• Korean Journal of Environmental Biology
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• v.42 no.3
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• pp.332-344
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• 2024

Phosphine (PH₃) fumigation has been widely used for controlling storedgrain insect pests, causing the development of resistance of stored-grain insect pests to phosphine. PH₃ resistance in Sitophilus oryzae has been reported in Korea. However, PH₃ resistance in Tribolium castaneum has not been reported yet. This study was conducted to determine susceptibilities of T. castaneum collected from five different domestic locations to PH₃. The susceptibility to PH₃ was investigated using the FAO fumigation method. All domestic T. castaneum individuals were controlled by PH₃ at 0.04 g m^-3 . At 0.01 g m^-3, T. castaneum collected from two domestic locations did not exhibit 100% mortality. A P45S point mutation in dihydrolipoamide dehydrogenase (dld) gene was found in a PH₃-resistant strain of T. castaneum (Aus07), but not in five domestic stains or a PH₃-susceptible strain (Aus10). No significant difference was found in dld or cyt-b5-r gene expression across all tested strains. However, the Gyeongju-collected strain of T. castaneum showed more than a 1.7-fold increase in cyt-b5-r expression compared to the Aus07 strain. cDNA sequence analysis revealed that P45S (C133T) in the dld gene was only present in Aus07. A characteristic single nucleotide polymorphism in the cyt-b5-r gene sequence was identified in the five domestic strains. This study suggests that it is necessary to continuously monitor PH₃-susceptibility of T. castaneum in Korea to quickly identify resistant individuals and prevent the spread of PH₃ resistance through rapid control.

• Molecules and Cells
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• v.25 no.4
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• pp.510-522
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• 2008

To construct the molecular systematics of the genus Megoura (Hemiptera: Aphididae), DNA based-identification was performed using four mitochondrial and three nuclear DNA regions: partial cytochrome c oxidase I (COI), partial tRNA-leucine + cytochrome c oxidase II (tRNA/COII), cytochrome b (CytB), partial 12S rRNA + tRNA-valine + 16S rRNA (12S/16S), elongation factor-1 alpha ($EF1{\alpha}$), and the internal transcribed spacers 1 and 2 (ITS1, ITS2). Pairwise sequence divergences between taxa were compared, and phylogenetic analyses were performed based on each DNA region separately, and the combined datasets. COI, CytB, $EF1{\alpha}$, ITS1, and ITS2 were relatively effective in determining species and resolving their relationships. By contrast, the sequences of tRNA/COII and 12S/16S were not able to separate the closely related species. CytB and $EF1{\alpha}$ gave better resolution with higher average sequence divergences (4.7% for CytB, 5.2% for $EF1{\alpha}$). The sequence divergence of COI (3.0%) was moderate, and those of the two ITS regions (1.8% for ITS1, 2.0% for ITS2) were very low. Phylogenetic trees were constructed by minimum evolution, maximum parsimony, maximum likelihood, and Bayesian phylogenetic analyses. The results indicated that the phylogenetic relationships between Megoura species were associated with their host preferences. Megoura brevipilosa and M. lespedezae living on Lespedeza were closely related, and M. nigra, monophagous on Vicia venosa, was rather different from M. crassicauda, M. litoralis, and M. viciae, which are oligophagous on Lathyrus and Vicia. The three populations of M. crassicauda formed a clade separated from M. litoralis and M. viciae. Nevertheless M. litoralis and M. viciae, which are morphologically similar, were not separated due to negligible sequence divergence. We discuss the phylogenetic relationships of the Megoura, and the usefulness of the seven DNA regions for determining the species level phylogeny of aphids.

• Journal of Food Hygiene and Safety
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• v.27 no.3
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• pp.317-324
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• 2012

In order to determine an authenticity of food ingredient, we used DNA barcode method by universal primers. For identification of animal food ingredients, LCO1490/HCO2198 and VF2/FISH R2 designed for amplifying cytochrome c oxidase subunit1 (CO1) region and L14724/H15915 for cytochrome b (cyt b) region on mitochondrial DNA were used. Livestock (cow, pig, goat, sheep, a horse and deer) was amplified by LCO1490/HCO 2198, VF2/FISH R2 and L14724/H15915 primers. Poultry (chicken, duck, turkey and ostrich) was amplified by LCO1490/HCO 2198 and VF2/FISH R2 primers. But, Fishes (walleye pollack, herring, codfish, blue codfish, trout, tuna and rockfish) were only amplified by VF2/FISH R2 primers. For plant food ingredients, 3 types of primers (trnH/psbA, rpoB 1F/4R and rbcL 1F/724R) have been used an intergenic spacer, a RNA polymerase beta subunit and a ribulose bisphosphate carboxylase region on plastid, respectively. Garlic, onion, radish, green tea and spinach were amplified by trnH/psbA, rpoB 1F/4R and rbcL 1F/724R. The PCR product sizes were same by rpoB 1F/4R and rbcL 1F/724R but, the PCR product size using trnH/psbA primer was different with others for plants each. We established PCR condition and universal primer selection for 17 item's raw materials for foods and determine base sequences aim to PCR products in this study. This study can apply to determine an authenticity of foods through making an comparison between databases and base sequences in gene bank. Therefore, DNA barcode method using universal primers can be a useful for species identification techniques not only raw materials but also processed foods that are difficult to analyze by chemical analysis.

• Animal cells and systems
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• v.3 no.1
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• pp.89-96
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• 1999

Nucleotide sequences of a 501 base-pair (bp) fragment in the mitochondrial cytochrome b (cyt b) gene were analyzed for 12 populations of Rana rugosa from Korea and Japan using a polymerase chain reaction (PCR) and direct silver sequencing. Two genetically distinct groups (type-A and type-B) were found in Korea. Type-A was found throughout most of South Korea and type-B was restricted to the mid-southeastern regions (Samchok, Yongdok, Chongsong and Pohang). But in the Tonghae population, both types were found. The level of mitochondrial DNA (mtDNA) sequence differences ranged from 0% to .0.8% among six populations of type-A, and 0 to 1.0% among 4 populations of type-B. However, sequence differences between type-A and type-B ranged from 5.4% to 6.6%, Using Kimura's two-parameter distance, the level of genetic sequence divergence between type-A and type-B was 6.7%. The Japanese R. rugosa was clustered very far from the Korean R. rugosa with 14.7%. In the neighbor-joining and UPGMA tree, all Korean samples were grouped, but subdivided into two types in 99% of the bootstrap iteration.