Cereal Research Communications

, Volume 33, Issue 2–3, pp 553–560 | Cite as

Influence of Potassium Dihydrogen Phosphate on Callus Induction and Plant Regeneration in Rice (Oryza sativa L.)

  • Mranali Chauhan
  • S. L. KothariEmail author
Open Access


The effect of potassium dihydrogen phosphate (KH2PO4) on callus induction and plant regeneration from callus cultures of indica rice (Oryza sativa cultivar Pusa Basmati-1) was investigated. Callus was induced on Murashige and Skoog (MS) medium supplemented with 11.31 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and subsequent plant regeneration was obtained on MS medium containing 2.68 μM α-naphthaleneacetic acid (NAA) and 8.87 μM 6-benzylaminopurine (BAP). Both the induction and the regeneration media were supplemented with different levels of KH2PO4 (0 to12.50 mM). The level of KH2PO4 in the induction medium influenced the percent water content of the callus. KH2PO4 was found to be essential for plant regeneration as no shoot regeneration occurred in the absence of KH2PO4. Highest number of shoots per expiant was obtained when KH2PO4 level in the callus induction and plant regeneration medium was 0.625 mM (half of MS level) and 1.25 mM (normal MS level) respectively. By making amendments and modifications of the KH2PO4 level in the induction medium, the regeneration increased 2.5 folds. Almost similar regeneration frequency was observed from callus induced on lower levels of KH2PO4. Regenerated shoots were rooted on MS medium supplemented with 2.68 μM NAA. Rooted plantlets were transferred to soil where they survived and set seeds.


potassium dihydrogen phosphate callus induction plant regeneration water content indica rice 


  1. Abdullah R, Cocking EC, Thompson JA (1986) Efficient plant regeneration from rice protoplasts through somatic embryogenesis. Bio/Technol 4:1087–1090Google Scholar
  2. Chen TH, Lam L, Chen SH (1985) Somatic embryogenesis and plant regeneration from cultured young inflorescences of Oryza sativa L. (rice). Plant Cell Tiss Org Cult 4:51–54CrossRefGoogle Scholar
  3. Dahleen LS (1995) Improved plant regeneration from barley callus cultures by improved copper levels. Plant Cell Tiss Organ Cult 43:267–269Google Scholar
  4. Dahleen LS, Bregitzer P (2002) An improved media system for high regeneration rates from barley immature embryo-derived callus cultures of commercial cultivars. Crop Sci 42:934–938CrossRefGoogle Scholar
  5. Grimes HD, Hodges TK (1990) The inorganic NO3-: NH4+ ratio influences plant regeneration and auxin sensitivity in primary callus derived from immature embryos of indica rice (Oryza sativa L.). J Plant Physiol 136:362–367CrossRefGoogle Scholar
  6. He DG, Yang YM, Scott KJ (1989) The effect of macronutrients in the induction of embryogenic callus from immature embryos of wheat (Triticum aestivum L.). Plant Sci 64:251–258CrossRefGoogle Scholar
  7. Jain RK, Jain S, Wu R (1996) Stimulatory effect of water stress on plant regeneration of aromatic indica rice varieties. Plant Cell Rep 15:1449–1459Google Scholar
  8. Khanna HK, Raina SK (1997) Enhanced plant regeneration in basmati rice (Oryza sativa L. cv. Karnal Local) embryo calli through modification of NO3- + NH4+ concentrations. J Biochem Biotech 6:75–80Google Scholar
  9. Khanna HK, Raina SK (1998) Genotype X culture media interaction effects on regeneration response of three indica rice cultivars. Plant Cell Tiss Org Cult 52:145–153CrossRefGoogle Scholar
  10. Koetje DS, Grimes HD, Wang YC, Hodges TK (1989) Regeneration of indica rice (Oryza sativa L.) from primary callus derived from immature embryos. J Plant Physiol 135:184–190CrossRefGoogle Scholar
  11. Kothari SL, Agarwal K, Kumar S (2004) Inorganic nutrient manipulation for highly improved in vitro plant regeneration in finger millet — Eleusine coracana (L.) Gaertn. In Vitro Cell Dev Biol (in press)CrossRefGoogle Scholar
  12. Lee K, Jeon H, Kim M (2002) Optimization of a mature embryo based in vitro culture system for high frequency somatic embryogenic callus induction and plant regeneration from japonica rice cultivars. Plant Cell Tiss Org Cult 17:237–244CrossRefGoogle Scholar
  13. Leifert C, Murphy KP, Lumsden PJ (1995) Mineral and carbohydrate nutrition of plant cell and tissue cultures. Crit Rev Plant Sci 14(2):83–109CrossRefGoogle Scholar
  14. Loneragan JF, Asher CJ (1982) Response of plants to phosphate concentration in solution culture: II. Rate of phosphate absorption and its relation to growth. Soil Sci 103:311–318.CrossRefGoogle Scholar
  15. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  16. Nirwan RS, Kothari SL (2003) High copper levels improve callus induction and plant regeneration in Sorghum bicolor (L.) Moench. In Vitro Cell Dev Bio 2:165–168Google Scholar
  17. Nishi T, Yamada Y, Takahashi E (1968) Organ differentiation and plant restoration in rice callus. Nature 219:508–509CrossRefGoogle Scholar
  18. Oinam GS, Kothari SL (1995) Totipotency of coleoptile tissue in indica rice (Oryza sativa L. cv. CH 1039). Plant Cell Rep 14:245–248CrossRefGoogle Scholar
  19. Poddar K, Vishoni RK, Kothari SL (1997) Plant regeneration from embryogenie callus of finger millet Eieusine coracana (L.) Gaertn. on high concentration of NH4NO3 as a replacement of NAA in the medium. Plant Sci 129:101–106CrossRefGoogle Scholar
  20. Preece JE (1995) Can nutrient salts partially substitute for plant growth regulators? Plant Tiss Cult Biotech 1:26–37Google Scholar
  21. Purnhauser L (1991) Stimulation of shoot and root regeneration in wheat Triticum aestivum callus cultures by copper. Cereal Res Comm 19:419–423Google Scholar
  22. Purnhauser L, Gyulia G (1993) Effect of copper on shoot and root regeneration in wheat, triticale, rape and tobacco tissue cultures. Plant Cell Tiss Organ Cult 35:131–139CrossRefGoogle Scholar
  23. Ramage CM, Williams RR (2002) Mineral nutrition and plant morphogenesis. In Vitro Cell Dev Biol 38:116–124CrossRefGoogle Scholar
  24. Rance IM, Tian W, Mathews H, Kochko A, Beachy RN, Fauquet C (1994) Partial desiccation of mature embryo-derived calli, a simple treatment that dramatically enhances the regeneration ability of indica rice. Plant Cell Rep 13:647–651CrossRefGoogle Scholar
  25. Raval M, Chattoo BB (1993) Role of media constituents and proline in callus growth, somatic embryogenesis and regeneration of Oryza sativa cv indica. Ind J Exp Bio 31:600–603Google Scholar
  26. Sahrawat AK, Chand S (2002) Influence of ammonium nitrate on plant regeneration in indica rice (Oryza sativa Linn.). Ind J Biotech 1:367–371Google Scholar
  27. Sahrawat AK, Chand S (1999) Stimulatory effect of copper on plant regeneration in indica rice. J Plant Physiol 154:517–522CrossRefGoogle Scholar
  28. Sticklen MB (1991) Direct somatic embryogenesis and fertile plants from rice root cultures. J Plant Physiol 138:577–580CrossRefGoogle Scholar
  29. Wernicke W, Brettel R, Wakizuma T, Potrykus I (1981) Adventitious embryoid and root formation from rice leaves. Z Planzenphysiol 103:361–365CrossRefGoogle Scholar
  30. Zhao J, Zhou C, Yang HY (1999) In vitro development of early proembryos and plant regeneration via microculture in Oryza sativa. Plant Cell Tiss Org Cult 55:167–174CrossRefGoogle Scholar

Copyright information

© Akadémiai Kiadó, Budapest 2005

This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (, which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

Authors and Affiliations

  1. 1.Department of BotanyUniversity of RajasthanJaipurIndia

Personalised recommendations