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Suitable hormone-free medium for in vitro mass propagation via bioreactor culture of ever-bearing strawberry

Bioreactor를 이용한 사계성 딸기 조직배양묘 대량증식을 위한 적정 무호르몬 배지

  • Kim, Hye-Jin (Highland Agricultural Research Center, National Institute of Crop Science, RDA) ;
  • Lee, Jong-Nam (Highland Agricultural Research Center, National Institute of Crop Science, RDA) ;
  • Kim, Ki-Deog (Highland Agricultural Research Center, National Institute of Crop Science, RDA) ;
  • Im, Ju-Sung (Highland Agricultural Research Center, National Institute of Crop Science, RDA) ;
  • Lim, Hak-Tae (Department of Bio-Health Technology, Kangwon National University) ;
  • Yeoung, Young-Rok (Department of Plant Science, Gangneung-Wonju National University)
  • 김혜진 (국립식량과학원 고령지농업연구센터) ;
  • 이종남 (국립식량과학원 고령지농업연구센터) ;
  • 김기덕 (국립식량과학원 고령지농업연구센터) ;
  • 임주성 (국립식량과학원 고령지농업연구센터) ;
  • 임학태 (강원대학교 생명건강공학) ;
  • 용영록 (강릉원주대학교 식물생명과학과)
  • Received : 2011.08.11
  • Accepted : 2011.08.30
  • Published : 2011.09.30

Abstract

This study was carried out to determine optimal medium conditions for mass propagation by bioreactor culture of ever-bearing strawberry (Fragaria ${\times}$ ananassa D.). Two different type of nitride were that mixed $NH_4NO_3$ and $KNO_3$ or added $KNO_3$ only. And nitride concentrations were at the 4 levels of $1/2{\times}$, $1{\times}$, $2{\times}$ and $3{\times}$ that was included $NH_4NO_3$ and $KNO_3$. Sucrose content ranged at 3 levels of $10g{\cdot}L^{-1}$, $20g{\cdot}L^{-1}$ and $30g{\cdot}L^{-1}$ and medium pH were at the 3 levels of 4.6, 5.6 and 6.6. In bioreactor culture, medium that are included $NH_4NO_3$ and $KNO_3$ together in MS medium was suitable for mass propagation. Medium EC rose rapidly when the nitride concentration was increased. For that reason, plantlet growth was inhibited. Shoots of nitride $1/2{\times}$ concentration was 10.8 ea and $1/2{\times}$ concentration was suitable for shoot propagation. Fresh weight of sucrose $30g{\cdot}L^{-1}$ was 3,101 mg which was heaviest and aerial and ground part were higher than the other concentration. Shoots were increased in proportion to the increasing concentration of sucrose. In the pH condition, from pH 5.6 to 6.8 were appropriate for the optimum growth of aerial and ground part of plant. From the results, in bioreactor culture for mass propagation, MS medium was suitable $1/2{\times}$ concentration that was included $NH_4NO_3$ and $KNO_3$ together, and added $30g{\cdot}L^{-1}$ of sucrose, and then adjusted pH between 5.6 and 6.6.

본 실험은 사계성 딸기의 무병묘 대량생산을 위한 bioreactor 배양 시 배지조건을 구명하기 위해 실시하였다. 배지조건은 MS배지 내 질소질 종류 2처리 ($NH_4NO_3-KNO_3$혼합, $KNO_3$ 단독), MS배지 내 질소농도 4처리 (1/2배, 1배, 2배, 3배), sucrose농도 3처리 ($10g{\cdot}L^{-1}$, $20g{\cdot}L^{-1}$, $30g{\cdot}L^{-1}$)와 배지 pH 3처리 (pH 4.6, 5.6, 6.6) 등을 실험하였다. 사계성 딸기의 bioreactor 배양 시 MS배지 내에 $NH_4NO_3$$KNO_3$를 혼합처리한 배지가 생육량이 많았다. 질소농도가 높아질수록 배지 EC가 급격히 높아지고, 그에 따라 유식물체 생육이 억제되었다. 질소질 1/2배에서 생육량이 많고 신초가 10.8개로 가장 많이 발생하였다. Bioreactor 배양 시 sucrose농도는 $30g{\cdot}L^{-1}$에서 생체중이 3,101 mg으로 가장 무거웠고, 지상부와 지하부 생육량 모두 다른 농도에 비해 많았다. 또한 sucrose농도가 높아질수록 신초수도 증가되는 경향을 보였다. 배지 pH는 5.6-6.6 범위에서 지상부와 지하부 생육량이 모두 많았다. 따라서 사계성 딸기 조직배양묘의 bioreactor를 이용한 대량증식 및 뿌리발달에 알맞은 배지 조건은 MS 무기염을 기본으로 하여 배지 내 질소질은 $NH_4NO_3$$KNO_3$ 혼합, 농도는 1/2배, sucrose농도는 $30g{\cdot}L^{-1}$과 배지 pH는 5.6-6.6 범위가 가장 적당하였다.

Keywords

References

  1. Abdullah MA, Ariff AB, Marziah M, Ali AM, Lajis NH (2000) Strategies to overcome foaming and wall-growth during the cultivation of Morinda elliptica cell suspension culture in a stirred-tank bioreactor. Plant Cell Tiss Org Cult 60:205-212 https://doi.org/10.1023/A:1006495107778
  2. Akita M (2000) Bioreactor culture of plant organs In: Spier RE, Griffiths B, Scragg AH (eds), The Encyclopaedia of cell technology. John Wiley & Sons, Inc., New York, pp 129-138
  3. Aloni R (1980) Role of auxins and sucrose in the differentiation of sieve and trachery elements in plant tissue cultures. Plant 150: 255-263 https://doi.org/10.1007/BF00390835
  4. Bennett IJ, McDavid DAJ, McComb JA (2003) The influence of ammonium nitrate, pH and indole butyric acid on root induction and survival in soil of micropropagated Eucalyptus plobulus. Biol Plant 47:355-360
  5. Bhatia P, Ashwath N (2005) Effect of medium pH on shoot regeneration from the cotyledonary explants of tomato. Biotechnology 4:7-10 https://doi.org/10.3923/biotech.2005.7.10
  6. Bhatt ID, Dhar U (2000) Micropropagation of Indian wild strawberry. Plant Cell Tiss Org Cult 60:83-88 https://doi.org/10.1023/A:1006471815566
  7. Bofunia H, Przywara L (1990) Rola cukrowców roślinnych kulturach in vitro. Wiad Bot [Sugars in plant tissue culture] 43:25-36
  8. Boxus PH (1974) The production of strawberry plants by in vitro micropropagation. J Hort Sci 49:209-210
  9. Boxus PH, Quoirin M, Laine JM (1977) Applied and fundamental aspects of plant cell, tissue and organ culture. In: Reinert J, Bajaj YPS (eds), Large scale propagation of strawberry plants from tissue culture. Springer-Verlag, Heidelberg, pp 130-143
  10. Brown DCW, Leung DWM, Thorpe TA (1979) Osmotic requirements for shoot formation in tobacco callus. Physiol Plant 46: 36-41 https://doi.org/10.1111/j.1399-3054.1979.tb03182.x
  11. Chattopadhyay S, Datta SK, Ray M (1992) In vitro effect of $NH_4NO_3$ on growth and alkaloid content of Tylophora indica Merr. Phytomorphism 42:139-144
  12. Correll MJ, Wu Y, Weathers PJ (2000) Controlling hyperhydration of carnations (Dianthus caryophyllus L.) grown in a mist reactor. Biotechnol Bioeng 71:307-314 https://doi.org/10.1002/1097-0290(2000)71:4<307::AID-BIT1019>3.0.CO;2-9
  13. Damiano C (1980) Proceedings of the conference on nursery production of fruit plants through tissue culture. Applications and feasibility. In: Planning and building a tissue culture laboratory. USDA, pp 93-101
  14. Escalona M, Lorenzo JC, Gonzalez B, Daquinta M, Gonzalez JL, Desjarins Y, Borroto CG (1999) Pineapple (Ananas comosus L. Merr) micropropagation in temporary immersion systems. Plant Cell Rpt 18:743-748 https://doi.org/10.1007/s002990050653
  15. Gao WY, Fan L, Paek KY (2000) Yellow and red pigment production by cell cultures of Carthamus tinctoris in a bioreactor. Plant Cell Tiss Org Cult 60:95-100 https://doi.org/10.1023/A:1006404724055
  16. Gaspar T, Kevers C, Debergh P, Maeng L, Paques M, Boxus P (1987) Vitrification: Morphological, Physiological and ecological aspects. In: Bonga JM, Durzan J (eds), General principles and biotechnology. Cell and Tissue Culture in Forestry. The Netherlands
  17. Gibson SI (2000) Plant sugar-response pathways. Part of a complex regulatory web. Plant Physiol 124:1532-1539 https://doi.org/10.1104/pp.124.4.1532
  18. Grimes HD, Hodge TK (1990) The inorganic $NO_3^-:NH_4^+$ ratio influences plant regeneration and auxin sensitivity in primary callus derived from immature embryos of indica rice (Oryza sativa L.). J Plant Physiol 136:362-367 https://doi.org/10.1016/S0176-1617(11)80063-5
  19. Honda H, Liu C, Kobayashi T (2001) Large-scale plant micropropagation. Adv Biochem Eng Biotechnol 72:157-182
  20. Hussey G (1986) Vegetative propagation of plants by tissue culture. In: Yeoman MM (ed), Plant Cell Culture Technology, Blackwell Scientific Publications, Oxford
  21. Ingram B, Mavituna F (2000) Effect of bioreactor configuration on the growth and maturation of Picea sitchensis somatic embryo cultures. Plant Cell Tiss Org Cult 61:87-96 https://doi.org/10.1023/A:1006333907907
  22. Kirschbaum DS, Candtliffe DJ, Shaw NL, Liu JR (2004) Direct adventitious shoot formation on seedling radicles in seed cultures of strawberry. J Plant Biol 47:160-162 https://doi.org/10.1007/BF03030647
  23. Lee JN (2006) Physiological and ecological response of ever-bearing strawberry in the highlands cultivation for off-season production. PhD. thesis. Gangneung National University. Gangneung
  24. Lee JN, Kim HJ, Kim KD, Kwon YS, Lim HT, Yeoung YR (2010a) In vitro mass propagation and economic effects of bioreactor culture in ever-bearing strawberry 'Goha'. Kor J Hort Sci Technol 28:845-849
  25. Lee JN, Kim HJ, Kim KD, Kwon YS, Yeoung YR, Lim HT (2010b) Appropriate in vitro culture conditions of growing medium for new ever-bearing strawberry 'Goha'. Kor J Hort Sci Technol 28:1051-1056
  26. Leifert C, Pryce S, Lumsden P, Waites P (1992) Effects of medium acidity on growth and rooting of different plant species growing in vitro. Plant Cell Tiss Org Cult 30:171-179 https://doi.org/10.1007/BF00040019
  27. Lipavska H, Vreugdenhil D (1996) Uptake of mannitol from the media by in vitro grown plants. Plant Cell Tiss Org Cult 45: 103-107 https://doi.org/10.1007/BF00048751
  28. Le Roux JJ, Van Staden J (1991) Micropropagation and tissue culture of Eucalyptus -a review. Tree Physiol 9:435-477 https://doi.org/10.1093/treephys/9.4.435
  29. Leva AR, Bartolini G, Muleo R, Biricoliti S, Benelli A (1990) Micromorphological studies on Actinidia cv. Harvard calli as influenced by different sugars. In: X X III International Horticultural Congress (ed) Abstracts of Contributed Papers: 3123. Firenze. Italy
  30. Lorenzo JC, Ojeda E, Espinosa A, Borroto C (2001) Field performance of temporary immersion bioreactor-derived sugarcane plants. In vitro Cellular Dev Biol Plant 37:803-806 https://doi.org/10.1007/s11627-001-0133-8
  31. McComb JA, Bennett IJ, Tonkin CM (1996) In vitro propagation of Eucalyptus species. In: Taji A, Williams R (eds), Tissue culture of Australian native plants, University of New England Press, Armidale, pp 112-156
  32. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue culture. Physiol Plant 15:473-479 https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
  33. Paek KY, Hahn EJ, Son SH (2001) Application of bioreactors for large-scale micropropagation systems of plants. In vitro Cellular Dev Biol Plant 37:284-292
  34. Passey AJ, Barrett KJ, James DJ (2003) Adventitious shoot regeneration from seven commercial strawberry cultivars (Fragaria ${\times}$ ananssa Duch.) using a range of explants types. Plant Cell Rpt 21:397-401 https://doi.org/10.1007/s00299-002-0530-4
  35. Sansavini S, Rosati P, Gaggioli D, Toshi MF (1990) Inheritance and stability of somaclonal variation in micropropagated strawberry. Acta Hortic 280:375-384
  36. Sathyanarayana BN, Blake J (1994) The effect of nitrogen sources and initial pH of the media with or without buffer on in vitro rooting of jackfruit (Artocarpus heterophyllus Lam). Physiololgy, Growth and Development of Plants in Culture. Kluwer Academic Publishers, Netherlands, pp 77-78
  37. Selby C, Harvey BMR (1990) The influence of composition of the basal medium on the growth and morphogenesis of cultured sitka spruce (Picea sitchensis) tissues. Ann Bot 65:395-407 https://doi.org/10.1093/oxfordjournals.aob.a087950
  38. Sriskandarajah S, Skirvin RM, Abu-Qaoud H (1990) The effect of some macronutrients on adventitious root development on scion apple cultivars in vitro. Plant Cell Tiss Org Cult 21:185-189 https://doi.org/10.1007/BF00033440
  39. Steinitz B (1999) Sugar alcohols display non osmotic roles in regulating morphogenesis and metabolism in plants that do not produce polyols as primary photosynthetic products. J Plant Physiol 155:1-8 https://doi.org/10.1016/S0176-1617(99)80133-3
  40. Takayama S (1991) Mass propagation of plants through shake and bioreactor culture techniques. In: Bajaj, YPS (ed.) Biotechnology in Agriculture and Forestry. Vol. 17. Springer-Verlag, Berlin, pp 495-515
  41. Takayama S (2002) Practical aspects of bioreactor application in mass propagation of plants. Abst. 1st Int. Symp. Liquid systems for in vitro mass propagation of plants. Norway, pp 60-62
  42. Takayama S, Misawa M (1981) Mass propagation of Begonia hiemalis plantlets by shake culture. Plant Cell Physiol 22:461-468
  43. Vinocur B, Carmi T, Altman A, Ziv M (2000) Enhanced bud regeneration in aspen (Populus tremula L.) roots cultured in liquid media. Plant Cell Rpt 19:1146-1154 https://doi.org/10.1007/s002990000243
  44. Williams RR, Taji AM, Bolton JA (1985) Specificity and interaction among auxins, light, and pH in rooting of Australian woody species in vitro. HortiScience 20:1052-1053
  45. Woodward AJ, Bennett IJ, Pusswonge S (2006) The effect of nitrogen source and concentration, medium pH and buffering on in vitro shoot growth and rooting in Eucalyptus marginata. Scientia Hort 110:208-213 https://doi.org/10.1016/j.scienta.2006.07.005

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