Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 107, Issue 2, pp 243–250 | Cite as

Encapsulation of cauliflower (Brassica oleracea var botrytis) microshoots as artificial seeds and their conversion and growth in commercial substrates

  • Hail Z. Rihan
  • Mohammed Al-Issawi
  • Stephen Burchett
  • Michael P. FullerEmail author
Original Paper


An effective protocol for the mass production of cauliflower microshoots was refined using the meristematic layer of cauliflower curd. After the meristematic layer was surface sterilized and shaved off, a commercial blender was used for homogenization and several blending treatments were tested in the range 15–120 s and 30 s was found to be optimal in terms of the amount explants produced and their subsequent growth ability. Explants were cultivated in S23 liquid medium (4.4 g L−1 MS (Murashige and Skoog) and 3% v/w sucrose) supplemented with several combinations of plant growth regulators (PGRs) including 1 and 2 mg L−1 of Kinetin in combination with three types of auxins (indole butyric acid (IBA), Naphthaleneacetic acid (NAA) and Indole-3-acetic acid (IAA)), each at 1 and 2 mg L−1 concentration. The use of 2 mg L−1 Kinetin and 1 mg L−1 IBA gave the best results in terms of its effects on explant induction. Microshoots of different sizes were encapsulated in a sodium alginate matrix and the optimal stage suitable for the production of artificial seeds was assessed in terms of both subsequent conversion and plantlet viability. The feasibility of cultivating cauliflower artificial seeds in commercial substrates (compost, vermiculite, perlite and sand) irrigated with different solution mixtures including sterilized distilled water (SDW), PGRs-free S23 medium and S23 medium supplemented with Kinetin (1 and 2 mg L−1) and IBA or NAA at (1 and 2 mg L−1) was investigated. The use of 2 mg L−1 Kinetin and 2 mg L−1 NAA applied with S23 gave the optimal response with both perlite and compost. This study showed high growth capacity of cauliflower artificial seeds in commercial substrates which is considered a promising step for their direct use in vivo.


Sodium alginate Encapsulation Curd Meristems Artificial seeds 



The authors gratefully acknowledge the provision of a PhD grant from the University of Damascus, Syria.


  1. Antonietta G, Ahmad H, Maurizio M, Alvaro S (2007) Preliminary research on conversion of encapsulated somatic embryos of Citrus reticulata Blanco, cv. Mandarino Tardivo di Ciaculli. Plant Cell Tiss Organ Cult 88(1):117–120CrossRefGoogle Scholar
  2. Böttgor M (1974) Apical dominance in roots of Pisum sativum L. Planta (Ber1) 121:253–261CrossRefGoogle Scholar
  3. Chand S, Singh AK (2004) Plant regeneration from encapsulated nodal segments of Dalbergia sissoo Roxb. a timber-yielding leguminous tree species. J Plant Physiol 161(2):237–243PubMedCrossRefGoogle Scholar
  4. Cooke RC (1979) Homogenization as an aid in tissue culture propagation of Platycerium and Davallia. Hortic Sci 14:21–22Google Scholar
  5. Eriksen EN (1974) Root Formation in Pea Cuttings. Physiol Plant 30(2):163–167CrossRefGoogle Scholar
  6. Fujii J, Slade D, Redenbaugh K (1989) Maturation and greenhouse planting of alfalfa artificial seeds. In Vitro Cell Dev Bio Plant 25(12):1179–1182CrossRefGoogle Scholar
  7. Germanà M, Micheli M, Chiancone B, Macaluso L, Standardi A (2011) Organogenesis and encapsulation of in vitro-derived propagules of Carrizo citrange. Citrus sinensis (L.);Poncirius trifoliata (L.). Plant Cell Tiss Organ Cult 78:1–9Google Scholar
  8. Goodwin PB, Morris SC (1979) Application of phytohormones to pea roots after removal of the apex: effect on lateral root production. Aust J Plant Physiol 6:195–200CrossRefGoogle Scholar
  9. Grout BWW, Crisp P (1977) Practical aspects of the propagation of cauliflower by meristem culture. Acta Hort 78:289–296Google Scholar
  10. Hinchee MAW, Rost TL (1986) The Control of Lateral Root Development in Cultured Pea Seedlings. I. The Role of Seedling Organs and Plant Growth Regulators. Bot Gaz 147(2):137CrossRefGoogle Scholar
  11. Janssens J, Sepelie M (1989) Rapid multiplication of ferns by homogenization. Sci Hortic 38:161–164CrossRefGoogle Scholar
  12. Kieffer M, Simkins N, Fuller MP, Jellings AJ (2001) A cost effective protocol for in vitro mass propagation of cauliflower. Plant Sci 160(5):1015–1024PubMedCrossRefGoogle Scholar
  13. Knauss JF (1976) A partial tissue culture method for pathogen-free propagation of selected ferns from spores. Proc Fla State Hort Soc 89:63–365Google Scholar
  14. Komor E, Liegl I, Schobert C (1993) Loading and translocation of various cytokinis in phloem and xylem of the seedlings of Ricinus-communis L. Planta 191(2):252–255CrossRefGoogle Scholar
  15. Kuiper D (1988) Growth Responses of Plantago major L. ssp. pleiosperma (Pilger) to Changes in Mineral Supply: Evidence for Regulation by Cytokinins. Plant Physiol 87(3):555–557PubMedCrossRefGoogle Scholar
  16. Kuiper D, Kuiper PJC, Lambers H, Schuit J, Staal M (1989) Cytokinin concentration in relation to mineral nutrition and benzyladenine treatment in Plantago major ssp. pleiosperma. Physiol Plant 75(4):511–517CrossRefGoogle Scholar
  17. Kumar A, Kumar VA, Kumar J (1993) Rapid in vitro propagation of cauliflower. Plant Sci 90:175–178CrossRefGoogle Scholar
  18. Machii H, Yamanouchi H (1993) Growth of mulberry synthetic seeds on vermiculite, sand and soil media. J SericSci Jpn 62:85–87Google Scholar
  19. Malabadi RB, Staden Jv (2005) Storability and germination of sodium alginate encapsulated somatic embryos derived from the vegetative shoot apices of mature Pinus patula trees. Plant Cell Tiss Organ Cult 82(3):259–265CrossRefGoogle Scholar
  20. Mandal J, Pattnaik S, Chand P (2000) Alginate encapsulation of axillary buds of;Ocimum americanum L. (hoary basil), Basilicum L. (sweet basil), Gratissimum L. (shrubby basil), and Sanctum L. In Vitro Cell Dev Biol Plant 36(4):287–292CrossRefGoogle Scholar
  21. Micheli M, Pellegrino S, Piccioni E, Standardi A (2002) Effects of double encapsulation and coating on synthetic seed conversion in M.26 apple rootstock. J Microencapsul 19:347–356PubMedCrossRefGoogle Scholar
  22. Micheli M, Hafiz IA, Standardi A (2007) Encapsulation of in vitro-derived explants of olive (Olea europaea L. cv. Moraiolo): II. Effects of storage on capsule and derived shoots performance. Sci Hortic 113(3):286–292CrossRefGoogle Scholar
  23. Mishra J, Singh M, Palni L, Nandi S (2010) Assessment of genetic fidelity of encapsulated microshoots of Picrorhiza kurrooa. Plant Cell Tiss Organ Cult :1–6Google Scholar
  24. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15(3):473–497CrossRefGoogle Scholar
  25. Naik SK, Chand PK (2006) Nutrient-alginate encapsulation of in vitro nodal segments of pomegranate (Punica granatum L.) for germplasm distribution and exchange. Sci Hortic 108(3):247–252CrossRefGoogle Scholar
  26. Nakashimada Y, Uozumi N, Kobayashi T (1995) Production of plantlets for use as artificial seeds from horseradish hairy roots fragmented in a blender. J Ferment 79(5):458–464CrossRefGoogle Scholar
  27. Nyende A, Schittenhelm S, Mix-Wagner G, Greef J-M (2003) Production, storability, and regeneration of shoot tips of potato (Solanum tuberosum L.) encapsulated in calcuim alginate hollow beads. In Vitro Cell Dev Biol Plant 39(5):540–544CrossRefGoogle Scholar
  28. Pintos B, Bueno M, Cuenca B, Manzanera J (2008) Synthetic seed production from encapsulated somatic embryos of cork oak (Quercus suber L.) and automated growth monitoring. Plant Cell Tiss Organ Cult 95(2):217–225CrossRefGoogle Scholar
  29. Pow JJ (1969) Clonal propagation in vitro from cauliflower curd. Hortic Sci 9:151–152Google Scholar
  30. Rai MK, Jaiswal VS, Jaiswal U (2008) Encapsulation of shoot tips of guava (Psidium guajava L.) for short-term storage and germplasm exchange. Sci Hortic 118(1):33–38CrossRefGoogle Scholar
  31. Rani Debi B, Taketa S, Ichii M (2005) Cytokinin inhibits lateral root initiation but stimulates lateral root elongation in rice (Oryza sativa). J Plant Physiol 162(5):507–515PubMedCrossRefGoogle Scholar
  32. Roy B, Mandal AB (2008) Development of synthetic seeds involving androgenic and pro-embryos in elite indica rice. Ind J Biotechnol 7(4):515–519Google Scholar
  33. Sandoval-Yugar EW, Dal Vesco LL, Steinmacher DA, Stolf EC, Guerra MP (2009) Microshoots encapsulation and plant conversion of Musa sp. cv. ‘Grand Naine’. Ciencia Rural 39(4)Google Scholar
  34. Singh S, Rai M, Asthana P, Pandey S, Jaiswal V, Jaiswal U (2009) Plant regeneration from alginate-encapsulated shoot tips of Spilanthes acmella L. Murr., a medicinally important and herbal pesticidal plant species. Acta Physiologiae Plantarum 31(3):649–653CrossRefGoogle Scholar
  35. Soneji JR, Rao PS, Mhatre M (2002) Germination of synthetic seeds of pineapple (Ananas comosus L. Merr.). Plant Cell Rep 20:891–894CrossRefGoogle Scholar
  36. Standardi A, Piccioni E (1998) Recent Perspectives on Synthetic Seed Technology Using Nonembryogenic In vitro-Derived Explants. Plant Sci 159:968–978Google Scholar
  37. Sundararaj SG, Agrawal A, Tyagi RK (2010) Encapsulation for in vitro short-term storage and exchange of ginger (Zingiber officinale Rosc.) germplasm. Sci Hortic 125(4):761–766CrossRefGoogle Scholar
  38. Taha RM, Hasbullah NA, Awal A (2009) Production of synthetic seeds from micro shoots and somatic embryos of Gerbera jamesoni bolus ex. hook f. Acta Hort 829:91–98Google Scholar
  39. Teng WL, Ngai YW (1999) Regeneration of Oxalis triangularis ssp. triangularis from suspension cells cultured in three different systems (solid, liquid-flask and bioreactor cultures). Plant Cell Rep 18(9):701–706CrossRefGoogle Scholar
  40. Teng WL, Teng MC (1997) In vitro regeneration patterns of Platycerium bifurcatum leaf cell suspension culture. Plant Cell Rep 16(12):820–824CrossRefGoogle Scholar
  41. Vandemoortele J-L, Kevers C, Billard J-P, Gaspar T (2001) Osmotic pretreatment promotes axillary shooting from cauliflower curd pieces by acting through internal cytokinin level modifications. J Plant Physiol 158(2):221–225CrossRefGoogle Scholar
  42. Ziv M, Ariel T (1991) Bud proliferation and plant regeneration in liquid-cultured philodendron treated with ancymidol and paclobutrazol. J Plant Growth Regul 10(1):53–57CrossRefGoogle Scholar
  43. Ziv M, Ronen G, Raviv M (1998) Proliferation of meristematic clusters in disposable presterilized plastic bioreactors for the large-scale micropropagation of plants. In Vitro Cell and Dev Biol Plant 34:152–158CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Hail Z. Rihan
    • 1
  • Mohammed Al-Issawi
    • 1
  • Stephen Burchett
    • 1
  • Michael P. Fuller
    • 1
    Email author
  1. 1.School of Biomedical and Biological SciencesFaculty of Science and Technology, University of PlymouthPlymouthUK

Personalised recommendations