Great molecular variation within the species Phytoseius finitimus (Acari: Phytoseiidae): implications for diagnosis decision within the mite family Phytoseiidae Marie-Stéphane Tixier, Victor dos Santos Vicent, Martial Douin, Carlo Duso, Serge Kreiter To cite this version: Marie-Stéphane Tixier, Victor dos Santos Vicent, Martial Douin, Carlo Duso, Serge Kreiter. Great molecular variation within the species Phytoseius finitimus (Acari: Phytoseiidae): implications for diagnosis decision within the mite family Phytoseiidae. Acarologia, Acarologia, 2017, 57 (3), pp.493- 515. 10.1051/acarologia/20174168. hal-01520212 HAL Id: hal-01520212 https://hal.archives-ouvertes.fr/hal-01520212 Submitted on 10 May 2017 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. 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Acarologia 57(3): 493–515 (2017) DOI: 10.1051/acarologia/20174168 Great molecular variation within the species Phytoseius finitimus (Acari: Phytoseiidae): implications for diagnosis decision within the mite family Phytoseiidae Marie-Stéphane TIXIER1 B, Victor DOS SANTOS VICENTE1, Martial DOUIN1, Carlo DUSO2 and Serge KREITER1 (Received 23 August 2016; accepted 30 November 2016; published online 05 May 2017; edited by Farid FARAJI) 1 Montpellier SupAgro, UMR CBGP, INRA/ IRD/ CIRAD/ Montpellier SupAgro, 755 Avenue du Campus Agropolis, CS 30016, 34988 Montferrier-sur-Lez cedex, France. [email protected] (B), [email protected], [email protected]; [email protected] 2 Department of Agronomy, Food, Natural resources, Animals and Environment, University of Padova, Viale dell’Università 16, 35020 Legnaro (PD), Italy. [email protected] ABSTRACT — Molecular markers are increasingly used for species identification and new taxa description. However, rules to determine frontiers between populations and species are not clear depending on taxa considered. For mites, few studies deal with molecular diagnoses, making associated decision’ rules difficult. The present study focuses on a species of the predatory mite family Phytoseiidae (Phytoseius finitimus), considered for biological control of mites and small in- sect pests in fruit orchards and vineyards in the Mediterranean basin. This paper aims to elucidate the causes of great molecular variations and questions the occurrence of cryptic species. Molecular (12S rRNA, CytB mtDNA, ITSS) and mor- phological analyses were performed on four populations collected in Corsica and Italy in crops (vine and kiwi) and in an uncultivated environment (Viburnum lantana). Different methods for identifying species have been used (tree approaches, distances and ABGD algorithms). A reference database of distances within and between Phytoseiidae species has been elaborated to inform the present question and to assist with further diagnosis within Acari. Mitochondrial DNA analyses show that specimens from V. lantana were well separated from the three other populations with high genetic distances, suggesting the existence of a cryptic species. Molecular ITSS analyses coupled with morphological features show how- ever that the four populations seem to belong to the same species. The great mitochondrial polymorphism is discussed in regards to: (i) genetic distances reported for Phytoseiidae species and (ii) potential biological differences between populations (cultivated versus uncultivated areas). This study clearly emphasizes the necessity of integrative taxonomy approaches for diagnosis decisions. Furthermore, based on the polymorphism herein detected, maximal intraspecific dis- tances are proposed (9, 23 and 2.8 % for 12S rRNA, CytB mtDNA and ITSS) for diagnosis decisions within Phytoseiidae. Further statistical analyses are however clearly required to determine statistical error for general and reliable decision making. KEYWORDS — Acari; molecular taxonomy; morphometrics; species delineation; integrative taxonomy http://www1.montpellier.inra.fr/CBGP/acarologia/ 493 ISSN 0044-586-X (print). ISSN 2107-7207 (electronic) Tixier M.-S. et al. INTRODUCTION DeSalle et al. 2005; Collins and Cruickshank 2013). When elaborating such a database for Phytoseiidae Molecular markers for differentiating microorgan- species, high molecular variations were observed ism, plant and animal species are currently used in within specimens identified as P. finitimus during taxonomy. The rapid technical developments (i.e. samplings carried out in Italy and France, two areas DNA extraction, PCR and Sanger sequencing) dur- where this species is most commonly found. ing these last decades as well barcoding approaches and associated conceptual definitions have led to the development of molecular diagnosis (i.e. Hebert et al. 2003a,b; 2004a,b; DeSalle et al. 2005; Gold- Phytoseius finitimus is a generalist predator and is stein and DeSalle 2011). Molecular markers were quite common in Mediterranean vineyards (i.e. Ra- traditionally used when doubts on species identity gusa and Ciulla 1991; Duso and Vettorazzo 1999; arise from morphological comparisons and some- Duso and Fontana 2002; Peverieri et al. 2009; times from biological features between populations. Miñarro and Kreiter 2012). Some studies have However, cryptic species are more and more re- shown its ability to control phytophagous mites, vealed via comparison of DNA sequences of appar- whiteflies and thrips and to develop when fed with ently morphologically similar specimens (i.e. Pons pollen (Pappas et al. 2013) and Ahmad et al. (2015) et al. 2006; Witt et al. 2006; Bickford et al. 2007; recently studied its interspecific competition fea- Valentini et al. 2009; Foottit et al. 2009; Pauls et tures. The present study aims to elucidate the al. 2010; Jorger et al. 2012; Kekkonen and Hebert, great molecular distances observed between four P. 2014; Kekkonen et al. 2015). The present study fo- finitimus populations and to determine how these cuses on a species of the mite family Phytoseiidae: great variations are related or not to cryptic species Phytoseius finitimus Ribaga. This family, contain- occurrence. Answering this question is clearly im- ing 2,479 valid species (Demite et al. 2016), is one portant for biological control applications, as imple- of the most well-known within the Mesostigmata, mentations and advice for agro-environmental man- due to the predatory abilities of numerous species agement would be different if one or two species are to control mite and small insect pests (McMurtry named under P. finitimus. To determine if one or sev- and Croft 1997; Gerson et al. 2003; McMurtry et eral species were present, an integrative taxonomic al. 2013). Correct identification of these predators approach based on three molecular fragments and is clearly of huge importance for ensuring the suc- morphological analyses has been carried out. The cess of biological control, as different species have number of analytical developments for assessing different biological features (i.e. Tixier et al. 2006; species within a DNA sequence dataset is increas- McMurtry et al. 2013). Phytoseiidae species identifi- ing for ensuring diagnosis decision (i.e. Kekkonen cation is essentially based on female morphological et al. 2015). Traditional cluster tree-based and dis- characters. However, the reliability of those char- tance threshold-based approach as well as the over- acters as well as the low number of morphological lapping ABGD (Automatic Barcode Gap Discovery) features potentially available (due to the mite small algorithm have been herein used to determine their size, 500 µm maximum in length) have led to the respective performance and congruence. Further- development of molecular studies to resolve ques- more, a compilation of genetic distances between tions concerning synonymies (i.e. Jeyaprakash and Phytoseiidae species retrieved from already pub- Hoy 2002; Noronha et al. 2003; Tixier et al. 2006, lished works is also provided