Journal of Plant Biotechnology

  • 제50권
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  • Pages.19-26
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  • 2023
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  • 1229-2818(pISSN)
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  • 2384-1397(eISSN)
한국식물생명공학회 (The Korean Society of Plant Biotechnology)
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인삼 약 배양을 통한 배 발생과 식물 재분화 및 유세포 분석기를 이용한 배수성 검정

Embryogenesis and plant regeneration of Panax ginseng Meyer via anther culture and ploidy assessment using flow cytometry

  • 이정우 (농촌진흥청 국립원예특작과학원 인삼특작부) ;
  • 방경환 (농촌진흥청 국립원예특작과학원 인삼특작부) ;
  • 김동휘 (농촌진흥청 국립원예특작과학원 인삼특작부) ;
  • 김장욱 (농촌진흥청 국립원예특작과학원 인삼특작부) ;
  • 김영창 (농촌진흥청 연구정책국 융복합혁신전략팀) ;
  • 조익현 (단국대학교 생명공학대학 생명자원학부)
  • Jung-Woo Lee (Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, Rural Development Administration) ;
  • Kyong-Hwan Bang (Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, Rural Development Administration) ;
  • Dong-Hwi Kim (Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, Rural Development Administration) ;
  • Jang-Uk Kim (Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, Rural Development Administration) ;
  • Young-Chang Kim (Research Policy Bureau, Rural Development Administration) ;
  • Ick-Hyun Jo (Department of Crop Science and Biotechnology, Dankook University)
  • 투고 : 2023.02.20
  • 심사 : 2023.03.21
  • 발행 : 2023.03.29
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초록

이번 연구에서 인삼의 약 배양을 통한 배 형성과 식물체의 재분화가 확인되었다. 인삼의 약 배양에서 캘러스 유도에 적합한 기본 배지는 MS 배지였고 sucrose 농도는 3%였으며, 배 형성에 적합한 호르몬 농도는 2,4-D 1.0 mg/L 또는 3.0 mg/L이었다. 저온 전처리에 의한 인삼 약 배양의 캘러스 유도와 배 형성의 효율성 차이는 확인할 수 없었다. 인삼의 품종별 약 배양의 효율성을 검정한 결과, 청선 품종의 배 형성률이 가장 우수하였으며 선운 품종은 배 형성이 관찰되지 않았다. 약 배양으로 유도된 캘러스의 배수성을 검정한 결과, 반수체 캘러스가 확인되었지만 이 캘러스에서 배는 형성되지 않았다. 약 배양 유래의 배를 발아배지에 치상하였을 때 정상적으로 식물체가 재분화 하였으며, 이 식물체의 배수성을 검정한 결과, 모두 2배체로 확인되었다. 추후 DNA 마커를 이용한 약 배양 유래 식물체의 동형접합성의 확인이 필요하다고 판단된다. 인삼 약 배양을 통하여 동형접합성 식물체를 생산할 수 있다면 인삼 육종 효율성을 증진시키는데 크게 기여할 수 있을 것이다.

Korean ginseng (Panax ginseng Meyer) is an economically important plant because of it is rich in saponins. It is mainly cultivated in Asia, including Korea and China. Since ginseng requires a long breeding period due to juvenility, homozygote production techniques, such as anther culture, must be urgently established. In the present study, callus induction and embryogenesis through anther culture were observed in P. ginseng. Murashige and Skoog medium was used as the basal medium suitable for callus induction. When the medium was supplemented with 3% sucrose, the callus induction rate was high and the callus size was large. Cold pretreatment did not significantly affect callus induction and embryogenesis. Embryogenesis was the most efficient when the embryo-formation medium was supplemented with 1.0 or 3.0 mg/L 2,4-dichlorophenoxyacetic acid. Cultivar significantly affected anther culture efficiency. Specifically, 'Cheongseon' showed the highest embryo-formation efficiency, whereas no embryogenesis occurred in 'Sunun'. Ploidy assessment revealed the haploid status of the induced calli. Embryos derived from anther culture formed shoots upon transfer to germination medium, although no difference in ploidy was noted between the induced callus and control. Overall, the anther culture conditions established in the present study may contribute to the production of homozygous P. ginseng plants in the future.

키워드

과제정보

본 연구는 농촌진흥청에서 주관하는 인삼 육종효율 증진을 위한 기반기술 개발(과제번호: PJ01018702)의 연구비 지원에 의해 이루어진 결과로 이에 감사드립니다.

참고문헌

  1. Ardebili SH, Shariatpanahi ME, Amiri R, Emamifar M, Nematzadeh G, Noori SAS, Oroojloo M and Heberle-Bors E (2011) Effect of 2,4-D as a novel inducer of embryogenesis in microspores of Brassica napus L. Czech J Genet Plant Breed 47:114-122 https://doi.org/10.17221/4/2011-CJGPB
  2. Bajaj YPS (1990) In vitro production of haploids and their use in cell genetics and plant breeding. Haploids in crop improvement 12:1-4 https://doi.org/10.1007/978-3-642-61499-6_1
  3. Bang KH, Kim YC, Lee JW, Cho IH, Hong CE, Hyun DH, Kim JU (2020) Major achievement and prospect of ginseng breeding in Korea. Korean J Breed 52:170-178 https://doi.org/10.9787/KJBS.2020.52.S.170
  4. Blasco M, Badenes ML, del Mar Naval M (2015) Embryogenic response from anther culture of cultivars of loquat (Eriobotrya japonica (Thunb.) Lindl.) from different origins. Euphytica 205:337-348
  5. Buyser JD, Henry Y, Taleb G (1985) Wheat androgenesis: Cytogenetical analysis and agronomic performance of doubled haploids. Z Pflanzenzucht 95:23-34
  6. Choi YE, Yang DC, Yoon ES, Choi KT (1999) High-efficiency plant production via direct somatic single embryogenesis from preplasmolysed cotyledons of Panax ginseng and possible dormancy of somatic embryos. Plant Cell Rep 18:493-499 https://doi.org/10.1007/s002990050610
  7. Chu CC (1978) The N6 medium and its applications to anther culture of cereal crops. Proc Symp On Plant Tissue Culture 43-50
  8. Datta SK (2005) Androgenic haploids: Factors controlling development and its application in crop improvement. Current Sci 11: 1870-1878
  9. Doi H, Takahashi R, Hikage T, Takahata Y (2010) Embryogenesis and doubled haploid production from anther culture in gentian (Gentiana triflora). Plant Cell Tiss Organ Cult 102:27-33 https://doi.org/10.1007/s11240-010-9700-1
  10. Dunwell JM (2010) Haploids in flowering plants: Origins and exploitation. Plant biotech J 8:377-424 https://doi.org/10.1111/j.1467-7652.2009.00498.x
  11. Gamborg OL, Miller R, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151-158 https://doi.org/10.1016/0014-4827(68)90403-5
  12. Germana MA (2006) Doubled haploid production in fruit crops. Plant Cell Tiss Organ Cult 86:131-146 https://doi.org/10.1007/s11240-006-9088-0
  13. Germana MA (2011) Anther culture for haploid and doubled haploid production. Plant Cell Tiss Organ Cult 104:283-300 https://doi.org/10.1007/s11240-010-9852-z
  14. Jayakodi M, Choi BS, Lee SC, Kim NH, Park JY, Jang W, Lakshmanan M, Mohan SVG, Lee DY, Yang TJ (2018) Ginseng Genome Database: An open-access platform for genomics of Panax ginseng. BMC plant Biol 18:1-7 https://doi.org/10.1186/s12870-018-1282-9
  15. Kadota M, Niimi Y (2004) Production of triploid plants of Japanese pear (Pyrus pyrifolia Nakai) by anther culture. Euphytica 138:141-147 https://doi.org/10.1023/B:EUPH.0000046756.01934.34
  16. Kim JY, Adhikari PB, Ahn CH, Kim DH, Kim YC, Han JY, Kondeti S, Choi YE (2019) High frequency somatic embryogenesis and plant regeneration of interspecific ginseng hybrid between Panax ginseng and Panax quinquefolius. J Ginseng Res 43:38-48 https://doi.org/10.1016/j.jgr.2017.08.002
  17. Kim YJ, Jang MG, Zhu L, Silva J, Zhu X, Sukweenadhi J, Kwon WS, Yang DC, Zhang D (2016) Cytological characterization of anther development in Panax ginseng Meyer. Protoplasma 253:1111-1123 https://doi.org/10.1007/s00709-015-0869-3
  18. Kim YJ, Lee OR, Kim KT, Yang DC (2012) High frequency of plant regeneration through cyclic secondary somatic embryogenesis in Panax ginseng. J Ginseng Res 36:442-448 https://doi.org/10.5142/jgr.2012.36.4.442
  19. Kiviharju E, Pehu E (1998) The effect of cold and heat pretreatments on anther culture response of Avena sativa and A. sterilis. Plant Cell Tiss Organ Cult 54:97-104 https://doi.org/10.1023/A:1006167306638
  20. Kozak K, Galek R, Waheed, MT, Sawicka-Sienkiewicz E (2012) Anther culture of Lupinus angustifolius: callus formation and the development of multicellular and embryo-like structures. Plant Growth Regul 66:145-153 https://doi.org/10.1007/s10725-011-9638-2
  21. Kruczkowska H, Pawlowska H, Skucinska B (2005) Effect of 2,4-D concentration on the androgenic response in anther culture of barley. Cereal Res Commun 33:727-732 https://doi.org/10.1556/CRC.33.2005.2-3.141
  22. Lee HY, Khorolragchaa A, Sun MS, Kim YJ, Kim YJ, Kwon WS, Yang DC (2013) Plant Regeneration from Anther Culture of Panax ginseng. Korean J Plant Resour 26:383-388 https://doi.org/10.7732/kjpr.2013.26.3.383
  23. Lee JH, Kim YJ, Jung DY, Shim JS, Kim IH, Yang DC (2009) In vitro induction of tetraploid roots by various pretreatments from anther of Panax ginseng C. A. Meyer. J Ginseng Res 33:65-71
  24. Lee JW, Jo IH, Kim JU, Hong CE, Bang KH, Park YD (2019) Determination of mutagenic sensitivity to gamma rays in ginseng (Panax ginseng) dehiscent seeds, roots, and somatic embryos. Hortic Environ Biotechnol 60:721-731 https://doi.org/10.1007/s13580-019-00164-2
  25. Lichter R (1982) Induction of haploid plants from isolated pollen of Brassica napus. Pflanzenphysiol 105:427-434 https://doi.org/10.1016/S0044-328X(82)80040-8
  26. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue. Physiol Plant 15:473-497 https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
  27. Rodrigues LR, Terra TF, Bered F, Bodanese-Zanettini MH (2004) Origin of embryo-like structures in soybean anther culture investigated using SSR marker. Plant Cell Tiss Organ Cult 77:287-289 https://doi.org/10.1023/B:TICU.0000018397.26315.17
  28. Schenk RU, Hildebrandt AC (1972) Medium and techniques for induction and growth of monocotyledonous and dicotyledonous plant cell cultures. Canadian J Bot 50:199-204 https://doi.org/10.1139/b72-026
  29. Talebi R, Rahemi MR, Arefi H, Nourozi M, Bagheri N (2007) In vitro plant regeneration through anther culture of some Iranian local rice (Oryza sativa L.) cultivars. Pakistan J Biol Sci 10:2056-2060 https://doi.org/10.3923/pjbs.2007.2056.2060
  30. Waminal NE, Park HM, Ryu KB, Kim JH, Yang TJ, Kim HH (2012) Karyotype analysis of Panax ginseng CA Meyer, 1843 (Araliaceae) based on rDNA loci and DAPI band distribution. Comp Cytogenet 6:425-441 https://doi.org/10.3897/compcytogen.v6i4.3740
  31. Wang GF, Qin HY, Sun D, Fan ST, Yang YM, Wang ZX, Xu PL, Zhao Y, Liu YX, Ai J (2018) Haploid plant regeneration from hardy kiwifruit (Actinidia arguta Planch.) anther culture. Plant Cell Tiss Organ Cult 134:15-28 https://doi.org/10.1007/s11240-018-1396-7
  32. Wen J, Plunkett GM, Mitchell AD, Wagstaff SJ (2001) The evolution of Araliaceae: A phylogenetic analysis based on ITS sequences of nuclear ribosomal DNA. Syst Bot 26:144-167
  33. Zhang C, Tsukumi T, Ikeda M, Sato M, Okada H, Ohashi Y, Matsuno H, Yamamoto T, Wada M, Yoshikawa N, Matsumoto S, Li J, Mimida N, Watanabe M, Suzuki A, Komori S (2013) Effects of the microspore development stage and cold pre-treatment of flower buds on embryo induction in Apple (Malus x domestica Borkh.) anther culture. J Jan Soc Hort Sci 82:114-124 https://doi.org/10.2503/jjshs1.82.114
  34. Zhang FL, Takahata Y (2001) Inheritance of microspore embryogenic ability in Brassica crops. Theor Appl Genet 103:254-258 https://doi.org/10.1007/s001220100602
  35. Zhang JY, Sun HJ, Song IJ, Bae TW, Kang HG, Ko SM, Kwon YI, Kim IW, Lee JC, Park SY, Lim PO, Kim YH, Lee HY (2014) Plant regeneration of Korean wild ginseng (Panax ginseng Meyer) mutant lines induced by γ-irradiation (60Co) of adventitious roots. J Ginseng Res 38:220-225 https://doi.org/10.1016/j.jgr.2014.04.001
  36. Zhang SC, Wei AZ, Yang TX (2011) Advances in the research and application of haploid and doubled haploid technologies in fruit trees. J Fruit Sci 28:869-874
  37. Zhao G, Liu Y, Li J (1998) Germination of embryo in soybean anther culture. Chin Sci Bull 43:1991-1995 https://doi.org/10.1007/BF03186991
  38. Zheng MY, Konzak CF (1999) Effect of 2,4-dichlorophenoxyacetic acid on callus induction and plant regeneration in anther culture of wheat (Triticum aestivum L.). Plant Cell Rep 19:69-73