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Streito: a Barcode Database to Identify the Vectors of Xylella Fastidiosa in Europe

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A barcode database to identify the vectors of Xylella fastidiosa in Europe. By Streito Jean-Claude, Pierre Eric, Genson Guénaëlle, Bellifa Maxime, Chartois Marguerite, †Germain Jean-François, Astrid Cruaud & Jean-Yves Rasplus. Ajaccio 29/10/2019 _01 Introduction Why DNA barcoding of vectors of Xf is important? .02 Streito et al. / Barcoding of Xylella fastidiosa vectors Ajaccio 29/10/2019 Why barcoding? 1) Potential vectors of Xf are diversified 4 Hemiptera famillies, 120 spp. for Europe Aphrophoridae (29 spp.) Cercopidae (7 spp.) Cicadellidae (9 spp.) Cicadidae (79 spp.) For an overview of Xf vectors I invite you to go and see the poster P58 by Wilson et al. .03 Streito et al. / Barcoding of Xylella fastidiosa vectors Ajaccio 29/10/2019 Why barcoding? 2) Main vectors must be dissected to be identified It is possible and reliable but time-consuming and not suitable for mass screening of Xf in insects .04 Streito et al. / Barcoding of Xylella fastidiosa vectors Ajaccio 29/10/2019 Why barcoding? .05 Streito et al. / Barcoding of Xylella fastidiosa vectors Ajaccio 29/10/2019 Why barcoding? 1) Potential vectors of Xf are diversified 2) Main vectors must be dissected to be identified 3) The taxonomy of Xf’s vectors is not 100% satisfactory Barcoding is known to be adequate to check for taxonomical problems: cryptic species and synonymies This is why we started to barcode the vectors of Xf when the bacterium was detected in Corsica in summer 2015 .06 Streito et al. / Barcoding of Xylella fastidiosa vectors Ajaccio 29/10/2019 _02 Materials & Methods Our main concern is to avoid misidentification and to provide reliable sequences .07 Streito et al. / Barcoding of Xylella fastidiosa vectors Ajaccio 29/10/2019 Barcoding methods: multiple quality controls Species to be Sampling conditioning 1) IdentificationIdentification by Databases included storage a taxonomist Web portal Philaenus italosignus Philaenus lukasi Philaenus maghresignus Philaenus signatus Philaenus spumarius Philaenus tarifa Philaenus tesselatus 2)DNA Non extraction destructive and Collections extractionstorage ADN 3) Vouchers PCR amplification COI Sequencing Phylogenetic analysis 1st quality control: sequences 2nd quality control .08 Streito et al. / Barcoding of Xylella fastidiosa vectors Ajaccio 29/10/2019 Barcoding methods: Standard Barcode: COI 658 bp (Herbert et al., 2003) First: Classical PCR and Sanger sequencing Now: two-step PCR and Miseq sequencing .09 Streito et al. / Barcoding of Xylella fastidiosa vectors Ajaccio 29/10/2019 Barcoding methods: multiple quality controls Species to be Sampling conditioning 1) IdentificationIdentification by Databases included storage a taxonomist Web portal Philaenus italosignus Philaenus lukasi Philaenus maghresignus Philaenus signatus Philaenus spumarius Philaenus tarifa Philaenus tesselatus 5) Consistency 2)DNA Non extraction destructive and Collections extractionstorage ADN morpho/molecular 3) Vouchers identification PCR amplification COI Sequencing 4) Pipeline to check Phylogenetic analysis 1st quality control: sequences sequences (non-coding, 2nd quality control contaminants, numts…) .010 Streito et al. / Barcoding of Xylella fastidiosa vectors Ajaccio 29/10/2019 Photography of all species available Help for morphological identification On the CBGP collection platform: with a Keyence VHX5000 microscope .011 Streito et al. / Barcoding of Xylella fastidiosa vectors Ajaccio 29/10/2019 Database : validated sequences and photos Data are stored in the Database Arthemis http://arthemisdb.supagro.inra.fr/DefaultInfo.aspx?Page=Home Freely available in Arthemis : - Pictures - Sequences validated Arthemis DB@se focuses on pests and their natural enemies. To date it contains 21 169 COI sequences of Arthropods .012 Streito et al. / Barcoding of Xylella fastidiosa vectors Ajaccio 29/10/2019 _03 Results .013 Streito et al. / Barcoding of Xylella fastidiosa vectors Ajaccio 29/10/2019 Sampling North American vectors European vectors Total putative vectors : 22 849 - Aphrophoridae : 11 199 - Cercopidae : 165 Xylella samples - Cicadellidae : 11 364 QBOL samples - Cicadidae : 121 .014 Streito et al. / Barcoding of Xylella fastidiosa vectors Ajaccio 29/10/2019 Number of barcodes and pictures Number of Xf vectors included: http://arthemisdb.supagro.inra.fr/DefaultInfo.aspx?Page=Home European vectors: 21(+16) spp. 234 (+151) COI Non European vectors: 23 spp. 173 COI Total number of spp. 44(+32) 407(+151) COI Number of pictures : 271 (24 European spp. and 5 non European spp.) .015 Streito et al. / Barcoding of Xylella fastidiosa vectors Ajaccio 29/10/2019 Results: Philaenus 7 spp. in Europe Number of Bold Espèces / sous-espèces Pictures Séquences 5 spp. sequenced and GenBank (in process) imaged Philaenus italosignus 7 4 1 Philaenus loukasi 0 0 1 + Philaenus maghresignus 9 1 2 few sequences available Philaenus signatus (7) (7) 1 Philaenus spumarius 32 39 (11) > 100 in international Philaenus tarifa 0 0 2 databases Philaenus tesselatus 11 29 (8) 2 Philaenus arslani (Liban) 0 0 1 Philaenus elbursianus (Iran) 0 0 0 All European spp. Philaenus iranicus (Iran) 0 0 0 sequenced .016 Streito et al. / Barcoding of Xylella fastidiosa vectors Ajaccio 29/10/2019 Results: taxonomic concerns Philaenus COI separates all spp. studied except spumarius/tesselatus .017 Streito et al. / Barcoding of Xylella fastidiosa vectors Ajaccio 29/10/2019 Results: taxonomic concerns spumarius/tesselatus Philaenus Philaenus spp. : - same variable habitus - aedeagus diagnostic but difficult between spumarius/tesselatus but Is P. tesselatus a good species ? For more details I invite you to go and see the poster P60 by Seabra et al. !! .018 Streito et al. / Barcoding of Xylella fastidiosa vectors Ajaccio 29/10/2019 Results Neophilaenus Number of Bold 11 spp. in Europe Espèces / sous-espèces Pictures Séquences GenBank (in process) 2 spp. sequenced Neophilaenus albipennis 2 (1) 0 5 imaged Neophilaenus angustipennis 0 0 0 Neophilaenus campestris 9 31 (8) 3 + Neophilaenus exclamationis 9 (1) 0 Nearly no Neophilaenus infumatusA difficult genus0 , more0 0 barcodes Neophilaenus limpidusbarcodes are 0needed0 0 Neophilaenus lineatus 9 8 (6) >100 in international db Neophilaenus longiceps 0 0 0 Neophilaenus minor 2 (2) 0 Neophilaenus modestus 0 0 0 <50% European Neophilaenus pallidus 0 0 0 spp. sequenced .019 Streito et al. / Barcoding of Xylella fastidiosa vectors Ajaccio 29/10/2019 Results Aphrophora 7 spp. in Europe Number of Bold Espèces / sous-espèces Pictures Séquences 4 spp. sequenced GenBank (in process) and imaged Aphrophora alni 7 11 (6) >100 Aphrophora corticea 9 5 (5) 3 AphrophoraA lot ofmajor taxonomic problems0 0 remains0 In international db Aphrophorabeforepectoralisreliable barcode13 -3based 1 confusion between Aphrophora salicina 13 3 (13) 7 Aphrophoraidentificationsimilis can0 be performed0 0 alni/paralella and Aphrophora willemsi 0 0 0 salicina/pectoralis .020 Streito et al. / Barcoding of Xylella fastidiosa vectors Ajaccio 29/10/2019 Results Lepyronia and other genus Number of Bold Espèces / sous-espèces Pictures Séquences GenBank (in process) Lepyronia coleoptrata 15 7 (9) 14 Lepyroniella petrovi (Pologne, Russie…) 0 0 0 Paraphilaenus notatus [F, Ukraine, Russie…] 0 0 0 Peuceptyelus coriaceus 0 0 0 Lepyronia coleoptrata : COI and morpho ok Other genus are missing .021 Streito et al. / Barcoding of Xylella fastidiosa vectors Ajaccio 29/10/2019 Results Cercopidae Number of Bold Espèces / sous-espèces Pictures Séquences 7 spp. in Europe GenBank (in process) 5 spp. sequenced Cercopis arcuata (15) (7) 0 Cercopis intermedia 7 15 0 and 6 imaged Cercopis sabaudiana 0 (1?) 0 + Cercopis sanguinolenta 7 2(12) 5 Cercopis vulnerata 9 19 8 misidentifications in Haematoloma dorsata 5 15 (3?) 2 international db Triecphorella geniculata (10) 0 0 .022 Streito et al. / Barcoding of Xylella fastidiosa vectors Ajaccio 29/10/2019 Results: taxonomic concerns Cercopidae C. intermedia C. sanguinolenta COI discriminates all spp. studied Sequences of C. vulnerata C. sabaudiana and Triecphorella Only one taxonomic problem remains : are missing is C. sabaudiana a good speciesH. dorsata ? C. arcuata .023 Streito et al. / Barcoding of Xylella fastidiosa vectors Ajaccio 29/10/2019 Results Cicadellidae putative vectors of Xf Number of Bold 9 spp. listed for Espèces / sous-espèces Pictures Séquences GenBank (in process) Europe Ledra aurita 8 11 0 5 spp. sequenced Cicadella viridis 13 14 81 Cicadella lasiocarpae 0 0 0 and 6 imaged Graphocephala fennahi 1 6 21 + Evacanthus acuminatus 2 2 (1) 10 Evacanthus interruptus 7 5 (4) 11 G. fennahi/G. Evacanthus rostagnoi 0 0 0 coccinea confused Anoterostemma ivanoffi 0 0 0 in international db Errhomenus brachypterus (4) 0 0 .024 Streito et al. / Barcoding of Xylella fastidiosa vectors Ajaccio 29/10/2019 Cicadidae (1/79 spp. Europe ; 1/20 ssp. France) Aestuansella aestuans (Fabricius 1794) Cicada barbara ssp. barbara (Stål, 1866) Cicada barbara ssp. lusitanica Boulard, 1982 Cicada cretensis Quartau & Simões, 2005 Cicada mordoganensis Boulard, 1979 Results Cicada orni Linné, 1758 [F, C] Number of Cicadatra alhageos (Kolenati, 1857) Cicadatra atra (Olivier, 1790) [F] Bold Cicadatra glycyrrhizae (Kolenati 1857) Espèces / sous-espèces Pictures Sequences Cicadatra hyalina (F., 1798) Cicadatra icari Simões, Sanborn & Quartau, 2013 GenBank Cicadatra karpathoensis Simões, Sanborn & Quartau, 2013 (in process) Cicadatra persica Kirkaldy, 1909 Cicadatra platyptera Fieber, 1876 Cicadetta albipennis Fieber, 1876 Cicadetta anapaistica ssp. anapaistica Hertach 2011 Cicadidae
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  • The Occurrence of Froghoppers in Belgium (Homoptera, Cercopidae)

    The Occurrence of Froghoppers in Belgium (Homoptera, Cercopidae)

    VERHANDELINGEN VAN HET SYMPOSIUM "INVERTEBRATEN VAN BELGIE", 1989, p. 273-278 COMPTES RENDUS DU SYMPOSIUM "INVERTEBRES DE BELGIQUE", 1989, p. 273-278 The occurrence of froghoppers in Belgium (Homoptera, Cercopidae) by Jan VAN STALLE Summary many species has been clarified. In the light of this progress we have re-examined all collections ; accor• The Belgian species of Cercopidae are revised. In all 13 species are dingly, the list of Belgian species given below is only presently recognised in Belgian collections. No specimens were found of based on specimens, and old literature data are Neophilaenus exclamalionis (THUNBERG, 1784) and Aphrophora corticea GERMAR, 1821, which, accordingly, have to be removed from considered as insufficient to prove the occurrence of a the list of Belgian species. Aphrophora wittemsi LALLEMAND, 1946 is species in Belgium. synonymized with Aphrophora alpina MELICHAR, 1900. A key is given for all Aphrophora species occurring in Belgium. Key-words : Cercopidae, Belgium. Material and methods Samenvatting The present study is based on material deposited in the collections of the Koninklijk Belgisch Instituut voor Er wordt een revisie gegeven van de Belgische Cercopidae. In totaal werden 13 soorten herkend in de Belgische collecties. Er werd geen Natuurwetenschappen. These data are completed with enkel exemplaar gevonden van Neophilaenus exclamationis (THUN• own observations and collections submitted to me for BERG, 1784) and Aphrophora corticea GERMAR, 1821, die bijgevolg identification. For this purpose several works have verwijderd moeten worden van de lijst der Belgische soorten. Aphro• been used since there is no comprehensive study which phora willemsi LALLEMAND, 1946 is een nieuwe synoniem van Aphrophora alpina MELICHAR, 1900.
  • An Appraisal of the Higher Classification of Cicadas (Hemiptera: Cicadoidea) with Special Reference to the Australian Fauna

    An Appraisal of the Higher Classification of Cicadas (Hemiptera: Cicadoidea) with Special Reference to the Australian Fauna

    © Copyright Australian Museum, 2005 Records of the Australian Museum (2005) Vol. 57: 375–446. ISSN 0067-1975 An Appraisal of the Higher Classification of Cicadas (Hemiptera: Cicadoidea) with Special Reference to the Australian Fauna M.S. MOULDS Australian Museum, 6 College Street, Sydney NSW 2010, Australia [email protected] ABSTRACT. The history of cicada family classification is reviewed and the current status of all previously proposed families and subfamilies summarized. All tribal rankings associated with the Australian fauna are similarly documented. A cladistic analysis of generic relationships has been used to test the validity of currently held views on family and subfamily groupings. The analysis has been based upon an exhaustive study of nymphal and adult morphology, including both external and internal adult structures, and the first comparative study of male and female internal reproductive systems is included. Only two families are justified, the Tettigarctidae and Cicadidae. The latter are here considered to comprise three subfamilies, the Cicadinae, Cicadettinae n.stat. (= Tibicininae auct.) and the Tettigadinae (encompassing the Tibicinini, Platypediidae and Tettigadidae). Of particular note is the transfer of Tibicina Amyot, the type genus of the subfamily Tibicininae, to the subfamily Tettigadinae. The subfamily Plautillinae (containing only the genus Plautilla) is now placed at tribal rank within the Cicadinae. The subtribe Ydiellaria is raised to tribal rank. The American genus Magicicada Davis, previously of the tribe Tibicinini, now falls within the Taphurini. Three new tribes are recognized within the Australian fauna, the Tamasini n.tribe to accommodate Tamasa Distant and Parnkalla Distant, Jassopsaltriini n.tribe to accommodate Jassopsaltria Ashton and Burbungini n.tribe to accommodate Burbunga Distant.
  • Arthropods As Vector of Plant Pathogens Viz-A-Viz Their Management

    Arthropods As Vector of Plant Pathogens Viz-A-Viz Their Management

    Int.J.Curr.Microbiol.App.Sci (2018) 7(8): 4006-4023 International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 7 Number 08 (2018) Journal homepage: http://www.ijcmas.com Review Article https://doi.org/10.20546/ijcmas.2018.708.415 Arthropods as Vector of Plant Pathogens viz-a-viz their Management Ravinder Singh Chandi, Sanjeev Kumar Kataria* and Jaswinder Kaur Department of Entomology, Punjab Agricultural University, Ludhiana-141 004, Punjab, India *Corresponding author ABSTRACT An insect which acquires the disease causing organism by feeding on the diseased plant or by contact and transmit them to healthy plants are known as insect vectors of plant diseases. Most of the insect vectors belong to the order Hemiptera, Thysonaptera, Coleoptera, Orthoptera and Dermaptera. Homopteran insects alone are known to transmit K e yw or ds about 90 per cent of the plant diseases. About 94 per cent of animals known to transmit plant viruses are arthropods. On the basis of the method of transmission and persistence in Insect vectors, Plant pathogens, the vector, viruses may be classified into three categories viz. non-persistent, semi persistent and persistent viruses. Irrespective of the type of transmission, virus-vector Management relationship is highly specific and spread of vector borne diseases also depends upon Article Info potential of vector to spread the disease. Also for transmission of virus, activity of insect vectors is more important rather than their number. There is a high degree of specificity of Accepted: 22 July 2018 phytoplasma to insects and interaction between these two is complex and variable.