DNA Barcoding    

processed species at   march 17, 2012

Agelastica alni
Altica brevicollis
Altica lythri
Altica oleracea
Altica pusilla
Aphthona cyparissiae
Aphthona flava
Aphthona herbigrada
Aphthona lutescens
Aphthona nonstriata
Aphthona sp.
Aphthona sp.
Argopus ahrensii
Arima marginata
Arrhenocoela lineata
Bruchus sp.
Bruchus sp.
Bruchus sp.
Calomicrus circumfusus
Calomicrus sp.
Cassida algirica
Cassida alpina
Cassida denticollis
Cassida ferruginea
Cassida inopinata
Cassida inquinata
Cassida margaritacea
Cassida prasina
Cassida rubiginosa
Cassida sanguinolenta
Cassida subreticulata
Cassida vibex
Cassida viridis
Chaetocnema conducta
Chaetocnema hortensis
Chrysochus asclepiadaeus
Chrysolina americana
Chrysolina fastuosa
Chrysolina graminis
Chrysolina haemoptera
Chrysolina herbacea
Chrysolina polita
Chrysolina rossia
Chrysolina sp.
Chrysolina sturmi
Chrysolina varians
Chrysolina viridana
Chrysomela populi
Chrysomela vigintipunctata
Clytra laeviuscula
Clytra quadripunctata
Colaspidea sp.
Colasposoma sp.
Coptocephala scopolina
Coptocephala sp.
Crepidodera aurata
Crepidodera aurea
Crepidodera fulvicornis
Crepidodera lamina
Crepidodera plutus
Cryptocephalus albzorensis
Cryptocephalus aquitanus
Cryptocephalus aureolus
Cryptocephalus bameuli
Cryptocephalus barii
Cryptocephalus biguttatus
Cryptocephalus bilineatus
Cryptocephalus bimaculatus
Cryptocephalus bipunctatus
Cryptocephalus blanduloides
Cryptocephalus cantabricus
Cryptocephalus chrysopus
Cryptocephalus connexus
Cryptocephalus convergens
Cryptocephalus cordiger
Cryptocephalus cristula
Cryptocephalus cyanipes
Cryptocephalus czwalinae
Cryptocephalus dumonti
Cryptocephalus elegantulus
Cryptocephalus eridani
Cryptocephalus flavipes
Cryptocephalus fulvus
Cryptocephalus globicollis
Cryptocephalus hennigi
Cryptocephalus hypochaeridis
Cryptocephalus imperialis
Cryptocephalus informis
Cryptocephalus janthinus
Cryptocephalus labiatus
Cryptocephalus leonhardi
Cryptocephalus loreyi
Cryptocephalus marginatus
Cryptocephalus marginellus
Cryptocephalus mariae
Cryptocephalus molossus
Cryptocephalus moraei
Cryptocephalus nitidus
Cryptocephalus ocellatus
Cryptocephalus octoguttatus
Cryptocephalus pini
Cryptocephalus primarius
Cryptocephalus pusillus
Cryptocephalus pygmaeus
Cryptocephalus pygmaeus vittula
Cryptocephalus quadripunctatus
Cryptocephalus renatae
Cryptocephalus rugicollis
Cryptocephalus samniticus
Cryptocephalus scapularis
Cryptocephalus sericeus
Cryptocephalus sexpunctatus
Cryptocephalus signatifrons
Cryptocephalus strigosus
Cryptocephalus tetraspilus
Cryptocephalus therondi
Cryptocephalus transcaucasicus
Cryptocephalus transiens
Cryptocephalus trimaculatus
Cryptocephalus turcicus
Cryptocephalus variegatus
Cryptocephalus violaceus
Cryptocephalus violaceus scaffaiolus
Cryptocephalus vittula
Cryptocephalus zoiai
Derocrepis rufipes
Derocrepis sodalis
Diabrotica virgifera
Dibolia sp.
Dicladispa testacea
Donacia brevitarsis
Donacia crassipes
Donacia obscura
Entomoscelis rumicis
Epitrix pubescens
Exosoma lusitanicum
Exosoma sp.
Galeruca laticollis
Galeruca pomonae
Galeruca sp.
Galeruca tanaceti
Galerucella lineola
Galerucella nymphaeae
Galerucella pusilla
Gastrophysa viridula
Gonioctena quinquepunctata
Gonioctena sp.
Hermaeophaga mercurialis
Hippuriphila modeeri
Hispa atra
Hypocassida cornea
Hypocassida subferruginea
Labidostomis longimana
Labidostomis sp.
Labidostomis sp.
Labidostomis taxicornis
Lachnaia italica
Lema cyanella
Lilioceris merdigera
Linaeidea aenea
Lochmaea caprae
Lochmaea crataegi
Longitarsus aeneicollis
Longitarsus apicalis
Longitarsus exoletus
Longitarsus luridus
Longitarsus melanocephalus
Longitarsus pinguis
Longitarsus refugiensis
Longitarsus sp.
Longitarsus succineus
Luperus flavipes
Luperus leonardii
Luperus longicornis
Luperus luperus
Luperus sp.
Luperus viridipennis
Lythraria salicariae
Macrolenes dentipes
Melasoma populi
Neocrepidodera cyanipennis
Neocrepidodera ferruginea
Neocrepidodera melanostoma
Neocrepidodera peirolerii
Neocrepidodera sp.
Neocrepidodera transversa
Nimphius sp.
Oomorphus concolor
Oreina bifrons
Oreina cacaliae
Oreina elongata
Oreina liturata
Oreina sp.
Oreina sp.
Oreina sp.
Oreina speciosa
Oreina speciosissima
Oreina virgulata
Orsodacne cerasi
Orsodacne variabilis
Oulema melanopus
Oulema duftschmidi
Pachybrachis burlinii
Pachybrachis exclusus etruscus
Pachybrachis hieroglyphicus
Pachybrachis hippophaes
Pachybrachis karamani
Pachybrachis sp.
Pachybrachis sp.
Pachybrachis tessellatus
Phaedon cochleariae
Phratora vitellinae
Phyllotreta ochripes
Phyllotreta sp.
Phyllotreta vittula
Plagiodera versicolora
Plateumaris consimilis
Podagrica malvae
Prasocuris phellandrii
Psylliodes affinis
Psylliodes brisouti
Psylliodes dulcamarae
Psylliodes gibbosus
Psylliodes kuntzei
Psylliodes sp.
Psylliodes thlaspis
Sermylassa halensis
Smaragdina affinis
Smaragdina chloris
Smaragdina concolor
Smaragdina flavicollis
Smaragdina limbata
Smaragdina salicina
Smaragdina sp.
Sphaeroderma rubidum
Sphaeroderma testaceum
Stylosomus minutissimus
Timarcha latipes
Timarcha nicaeensis
Tituboea biguttata
Tituboea sp.
Tituboea sp.

DNA barcoding of the Chrysomelidae of the Mediterranean Region

Matteo Montagna
Davide Sassi
Giuseppe Carlo Lozzia
Renee Labee

      The C-Bar (Chrysomelidae Barcoding) project, developed within the consortium of Barcode of Life Database (BOLD), has as goal the creation of sequences database for the Leaf Beetles of the Mediterranean region (MEDLB). The project, activated at the Molecular Entomology Laboratory (LEM DIPSA, University of Milan) by prof. Giuseppe Lozzia, was founded by the idea of Matteo Montagna and Davide Sassi. The project aims to obtain a fragment of approximately 650 base pairs of the gene coding for cytochrome oxidase I for all the chrysomelid species present in the Mediterranean Region. The purpose is to realize a tool able to identify at species level adult, pre-imaginal stages and fragments of biological samples.
        The molecular data obtained (650 bp of COI gene) allow also the identification of criptospecies undetectable morphologically and, ultimately, when laboratory practices and technology allow routine molecular analysis, the species identification even in the absence of expertise. These data permit to obtain valuable information on the intraspecific genetic variability (analyzing samples from different localities of the species distribution area), the evolutionary dynamics and timing of speciation of the taxa analyzed. A further possibility provided by C-Bar project, quite innovative even in the context of the revolutionary applications of molecular biosystematics is the molecular characterization of the gut content of the specimens. In this way you could get an overwhelming amount of information on the host plants, greatly increasing the modest knowledge of the biology of almost all of our species. Finally, the collected data can be used in the census on the fauna of a particular ecosystem or of entire nations, as well as used in biogeographic studies to define and/or complete information on the species area and possibly make new knowledge on species phenology.
         The approach used by the Laboratory of Molecular Entomology (LEM DIPSA, University of Milan) provides integration between the traditional taxonomy, based on morphology, and the support provided by molecular analysis of intraspecific genetic variability. The specimens collected during the campaigns are preserved in absolute alcohol for subsequent DNA extraction and are accompanied by the collection information (i.e. collection date, location, latitude, longitude, altitude, collector and ecological notes).
        Since the molecular approach will not and can not replace the traditional morphological study, but must accompany the traditional taxonomy by integrating information, DNA extraction is performed using a technique that preserves the insect morphology. Then the samples are prepared and dry stored in the collections of the LEM DIPSA for all the necessary comparisons.
        The molecular data used in research are deposited in the NCBI database and BOLD and, once published, will be used freely by any registered user.
To date, at the end of the preliminary study, we have obtained molecular sequences for 582 specimens belonging to 208 species (alphabetical list) from different localities in the Mediterranean (Italy, France, Tunisia, Greece, Turkey). By the summer of 2012 the database will include, therefore, sequences belonging to 240 taxa with a minimum of 4 analyzed samples from different locations.
        At present the project does not present a closing date. Although the great cooperation with the BOLD consortium is close to its ending, we plan to continue the research and we are working to raise founds needed in order to run the expensive laboratory procedures.

        How to get involved
        The initiative in which we are involved needs help in to collect specimens from different localities, to increase the number of processed species within the database in order to have a good representation of intraspecific genetic variability by analyzing samples of different origin.
        The collaboration, even occasional, of friends and entomologists is desirable and in fact essential for the success of the project. We therefore call on all the entomologists available to collaborate, the possibility to give a valuable contribution to the project by collecting the material.

        If you are interested in helping us
        please contact Matteo Montagna (e-mail) or Davide Sassi (e-mail) and we will send you the DNA Barcoder kit (test tubes, tweezers and absolute ethanol for the samples storage) and the manual for the manipulation of the samples. Even few specimens of relatively common species, can be very interesting if they come from areas where we have only few samples. For this reason, everyone is able to provide a very important contribution.


        We take this opportunity to publicly thank the friends who in the past have provided interesting material, namely Fernando Angelini, Maurizio Bollino, Andrea Carlin, Juan de la Rosa, Francesco Izzillo, Carlo Leonardi, Gianluca Magnani.

        Herbert PDN, 2005. The Promise of DNA Barcoding for Taxonomy. Systematic Biology, 54: 852-859.
        Hebert PD, Cywinska A, Ball SL, deWaard JR, 2003a. Biological identifications through DNA barcodes. Proc Biol Sci, 270: 313-321.
        Hebert PD, Ratnasingham S, deWaard JR, 2003b. Barcoding animal life: cytochrome c oxidase subunit 1 divergences among closely related species. Proc Biol Sci, 270 Suppl 1: S96-S99.
        Will KW, & Rubinoff D, 2004. Myth of the molecule: DNA barcodes for species cannot replace morphology for identification and classification. Cladistics 20: 47–55.
        Kubisz D, Kajtoch L, Mazur MA, Rizun V, 2012. Molecular barcoding for central-eastern European Crioceris leaf-beetles (Coleoptera: Chrysomelidae). Cent Eur J Biol, 7:69-76