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An improved plant regeneration system and ex vitro acclimatization of Ocimum basilicum L.

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Abstract

An efficient, rapid and large scale propagation of a multipurpose herb, Ocimum basilicum through in vitro culture of nodal segments with axillary buds from mature plants has been accomplished. Among the cytokinins, 6-benzyladenine (BA), thidiazuron (TDZ), kinetin (Kin) and 2-isopentenyl adenine (2-iP) tested as supplements to Murashige and Skoog (MS) medium, 5.0 μM BA was optimum in inducing bud break. The highest rate of shoot multiplication was achieved on half-strength MS medium supplemented with 2.5 μM BA and 0.5 μM indole-3-acetic acid (IAA) combination. The shoots regenerated from TDZ supplemented medium when subcultured to hormone-free MS medium considerably increased the rate of shoot multiplication and shoot length by the end of third subculture. For rooting, MS medium supplemented with 1.0 μM indole-3-butyric acid (IBA) proved to be better than that supplemented with IAA or α-naphthalene acetic acid (NAA). The in vitro raised plantlets with well developed shoots and roots were successfully established in earthen pots containing garden soil and were grown in greenhouse with 90% survival rate. Chlorophyll a and b, carotenoids and net photosynthetic rate were measured in leaves during ex vitro acclimatization at 0, 7, 14, 21 and 28 days. Firstly these parameters showed a decreasing trend but subsequently increased after 7 days of acclimatization. These findings indicate that the adaptation of micropropagated plants to ex vitro conditions is more extended in time than generally accepted.

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Abbreviations

BA:

6-Benzyladenine

IAA:

Indole-3-acetic acid

IBA:

Indole-3-butyric acid

2-iP:

2-Isopentenyl adenine

Kin:

Kinetin

NAA:

α-Naphthalene acetic acid

PN :

Net photosynthetic rate

TDZ:

Thidiazuron

References

  • Anonymous (1966) The wealth of India: a dictionary of Indian raw materials and industrial products, vol 7. CSIR, New Delhi, pp 79–89

    Google Scholar 

  • Anonymous (2001) The wealth of India: a dictionary of Indian raw materials and industrial products, vol 7. CSIR, New Delhi, pp 217–218

    Google Scholar 

  • Babu KN, Anu A, Remashree AB, Praveen K (2000) Micropropagation of curry leaf tree. Plant Cell Tissue Organ Cult 61:199–203

    Article  CAS  Google Scholar 

  • Begum F, Amin MN, Azad MAK (2002) In vitro rapid clonal propagation of Ocimum basilicum L. Plant Tissue Cult 12:27–35

    Google Scholar 

  • Estrada-Luna AA, Davies FT Jr, Egilla JN (2001) Physiological changes and growth of micropropagated chile ancho pepper plantlets during acclimatization and post acclimatization. Plant Cell Tissue Organ Cult 66:17–24

    Article  CAS  Google Scholar 

  • Faisal M, Anis M (2003) Rapid mass propagation of Tylophora indica Merrill via leaf callus culture. Plant Cell Tissue Organ Cult 75:125–129

    Article  CAS  Google Scholar 

  • Faisal M, Ahmad N, Anis M (2005) Shoot multiplication in Rauvolfia tetraphylla L. using thidiazuron. Plant Cell Tissue Organ Cult 80:187–190

    Article  CAS  Google Scholar 

  • Faisal M, Siddique I, Anis M (2006) An efficient plant regeneration system for Mucuna pruriens L. (DC.) using cotyledonary node explants. In Vitro Cell Dev Biol Plant 42:59–64

    Article  CAS  Google Scholar 

  • Heywood VH (1978) Flowering plants of the world. Oxford University Press, Oxford, pp 239–240

    Google Scholar 

  • Husain MK, Anis M (2006) Rapid in vitro propagation of Eclipta alba (L.) Hassk. through high frequency axillary shoot proliferation. Acta Physiol Plant 28:325–330

    Article  CAS  Google Scholar 

  • Jang GW, Kim KS, Park RD (2003) Micropropagation of Venus fly trap by shoot culture. Plant Cell Tissue Organ Cult 72:95–98

    Article  CAS  Google Scholar 

  • Jeon MW, Ali MB, Hahn EJ, Paek KY (2005) Effects of photon flux density on the morphology, photosynthesis and growth of a CAM orchid, Doritaenopsis during post micropropagation acclimatization. Plant Growth Regul 45:139–147

    Article  CAS  Google Scholar 

  • Kadlecek P, Ticha I, Capkova V, Schafer C (1998) Acclimatization of micropropagated tobacco plantlets. In: Garab G (ed) Photosynthesis: mechanism and effects, Vol. 5. Kluwer Academic Publishers, Dordrecht, Boston, London, pp 3853–3856

    Google Scholar 

  • Kozai T (1991) Micropropagation under autotrophic conditions. In: Debergh PC, Zimmermann R (eds) Micropropagation: technology and application. Kluwer Academic Publisher, Dordrecht, The Netherlands, pp 447–469

    Chapter  Google Scholar 

  • Kumar AD, Seeni S (1998) Rapid clonal multiplication through in vitro axillary shoot proliferation of Aegle marmelos (L.) Corr., a medicinal tree. Plant Cell Rep 17:422–426

    Article  Google Scholar 

  • Kumar R, Sharma K, Agrawal V (2005) In vitro clonal propagation of Holarrhena antidysenterica L. (wall.) through nodal explants of mature trees. In Vitro Cell Dev Biol Plant 41:137–144

    Article  CAS  Google Scholar 

  • Mac Kinney G (1941) Absorption of light by chlorophyll solution. J Biol Chem 140:315–322

    CAS  Google Scholar 

  • Maclachlan S, Zalik S (1963) Plastid structure, chlorophyll concentration and free amino acid composition of chlorophyll mutant by barley. Can J Bot 1053–1062

  • Martinez-Pulido C, Harry IS, Thorpe TA (1992) Optimization of bud induction in cotyledonary explants of Pinus canariensis. Plant Cell Tissue Organ Cult 29:247–255

    Article  CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue culture. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Murthy BNS, Victor J, Singh RPS, Fletcher RA, Saxena PK (1996) In vitro regeneration of chick pea (Cicer arietinum L.) : stimulation of direct organogenesis and somatic embryogenesis by thidiazuron. Plant Growth Regul 19:233–240

    Article  CAS  Google Scholar 

  • Preece JE, Sutter EG (1991) Acclimatization of micropropagated plants to the greenhouse and field. In: Debergh PC, Zimmerman RM (eds) Micropropagation: technology and applications. Kluwer Academic Publisher, Dordrecht, The Netherlands, pp 71–93

    Chapter  Google Scholar 

  • Pospisilova J, Ticha I, Kadlecek KP, Haisel D, Plzakova S (1999) Acclimatization of micropropagated plants to ex vitro conditions. Biol Plant 42:481–497

    Article  Google Scholar 

  • Sahoo Y, Pattnaik SK, Chand PK (1997) In vitro clonal propagation of a medicinal plant Ocimum basilicum L. (Sweet basil) by axillary shoot proliferation. In Vitro Cell Dev Biol Plant 33:293–296

    Article  Google Scholar 

  • Siddique I, Anis M (2007a) In vitro shoot multiplication and plantlet regeneration from nodal explants of Cassia angustifolia (Vahl.): a medicinal plant. Acta Physiol Plant 29:233–238

    Article  CAS  Google Scholar 

  • Siddique I, Anis M (2007b) Rapid micropropagation of Ocimum basilicum using shoot tip explants pre-cultured in thidiazuron supplemented liquid medium. Biol Plant 51:787–790

    Article  CAS  Google Scholar 

  • Singh NK, Sehgal CB (1999) Micropropagation of Holy Basil (Ocimum sanctum Linn.) from young inflorescences of mature plants. Plant Growth Regul 29:161–166

    Article  CAS  Google Scholar 

  • Van Huylenbroeck JM, Debergh PC (1996) Impact of sugar concentration in vitro on photosynthesis and carbon metabolism during ex vitro acclimatization of Spathiphyllum plantlets. Physiol Plant 96:298–304

    Article  Google Scholar 

  • Van Huylenbroeck JM, Piqueras A, Debergh PC (1998) Photosynthesis and carbon metabolism in leaves formed prior and during ex vitro acclimatization of micropropagated plants. Plant Sci 134:21–30

    Article  Google Scholar 

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Acknowledgments

Financial support from the DST-FIST Programme (2005), Govt. of India, New Delhi is highly appreciated.

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Authors and Affiliations

  1. Plant Biotechnology Laboratory, Department of Botany, Aligarh Muslim University, Aligarh, 202002, India

    Iram Siddique & M. Anis

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  1. Iram Siddique
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  2. M. Anis
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Correspondence to M. Anis.

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Communicated by E. Lojkowska.

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Siddique, I., Anis, M. An improved plant regeneration system and ex vitro acclimatization of Ocimum basilicum L.. Acta Physiol Plant 30, 493–499 (2008). https://doi.org/10.1007/s11738-008-0146-6

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  • DOI: https://doi.org/10.1007/s11738-008-0146-6

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