Dec 30, 2022
Population relationships of Oomyzus scaposus (Hymenoptera: Eulophidae)
- nedved1,2,
- Lida Fekrat3
- 1University of South Bohemia, Ceske Budejovice, Czechia;
- 2Biology Center, Academy of Sciences, Ceske Budejovice, Czechia;
- 3Ferdowsi University of Mashhad, Mashhad, Iran
Protocol Citation: nedved, Lida Fekrat 2022. Population relationships of Oomyzus scaposus (Hymenoptera: Eulophidae). protocols.io https://dx.doi.org/10.17504/protocols.io.81wgbynnqvpk/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: December 28, 2022
Last Modified: December 30, 2022
Protocol Integer ID: 74533
Funders Acknowledgement:
Grant Agency of Czech Republic
Grant ID: 20-10003S
Abstract
Comparison of COI DNA barcodes from Iranian and other populations of Oomyzus scaposus and other species of Oomyzus.
Population relationships of Oomyzus scaposus (Thomson) (Hymenoptera: Eulophidae)
Population relationships of Oomyzus scaposus (Thomson) (Hymenoptera: Eulophidae)
We extracted DNA from the wasps using Tissue DNA genomic DNA extraction mini kit (Favorgen) according to the manufacturer’s instructions.
Partial sequences of the mitochondrial gene Cytochrome Oxidase I (COI) were amplified using the primers HCO and LCO (Folmer et al., 1994). PCR cycling conditions consisted of an initial denaturation of 3 min at 94°C followed by 35 cycles, each of 30 s at 94°C, 40 s at 52°C and 30 s at 72°C, and a final extension of 72°C for 10 minutes. After a check through electrophoresis, all the PCR products were sequenced by BioneerCorporation (Korea).
Forward and reverse sequences were assembled using Geneious, version 2019.
Sequence similarities were checked using the BLAST program of the GenBank database.
The sequences were submitted to GenBank under accession numbers OP784016 and OP784017. All specimens used for sequencing are represented by voucher specimens belonging to the same populations and collected at the same time from the same host and are deposited in the private collection of the first author (AB). For phylogenetic analysis, 15 COI sequences of Oomyzus species available in the GenBank database (NCBI) were retrieved (see accession numbers in Fig. 3). A parasitoid wasp, Baryscapus, from another genus of the same subfamily (Tetrastichinae) was included as an outgroup for phylogenetic analysis.
Phylogenetic relationships were reconstructed using MEGAX with the Kimura 2-parameter (K2) distance model, and the Maximum Likelihood (ML) method.One thousand bootstrap replicates were performed to assess branch support in the given tree.
Pairwise Kimura-2-parameter (K2P) distance was measured to understand the evolutionary divergence rate among the species based on the COI gene – estimating the pairwise intra- and inter-species K2P distances among the sequences.