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Razlikovanje med gosenicami vešč sovk na osnovi črtne kode DNA
ID Ruparčič, Matija (Author), ID Dolinar, Marko (Mentor) More about this mentor... This link opens in a new window

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Abstract
Sovke (Noctuidae) so številčna družina nočnih metuljev, med katerimi najdemo tudi vrste, katerih gosenice povzročajo precejšnjo škodo na pridelkih. Gosenice različnih vrst teh metuljev so si med seboj po izgledu zelo podobne, zato jih je morfološko skoraj nemogoče ločiti med seboj. Pri tem nam lahko pomagajo molekularnobiološke značilnosti organizmov, med njimi črtna koda DNA. Ta temelji na ideji, da v genomu vseh živih bitij obstaja univerzalen segment, preko katerega lahko organizmu določimo vrsto. Univerzalnega segmenta DNA trenutno ne poznamo, saj je kriterij za naziv črtne kode zelo strog, zato se za posamezne skupine organizmov uporabljajo različne črtne kode. Kljub visoki uspešnosti in široki uporabnosti črtne kode DNA ne moremo uporabljati kot edinega taksonomskega pristopa, zato je poleg nje priporočljivo uporabljati še druge, tradicionalne metode. Cilj diplomskega dela je bil preveriti, ali črtna koda DNA omogoča določevanje gosenic sovk do vrste natančno. Zaradi prepovedi opravljanja eksperimentov v povezavi z omejevanjem širjenja virusa SARS CoV 2 smo se osredotočili na bioinformatsko analizo, kjer smo primerjali nukleotidna zaporedja črtnih kod iz štirih sovk ter enega modrina (Lycaenindae), ki je predstavljal negativno kontrolo. Ugotovili smo, da bazi podatkov GenBank in BOLD vsebujeta nekaj nepravilno pripisanih nukleotidnih zaporedij, zato je treba zaporedja iz vsake uporabljene baze ročno preveriti. Ugotovili smo tudi, da je črtna koda DNA primerna za razlikovanje med vrstami sovk, z izjemo sestrskih vrst Chrysodeixis chalcites in Chrysodeixis eriosoma, ki sta si po nukleotidnih zaporedjih preveč podobni, da bi ju lahko samo preko črtne kode DNA ločili na dve različni vrsti. Po drugi strani smo ugotovili, da se zaporedja črtnih kod predstavnikov Trichoplusia ni iz Evrazije precej razlikujejo od tistih iz Severne Amerike, čeprav gre za isto vrsto. Za konec smo opravili še meteorološko analizo zadnjih petih zim v Zgornji Vipavski dolini. Ugotovili smo, da so se zime v zadnjih petih letih sicer iz leta v leto rahlo otoplile, vendar trenutno stanje še ne predstavlja nevarnosti, da bi Chrysodeixis chalcites uspela prezimiti v notranjosti Primorske, razen na zaščitenih mikrolokacijah, in se s tem razširiti v notranjost celine.

Language:Slovenian
Keywords:Noctuidae, črtna koda DNA, COI
Work type:Bachelor thesis/paper
Typology:2.11 - Undergraduate Thesis
Organization:FKKT - Faculty of Chemistry and Chemical Technology
Year:2020
PID:20.500.12556/RUL-119527 This link opens in a new window
COBISS.SI-ID:28646403 This link opens in a new window
Publication date in RUL:09.09.2020
Views:1766
Downloads:298
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Secondary language

Language:English
Title:Discrimination between owlet moth caterpillars based on DNA barcode
Abstract:
The family of owlet moths (Noctuiade) is comprised of numerous species, many of which are known as pests. To prevent as much damage as possible, it is crucial to distinguish between them while still in their larval state. Since many owlet moth larvae of different species look very much alike, it is almost impossible to tell them apart morphologically. Molecular methods have thus been implemented to aid in this problem, among them DNA barcoding. DNA barcoding is based on the idea that every living being contains a short universal segment in their DNA which can be used to discriminate them to the species level. However, the requirements for a region to be considered a barcode are very strict, which is why we still haven't found a truly universal segment of DNA. Therefore, different regions of DNA are used as barcodes for different groups of organisms. While DNA barcoding may have a very high success rate and can be used in many fields of science, it is insufficient when used alone. It yields the best results when combined with traditional methods. The purpose of this thesis was to test the effectivenes of DNA barcoding for distinguishing between different species of noctuid larvae. Since we were unable to complete the experiments due to COVID 19 epidemic measures, we focused on a bioinformatic analysis, in which we compared barcode sequences from four noctuid species and one from the family Lycaenidae, the latter used as negative control. We noticed that both databases used in the analysis (GenBank and BOLD) contained sequences that had been annotated to a wrong species, so caution is advised when using downloaded sequences. We found DNA barcoding to be an effective method for distinguishing between noctuid species, with the exception of the sister species Chrysodeixis chalcites and Chrysodeixis eriosoma, as their sequences differ too little to be delineated by barcoding alone. On the other hand, we noticed that in the case of Trichoplusia ni sequences from samples caught in North America differ significantly from the ones caught in Eurasia. We concluded the research by a meteorological analysis where we analyzed the past five winters in the Upper Vipava valley. We found out that eventhough winters have become warmer in the last five years, they do not yet allow Chrysodeixis chalcites a successful overwintering and thus its spread to the inner area of Primorska, except in sheltered microlocations.

Keywords:Noctuidae, DNA barcoding, COI

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