Plant regeneration from suspension-cultured cell clusters of Arabidopsis thaliana

애기장대(Arabidopsis thaliana)의 현탁배양세포괴로부터 식물체 재분화

  • 김명덕 (강원대학교 자연과학대학교 생명과학부) ;
  • 김준철 (강원대학교 자연과학대학교 생명과학부) ;
  • 진창덕 (강원대학교 자연과학대학교 생명과학부) ;
  • 임창진 (강원대학교 자연과학대학교 생명과학부) ;
  • 한태진 (한림대학교 자연과학대학 생물학과)
  • Published : 1998.05.01

Abstract

Callus induction from leaf and stem explants of Arabidopsis thaliana was successfully obtained when leaf explants were cultured on MS medium containing 2.0 mg/L 2,4-D in the dark and also, when stem explants were cultured on CP medium containing 0.5 mg/L 2,4-D and 0.1 mg/L BAP. Explant-derived sliced calli were suspension-subcultured every week in CP liquid medium with 0.5 mg/L 2,4-D and 0.1 mg/L BAP in the dark, and shoot-forming cell clusters of nodular, pale yellow and knobby type were selected after 7-8 weeks of culture. Shoots were initiated from the green spots of the selected shoot forming calli cultured on MS regeneration medium containing 0.05 mg/L IAA, 7.0 mg/L 2-iP and 30 g/L sucrose under continous illumination for four weeks. Shoot regeneration frequency (calli regenerating at least one shoot) was more than 50%. For plant regeneration, excised shoots were trnasferred to hormone free medium for root initiation after 4 weeks of culture. The regenerants were bolting after 2 weeks of culture and formed in vitro flowering buds within bracts after 4 weeks of culture.

기내에서 발아시킨 애기장대(Arabidopsis thaliana)의 잎과 줄기 절편체로부터 캘러스 유도는, 잎절편체의 경우 2mg/L 2,4-D가 첨가된 MS 고체배지에서 유도되었고, 줄기절편체의 경우는 0.5mg/L 2,4-D와 0.1mg/L BAP가 첨가된 CP 고체배지에서 배양 4주 후 다량의 캘러스가 유도되었다. 유도된 캘러스를 잘게 자르고 0.5mg/L 2,4-D와 0.1mg/L BAP가 첨가된 CP 액체배지에서 7일 간격으로 암조건에서 4개월간 배양하였을 때 균일한 shoot-forming(SF)캘러스를 얻을 수 있었다. 액체배지에서 유도된 SF 세포괴는 0.05 mg/L IAA, 7.0mg/L 2-iP, 30g/L sucrose가 첨가된 MS 재분화배지에서 광조건으로 배양하였을 때 캘러스를 거쳐 녹화되기 시작하였으며 배양 4주 후부터는 전체적으로 녹화된 SF캘러스로부터 shoot가 형성되어 식물체 재분화가 가능하였다. 또한 재분화 배지에 옮겼을 때 IAA와 2-iP가 첨가된 배지에서 50% 이상의 shoot 형성률(SF 캘러스당 한 개 이상의 shoot가 형성된 캘러스의 백분율)을 보였다. 절단된 shoot는 호르몬이 첨가되지 않은 배지에서 4주 후 뿌리를 형성하였으며, 재분화된 식물체는 기내에서 6주 후부터 화뢰가 형성되고 꽃이 피기 시작하였다.

Keywords

References

  1. Plant Physiol v.136 Restoration of the regeneration potential of long term cell culture in rice(Oryza sativa) by salt pretreatment Binnh OQ;Heszky LE
  2. Plant Cell Rep v.8 Relative regerneration proficiency of Arabidopsis thaliana ecotypes Chaudhury AM;Singer ER
  3. Z Pflanzenphysiol v.100 Embryoids from cultured immature embryos of Lolium multiflorum Dale PJ
  4. Plant Sci v.67 Embryogenesis and regeneration from microspores of both indica and japonica rice(Oryza sativa) Data SK;Data K;Potrykus I
  5. Z Pflanzenphysiol v.91 Induction, culture and differentiation of callus from immature rachises, seeds and embroys of Triticum Gosch-Wackerle G;Avivi L;Galum E
  6. Z Pflanzenphysiol v.107 Long term plant regeneration somatic embroygenesis and green spot formation in secondary oat(Avena sativa) callus Heyser JW;Nabors MW
  7. Plant Cell Rep v.9 Arabidopsis thaliana: protocol for plant regeneration from protoplast Karesch H;Bilang R;Potrykus I
  8. Agricultural application of plant culture and is industrialization Rapid multiplication of plants through micropropagation Kim KW
  9. Method in Arabidopsis research Koncz C;Chua NH;Schell J
  10. Plant Mol Biol v.21 cDNA cloning of ECP 40, an embryogenic-cell protein in carrot and its expression during somatic and embryogenesis Kiyosue T;Yamaguchi K;Shinozaki K;Kamada H;Harada H
  11. Mol Gen Gent v.194 The DNA of Arabidopsis thaliana Leutwiler LS;Hough EB;Meyerowitz EM
  12. Cell v.56 Arabidopsis, a useful weed Meyerowitz EM
  13. Physiol Plant v.15 A revised medium for rapid growth and bioassays with tobocco tissue cultures Murashige T;Skoog F
  14. Planta v.157 Long duration high frequency of plant generation from cereal tissue culture Narbors MW;Heyser TA;Dykes Demott KJ
  15. Plant Cell Rep v.7 High-frequency plant regeneration from cultured cotyledons of Arabidopsis thaliana Pattom DA;Meinke DW
  16. J Mol Biol v.187 Characterization of the genome of Arabidopsis thaliana Pruitt RE;Meyerowitz EM
  17. Z. Pflazenphysiol v.113 Cytokinin mediated long-term, high-frezuency plant regeneration in rice tissue culture Raghaven Ram NV;Nabors MW
  18. Plant Cell v.4 Effect of inozing radiation on a plant genome: Analysis of two Arabidopsis transparent testa mutations Shirley BW;Hanley S;Goodman HM
  19. Ann Bot v.38 The decline of embryogenic potential as callus supension cultures of carrot(Daucus aerota L.) are serially subcultured Smith SM;Street HE
  20. Crop Science v.24 Callus induction, plant regeneration and evidence of somatic embryogenesis in Red fescue Torello WA;Symington AG;Rufner R
  21. Plant Cell Tiss Org Cult v.12 Somatic embrypgenesis in cotton(Gossypium) 1. Effect of source of explant and hormone regime Trlinder N;Goodin JR