Journal of Plant Biotechnology

  • Volume 50
  • /
  • Pages.176-182
  • /
  • 2023
  • /
  • 1229-2818(pISSN)
  • /
  • 2384-1397(eISSN)
The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Optimal culture methods for plant regeneration via shoot organogenesis in the 'Fuji' apple

사과 '후지'의 기관형성을 통한 식물체 재생에 효율적인 배양방법

  • Yoon Kyung Lee (Department of Plant and Food Sciences, Sangmyung University) ;
  • Youngju Kwon (Department of Plant and Food Sciences, Sangmyung University) ;
  • Yong Joon Yang (Department of Plant and Food Sciences, Sangmyung University)
  • 이윤경 (상명대학교 식물식품공학과) ;
  • 권영주 (상명대학교 식물식품공학과) ;
  • 양용준 (상명대학교 식물식품공학과)
  • Received : 2023.09.14
  • Accepted : 2023.09.18
  • Published : 2023.10.13
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Abstract

Plant regeneration protocols for adventitious shoot organogenesis from apple (Malus domestica 'Fuji') leaf explants were developed in the present study. The effects of dark incubation periods in the early stages of culture, pre-treatment methods, the number of explants per culture container, the type of culture containers, and the orientation of the explants on culture media were evaluated to determine the optimal shoot regeneration conditions for 'Fuji' apple leaf explants. Light incubation of explants produced minimal response. However, dark incubation of explants for 4 weeks during the initial culture period enhanced shoot regeneration frequency. Comparing the number of explants per container, a higher percentage of shoot regeneration was obtained with nine explants per container compared with four explants per container. Pre-treatment, before culture, by dipping explants in a liquid regeneration medium containing 40 g/L of sorbitol for 2 hours produced the highest shoot formation rate, and the time of shoot formation was accelerated. The percentage of shoot regeneration and number of shoots per regenerating explant reached a maximum of 87.5% and 4.7, respectively. The regenerated shoots were elongated and rooted on a rooting medium of 1/4 MS with 0.2 mg/L IBA. The plantlets were successfully acclimatized, and the regenerated plants produced normal phenotypes.

사과 '후지'의 잎 절편체로부터 신초 기관형성을 통한 효율적인 식물체 재생 시스템을 확립하기 위하여 암처리 기간, 전처리 방법, 배양 용기당 치상 절편체 수, 배양 용기의 종류와 배지 위의 절편체 치상방향 등을 달리하여 실험을 실시하였다. 배양 초기 암조건에서의 배양은 신초 형성에 필수적이며, 4주 동안 배양한 후 명조건으로 옮겨 배양하는 것이 신초 재생에 가장 효과적이었다. 배양 전 전처리로는 sorbitol 40 g/L가 포함된 액체 재생배지에 2시간 동안 침지하였을 때 신초 형성율이 87.5%까지 증가하였으며, 신초 형성 시기가 빨라지는 것으로 나타났다. 배양 용기당 치상 절편체 수는 9개를 치상하는 것이 신초 재생에 효과적이었으며, 배양 용기는 100 ml 삼각플라스크를 사용하는 것이 petri dish를 사용하는 것에 비해 신초 형성율이 약 3배 정도 증가하였다. 잎절편체의 배지 위 치상방향은 배축면이 배지에 닿게 치상했을 때, 신초 형성율과 재생된 신초수가 다소 높게 나왔다. 암조건에서 4주간 배양한 후 명조건으로 옮겨총8주간 배양하여 재생된 신초는 1/4 MS에 0.2 mg/L의 IBA가 첨가된 배지에서 발근을 유도한 후 활착시켜 온실에서 재배하였을 때 정상적인 표현형을 보여주었다.

Keywords

Acknowledgement

본 연구는 고인이 되신 형남인 교수님의 지도로 이루어진 것이며 이에 깊은 감사와 존경을 표합니다.

References

  1. Bai L, Guo S, Liu Q, Cui X, Zhang X, Zhang L, Yang X, Hou M, Ho CT, Bai N (2016) Characterization of nine polyphenols in fruits of Malus pumila Mill. by high-performance liquid chromatography. J Food Drug Anal 43:1071-1081  https://doi.org/10.1016/j.jfda.2015.10.002
  2. Bhatti S, Jha G (2010) Current trends and future prospects of biotechnological interventions through tissue culture in apple. Plant Cell Rep 29:1215-1225  https://doi.org/10.1007/s00299-010-0907-8
  3. Buddendorf-Joosten JMC, Woltering EJ (1994) Components of the gaseous environment and their effects on plant growth and development in vitro. Plant Growth Regul 15:1-16  https://doi.org/10.1007/BF00024671
  4. Chi GL, Pua EC, Goh CJ (1991) Role of ethylene on de novo shoot regeneration from cotyledons of Brassica campestris ssp pekinensis (Lour) Olsson in vitro. Plant Physiol 96:178-183  https://doi.org/10.1104/pp.96.1.178
  5. Chu CC, Wang CC, Sun CS, Hsu C, Yin KC, Chu CY (1975) Establishment of an efficient medium for anther culture of rice through comparative experiments on the nitrogen sources. Sci Sin 18:659-668 
  6. Famiani F, Ferradini N, Staffolani P, standardi A (1994) Effect of leaf excision time and age, BA concentration and dark treatments on in vitro shoot regeneration of M. 26 apple rootstock. J Hort Sci 69:679-685  https://doi.org/10.1080/14620316.1994.11516500
  7. Fasolo F, Zimmerman RH, Fordham I (1989) Adventitious shoot formation on excised of in vitro grown shoot of apple cultivars. Plant Cell Tiss Org Cult 16:75-87  https://doi.org/10.1007/BF00036516
  8. Guo B, Abbasi BH, Zeb A, Xu LL, Wei YH (2011) Thidiazuron: a multi-dimensional plant growth regulator. Afr J Biotech 10: 8984-9000  https://doi.org/10.5897/AJB11.636
  9. Hanke MV, Flachowsky H, Peil A, Emeriewen OF (2020) "Malus x domestica apple," in Biotechnology of Fruit and Nut Crops 
  10. Huxter TJ, Thorpe TA, Reid DM (1981) Shoot initiation in light-and dark-grown tobacco callus: the role of ethylene. Physiol Plant 53:319-326  https://doi.org/10.1111/j.1399-3054.1981.tb04507.x
  11. Kevers C, Boyer N, Courduroux JC, Gaspar T (1992) The influence of ethylene on proliferation and growth of rose shoot cultures. Plant Cell Tiss Org Cult 28:175-181  https://doi.org/10.1007/BF00055514
  12. Kim MJ, Kim YG, Kim HS, Cheong C, Jang KH, Kang SA (2014) Effects of antioxidant activities in ethanol extract of apple peel, grape peel, and sweet potato peel as natural antioxidant. J Kor Acad-Ind Coop Soc 15:3766-3733 
  13. Lee IK, Ko BN, Woo SG (2008) Competitiveness and enhancing strategy for Korean apples. Kor J Intl Agri 20:184-189 
  14. Lee KW, Lee SJ, Kang NJ, Lee CY, Lee HJ (2004) Effects of phenolics in Empire apples on hydrogen peroxide-induced inhibition of gap-junctional intercellular communication. Biofactors 21:361-365  https://doi.org/10.1002/biof.552210169
  15. Li BQ, Feng CH, Hu LY, Wang MR, Chen L, Wang QC (2014) Shoot regeneration and cryopreservation of shoot tips of apple (Malus) by encapsulation-dehydration. In Vitro Cell Dev Biol-Plant 50:357-368  https://doi.org/10.1007/s11627-014-9616-2
  16. Linsmaier EM, Skoog F (1965) Organic growth factor requirements of tobacco tissue cultures. Physiol Plant 18:100-127  https://doi.org/10.1111/j.1399-3054.1965.tb06874.x
  17. Magyar-Tabori K, Dobranszki J, da Silva JAT, Bulley SM, Hudak I (2010) The role of cytokinins in shoot organogenesis in apple. Plant Cell Tiss Org Cult 101:251-267  https://doi.org/10.1007/s11240-010-9696-6
  18. Mitic N, Stanisic M, Milojevic J, Tubic L, Cosic T, Nikolic R, Miletic R (2012) Optimization of in vitro regeneration from leaf explants of apple cultivars Golden Delicious and Melrose. HortScience 47:1117-1122  https://doi.org/10.21273/HORTSCI.47.8.1117
  19. Murthy BNS, Murch SJ, Saxena PK (1998) Thidiazuron: A potent regulator of in vitro plant morphogenesis. In Vitro Cell Dev Biol-Plant 34:267-275  https://doi.org/10.1007/BF02822732
  20. Pawlicki N, Welander M (1994) Adventitious shoot regeneration from leaf segments of in vitro cultured shoots of the apple rootstock Jork 9. Hortsicence 69:687-696
  21. Statistical Korea (KOSTAT) (2022) Statistics annual report Daejeon, Korea 
  22. Welander M (1988) Plant regeneration from leaf and stem segments of shoots raised in vitro from mature apple trees. J Plant Physiol 132:738-744  https://doi.org/10.1016/S0176-1617(88)80238-4
  23. Yepes LM, Aldwinckle HS (1994) Factors that leaf regeneration efficiency in apple, and effect of antibiotics in morphogenesis. Plant Cell Tiss Org Cult 37:257-269  https://doi.org/10.1007/BF00042339