Tracking mite trophic interactions by multiplex PCR

Abstract

BACKGROUND: A thorough knowledge of trophic webs in agroecosystems is essential to achieve successful biological pest control. Phytoseiid mites are the most efficient natural enemies of tetranychid mites, which include several important pests worldwide. Nevertheless, phytoseiids may feed on other food sources including other microarthropods, plants and even other phytoseiids (intraguild predation), which can interfere with biological control services. Molecular gut content analysis is a valuable tool for characterizing trophic interactions, mainly when working on microarthropods such as mites. We have designed new primers for Phytoseiidae, Tetranychidae and Thysanoptera identification and they have been multiplexed in a polymerase chain reaction (PCR) together with universal plant primers. Additionally, we have estimated prey DNA detectability success over time (DS 50 ) considering the most probable events in Spanish citrus orchards: the phytoseiid Euseius stipulatus as a predator, the phytoseiid Phytoseiulus persimilis as intraguild prey, and the thrips Frankliniella occidentalis and Anaphothrips obscurus as alternative prey to Tetranychus urticae. gut of the phytoseiid predator. DS 50 for E. stipulatus as the predator were 1.3, 2.3 and 18.7 h post feeding for F. occidentalis, A. obscurus and P. persimilis as prey, respectively. CONCLUSION: Tracking of the trophic relationships within the citrus acarofauna, and the unveiling of the role of alternative food sources will pave the way for enhancing T. urticae biological control. This multiplex PCR approach could be applicable for these purposes in similar agroecosystems. RESULTS: The designed multiplex PCR allows the identification of phytoseiids (both predator and intraguild prey) and detects alternative food sources mentioned above in the DNA extractionDNA of Phytoseiidae, Tetranychidae, Thysanoptera and other potential food sources for Phytoseiidae mites in citrus orchards and those provided in the laboratory rearings was extracted following different protocols depending on the organism and the objective of the study.Acari and Hemiptera from the UJI-IVIA DNA collection and new Acari DNA used for feeding trials (Tables 1 and 3), were previously Pest Manag Sci 2020; 76: 597-608

Plant-feeding may explain why the generalist predator Euseius stipulatus does better on less defended citrus plants but Tetranychus-specialists Neoseiulus californicus and Phytoseiulus persimilis do not

“…These induced genotypedependent responses closely match those triggered by E. stipulatus (Cruz-Miralles et al 2019). Together with results by Gómez-Martínez et al (2020) showing that this phytoseiid is able to obtain liquids by piercing a parafilm membrane, these results could be taken as an indirect evidence of the plant cell-sap feeding of E. stipulatus. Indeed, the cheliceral morphology typical of phytoseiid plant cell-sap feeders has been observed in different species of the genus Euseius (Adar et al 2012).…”

DNA Barcoding and Phylogeny of Acari Species Based on ITS and COI Markers

“…With the results here provided, increasing the number of sequences of ITS and by determining the intraspecific p-distances, we corroborate its usage as another DNA barcoding region that allows for reliable species identification based on nucleotide p-distances or phylogenetic closeness to known voucher specimens' sequences. These two fragments have shown enough contiguous sequence to develop species-specific primers to set up a PCR marker which is easily implemented in pest management programs, as many of the pest management officers have access to routine low-cost molecular biology laboratories (see [9,11]). In this work, we were able to infer DNA barcodes from ITS sequences, as they show high interspecies and low intraspecific nucleotide distances that altogether with phylogenetic relationships allow the assignment to species (see Table S6).…”

“…In this sense, the taxonomy and systematics of Tetranychidae and Phytoseiidae have been traditionally based on morphology [8], but their minute size and reduced number of morphological taxonlinked structures make it a difficult task. These drawbacks have driven acarologists into the world of DNA markers in the past decades, which jointly with the use of sequence identity percentage and/or phylogenetic neighborhood allow species identification [4,[9][10][11]. Over the last years, several kinds of molecular markers have been used to study the Acari species with different purposes, from clarification of taxonomic problems [12][13][14][15][16][17][18], to the understanding of the dynamic of the pest [19], the establishment of dispersal patterns [19,20], the determination of population structure [21][22][23][24][25][26][27], their food preferences [9,11], and even to the determination of geographical origin of invasive species producing an outbreak [28][29][30][31][32].…”

“…These drawbacks have driven acarologists into the world of DNA markers in the past decades, which jointly with the use of sequence identity percentage and/or phylogenetic neighborhood allow species identification [4,[9][10][11]. Over the last years, several kinds of molecular markers have been used to study the Acari species with different purposes, from clarification of taxonomic problems [12][13][14][15][16][17][18], to the understanding of the dynamic of the pest [19], the establishment of dispersal patterns [19,20], the determination of population structure [21][22][23][24][25][26][27], their food preferences [9,11], and even to the determination of geographical origin of invasive species producing an outbreak [28][29][30][31][32]. Moreover, one species, Tetranychus urticae Koch (Trombidiformes: Tetranychidae), has been the focus of an International Genome consortium that has achieved the first full genome and transcriptome record for an economically important Acari agricultural pest [33].…”

Identification of Glycycometus malaysiensis (for the first time in Brazil), Blomia tropicalis and Dermatophagoides pteronyssinus through multiplex PCR

“…Due to these drawbacks, molecular techniques based on ribosomal DNA (rDNA) and cytochrome oxydase subunit 1 ( coxI ) have been used as important tools on mite identification and phylogenetic analysis (Lava Kumar et al 1999 ; Navajas and Fenton 2000 ; Vargas et al 2005 ; Ben-David et al 2007 ; De Rojas et al 2007 ; Osakabe et al 2008 ; Wong et al 2011 ; Yang et al 2011 ; Matsuda et al 2012 ; Thet-Em et al 2012 ; Arimoto et al 2013 ; Roussel et al 2013 ; Stephenson et al 2013 ; Navia et al 2014 ; Beroiz et al 2014 ; Khaing et al 2014 ; Lv et al 2014 ; Juan et al 2015 ; Li et al 2015 ; Stalažs and Moročko-Bičevska 2016 ; Peltier et al 2018 ; Syromyatnikov et al 2018 ; Zélé et al 2018 ; Zhou et al 2020 ; Gómez-Martínez et al 2020 ; Oliveira et al 2021 ; Zhang et al 2021 ).…”