrometric techniques indicated complete agreement of genera classification, although cluster analysis revealed distinct taxonomic grouping patterns. 4 MALDI-TOF MS provided reliable identification of culturable gut bacteria, demonstrating similar efficacy, with cheaper and faster results relative to partial 16S rRNA gene sequencing, and also showed an interesting and unexpected phyloproteomic correlation.

Characterization of Terpene Synthase from Tea Green Leafhopper Being Involved in Formation of Geraniol in Tea (Camellia sinensis) Leaves and Potential Effect of Geraniol on Insect-Derived Endobacteria

“…The similar results were found in phylogenetic analysis (Figure 7). These endobacteria were also found in other insects [26][27][28][29]. Acinetobacter group was reported to be the major endobacteria in other insects [26].…”

“…These endobacteria were also found in other insects [26][27][28][29]. Acinetobacter group was reported to be the major endobacteria in other insects [26]. Acinetobacter johnsonii is a common endosymbiont in insect [27,28].…”

Survey of bacteria associated with western corn rootworm life stages reveals no difference between insects reared in different soils

“…Available microbial sampling and survey methods have known and unknown flaws. Most early publications of Diabrotica associated bacteria relied on bacterial culture followed by morphological and sequence identification 36–38 . With soil and rhizosphere bacteria, the percentage of culturable bacteria is believed to be ~1% 26 .…”

How Hosts Taxonomy, Trophy, and Endosymbionts Shape Microbiome Diversity in Beetles

“…Despite such complexity and diversity, knowledge of microbiota in this group of insects is limited. So far, microbiome has only been examined in single species belonging to one of the trophic guilds: detritivores–coprophages [31, 32], scavengers [33], xylophages or cambiophages [34–38], herbivores [39–42], as well as carnivores [43–45]. Complex analyses concerning diversity of microbiota in relation to beetle phylogeny (taxonomy) and ecology (e.g., trophy) has yet to be conducted.…”