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A kutatási téma leírása:
Native to East Asia (China, Korea, Japan, Taiwan and northern Vietnam) (Josifov and Kerzhner, 1978; Rider et al. 2002), the brown marmorated stink bug, Halyomorpha halys (Stål, 1855) (Hemiptera: Heteroptera: Pentatomidae), has emerged as a harmful invasive insect pest in North America and Europe in the 1990s and 2000s, respectively (Haye et al. 2015). This pest has been identified in 44 states of the USA (Northeastern IPM Center, 2017). In Europe, it was first found more than 10 years ago in Switzerland (Zurich, 2004) (Wermelinger et al. 2008). In recent years, it has spread to additional countries (Leskey and Nielsen, 2018). The first specimens of H. halys in Hungary were identified in 2013 at two locations in Budapest namely Péterimajor and Buda Arboretum (Vétek et al. 2014). In 2014, this species was found in additional locations in the capital (Papp et al. 2014; Korányi et al. 2015). One year later, further detections were reported from other sites (Budakalász and Martonvásár) (Vétek, 2016). In 2016, the pest was reported to be present in several other parts of Hungary and mass occurrence was observed at Pécs, in the southern part of Hungary. (Mészáros, 2016).
Halyomorpha halys has become one of the most harmful invasive insect pests in the world. This polyphagous species feeds on over 170 plants, comprising many agricultural plants that have a great economic importance (e.g., soybeans and sweet-corn), various fruits (e.g., apples and peaches), vegetables (e.g. tomato and pepper) and ornamentals (Leskey and Nielsen, 2018). In the mid-Atlantic region of the USA, it has become the dominant Pentatomidae species (Nielsen and Hamilton, 2009; Nielsen et al. 2011), inducing $37 million in losses to the apple growers, as well as unreported losses to a variety of other fruits, vegetables, row crops and ornamentals including peaches, nectarines, tomatoes, peppers, sweet corn, and soybeans (Leskey and Nielsen, 2018). In Switzerland, H. halys has been recorded on more than 51 host plants in 32 families, which include many exotic and native plants (Haye et al. 2014b). In comparison, 106 host plants in 45 families have been reported in Asia (Lee et al. 2013). Damage by H. halys in Hungary was first detected in dry bean („Etna‟) and forced green hot pepper („Daras‟) in 2016 (Vétek and Korànyi, 2017).
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The molecular genetic markers (mitochondrial and nuclear) are useful tools to examine
diversity, to determine the source of an invasive pest, and to understand the invasion
processes (Ficetola et al. 2008). There are only a few records worldwide of molecular
characterization of H. halys populations (Lee et al. 2009; Xu et al. 2014; Gariepy et al. 2014;
Cesari et al. 2014; Gariepy et al. 2015; Morrison III et al. 2017). Despite its presence in
Hungary, the Hungarian populations of H. halys occurring in different parts of the country
have not been genetically characterized yet. For these reasons, this study aims to reveal the
genetic diversity of the Hungarian populations of H. halys and to identify its pathways of
entry and dispersal in Hungary as parts of the development of an effective management
strategy.
Based on previous studies (Xu et al. 2014; Cesari et al. 2014; Gariepy et al. 2014), we
intend to analyze three portions of mitochondrial gene: Cytochrome Oxydase I (COI),
Cytochrome Oxydase II (COII) and Cytochrome B, of the Hungarian populations of H. halys.
These genes have been successfully used to evaluate the geographic origin and colonization
history of a number of introduced invertebrates (Franks et al. 2011; Teske et al. 2011; Kanuch
et al. 2012). In particular, the COI will be used to evaluate the genetic diversity of the
Hungarian populations of H. halys. To carry out this study, field and laboratory work will
both be necessary.
Samplings in the different locations in Hungary where the species is known to occur
will take place. From each specimen, the Genomic DNA will be extracted from a single leg
using a Chelex extraction method (Walsh et al. 1991). Using polymerase chain reaction (PCR)
techniques, the Cytochrome Oxydase I, Cytochrome Oxydase II and Cytochrome B will be
amplified. We will employ specific PCR primers used in previous studies. The Cytochrome
Oxidase I will be amplified using Primes LCO-1490 (5‟-GGT CAA CAA ATC ATA AAG
ATA TTGG-3‟) and HCO-2198 (5‟-TAA ACT TCA GGG TGA CCA AAA AATCA- 3‟)
(Folmer et al. 1994). The COII gene will be amplified with primers HhalysCO2F2 (5‟-TAA
CCC AAG ATG CAA ATT CT-3‟) and HhalysCO2R2 (5‟-CCA TAT ATA ATT CCT GGA
CGA-3‟) (Xu et al. 2014; Cesari et al. 2014). The Cytochrome B will be amplified using
primers described by Muraji et al. 2000 in Gariepy et al. 2014 (forward: 5‟-TAG GAT ATG
TTT TAC CTT GAG GACA-3‟; reverse: 5‟-CTC CTC CTA ATT TAT TAG GAA TTG-3‟).
The amplified products will be subject to electrophoresis on agarose gel. The PCR products
will be purified with ExoSAP-IT and will be sequenced using ABI 3730 DNA Analyzer.
Nucleotide Sequences will be aligned with the Clustal W algorithm and will be edited using
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Codon Code Aligner program (Gariepy et al. 2014; Gariepy et al. 2015). For appropriate
molecular comparisons, we will include in our analysis COI, COII and Cytochrome B
sequences from GenBank.
To provide better qualitative information for the potential pathways of invasion, the
Parsimony Cladogram network analysis between haplotypes will be performed using TCS
1.21 according to the study of Cesari et al. 2014. Measures of genetic diversity were
calculated using DnaSP v5 (Librado and Rozas, 2009), including number of haplotypes,
haplotype diversity and nucleotide diversity. The Measures of overall haplotype and
nucleotide diversity between the COI, COII and Cytochrome B will be generated using
DnaSP v5.10.01 and ARLEQUIN v3.1 (Gariepy et al. 2014). Further related research topics
may complete this work depending on the results obtained.
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