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ITS2 Ribosomal DNA Sequence Variation of the Bumblebee, Bombus ardens (Hymenoptera: Apidae)
Hyung Keun Oh,Hyung Joo Yoon,Min Jee Kim,Iksoo Kim 한국응용곤충학회 2009 한국응용곤충학회 학술대회논문집 Vol.2009 No.05
The bumblebees, Bombus species are valuable natural resources being utilized for greenhouse pollination. Low level of genetic variation of Bombus species has been reported previously. In this study, we sequenced complete internal transcribed spacer 2 (ITS2) of the nuclear rDNA from 100 individuals of B. ardens collected from seven localities in Korean peninsula. The ITS2 sequence of B. ardens is longest, ranging in size from 1,940 bp-1,954 bp among known in insects, which ranges approximately from 241 bp-1,728 bp. The ITS2 sequences have ~51% of C+C content and contain each two 27 bp repeats, 20 bp repeats, 33 bp repeats, and 34 bp repeats at the beginning. Such repeats were not found in other insects. Uncorrected pairwise distance among 96 sequences that were obtained from 100 individuals revealed a maximum sequence divergence of 1.03%. Genetic diversity (π) of each population ranged from 0.007801 to 0.009627, and the lowest diversity was obtained from islet population of Ulleungdo, indicating possibly small, isolation of the population. Significant level of genetic distance was only found when Ulleungdo population was compared to two other mainland populations. Except for this, overall, a very high rate of per generation migration ratio (Nm=7.1-infinite) and a very low level of genetic fixation (FST=0-0.06546) were detected between pairs of localities. Analysis of hierarchical relationships among localities consistently revealed no statistically significant structure among populations. Taken these together, the B. ardens populations on the Korean peninsula are panmictic this is consistent with our understanding of the dispersal capability.
Hyung Keun Oh,Hyung Joo Yoon,Min Jee Kim,Xinlong Wan,Iksoo Kim 한국응용곤충학회 2009 한국응용곤충학회 학술대회논문집 Vol.2009 No.10
The bumblebee, Bombus ignitus (Hymenoptera: Apidae), is a valuable natural resource that is one of the most notably utilized for greenhouse pollination in Korea. In order to understand the nature of genetic relationships, gene flow, and population structure of the species we sequenced a partial COI gene of mitochondrial DNA (mtDNA) corresponding to “animal barcode” region and the complete internal transcribed spacer 2 (ITS2) of the nuclear ribosomal DNA (nrDNA) with the individuals collected from Korean localities. Although the 658-bp long mtDNA sequence provided only six haplotypes with the maximum sequence divergence of 0.456% (3 bp, from 91 individuals), but the ITS sequences provided 33 sequence types with the maximum sequence divergence of 0.78% (16 bp, from 35 individuals), confirming better applicability of the ITS sequences to the study of intraspecific variation. The ITS2 sequences of B. ignitus were shown to be the longest among known insects, ranging in size from 2,034 bp ~ 2,045 bp and harbored two duplicated repeats, indicating unusual structure of B. ignitus ITS2 sequences compared with other insect ITS sequences. Overall, a very high per generation migration ratio, a very low level of genetic fixation, and no discernable hierarchical population/population group were noted to exist among populations of B. ignitus on the basis of both molecules, thus suggesting that the B. ignitus populations on the Korean peninsula are panmictic, which is consistent with our understanding of the dispersal capability.
Hyung Keun Oh,Hyung Joo Yoon,Min Jee Kim,Xinlong Wan,Seong Ryul Kim,Yeon Soo Han,Iksoo Kim 한국응용곤충학회 2010 한국응용곤충학회 학술대회논문집 Vol.2010 No.05
The bumblebee, Bombus ignitus (Hymenoptera: Apidae), is a valuable natural resource that is one of the most notably utilized for greenhouse pollination in Korea. In order to understand the nature of genetic relationships, gene flow, and population structure of the species we sequenced a partial COI gene of mitochondrial DNA (mtDNA) corresponding to “animal barcode” region and the complete internal transcribed spacer 2 (ITS2) of the nuclear ribosomal DNA (nrDNA) collected from Korean localities. Although the 658-bp long mtDNA sequence provided only six haplotypes with the maximum sequence divergence of 0.61% (4 bp), the ITS sequences provided 84 sequence types with the maximum sequence divergence of 1.02% (21 sites), confirming better applicability of the ITS sequences to the study of intraspecific variation. The complete ITS2 sequences of B. ignitus were shown to be longest among known insects, ranging in size from 2,034 bp ~ 2,052 bp, harboring two duplicated repeats. Overall, a very high per generation migration ratio, a very low level of genetic fixation, and no discernable hierarchical population/ population group were noted to exist among populations of B. ignitus on the basis of both molecules, thus suggesting that the B. ignitus populations on the Korean peninsula are panmictic, which is consistent with our understanding of the dispersal capability of the species
ITS2 Ribosomal DNA Sequence Variation of the Bumblebee, Bombus ardens (Hymenoptera: Apidae)
Hyung Keun Oh,Hyung Joo Yoon,Min Jee Kim,Hyung Uk Jeong,Seong Ryul Kim,Jae Sam Hwang,Chang Hwan Bae,Ik Soo Kim 한국유전학회 2009 Genes & Genomics Vol.31 No.4
The bumblebee species, Bombus, is an invaluable natural resource for greenhouse pollination. Low levels of genetic variation of Bombus ardens have been reported in a previous mitochondrial (mt) gene study. In this study, we sequenced the complete internal transcribed spacer 2 (ITS2) of the nuclear rDNA obtained from 100 B. ardens individuals collected from several Korean localities, in an effort to assess its usefulness in characterizing the genetic diversity and relationships among populations of B. ardens. The ITS2 sequences of B. ardens were shown to be longest among known insects, ranging in size from 1,971-1,984 bp. The sequences harbor four duplicated repeats-~27 bp repeats, ~20 bp repeats, ~33 bp repeats, and ~34 bp repeats?which have never before been reported in other insect ITS2 rDNA. The maximum sequence divergence of 1.01% among 96 sequence types confirmed the applicability of this molecule to the study of intraspecific variation, revealing higher sequence variation as compared to the previously studied mt COI gene. Overall, a very high per generation migration ratio (Nm=5.83~infinite) and a very low level of genetic fixation (F(ST)=0-0.08) were noted to exist among populations of B. ardens. The high estimation of gene flow among most populations-in particular, between the remote island Ulleungdo and several inland populations-suggest that historical events may be more responsible for the contemporary population structure of B. ardens. The finding of the lowest genetic diversity (π) in the population on Ulleungdo Island (π=0.007434) may be reflective of a relatively small population size and the geographical isolation of the population as compared with other inland populations.
Min Jee Kim,Hyung Joo Yoon,Dong Young Kim,Hyung Wook Jeong,Hyung Keun Oh,Iksoo Kim 한국응용곤충학회 2008 한국응용곤충학회 학술대회논문집 Vol.2008 No.05
To understand geographic genetic variation of the species and relationships among populations of the bumble bee, Bombus ardens, we sequenced a portion of mitochondrial COI gene, which corresponds to "DNA Barcode" region (658 bp) from 101 B. ardens individuals collected over 11 localities in Korea. The sequence data were used to investigate genetic diversity within populations and species, geographic variation within species, phylogeographic relationship among populations, and phylogenetic relationship among haplotypes. A total of nine haplortpes were found, but they very close to each other (a maximum sequence divergence of 0. 304%). Summarized, overall moderate to low genetic diversity within populations and species was characteristic, concordant with the high potential to disperse of B. ardens in Korea. There was no clear regional subdivision was observed and relatively high rate of gene flow among localities and low FST was characteristic.
Min Jee Kim,Man Il Kim,Xinlong Wan,Hyung Keun Oh,Ki-Gyoung Kim,Iksoo Kim 한국응용곤충학회 2009 한국응용곤충학회 학술대회논문집 Vol.2009 No.10
Eumenis autonoe, a member of the lepidopteran family Nymphalidae (superfamily Papilionoidea) is an endangered species, and is found only on one isolated remote island, Jeju in South Korea, on Halla Mt., at altitudes higher than 1,400 meters. In this study, we report the complete mitochondrial genome (mitogenome) of E. autonoe. The 15,489-bp long E. autonoe genome evidenced the typical gene content found in animal mitogenomes, and harbors the gene arrangement identical to all other sequenced lepidopteran insects, which differs from the most common type found in insects, due to the movement of tRNAMet to a position 5’-upstream of tRNAIle. As has been observed in many other lepidopteran insects, no typical ATN codon for the COI gene is available. Thus, we also designated the CGA (arginine) found at the beginning of the COI gene as a lepidopteran COI starter, in accordance with previous suggestions. The 678-bp long A+T-rich region, which is second longest in sequenced lepidopteran insects, harbors 10 identical 27-bp long tandem repeats plus one 13-bp long incomplete final repeat. Such a repeat sequence has been, thus far, only rarely detected in lepidopteran mitogenomes. The E. autonoe A+T-rich region harbors a poly-T stretch of 19 bp and a conserved ATAGA motif located at the end of the region, which have been suggested to function as structural signals for minor-strand mtDNA replication.
Xinlong Wan,Jianhong Li,Hyung Uk Jeong,Hyung Keun Oh,Min Jee Kim,Iksoo Kim 한국응용곤충학회 2009 한국응용곤충학회 학술대회논문집 Vol.2009 No.10
In order to understand the nature of population genetic structure of the notorious pest insect, Spodoptera litura (Lepidoptera: Noctuidae), a total of 159 individuals of the tobacco cutworm collected from six Korean and five Chinese localities were cloned and sequenced their mitochondrial A+T-rich region. A total of 90 haplotypes ranged 324 bp ~ 372 bp in length and 0.30% ~ 4.85% in sequence divergence. Compared to the preliminary mitochondrial COI gene sequence data that provided nearly identical sequences in all samples, the A+T-rich region provided substantially high sequence variation, validating the applicability of this molecule to the study of intraspecific variation. Overall, a very high per generation migration ratio (Nm = 5.91 ~ infinite), a very low level of genetic fixation (FST = 0 ~ 0.077796), and no discernable isolated population were noted to exist among populations of S. litura, although some significant genetic differentiations were found between several pairs of populations. These results collectively suggest that the S. litura populations in both Korea and China are very well interrelated to each other. The structural analysis revealed that the S. litura A+T-rich region harbored two stretches of the [TA(A)]n sequence, the poly-A stretch, and a stem-and-loop structure that are well found in other lepidopteran A+T-rich region. Furthermore, the region harbors the 13 ~ 19-bp long T stretch and 5-bp long motif that have been suggested to function as a possible recognition site for the initiation of replication of the minor strand of mtDNA also were found.