Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
권호 표지
DOI QR코드

DOI QR Code

Optimal Medium Compositions for Plant Regeneration via Adventitious Shoot Formation Using 'Fuji' Apple Leaf Explants

사과 '후지'의 잎 절편체로부터 신초 기관형성을 통한 식물체 재생에 적합한 배지조성

  • Lee, Yoon Kyung (Department of Plant and Food Sciences, Sangmyung University) ;
  • ;
  • Hyung, Nam-In (Department of Plant and Food Sciences, Sangmyung University)
  • 이윤경 (상명대학교 식물식품공학과) ;
  • 권영주 (상명대학교 식물식품공학과) ;
  • 형남인 (상명대학교 식물식품공학과)
  • Received : 2019.10.01
  • Accepted : 2019.10.28
  • Published : 2019.12.31
Download PDF
⟨ Previous Next ⟩

Abstract

Plant regeneration protocols for adventitious shoot organogenesis from apple (Malus domestica 'Fuji') leaf explants were developed in the present study. The effects of different basal media, types and concentrations of carbon sources, and concentrations of plant growth regulators were evaluated to determine the optimal shoot regeneration conditions for 'Fuji' apple leaf explants. On different treatments involving combinations of basal media, LS and N6 media, and different types and concentrations of cytokinins, 6-benzyl-adenine (BA) and thidiazuron (TDZ), shoot regeneration rates were the highest in the N6 medium combined with BA. Among the plant growth regulator and carbon source combination treatments, 5.0 mg/L BA, and 0.1 mg/L α-naphthalene acetic acid (NAA) with 40 g/L sorbitol was the optimal combination for shoot regeneration. In addition, the optimal sorbitol concentrations for shoot regeneration were 40 g/L and 60 g/L. The highest regeneration (81.8%) was achieved using 40 g/L sorbitol. The regenerated shoots elongated and rooted on rooting medium, consisting of 1/4 MS medium with 0.2 mg/L indole-3-butyric acid (IBA). The plantlets were acclimatized and the regenerated plants exhibited normal phenotypes.

사과 '후지'의 잎 절편체로부터 신초 기관형성을 통한 효율적인 식물체 재생 시스템을 확립하기 위하여 배지조성 중 식물생장조절제, 기본배지와 당의 종류 및 농도를 달리하여 실험을 실시하였다. 식물생장조절제 NAA가 첨가된 상태에서 기본배지 중 N6와 LS, cytokinin 중 BA와 TDZ를 비교하였을 때, N6 배지에 BA 5.0 mg/L를 사용하는 것이 신초 형성율과 절편체당 재생 신초수에서 효과적이었다. BA보다 신초 형성에 효과적인 것으로 알려진 TDZ는 본 실험에서는 전반적으로 신초 형성율이 낮았다. 탄소 공급원과 식물생장조절제의 조합 처리에서는 sorbitol 40 g/L의 처리가 신초 형성율 67.3%, 절편체당 신초수 4.3개로 가장 좋았고, 식물생장조절제는 BA 5.0 mg/L와 NAA 0.1 mg/L의 혼합처리가 가장 효과적이었다. 암조건에서 3주간 배양한 후 명조건으로 옮겨 총 8주간 배양하여 재생된 신초는 1/4MS에 IBA 0.2 mg/L가 첨가된 배지에서 발근을 유도한 후 활착시켜 온실에서 재배하였을 때 정상적인 표현형을 보여주었다.

Keywords

References

  1. An G (1993) Current status and future perspectives of biotechnology using plant tissue culture techniques (III). Kor J Plant Tiss Cult 20:59-62
  2. Carl AH, Preece JE (1993) Thidiazuron: A potent cytokinin for plant tissue culture. Plant Cell Tiss Org Cult 33:105-119 https://doi.org/10.1007/BF01983223
  3. 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 Hort 18:659-668
  4. Defour M (1990) Improving yield of adventitious shoots in apple. Acta Hort 280:51-60 https://doi.org/10.17660/actahortic.1990.280.7
  5. 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 Biotechnol 69:679-685 https://doi.org/10.1080/14620316.1994.11516500
  6. Guo B, Abbasi BH, Zeb A, Xu LL, Wei YH (2011) Thidiazuron: A multi-dimensional plant growth regulator. Afr J Biotechnol 10:8984-9000 https://doi.org/10.5897/AJB11.636
  7. Kadota M, Niimi Y (2004) Influences of carbon sources and their concentrations on shoot proliferation and rooting of ‘Hosui’ Japanese pear. HortScience 39:1681-1683 https://doi.org/10.21273/HORTSCI.39.7.1681
  8. Karhu ST (1997) Sugar use in relation to shoot induction by sorbitol and cytokinin in apple. J Amer Sor Hort Sci 122:476-480 https://doi.org/10.21273/JASHS.122.4.476
  9. 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
  10. 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
  11. Loescher WH, Marlow GC, Kennedy RA (1982) Sorbitol metabolism and sink-source interconversions in developing apple leaves. Plant Physiol 70:335-339 https://doi.org/10.1104/pp.70.2.335
  12. 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
  13. Marino G, Magnanini E, Battistini S, Righetti B (1991) Effect of hormones and main carbon energy sources on in vitro propagation of apricot (Prunus armeniaca L.) cvs. 'San Castrese' and 'Portici'. Acta Hort 293:355-362 https://doi.org/10.17660/actahortic.1991.293.42
  14. Mitic N, Stanisic M, Milojevic J, Tubic L, Cosic T, Nikolic R, Ninkovic S, 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
  15. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio-assay with tobacco tissue cultures. Physiol Plant 15:473-497 https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
  16. 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
  17. Pawlicki N, Welander M (1994) Adventitious shoot regeneration from leaf segments of in vitro cultured shoots of the apple rootstock Jork 9. J Hort Sci 69:687-696 https://doi.org/10.1080/14620316.1994.11516501
  18. Predieri S, Fasolo F (1989) High-frequency shoot regeneration from leaves of the apple rootstock M26 (Malus pumila Mill.) Plant Cell Tiss Org Cult 17:133-142 https://doi.org/10.1007/BF00046858
  19. Seong ES, Song KJ, Jega S, Yu CY, Chung IM (2005) Silver nitrate and aminoethoxyvinylglycine affect Agrobacterium-mediated apple transformation. Plant Growth Reg 45:75-82 https://doi.org/10.1007/s10725-004-6126-y
  20. Sriskandarajah S, Goodwin PB, Speris J (1994) Genetic transformation of apple scion cultivar ‘Delicious’ via Agrobacterium tumefaciens. Plant Cell Tiss Org Cult 36:317-329 https://doi.org/10.1007/BF00046089
  21. Statistical Korea (KOSTAT) (2017) Statistics annual report. Daejeon Korea
  22. Wallart RA (1980) Distribution of sorbitol in Rosaceae. Phytochemistry 19:2603-2610 https://doi.org/10.1016/S0031-9422(00)83927-8
  23. 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
  24. Welander M, Maheswaran G (1992) Shoot regeneration form leaf explants of dwarfing apple rootstocks. J Plant Physiol 144:223-228 https://doi.org/10.1016/S0176-1617(11)80939-9
  25. 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