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ISSN : 1225-0171(Print)
ISSN : 2287-545X(Online)
Korean Journal of Applied Entomology Vol.63 No.3 pp.189-195
DOI : https://doi.org/10.5656/KSAE.2024.07.0.018

Revealing the Identity of Donacia aquatica (Linnaeus) and D. japana Chûjô & Goecke (Coleoptera: Chrysomelidae: Donaciinae) based on Literature Review, New Material, and DNA Barcodes

Yunji Jeong, Joong Youb Kim1, Jihwan Park2, Geonho Cho3, Jong Eun Lee4, Ki-Jeong Hong*
Department of Agriculture and Life Science, Sunchon National University, Suncheon 57922, Korea
1Research Center for Endangered Species, National Institute of Ecology, Yeongyang 36531, Korea
2Siheung Neunggok Elementary School, Siheung 14995, Korea
3Department of Forest Resources, Sunchon National University, Suncheon 57922, Korea
4Department of Biological Science, Andong National University, Andong 36729, Korea
*Corresponding author:curcul@scnu.ac.kr
April 24, 2024 July 11, 2024 July 29, 2024

Abstract


Members of the genus Donacia include six species, which belong to two subgenera found on the Korean Peninsula (An, 2019). Among them, Chujo (1941) reported the specimen collected from Ungi, Hamgyeongbuk-do, North Korea, as D. aquatica (Linnaeus, 1758), whereas Kimoto and Kawase (1966) reported and referred to the same specimen as D. japana. However, Hayashi (2020) stated that the D. japana recorded in Korea could have been a misidentification of D. aquatica. To solve this discrepancy, we compared specimens collected in South Korea (Pocheon and Ulsan), including morphological parts such male genitalia (Hayashi, 2020) and gene sequences of the cytochrome c oxidase subunit I (COI) region, with descriptions of D. japana, in this study. We conclude that the records of D. japana were replaced with those of D. aquatica in the Korean Peninsula, and D. japana is endemic to Japan.


Donacia japana , Donacia aquatica , misidentification , COI , Korea

문헌 고찰과 형태학적 차이, DNA 분석을 바탕으로 Donacia aquatica (Linnaeus)와 D. japana Chûjô & Goecke (딱정벌레목: 잎벌레과)의 재동정

정윤지, 김중엽1, 박지환2, 조건호3, 이종은4, 홍기정*
국립순천대학교 농생명과학과
1국립생태원 멸종위기종복원센터
2시흥능곡초등학교
3국립순천대학교 산림자원학과
4국립안동대학교 생명과학과

초록


한반도의 Donacia속에는 2아속에 속하는 6종이 있다(An, 2019). 이 중 Chujo(1941)는 북한 함경북도 웅기에서 채집한 표본을 D. aquatica (Linnaeus, 1758)로 보고하였고, Kimoto and Kawase (1966)는 D. japana로 표기하여 보고하였다. 그리고 Hayashi (2020)는 한국에 기록된 D. japana가 D. aquatica로 오인되었을 수 있다고 밝혔다. 이러한 혼란을 해결하기 위해 본 연구는 한국(포천과 울산)에서 채집한 표본과 D. japana에 대한 기재문, 수컷생식기와 같은 형태적인 부분(Hayashi, 2020), COI 유전자 염기서열을 비교하였다. 그 결과로 한반도에 기록된 D. japana은 D. aquatica로 대체되고, D. japana는 일본 고유종이라고 결론을 내리고자 한다.


Donacia japana , Doncia aquatica , 오동정 , COI , 한국

    The subfamily Donaciinae (reed beetles) is a relatively small group that belongs to the family Chrysomelidae. There are typically brightly metallic in color, dorsally and ventrally. The main morphological characters of the group are as following: sutural margin of elytron explanate apically, inner sutural bead sinuate far from apex and joined to outer bead, and elytral apex rounded; hypomeron of pronotum glabrous (or with few scattered setae); mandible with apical teeth approximate, lacking serrulate occlusal edge; median lobe without basal angulation, tegmen robust and tapered (Askevold, 1990).

    Reed beetles live in freshwater wetlands, such as ponds, marshes, bogs, fens, lakes, and streams, and their larvae feed on roots and/or subterranean stems of aquatic plants and are therefore considered aquatic beetles (Hayashi, 2020).

    The subfamily Donaciinae mainly inhabits in the temperate Holarctic zone excluding the Central America, the west India, Indonesia, northern Australia, and tropical and temperate Africa (An, 2019). Askevold (1990) recognized about 160 species in eight genera belonging to the subfamily Donaciinae worldwide.

    Korean members of the subfamily Donaciinae are reported until now three genera, Donacia Fabricius, 1775, Sominella Jacobson 1908 and Plateumaris Thomson, 1859 (An, 2019). The members of genus Donacia includes two and six species belonging to two subgenera, Cyphogaster Goecke and Donaciomina Medvedev, respectively in the Korean Peninsula (An, 2019).

    Among them, the distribution record of D. japana Chûjô & Goecke, 1956 from the Korean peninsula is questionable. Regarding three specimens collected on June 1, 1934 in Ungi, Hamgyeongbuk-do, North Korea, which Chûjô (1941) reported as D. aquatica (Linnaeus, 1758), Kimoto and Kawase (1966) reported that this should be referred as D. japana, but Hayashi (2020) stated that this may have been misidentification of D. aquatica.

    In this study, as a result of comparing specimens collected in South Korea (Pocheon and Ulsan), including morphological parts such male genitalia (Hayashi, 2020) and gene sequences of the cytochrome c oxidase subunit I (COI) region, with descriptions of D. japana, we conclude that the records of D. japana were replaced with those of D. aquatica in the Korean Peninsula, and D. japana is endemic to Japan.

    Materials and Methods

    Comparison of morphological characteristics

    We examined morphological characters for four specimens of Donacia aquatica collected from Pocheon (on May. 6, 2023) and Ulsan (on Apr. 23, 2022). Seven morphological characters of male used by Hayashi (2020) were selected to compare two species (D. aquatica and D. japana): antenna, longitudinal stripe on elytron, metafemur, metatibia, sternum VII, pygidial apex, and male genitalia. Specimens were taken pictures with a Dhyana 400DC microscope camera that is attached to Leica M125 Stereo microscope. The pictures were stacked by using the Mosaic 2.4 and Delta-Bio multifocus ver. 24. Specimens examined in this study are deposited in insect collections of the Department of Plant Medicine in Sunchon National University.

    Comparison of molecular data

    We sequenced and analyzed 680 bp fragments of the anterior part and 750 bp fragments of the posterior part from the mitochondrial cytochrome c oxidase subunit I (COI) of D. aquatica. Genomic DNA was extracted from each specimen using a DNeasy Blood and Tissue kit (Qiagen, Inc., Dusseldorf, Germany) following the manufacturer’s protocols. Primer sets LCO1490 (5'-GGT CAA ATC ATA AAG ATA TTG G-3') and HCO2198 (5'-TAA ACT TCA GGG TGA CCA AAA AAT CA-3') (Folmer et al., 1994) for comparison with the European species (of genus Donacia, and primer sets COA3107 (5'-TCT ATT ARD GGD GAD GCD CTA TCT TG-3') and COS2183N (5'-CAR CAY YTA TTY TGR TTY TTY GG-3') (Sota et al., 2004) for comparison with the Japanese species of genus Donacia were used to amplify the COI gene. The Primer sequences were removed from the resulting sequences, and the trimmed sequences were submitted to NCBI GenBank. A BLAST search tool in MEGA11 (Kumar et al., 2018) was used to identify the COI sequence data Donacia aquatica (Linnaeus). Phylogenetic tree was constructed by neighbor-joining method (Saitou and Nei, 1987) with the number of bootstrap replications was 1,000 times (Felsenstein, 1985) based on p-distances (Nei and Kumar, 2000). Plateumaris sericea (Linnaeus, 1758) (Coleoptera: Chrysomelidae: Donaciinae) was used as the outgroup.

    Results and Discussion

    Taxonomic account

    Family Chrysomelidae Latreille, 1802

    Subfamily Donaciinae Kirby, 1837

    Genus Donacia Fabricius, 1775

    Donacia Fabricius, 1775: 195. (Type species: Donacia crassipes Fabricius, 1775)

    Subgenus Donaciomima Medvedev, 1873

    Donaciomima Medvedev, 1873: 876 (Type species: Donacia clavareaui Jakobson, 1906)

    Donacia aquatica (Linnaeus, 1758) 홍날개뿌리잎벌레

    Leptura aquatica Linnaeus, 1758: 397.

    Leptura coccineofasciata Harrer, 1784: 226.

    Donacia concinna Weise, 1989: 179.

    Donacia cyanicollis Olsoufieff, 1903: 303.

    Donacia dentipes Fabricius, 1792: 116.

    Donacia fasciata Herbst, 1784: 100.

    Donacia latonae Gistel, 1857: 586.

    Donacia vittata Olivier, 1791: 292.

    Donacia waldaica Olsoufieff, 1903: 303.

    Records for Korean Donacia aquatica. Donacia aquatica - First recorded by Chûjô (1941: 451) based on specimens collected in Ungi, Hamgyeongbuk-do.

    Donacia japana - Misidentified by Kimoto and Kawase (1966: 40) based on same specimens reported by Chûjô (1941); after that, all the following records were simply cited from the report of Kimoto and Kawase (1966): ESK and KSAE (1994: 189), Lee and An (2001: 20), An (2019: 64), and KSAE and ESK (2021: 346).

    Comparison of morphological characteristics. Eight morphological characters of male used by Hayashi (2020) were selected to compare two species (Donacia aquatica and D. japana): antenna, longitudinal stripe on elytron, metafemur, metatibia, sternum VII, pygidial apex, and male genitalia. As a result of comparing specimens collected in South Korea (Pocheon and Ulsan) with descriptions of D. japana from Hayashi (2020), the longitudinal stripe on elytron continuous in D. aquatica and discontinuous in D. japana; male pygidial apex rounded in the former and shallowly emarginated in the latter; endophllus of D. aquatica basal supporting block (BSB) elongate, but D. japana BSB short; and a cap of tegmen broad subapically in the former and slender in the latter (Fig. 1).

    Comparison of molecular data. We sequenced and analyzed 680 bp fragments of the anterior part and 750 bp fragments of the posterior part from the mitochondrial COI of D. aquatica. Plateumaris sericea (Linnaeus, 1758) (Coleoptera: Chrysomelidae: Donaciinae) was used as the outgroup. A 680 bp of anterior region of the mtDNA COI gene was amplified from 1 individual Donacia beetles (Table 1) using a set of universal COI primers. Alignment of these sequences and five previously reported Donacia sequences from GenBank (Table 1) were used for phylogenetics analysis. And also, a 750 bp of posterior region of the mtDNA COI gene was amplified from 1 individual Donacia beetles (Table 2) using a set of COI primers designed by Sota et al. (2004). Alignment of these sequences and seven previously reported Donacia sequences from Gen- Bank (Table 2) were used for phylogenetics analysis. Analysis using both primer sets showed that the neighbor-joining tree grouped the four and seven morphologically identified Donacia species based on the COI gene sequences respectively, as well as the outgroup species Plateumaris sericea (Linnaeus, 1758) (Coleoptera: Chrysomelidae: Donaciinae) (Fig. 2 and 3). Ultimately, the results of these trees strongly support that the Korean Donacia species is same as the European D. aquatica, but is a different from Japanese D. japana.

    Comparison of geological distribution.Donacia aquatica is nearly distributed throughout the Palearctic region from Europe to the Russian Far East (Silfverberg, 2010). We were not examined to the North Korean specimens collected from Ungi, Hamgyeongbuk-do, which was first reported as the Korean D. aquatica in this study, while it is very likely that D. japana distributed in Japan which is isolated from the Northeast Asian continent, is an endemic species (Hayashi, 2020). Additionally, adults of the genus Donacia have poorly developed hind wings, so it is unlikely that they will naturally cross the sea and invade other countries.

    Biological notes. Adults of Donacia aquatica (L.) usually observed April to May on the host plants of Carex sp. and Sparganium fallax Graebn. in South Korea (Fig. 4). The host plant S. fallax Graebn. was known to be distributed only in Jeju Island (Kim et al., 2010), but additional distribution areas were confirmed by Kim (2024). Adults mainly feed on the leaves and flowers of the host plants mentioned above. Their oviposition pattern appears a cluster wrapped in a membrane between overlapping leaves of S. fallax Graebn. (Fig 5). Eggs hatched after about 7 days at room temperature.

    Acknowledgments

    This work was supported by a Research promotion program of SCNU in 2024.

    Statements for Authorship Positions and Contribution

    • Jeong, Y.: Sunchon National University, Researcher; Examined specimens and made manuscript.

    • Kim, J.Y.: Research Center for Endangered Species, Researcher; Collected and examined specimens.

    • Park. J.: Siheung Neunggok Elementary School, Researcher; Collected specimens.

    • Cho, G.: Sunchon National University, Professor, Ph.D; Oriented and advised the research.

    • Lee, J.E.: Andong National University, Professor, Ph.D; Advised the research.

    • Hong, K.-J.: Sunchon National University, Professor, Ph.D; Designed the research.

    All authors read and approved the manuscript.

    KJAE-63-3-189_F1.gif

    Morphological characters of Donacia aquatica (Linnaeus). Longitudinal stripes on elytra (A: mle; B: femlae), antennae (C), metafemur (D), metatibia (E), sternum (F), pygidium (G), and genitalia (H). Scale bar = 0.5mm.

    KJAE-63-3-189_F2.gif

    Neighbor-joining phylogenetic tree of Donacia species based on the cytochrome c oxidase subunit I (COI) gene sequences for comparison with European species.

    KJAE-63-3-189_F3.gif

    Neighbor-joining phylogenetic tree of Donacia species based on the cytochrome c oxidase subunit I (COI) gene sequences for comparison with Japanese species.

    KJAE-63-3-189_F4.gif

    Feeding of Donacia aquatica (Linnaeus) adults feeding on the leaves of Sparganium fallax Graebn. (A) and on the flower of Carex sp. (B).

    KJAE-63-3-189_F5.gif

    Egg cluster between leaves of Sparganium fallax Graebn. (A) and 1st instar larva of Donacia aquatica (Linnaeus).

    List of seven cytochrome c oxidase subunit I (COI) sequences of four species of genus Donacia and Plaeumaris sericea used in this study for comparison with European species

    List of nine cytochrome c oxidase subunit I (COI) sequences of seven species of genus Donacia and Plaeumaris sericea used in this study for comparison with Japanese species

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    Vol. 40 No. 4 (2022.12)

    Journal Abbreviation Korean J. Appl. Entomol.
    Frequency Quarterly
    Doi Prefix 10.5656/KSAE
    Year of Launching 1962
    Publisher Korean Society of Applied Entomology
    Indexed/Tracked/Covered By