Plant Cell, Tissue and Organ Culture

, Volume 46, Issue 3, pp 227–235 | Cite as

Somatic embryogenesis and plant regeneration of pepper in liquid media

  • Saadet Buyukalaca
  • Ferda Mavituna
Original Research Paper


A protocol was developed for regeneration of pepper (Capsicum annuum var. Ace) through somatic embryogenesis in liquid media. For embryogenic callus formation, mature zygotic embryo explants were used on basal Murashige and Skoog medium with 9.05 μM 2,4-dichlorophenoxyacetic acid and 3% sucrose. Embryogenic callus was transferred to liquid basal Murashige and Skoog medium with 4.52 μM 2,4-dichlorophenoxyacetic acid and 3% sucrose in order to increase the mass of the embryogenic culture. After pretreatment with potassium citrate, cells were placed into embryo initiation medium with 6 g l-1l-proline and a decreased (10 mM) ammonium concentration. Embryos were matured in 1.89 μM abscisic acid containing half-strength Murashige and Skoog medium and converted into plants bothin vivo andin vitro at up to a 97% efficiency.

Key words

Capsicum spp maturation zygotic embryo explant 



abscisic acid


Gamborg et al. (1968)




2,4-dichlorophenoxyacetic acid


indole 3-acetic acid


Murashige and Skoog (1962)


naphthaleneacetic acid


packed cell volume


Schenk and Hildebrandt (1972)


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. AgrawalS & ChandraN (1983) Differentiation of multiple shoot buds and plantlets in cultured embryos ofc. annuum var. Mathania. Curr. Sci. 52: 645–646Google Scholar
  2. AmmiratoPV (1983) Embryogenesis. In: EvansDA, SharpWR, AmmiratoPV & YamadaY (eds) Handbook of Plant Cell Culture, Vol 1 (pp 82–123). Macmillan, New YorkGoogle Scholar
  3. ArroyaR & RevillaMA (1991)In vitro plant regeneration from cotyledon and hypocotyl segments in two bell pepper cultivars. Plant Cell Rep. 10: 414–416Google Scholar
  4. CheeRP, LeskovarDI & CantliffeDJ (1992) Optimizing embryogenic callus and embryo growth of a synthetic seed system for sweetpotato by varying media nutrient concentrations. J. Amer. Soc. Hort. Sci. 117 (4): 663–667Google Scholar
  5. CheemaGS (1989) Somatic embryogenesis and plant regeneration inCarica papaya L. tissue culture derived from root explants. Plant Cell Rep. 6: 348–351Google Scholar
  6. ComptonME (1994) Statistical methods suitable for the analysis of plant tissue culture data. Plant Cell Tiss. Org. Cult. 37: 217–242Google Scholar
  7. Evans DA, Sharp WR & Flick CE (1981) Growth and behaviour of cell cultures: embryogenesis and organogenesis. In: Thorpe TA (ed) Plant Tissue Culture (pp 45–113), Academic PressGoogle Scholar
  8. FariM (1986) Pepper. In: BajajYPS (ed) Biotechnology in Agriculture and Forestry, Vol 2 (pp 345–362). Springer-Verlag, BerlinGoogle Scholar
  9. FariM & CzakoM (1981) Relationships between position and morphogenetic response of pepper hypocotyl explant culturein vitro. Sci. Hort. 15: 207–213Google Scholar
  10. FujiJ, SladeD, OlsenR, RuzinSE & RedenbaughK (1990) Alfalfa somatic embryo maturation and conversion into plants. Plant Sci. 72: 93–100Google Scholar
  11. GamborgOL, MillerRA & OjimaK (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp. Cell Res. 50: 151–158CrossRefPubMedGoogle Scholar
  12. GeKL, SasakumaT & TanakaM (1989) Callus induction and plant regeneration inCapsicum. Acta Bot. Sinica 31: 962–965Google Scholar
  13. GingasVM & LinebergerRD (1989) Asexual embryogenesis and plant regeneration inQuercus. Plant Cell Tiss. Org. Cult. 17: 191–203CrossRefGoogle Scholar
  14. GleddieS, KellerW & SetterfieldG (1983) Somatic embryogenesis and plant regeneration from leaf explants and cell suspensions ofSolanum melongena. Can. J. Bot. 61: 656–666Google Scholar
  15. GunayAL & RaoPS (1978)In vitro plant regeneration from hypocotyl and cotyledon explants of red pepper (Capsicum). Plant Sci. Lett. 11: 365–372Google Scholar
  16. HariniI & SitaGL (1993) Direct somatic embryogenesis and plant regeneration from immature embryos of chilli (Capsicum annuum L.). Plant Sci. 89: 107–112CrossRefGoogle Scholar
  17. KamadaH & HaradaH (1979) Studies on the organogenesis in carrot tissue cultures: I. Effects of growth regulators on somatic embryogenesis and root formation. Z. Pflanzenphysiol. 91: 225–266Google Scholar
  18. KaoAN & MichaylukMR (1981) Embryoid formation in alfalfa cell suspension cultures from different plants. In Vitro 17: 645–648Google Scholar
  19. KomamineA, MatsumotaM, TsukaharaM, FujiwaraA, KawaharaA, ItoM, SmithJ, NomuraK & FujimuraT (1990) Mechanism of somatic embryogenesis in cell cultures- Physiology, biochemistry and molecular biology. In: NijkampHJJ, van derPlasLHW & vanAartrijkJ (eds) Progress in Plant Cellular and Molecular Biology (pp 307–313). Kluwer Academic Publ., Dordrecht, the NetherlandsGoogle Scholar
  20. LentnerM & BishopT (1986) Experimental Design and Analysis. Valley Book Company, Blackburg, VAGoogle Scholar
  21. MomsonRA, KoningRE & EvansDA (1986) Pepper. In: EvansDA, SharpVR & AmmiratoPR (eds) Handbook of Plant Cell Culture, Vol 4 (pp 552–573). Macmillan, New York, NYGoogle Scholar
  22. MurashigeT & SkoogF (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473–497Google Scholar
  23. NicholJW, SladeD, VissP & StuartDA (1991) Effect of organic acid pretreatment on the regeneration and development (conversion) of whole plant from callus cultures of alfalfa,Medicago sativa L. Plant Sci. 79: 181–192CrossRefGoogle Scholar
  24. PhilipsGC & HustenbergerJF (1985) Organogenesis in pepper tissue cultures. Plant Cell Tiss. Org. Cult. 4: 261–269Google Scholar
  25. RaoKV, SuprasannaP & ReddyGM (1990) Somatic embryogenesis from immature glume calli ofZea mays L. Ind. J. Exp. Biol. 28: 531–533Google Scholar
  26. RedenbaughK, FujiiJ, SladeD, VissP & KosslerM (1991a) Artificial seeds-Encapsulated somatic embryos. In: BajajYPS (ed) Biotechnology in Agriculture and Forestry, Vol 17 (pp 395–416). Springer-Verlag, BerlinGoogle Scholar
  27. RedenbaughK, FujiiJ & SladeD (1991b) Synthetic seed technology. In: VasilIK (ed) Scale-up and Automation in Plant Propagation (pp 36–74). Academic Press Inc. OrlandoGoogle Scholar
  28. ReinertJ, TazawaM & SemenoffS (1967) Nitrogen compounds as factors of somatic embryogenesisin vitro. Nature 216: 1215–1216PubMedGoogle Scholar
  29. SchenkH & HildebrandtA (1972) Medium and techniques for induction and growth of monocotyledonous and dicotyledonous cell cultures. Can J. Bot. 50: 199–204Google Scholar
  30. SchullerA, ReutherG & GeigerT (1989) Somatic embryogenesis for seeds explant ofAbies alba. Plant Cell Tiss. Org. Cult. 17: 53–58Google Scholar
  31. ShettyK & McKersieBD (1993) Proline, thioproline and potassium mediated stimulation of somatic embryogenesis in alfalfa (Medicago sativa L.) Plant Sci. 88: 185–193CrossRefGoogle Scholar
  32. SkokutTA, ManchesterJ & ShaeferJ (1985) Regeneration in alfalfa tissue culture — Stimulation of somatic embryo production by amino acids and N-15 and NMR determination of nitrogen utilisation. Plant Physiol. 79: 579–583Google Scholar
  33. StuartD & StricklandS (1984) Somatic embryogenesis from cell cultures ofMedicago sativa L. I. The role of amino acid additions to the regeneration medium. Plant Sci. Lett. 34: 165–174Google Scholar
  34. StuartD, NelsenJ, StricklandS & NicholJ (1985) Factors affecting developmental processes in alfalfa cell culture. In: HenkeR, HughesK, ConstantinM & HollaenderA (eds) Tissue Culture in Forestry and Agriculture (pp 59–73). Plenum, New YorkGoogle Scholar
  35. Valera-MonteroLL & Ochao-AlejoN (1992) A novel approach for chilli peper (Capsicum annuum L.) plant regeneration: shoot induction in rooted hypocotyls. Plant Sci. 84: 215–219CrossRefGoogle Scholar
  36. van derValkP, ZaalMACM & Creemers-MolanaarJ (1989) Somatic embryogenesis and plant regeneration in inflorescence and seed derived callus cultures ofPoa pratensis L. (Kentucky bluegrass). Plant Cell Rep. 7: 644–647Google Scholar
  37. WetherellDF & DougallDK (1976) Sources of nitrogen supporting growth and embryogenesis in cultured wild carrot tissue. Physiol. Plant. 37: 97–103Google Scholar
  38. WhitePR (1963) The Cultivation of Animal and Plant Cells. 2nd edition, Ronald Press, New YorkGoogle Scholar

Copyright information

© Kluwer Academic Publishers 1996

Authors and Affiliations

  • Saadet Buyukalaca
    • 1
  • Ferda Mavituna
    • 1
  1. 1.Department of Chemical EngineeringUniversity of Manchester Institute of Science and TechnologyManchesterUK

Personalised recommendations