Korean journal of applied entomology (한국응용곤충학회지)

Korean Society of Applied Entomology (한국응용곤충학회)
권호 표지
DOI QR코드

DOI QR Code

Differentiation of Elytra Color Patterns in Multicolored Asian Ladybird Beetle, Harmonia axyridis (Coleoptera; Coccinellidae), using AFLP analyses

Amplified Fragment Length Polymorphism (AFLP)을 이용한 무당벌레(Harmonia axyridis : Coccinellidae)의 초시색상패턴의 변이 분석

  • Park, Cho Rong (Department of Agricultural Biology, Chungnam National University) ;
  • Kim, Jeong Hee (Department of Agricultural Biology, Chungnam National University) ;
  • Yu, Yong Man (Department of Agricultural Biology, Chungnam National University) ;
  • Youn, Young Nam (Department of Agricultural Biology, Chungnam National University)
  • 박초롱 (충남대학교 농업생명과학대학 응용생물학과) ;
  • 김정희 (충남대학교 농업생명과학대학 응용생물학과) ;
  • 유용만 (충남대학교 농업생명과학대학 응용생물학과) ;
  • 윤영남 (충남대학교 농업생명과학대학 응용생물학과)
  • Received : 2016.03.03
  • Accepted : 2016.06.23
  • Published : 2016.09.01
Download PDF
⟨ Previous Next ⟩

Abstract

Elytra of Harmonia axyridis exhibit varied color patterns. In the present study, we deciphered the genetic basis for intraspecific diversity of elytra color patterns in H. axyridis, using amplified fragment length polymorphism (AFLP). Twenty-eight AFLP reactions were performed to generate a total of 2,741 bands. Of these, 20 bands were polymorphic for each color pattern. The polymorphic bands showed differences of genetic character among different color patterns of H. axyridis. Among them, ten candidate AFLP markers were color-linked. S1, S2, and S20 markers were detected in Succinea 1 and 2 variants of H. axyridis, whereas S3 and S5 were specifically detected in the Conspicua variant. S15, S18, and S19 were specific to the Succinea 2 variant. Polymerase chain reaction (PCR) products of these ten AFLP markers were sequenced. BLAST analysis of these sequences against the GenBank database revealed their homology to DNA fragments of unknown function. Based on the color-linked AFLP markers, sequence characterized amplified region (SCAR) markers were designed for PCR amplification of genomic DNA. Of the ten AFLP markers, five were successfully converted into SCAR markers, which could discriminate elytra color polymorphism in H. axyridis.

무당벌레(Harmonia axyridis)는 종내에서 초시색상패턴이 매우 다양하게 존재한다. 본 논문에서는 서로 다른 색상패턴의 무당벌레를 대상으로 amplified fragment length polymorphism (AFLP)을 실시하여 무당벌레의 초시색상 패턴간 유전형질의 차이를 확인하고자 하였다. 총 28 개의 프라이머 조합으로 실험을 실시한 결과, 총 2,741 개의 밴드가 검출되었다. 그 중 20 개의 밴드(S1-S20)만이 특정 색상패턴에서 나타났다. 이들 가운데 9 개의 밴드를 색상에 연관된 AFLP 후보 지표로 선발하였다. 밴드 가운데 S1과 S2, S20은 Succinea 1, 2 변이형에 공통적으로 나타났으며, S3와 S5는 Conspicua 변이형에 특이적이었다. 또한 S13는 Spectabilis 변이형에, S15와 S18, S19는 Succinea 2 변이형에 특이적이었다. 특정 색상패턴에만 나타나는 9 개의 AFLP 지표들은 cloning을 통해 염기서열 분석을 실시하였고, GenBank를 이용해 다른 염기 서열과 비교를 해보았지만 아무런 상동성도 찾을 수가 없었다. 무당벌레 종 내 유전적 다양성을 평가한 결과, Spectabilis가 Conspicua보다 Succinea 변이형에 높은 유사성을 보였다. 색상에 연관된 AFLP 후보 지표를 기준으로 sequence characterized amplified region (SCAR) 지표로 변환하여 9 개의 AFLP 분자지표들 가운데에서 5 개만이 SCAR 지표로 전환될 수 있었으며, 이를 통해 AFLP 지표가 무당벌레의 색상과 연관되어 있는지 확인할 수 있었다.

Keywords

References

  1. Aarts, H.J., van Lith, L.A., Keijer, J., 1998. High resolution genotyping of Salmonella strains by AFLP-fingerprinting. Lett. Appl. Microbiol. 26, 131-135. https://doi.org/10.1046/j.1472-765X.1998.00302.x
  2. Belicek, J., 1976. Coccinellidae of Western Canada and Alaska with analyses of the transmontane zoogeographic relationship between the fauna of British Columbia and Alberta. Questiones Entomol. 12, 283-409.
  3. Blears, M.J., de Grandis, S.A., Lee, H., Trevors, J.T., 1998. Amplified fragment length polymorphism (AFLP): A review of the procedure and its applications. J. Ind. Microbiol. Biotechnol. 21, 99-114. https://doi.org/10.1038/sj.jim.2900537
  4. Bonin, A., Ehric,h D., Manel, S., 2007. Statistical analysis of amplified fragment length polymorphism data: a toolbox for molecular ecologists and evolutionists. Mol. Ecol. 16, 3737-3758. https://doi.org/10.1111/j.1365-294X.2007.03435.x
  5. Brown, M.W., Miller, S.S., 1998. Coccinellidae (Coleoptera) in apple orchards of eastern West Virginia and the impact of invasion by Harmonia axyridis. Entomol. News. 109, 136-142.
  6. Brown, P.M.J., Roy, H.E., Rothery, P., Roy, D.B., Ware, R.L., Majerus, M.E.N., 2008. Harmonia axyridis in Great Britain: Analysis of the spread and distribution of a non-native Coccinellidae. BioControl 53, 55-67. https://doi.org/10.1007/s10526-007-9124-y
  7. Carisio, L., Cervella, P., Palestrini, C., Delpero, M., Rolando, A., 2004. Biogeographically patterns of genetic differentiation in dung beetles of the genus Trypocopris (Coleoptera, Geotrupidae) inferred from mtDNA and AFLP analyses. J. Biogeography 31, 1149-1162. https://doi.org/10.1111/j.1365-2699.2004.01074.x
  8. Chapin, J.B., Brov, V.A., 1991. Harmonia axyridis (Pallas), the third species of the genus to be found in the United States (Coleoptera: Coccinellidae). Proc. Entomol. Soc. Wash. 93, 630-635.
  9. Day, W.H., Prokrym, D.R., Ellis, D.R., Chianer, R.J., 1994. The known distribution of the predator Propylea quatuordecimpunctata (Coleoptera: Coccinellidae) in the United States, and thoughts on the origin of this species and five other exotic lady beetles in Eastern North America. Entomol. News 105, 244-256.
  10. Dobzhansky, T. 1933. Geographical variation in ladybeetles. Am. Nat. 67, 97-126. https://doi.org/10.1086/280472
  11. ESK and KSAE., 1994. Check list of insects from Korea (eds). Kon-Kuk Univ. Press. Seoul. 774p.
  12. Hagen, K.S., 1962. Biology and ecology of predaceous Coccinellidae. Ann. Rev. Entomol. 7, 289-326. https://doi.org/10.1146/annurev.en.07.010162.001445
  13. Hawthorne, D.J., 2001. AFLP-based genetic linkage map of the Colorado potato beetle Leptinotarsa decemlineata: Sex chromosomes and a pyrethroid-resistance candidate gene. Genetics 158, 695-700.
  14. Hodek, I., 1973. Biology of Coccinellidae. Acad. Sci. Prague.
  15. Iperti, G., 1986. Les coccinellidae de France. Phytoma 377, pp. 14-22.
  16. Iperti, G., 1999. Biodiversity of predacious Coccinellidae in relation to bioindication and economic importance. Agricul. Ecosys. Environ. 74, 323-342. https://doi.org/10.1016/S0167-8809(99)00041-9
  17. Jiggins, C.D., Mavarez, J., Beltran, M., McMillan, W.O., Johnston. J.S., Bermingham, E., 2005. A genetic linkage map of the mimetic butterfly Heliconius melpomene. Genetics 171, 557-570. https://doi.org/10.1534/genetics.104.034686
  18. Jones, C.J., Edwards, K.J., Castaglione, S., 1997. Reproducibility testing of RAPD, AFLP and SSR markers in plants by a network of European laboratories. Mol. Breed. 3, 381-390. https://doi.org/10.1023/A:1009612517139
  19. Kazachkova, N., Fahleson, J., Meijer, J., 2004. Establishment of the amplified fragment length polymorphism (AFLP) technique for genotyping of pollen beetle (Meligethes aeneus)- A noxious insect pest on oilseed rate (Brassica napus). Mol. Biol. Rep. 31, 37-42. https://doi.org/10.1023/B:MOLE.0000013518.56517.be
  20. Kazachkova, N., Meijer, J., Exbom, B., 2007. Genetic diversity in pollen beetles (Meligethes aeneus) in Sweden: Role of spatial, temporal and insecticide resistance factors. Agricul. For. Entomol. 9, 259-269. https://doi.org/10.1111/j.1461-9563.2007.00345.x
  21. Kidd, K.A., Nalepa, C.A., Day, E.R., Waldvogel, M.G., 1995. Distribution of Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae) in North Carolina and Virginia. Proc. Entomol. Soc. Wash. 97, 729-731.
  22. Knorr, C., Cheng, H.H., Dodgson, J.B., 1999. Application of AFLP markers to genome mapping in poultry. Anim. Genet. 30, 28-35. https://doi.org/10.1046/j.1365-2052.1999.00411.x
  23. Koch, R.L., 2003. The multicolored Asian ladybeetle, Harmonia axyridis: a review of its biology, uses in biological control, and non-target impacts. J. Insect Sci. 3, 1-16.
  24. Komai, T., 1956. Genetics of ladybeetles. Adv. Genetics. 8, 155-189.
  25. Komai, T., Chino, M., Hosino, Y., 1950. Contributions to the evolutionary genetics of the lady-beetle, Harmonia. I. Geographic and temporal variations in the relative frequencies of the elytral pattern types and in the frequency of elytral ridge. Genetics 35, 589-601.
  26. Kuznetsov, V.N., 1997. Lady beetles of Russian Far East. The Sandhill Crane Press, Gainesville.
  27. Lamana, M.L., Miller, J.C., 1998. Temperature dependent development in an Oregon population of Harmonia axyridis (Coleoptera: Coccinellidae). Environ. Entomol. 27, 1001-1005. https://doi.org/10.1093/ee/27.4.1001
  28. Li, Z., Li, J., Wang, Q., He, Y., Liu, P., 2006. The effects of selective breeding on the genetic structure of shrimp Fenneropenaeus chinensis populations. Aquacul. 258, 278-282. https://doi.org/10.1016/j.aquaculture.2006.04.040
  29. Mackill, D.J., Zhang, Z., Renona, E.D., Colowit, P.M., 1996. Level of polymorphism and genetic mapping of AFLP markers in rice. Genome 39, 969-977. https://doi.org/10.1139/g96-121
  30. Majerus, M.E.N. 1994. Ladybirds. Harper Collins Publishers, UK.
  31. Mariette, S., Chagne, D., Lezier, C., Pastuszka, P., Raffin, A., Plomion. C., Kremer, A., 2001. Genetic diversity within and among Pinus pinaster populations: comparison between AFLP and microsatellite marker. Heredity 86, 469-479. https://doi.org/10.1046/j.1365-2540.2001.00852.x
  32. Moeller, E.M., Bahnweg, G., Sandermann, H., Geiger, H.H., 1992. A simple and efficient protocol for isolation of high molecular weight DNA from filamentous fungi fruit bodies, and infected plant tissues, Nucleic Acids Res. 20, 6115-6116. https://doi.org/10.1093/nar/20.22.6115
  33. Nalepa, C.A., Kidd, K.A., Ahlstron, K.R., 1996. Biology of Harmonia axyridis (Coleoptera: Coccinellidae) in winter aggregations. Ann. Entomol. Soc. Am. 89, 681-685. https://doi.org/10.1093/aesa/89.5.681
  34. Obata, S., 1986. Mechanisms of prey finding in the aphidophagous ladybird beetle, Harmonia axyridis (Coleoptera: Coccinellidae). Entomophaga 31, 303-311. https://doi.org/10.1007/BF02373340
  35. Park, H.C., 1993. Systematics and ecology of Coccinellidae (Insecta: Coleoptera) in Korea. PhD Thesis. Korea Univ. Korea.
  36. Parsons, Y.M, Shaw, K.L., 2001. Species boundaries and genetic diversity among Hawaiian crickets of the genus Laupala identified using amplified fragment length polymorphism. Mol. Ecol. 10, 1765-1772. https://doi.org/10.1046/j.1365-294X.2001.01318.x
  37. Reineke, A., Karlovsky, P., 2000. Simplified AFLP protocol: replacement of primer labeling by the incorporation of alpha-labeled nucleotides during PCR. Biotechniques 28, 622-623.
  38. Roa, A.C., Maya, M.M., Duque, M.C., Tohme, J., Allem, A.F., Bonierbale, M.W., 1997. AFLP analysis of relationships among cassava and other Manihot species. Theor. Appl. Genet. 95, 741-746. https://doi.org/10.1007/s001220050620
  39. Roy, H., Wajnberg, E., 2008. From biological control to invasion: The ladybird Harmonia axyridis as a model species. BioControl 53, 1-4. https://doi.org/10.1007/s10526-007-9127-8
  40. Rusell, J. R., Fuller, J.D., Macaulay, M., Hatz, B.G., Jahoor, A., Powell, W., Waugh, R., 1997. Direct comparison of levels of genetic variation among barley accessions detected by RFLPs, AFLPs, SSRs, and RAPDs. Theor. Appl. Genet. 95, 714-722. https://doi.org/10.1007/s001220050617
  41. Seo, M.J., 1999. Behavioral characteristics and effective control strategies to aphids with the Asian Ladybird Beetle, Harmonia axyridis (Coleoptera: Coccinellidae). Ms. Thesis, Chungnam Nat'l Univ., pp. 68.
  42. Seo, M.J., Kang, E.J., Kang, M.K., Lee, H.J., Seok, H.B., Lee, D.H., Park, S.N., Yu, Y.M., Youn, Y.N., 2007. Phenotypic variation and genetic correlation of elytra colored patterns of multicolored Asian lady beetles, Harmonia axyridis (Coleoptera: Coccinellidae) in Korea. Korean J. Appl. Entomol. 46, 235-249. https://doi.org/10.5656/KSAE.2007.46.2.235
  43. Seo, M.J., Kim, G.H., Youn, Y.N., 2008. Differences in biological and behavioural characteristics of Harmonia axyridis (Coleoptera: Coccinellidae) according to colour patterns of elytra. J. Appl. Entomol. 132, 239-247. https://doi.org/10.1111/j.1439-0418.2007.01241.x
  44. Seo, M.J., Youn, Y.N., 2000. The Asian Ladybird, Harmonia axyridis, as biological control agent: I. Predacious behavior and feeding ability. Kor. J. Appl. Entomol. 39, 50-71.
  45. Stewart, L.A., Dixon, A.F.G., 1989. Why big species of ladybird beetles are not melanic. Functional Ecol. 3, 165-171. https://doi.org/10.2307/2389297
  46. Sweetman, H.L., 1958. The principle of biological control. Brown WC, Dubuque Iowa.
  47. Tan, C.C., Li, J.C., 1934. Inheritance of the elytral color patterns of the ladybird beetle, Harmonia axyridis Pallas. Genetics 68, 252-265.
  48. Tedders, W.L., Schaefer, P.W., 1994. Release and establishment of Harmonia axyridis (Coleoptera: Coccinellidae) in the Southeastern United States. Entomol. News 105, 228-243.
  49. Vos, P., Hodgers, R., Bleeker, M., Reijans, M., van de Lee, T., Hornes, M., Frijters, A., Pot, J., Peleman, J., Kuiper. M., Zabeau, M., 1995. AFLP, a new technique for DNA fingerprinting. Nucl. Acids. Res. 23, 4407-4414. https://doi.org/10.1093/nar/23.21.4407
  50. Wichan, E., Yonemori, K., Kanzaki, S., 2000. Amplified fragment length polymorphism analysis for studying genetic relationships among Mangifera species in Thailand. J. Am. Soc. Hortic. Sci. 125, 160-164.
  51. Xiao, J.P., Chen, L.G., Xie, M., Liu, H.L., Ye, W.Q., 2009. Identification of AFLP fragments linked to seedlessness in Ponkan mandarin (Citrus reticulate Blanco) and conversion to SCAR markers. Sci. Horticul. 121, 505-510. https://doi.org/10.1016/j.scienta.2009.03.006
  52. Xu, D.H., Wahyuni, S., Sato, Y., Yamaguchi, M., Tsunematsu, H., Ban, T., 2006. Genetic diversity and relationships of Japanese peach (Prunus persica L.) cultivars revealed by AFLP and pedigree tracing. Gen. Res. Crop Evol. 53, 883-889. https://doi.org/10.1007/s10722-004-0575-z
  53. Yeon, I.J., Song, M.Y., Hwang, H.J., Sohn, M.H., Kim, J.B., Im, Y.J., Kim, Y.S., Kim, K.S., Bang, I.C., 2008. Population structure of the blue and purple types of the blue crab Portunus trituberculatus (Miers) from the west sea of Korea based on morphometric characteristics and AFLP analysis. J. Kor. Fish. Soc. 41, 94-101.
  54. Youn, Y.N., Seo, M.J., Shin, J.G., Jang, C., Yu, Y.M., 2003. Toxicity of greenhouse pesticides to multicolored Asian lady beetles, Harmonia axyridis (Coleoptera: Coccinellidae). Biol. Con. 28, 164-170. https://doi.org/10.1016/S1049-9644(03)00098-7
  55. Zhong, D., Pai, A., Yan, G., 2004. AFLP-based genetic linkage map for the red flour beetle (Tribolium castaneum). J. Heredity 95, 53-61. https://doi.org/10.1093/jhered/esh012