<scp>RNA</scp>‐sequencing analysis reveals abundant developmental stage‐specific and immunity‐related genes in the pollen beetle <i><scp>M</scp>eligethes aeneus</i>

Vogel, Heiko; Badapanda, Chandan; Knorr, Eileen; Vilcinskas, Andreas

doi:10.1111/imb.12067

Cited by 96 publications

(89 citation statements)

References 102 publications

(129 reference statements)

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“…Quality control measures, including the filtering of high-quality reads based on the score given in fastq files, the removal of reads containing primer/adapter sequences, and the trimming of read length, were carried out using CLC Genomics Workbench version 6.5. The de novo transcriptome assembly was carried out using the same software by comparing an assembly with standard settings and two additional CLC-based assemblies with different parameters and then selecting the presumed optimal consensus transcriptome, as described previously (20). Any conflicts among the individual bases were resolved by voting for the base with highest frequency.…”

Section: Methodsmentioning

confidence: 99%

Antimicrobial Peptides Expressed in Medicinal Maggots of the Blow Fly Lucilia sericata Show Combinatorial Activity against Bacteria

Pöppel

Vogel

Wiesner

et al. 2015

Antimicrob Agents Chemother

138

108

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The larvae of the common green bottle fly (Lucilia sericata) produce antibacterial secretions that have a therapeutic effect on chronic and nonhealing wounds. Recent developments in insect biotechnology have made it possible to use these larvae as a source of novel anti-infectives. Here, we report the application of next-generation RNA sequencing (RNA-Seq) to characterize the transcriptomes of the larval glands, crop, and gut, which contribute to the synthesis of antimicrobial peptides (AMPs) and proteins secreted into wounds. Our data confirm that L. sericata larvae have adapted in order to colonize microbially contaminated habitats, such as carrion and necrotic wounds, and are protected against infection by a diverse spectrum of AMPs. L. sericata AMPs include not only lucifensin and lucimycin but also novel attacins, cecropins, diptericins, proline-rich peptides, and sarcotoxins. We identified 47 genes encoding putative AMPs and produced 23 as synthetic analogs, among which some displayed activities against a broad spectrum of microbial pathogens, including Pseudomonas aeruginosa, Proteus vulgaris, and Enterococcus faecalis. Against Escherichia coli (Gram negative) and Micrococcus luteus (Gram positive), we found mostly additive effects but also synergistic activity when selected AMPs were tested in combination. The AMPs that are easy to synthesize are currently being produced in bulk to allow their evaluation as novel anti-infectives that can be formulated in hydrogels to produce therapeutic wound dressings and adhesive bandages.T he larvae of the common green bottle fly Lucilia sericata, a member of the blow fly family (Calliphoridae), are known as medical maggots or wound maggots, because they have been used to treat wounds as part of traditional medicine (1). This approach, commonly termed maggot therapy, is an approved and prospering alternative for the treatment of chronic and nonhealing wounds, e.g., those associated with diabetic ulcers (2-4). The therapeutic effects include the removal of necrotic tissue (debridement), the acceleration of wound healing, and wound disinfection (5, 6). The wound disinfection property has been attributed to antimicrobial components in the larval secretions, including small molecules with antibacterial activity (7, 8) and antimicrobial peptides (AMPs), such as the defensin-like lucifensin (9) and a recently reported AMP with potent antifungal activity, which accordingly was named lucimycin (10).The ability of L. sericata larvae to prosper on carrion and necrotic wounds suggests that they are adapted to colonize contaminated environments, e.g., by expressing a diverse spectrum of AMPs to protect them against the microbes they encounter (11). This theory is supported by the unique ability of rat-tailed maggots (larvae of the drone fly Eristalis tenax) to survive in highly contaminated aquatic habitats, such as liquid manure storage pits and cesspools, which also reflects their production of multiple diverse AMPs (12). Similarly, we found that the burying beetle Nicrophorus ...

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Section: Methodsmentioning

confidence: 99%

Antimicrobial Peptides Expressed in Medicinal Maggots of the Blow Fly Lucilia sericata Show Combinatorial Activity against Bacteria

Pöppel

Vogel

Wiesner

et al. 2015

Antimicrob Agents Chemother

138

108

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“…Tissue-specific transcriptome sequencing of four different mRNA pools was carried out on an Illumina HiSeq2000 Genome Analyzer platform. The transcriptome was annotated using BLAST, Gene Ontology and InterProScan searches using BLAST2GO PRO v2.6.1 (www.blast2go.de/) as described (34).…”

Section: Methodsmentioning

confidence: 99%

Evolution of moth sex pheromone composition by a single amino acid substitution in a fatty acid desaturase

Buček

Matoušková

Vogel

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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For sexual communication, moths primarily use blends of fatty acid derivatives containing one or more double bonds in various positions and configurations, called sex pheromones (SPs). To study the molecular basis of novel SP component (SPC) acquisition, we used the tobacco hornworm (Manduca sexta), which uses a blend of mono-, di-, and uncommon triunsaturated fatty acid (3UFA) derivatives as SP. We identified pheromone-biosynthetic fatty acid desaturases (FADs) MsexD3, MsexD5, and MsexD6 abundantly expressed in the M. sexta female pheromone gland. Their functional characterization and in vivo application of FAD substrates indicated that MsexD3 and MsexD5 biosynthesize 3UFAs via E/Z14 desaturation from diunsaturated fatty acids produced by previously characterized Z11-desaturase/conjugase MsexD2. Site-directed mutagenesis of sequentially highly similar MsexD3 and MsexD2 demonstrated that swapping of a single amino acid in the fatty acyl substrate binding tunnel introduces E/Z14-desaturase specificity to mutated MsexD2. Reconstruction of FAD gene phylogeny indicates that MsexD3 was recruited for biosynthesis of 3UFA SPCs in M. sexta lineage via gene duplication and neofunctionalization, whereas MsexD5 representing an alternative 3UFA-producing FAD has been acquired via activation of a presumably inactive ancestral MsexD5. Our results demonstrate that a change as small as a single amino acid substitution in a FAD enzyme might result in the acquisition of new SP compounds.fatty acid desaturase | Manduca sexta | sex pheromone biosynthesis | pheromone evolution | substrate specificity S ex pheromones (SPs) are a diverse group of chemical compounds that are central to mate-finding behavior in insects (1). Variation in SP composition between closely related species and among populations is well documented. Despite this variation, SPs are presumed to be under strong stabilizing selection, and thus the genetic mechanisms driving SP diversification represented an enigma (2). Research on SPs in moths (Insecta: Lepidoptera) helped establish the hypothesis of asymmetric tracking as a major driving force in SP diversification. In this scenario, abrupt changes in female SP composition via a shift in component ratio or the inclusion or loss of a component result in a distinct SP that attracts males with more broadly or differentially tuned SP preference (3). Assortative mating, the preferential mating of females producing a novel SP with males attracted to this SP, restricts gene flow between subpopulations with differing SP compositions. This can ultimately lead to speciation and fixation of novel communication channels (4). Work in insect models such as wasps (5), fruit flies (6), and especially moths (7-9) is helping uncover the genetic basis of SP diversification.In the majority of moth species, females use species-specific mixtures of SP components (SPCs) consisting of volatile fatty acid (FA) derivatives to attract conspecific males at long range. These SPCs are predominantly long-chain aliphatic (C12-C18) acetates, alcoh...

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“…Digital gene expression analysis was carried out by using QSeq Software (DNAStar Inc.) to remap the Illumina reads from all samples (each of the replicates of all samples was mapped individually) onto the reference backbone (Manduca Official GeneSet2) and then by counting the sequences to estimate expression levels, using previously described parameters for read mapping and normalization (Vogel et al, 2014a). Biases in the sequence datasets and different transcript sizes were corrected using the RPKM algorithm (reads per kilobase of transcript per million mapped reads) to obtain correct estimates for relative expression levels.…”

Section: Digital Gene Expression Analysismentioning

confidence: 99%

The plastic response of Manduca sexta to host and non-host plants

Koenig

Bretschneider

Heckel

et al. 2015

Insect Biochemistry and Molecular Biology

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RNA‐sequencing analysis reveals abundant developmental stage‐specific and immunity‐related genes in the pollen beetle Meligethes aeneus

Cited by 96 publications

References 102 publications

Antimicrobial Peptides Expressed in Medicinal Maggots of the Blow Fly Lucilia sericata Show Combinatorial Activity against Bacteria

Antimicrobial Peptides Expressed in Medicinal Maggots of the Blow Fly Lucilia sericata Show Combinatorial Activity against Bacteria

Evolution of moth sex pheromone composition by a single amino acid substitution in a fatty acid desaturase

The plastic response of Manduca sexta to host and non-host plants

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