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Genetic Diversity of Rehmannia glutinosa Genotypes Assessed by Molecular Markers

분자표지자에 의한 지황 유전집단의 유전적 다양성

  • Bang, Kyong-Hwan (Ginseng & Medicinal Plants Research Institute, RDA) ;
  • Chung, Jong-Wook (National Institute of Agricultural Biotechnology, RDA) ;
  • Kim, Young-Chang (Ginseng & Medicinal Plants Research Institute, RDA) ;
  • Lee, Jei-Wan (Ginseng & Medicinal Plants Research Institute, RDA) ;
  • Kim, Hong-Sig (College of Agriculture, Life and Environment Science, Chungbuk National University) ;
  • Kim, Dong-Hwi (Ginseng & Medicinal Plants Research Institute, RDA)
  • 방경환 (농촌진흥청 인삼약초연구소 인삼과) ;
  • 정종욱 (농업생명공학연구원 유전자원과) ;
  • 김영창 (농촌진흥청 인삼약초연구소 인삼과) ;
  • 이제완 (농촌진흥청 인삼약초연구소 인삼과) ;
  • 김홍식 (충북대학교 식물자원학과) ;
  • 김동휘 (농촌진흥청 인삼약초연구소 인삼과)
  • Published : 2008.04.30

Abstract

Random amplified polymorphic DNA (RAPD) markers were used to identify the genetic diversities among and within varieties and landraces of Rehmannia glutinosa. Polymorphic and reproducible bands were produced by 10 primers out of total 20 primers used in the experiment. In RAPD analysis of the 11 genotypes, 64 fragments out of 73 amplified genomic DNA fragments were polymorphic which represented an average 6.4 polymorphic fragments per primer. Number of amplified fragments with random primers ranged from 2 (OPA-1) to 13 (OPA-11) and varied in size from 200 bp to 1,400 bp. Especially, OPA-10, OPA-11 and OPA-19 primers showed specific bands for varieties of Korea Jiwhang and Jiwhang il ho, which could be useful for discriminating from other varieties and landraces of R. glutinosa. Percentage polymorphism ranged from a minimum of 50% (OPA-1) to a maximum of 100% (OPA-11), with an average of 87.7%. Similarity coefficients were higher in the genotypes of Korea Jiwhang and Jiwhang il ho than in other populations. In cluster analysis, genotypes of Korea Jiwhang, Jiwhang il ho, and Japanese accession were separated from those of other varieties and landraces. Average of genetic diversity within the population $(H_S)$ was 0.110, while average of total genetic diversity $(H_T)$ was 0.229. Across all RAPD makers the $G_{ST}$ value was 0.517, indicating that about 52% of the total genetic variation could be explained by RAPDs differences while the remaining 48% might be attributable to differences among samples. Consequently, RAPD analysis was useful method to discriminate different populations such as domestic varieties and other landraces. The results of the present study will be used to understand the population and evolutionary genetics of R. gllutinosa.

RAPD 분석을 이용하여 지황 육성 계통과 지역 수집종 들을 구분할 수 있는 분자표지자를 선발하고, 집단 간, 집단 내 유전적 다양성을 평가하기 위하여 본 실험을 수행하였다. 총 20개의 임의 primer를 이용하여 PCR 한결과, 육성 계통과 수집종 들을 구별할 수 있는 OPA-1 등 10개의 재현성과 다형성이 좋은 프라이머 들을 선발하였다. 특히 OPA-10, OPA-11 및 OPA-19는 고려지황과 지황1호를 다른 계통 및 수집종 들과 구별할 수 있었으며, 이들 프라이머를 이용하여 0.9 kb, 1.2 kb, 1.3 kb 및 1.4 kb등의 육성계통 특이적인 DNA 밴드들을 확보할 수 있었다. 한편 이들의 결과를 토대로 통계처리에 의한 유전분석 결과, 고려지황, 지황1호 및 일본지황은 집단 내 유사도가 높아 다른 집단들과 구별되었다. 결론적으로, RAPD 분석을 통한 결과는 지황의 유전적 다양성 이해와 특정 계통을 다른 계통 및 수집종 들과 구분할 수 있는 방법으로 이용될 수 있다.

Keywords

References

  1. Bornet, B., F. Goraguer, G. Joly and M. Branchard. 2002. Genetic diversity in European and Argentinean cultivated potatoes (Solanum tuberosum sub. tuberosum) detected by inter-simple sequence repeats (ISSRs). Genome 45, 481-484 https://doi.org/10.1139/g02-002
  2. Cao, Z. L., M. Zhu, H. X. Liu and Z. M. Xiao. 1988. Effects of radix Rehmanniae and processed radix Rehmanniae on some immune functions in mice. Zhong Yao Tong Bao 13, 22-24, 62-63
  3. Cheng, J. L., L. Q. Huang, A. J. Shao and S. F. Lin. 2002. RAPD analysis on different varieties of Rehmannia glutinosa. Zhongguo Zhong Yao Za Zhi 27, 505-508
  4. Dellaporta, S. L. 1983. A plant DNA minipreparation, version II. Plant Mol. Biol. Rep. 1, 19-25 https://doi.org/10.1007/BF02712670
  5. Huff, D. R., R. Peakall and P. E. Smouse. 1993. RAPD variation within and among natural populations of outcrossing buffalo grass (Buchloe dactyloides). Theor. Appl. Genet. 86, 927-934
  6. Korea Food & Drug Administration. 2002. The Korean herbal pharmacopoeia. pp. 129
  7. Lewontin, R. C. 1972. The apportionment of human diversity. Evo.l Biol. 6, 381-394
  8. Lynch, M. and B. G. Milligan. 1994. Analysis of population genetic structure with RAPD markers. Mol. Ecol. 3, 391-399
  9. Nei, M. 1978. Estimation of average heterozygosity and genetic distance from a small number of individuals. Genetics 89, 583-590
  10. Nei, M. and W. Li. 1979. Mathematical model for study the genetic variation in terms of restriction endonucleases. Proc. Natl. Acad. Sci. U.S.A. 74, 5267-5273
  11. Paterson, A. H., S. D. Tanksley and M. E. Sorrells. 1991. DNA markers in plant improvement. Adv. Agron. 46, 39-90 https://doi.org/10.1016/S0065-2113(08)60578-7
  12. Prevost, A. and M. J. Wilkinson. 1999. A new system of comparing PCR primers applied to ISSR fingerprinting of potato cultivars. Theor. Appl. Genet. 98, 107-112 https://doi.org/10.1007/s001220051046
  13. Soller, M. and J. S. Beckmann. 1993. Genetic polymorphism in varietal identification and genetic improvement. Theor. Appl. Genet. 67, 25-33 https://doi.org/10.1007/BF00303917
  14. Stewart, C. N. and L. Excoffier. 1996. Assessing population genetic structure and variability with RAPD data: application to Vaccinum macrocarpon (American cranberry). J. Evol. Biol. 9, 153-171 https://doi.org/10.1046/j.1420-9101.1996.9020153.x
  15. Tomoda, M., M. Tanaka and N. Komdo. 1971. Water soluble constituents of Rehmannia glutinosa Lib. Chem. Pharm. Bull. 19, 2411-2419 https://doi.org/10.1248/cpb.19.2411
  16. Tsumura, Y., K. Ohba and S. H. Strauss. 1996. Diversity and inheritance of inter-simple repeat polymorphism in Douglas fir (Pseudotsuga menziesii) and Sugi (Crypromeria japonica). Theor. Appl. Genet. 92, 40-45 https://doi.org/10.1007/BF00222949
  17. Vos, P., R. Hogers, M. Bleeker, M. Reijans and van de Lee T. Hornes. 1995. AFLP: A new technique for DNA fingerprinting. Nucleic Acids Res. 23, 4407-4414 https://doi.org/10.1093/nar/23.21.4407
  18. Williams, J. G. K., A. R. Kubelik, K. J. Livak, J. A. Rafalski and S. V. Tingey. 1990. DNA polymorphism amplified by arbitrary primers are useful as genetic markers. Nucl. Acids Res. 18, 6531-6535 https://doi.org/10.1093/nar/18.22.6531
  19. Wu, K. S. and S. D. Tanksley. 1993. Abundance, polymorphism and genetic mapping of microsatellites in rice. Mol. Gen. Genet. 24, 225-235
  20. Yeh, F. C., R. C. Yang, T. B. J. Boyle, Z. H. Ye and J. X. Mao. 1997. POPGENE, the user-friendly shareware for population genetic analysis. Version 1.21. Molecular Biology and Biotechnology Centre University of Alberta Canada
  21. Zietkiewicz, E., A. Rafalski and D. Labuda. 1994. Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification. Genomics 20, 176-183 https://doi.org/10.1006/geno.1994.1151
  22. Zhou, Y. Q., J. Z. Jing, B. H. Zhang and J. F. Jia. 2004. Assessment of genetic diversity of Rehmannia glutinosa germplasm detected by RAPDs and ISSRs. Yi Chuan 26, 922-928