Identification, structural characterisation and expression analysis of a defensin gene from the tiger beetle Calomera littoralis (Coleoptera: Cicindelidae)

Rodríguez-García, María Juliana; García-Reina, Andrés; Machado, Vilmar; Galián, José

doi:10.1016/j.gene.2016.05.030

Cited by 10 publications

(7 citation statements)

References 55 publications

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“…A defensin gene was tested in C . littoralis after an infection, which seemed to increase its expression as it was showed in a previous work 37 , being in accordance to other studies performed for defensin genes in insects 51 , 52 . On the other hand, the ubiquitin UBC9 was tested for the rest of the conditions in both species, showing significant differences in the body parts comparison, primarily in gonads, as was previously checked in a different work 38 and in other insects such as Drosophila 53 , which confirmed the relation of this gene with cicindelids reproductive system.…”

Section: Discussionsupporting

confidence: 92%

“…littoralis , with an additional new condition: infection of beetles with 1 μl of 1 mg/ml lipopolysaccharides (LPS) of Escherichia coli as previously performed by Rodríguez-García et al . 37 and harvested after 12 hours of infection. Control samples were injected with 1 μl of insect saline solution.…”

Section: Methodsmentioning

confidence: 99%

“…littoralis (sex: Myo and Myo-Alc; infection: NADH and B-t; body parts: NADH, 60Sa and ArgKin). Both target genes were amplified as described in previous works with previously described primers 37 , 38 . UBC9, an ubiquitin gene related to reproduction and sexual activity, was tested for all conditions and species with the exception of the infection in C .…”

Section: Methodsmentioning

confidence: 99%

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Validation of reference genes for quantitative real-time PCR in tiger beetles across sexes, body parts, sexual maturity and immune challenge

García-Reina

Rodríguez-García

Galián

2018

Sci Rep

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Reference genes are frequently used as normalizers for expression studies despite not being previously verified to present suitable stabilities. Considering the interest that tiger beetles have generated in the past years, resulting in a variety of studies, it is crucial to dispose of a validated reference gene panel for expression studies. Nine candidate genes were tested in Cicindela campestris and Calomera littoralis across several conditions and their transcription levels were assessed with geNorm, NormFinder, BestKeeper and ΔCT method algorithms. Results showed high stabilities across sexes, immune challenge and gonad developmental stages for all genes tested, while body parts comparison presented less constant expression values. Only two genes are sufficient to perform a proper normalization for most of the conditions tested, except for the body parts comparison in C. littoralis, which requires the use of at least three reference genes. On the whole, no universal gene is found to be suitable for all situations, but according to the acceptable range of values, NADH, B-t, Vatpase and ArgKin seem to present the most constant expression stability, indicating their suitability as reference genes in most of the conditions. This is the first report evaluating the stability of housekeeping genes in adephagan beetles.

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Section: Discussionsupporting

confidence: 92%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Validation of reference genes for quantitative real-time PCR in tiger beetles across sexes, body parts, sexual maturity and immune challenge

García-Reina

Rodríguez-García

Galián

2018

Sci Rep

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“…The signal peptide and propeptide cleavage sites were predicted using the ProP 1.0 Server (Duckert et al, 2004). The structure of the defensins was predicted as described by Rodríguez-García et al (2016). Briefly, protein model templates were identified based on sequence alignments generated with the Hhpred server (https://toolkit.tuebingen.mpg.de/tools/hhpred).…”

Section: Defensin Sequence Alignment and Protein Structure Predictionmentioning

confidence: 99%

Differential Expression of Putative Ornithodoros turicata Defensins Mediated by Tick Feeding

Armstrong

Kneubehl

Mitchell

et al. 2020

Front. Cell. Infect. Microbiol.

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Additional research on soft ticks in the family Argasidae is needed to bridge the knowledge gap relative to hard ticks of the family Ixodidae; especially, the molecular mechanisms of Ornithodoros biology. Ornithodoros species are vectors of human and animal pathogens that include tick-borne relapsing fever spirochetes and African swine fever virus. Soft tick vector-pathogen interactions involving components of the tick immune response are not understood. Ticks utilize a basic innate immune system consisting of recognition factors and cellular and humoral responses to produce antimicrobial peptides, like defensins. In the present study, we identified and characterized the first putative defensins of Ornithodoros turicata, an argasid tick found primarily in the southwestern United States and regions of Latin America. Four genes (otdA, otdB, otdC, and otdD) were identified through sequencing and their predicted amino acid sequences contained motifs characteristic of arthropod defensins. A phylogenetic analysis grouped these four genes with arthropod defensins, and computational structural analyses further supported the identification. Since pathogens transmitted by O. turicata colonize both the midgut and salivary glands, expression patterns of the putative defensins were determined in these tissues 1 week post engorgement and after molting. Defensin genes up-regulated in the tick midgut 1 week post blood feeding were otdA and otdC, while otdD was up-regulated in the midgut of post-molt ticks. Moreover, otdB and otdD were also up-regulated in the salivary glands of flat post-molt ticks, while otdC was up-regulated within 1 week post blood-feeding. This work is foundational toward additional studies to determine mechanisms of vector competence and pathogen transmission from O. turicata.

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“…The members of this family differ in size, being known, currently, the subfamilies α, β and θ defensin. In addition, the family defensin have members with action against bacteria, fungi and protozoa [18,21,51] .…”

Section: Classes and Familiesmentioning

confidence: 99%

Technological Potential of Antimicrobial Peptides: a Systematic Review

Machado¹,

Gelinski²,

Baratto³

et al. 2019

ijps

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Machado et al.: Review of Antimicrobial Peptides Resistance to antibiotics has being considered one of the greatest public health problems worldwide. The objective of this systematic review was to compile important bibliographical references that support the studies related to the biotechnological potential of antimicrobial peptides. Antimicrobial peptides are considered the major components of the innate immune system and work in defence against infections caused by different microorganisms. Many researchers argue that the studies associated with the discoveries of antimicrobial peptides that are more effective in the treatment of infections caused by microorganisms be also directed to the substances produced by insects. The major aspects associated to the technological potential of antimicrobial peptides are, general characteristics, classification, mode of action and prospects for applying, mainly in the pharmaceutical industry. It can be concluded that research for new substances with action against pathogenic microorganisms aim mainly to seek alternatives to the use of antibiotics. In addition, anticancer peptides are small cationic molecules with promising pharmacological use. However, the products for application in the protection of food, treatment of skin infections and its use in the cosmetics industry are those with the greatest potential.

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Identification, structural characterisation and expression analysis of a defensin gene from the tiger beetle Calomera littoralis (Coleoptera: Cicindelidae)

Cited by 10 publications

References 55 publications

Validation of reference genes for quantitative real-time PCR in tiger beetles across sexes, body parts, sexual maturity and immune challenge

Validation of reference genes for quantitative real-time PCR in tiger beetles across sexes, body parts, sexual maturity and immune challenge

Differential Expression of Putative Ornithodoros turicata Defensins Mediated by Tick Feeding

Technological Potential of Antimicrobial Peptides: a Systematic Review

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