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In vitro plant regeneration from cotyledon-derived callus cultures of leguminous tree Gleditsia caspica Desf.

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Abstract

An in vitro regeneration system was developed using organogenic callus derived from in vitro grown cotyledonary explants of Gleditsia caspica Desf., an important leguminous tree. Murashige and Skoog (MS) basal medium augmented with 0.2 g L−1 myo-inositol and various concentrations of either 2,4-dichlorophenoxyacetic acid (2,4-D), naphthaleneacetic acid, or indole-3-butyric acid (IBA) alone as well as combined with cytokinins was used for callus induction. The highest frequency of organogenic yellowish-white and nodular callus (93 %) was obtained from explants grown on medium supplemented with 13.5 μM 2,4-D and 4.4 μM benzyladenine (BA). The yellowish-white and nodular callus when transferred to MS medium supplemented with BA (2.2–17.7 μM) or kinetin (KT; 2.3–18.8 μM) solely or in combination with 2.3 μM 2,4-D produced several microshoots after 5 weeks culture. The calli cultured on MS medium with 4.4 μM BA singly showed superior growth response and produced both maximum shoot regeneration (94 %) and the highest mean number (4.3) of microshoots per callus. Transfer of regenerated microshoots onto modified MS basal medium fortified with 5.8 μM gibberellic acid and 4.4 μM BA resulted in the maximum number of internodes per shoot and the highest shoot elongation after a period of 6 weeks. Optimum rooting of 90 %, an average 6.1 roots per shoot, and a mean root length of 3.6 cm was observed when half-strength MS medium was supplemented with 9.8 μM IBA and 0.92 μM KT. The regenerated healthy plants with well-developed shoots and roots showed a survival rate of 77 % after acclimatization and transplanting to garden soil for a 10-week hardening period under ex vitro conditions.

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Abbreviations

2,4-D:

2,4-Dichlorophenoxyacetic acid

BA:

N6-Benzyladenine

GA3 :

Gibberellic acid

IBA:

Indole-3-butyric acid

KT:

Kinetin

MI:

Myo-inositol

MS:

Murashige and Skoog

NAA:

Naphthalene acetic acid

PGR:

Plant growth regulator

WPM:

Woody plant medium

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Acknowledgments

The authors would like to thank Dr. Paula M. Pijut, USDA Forest Service, for her helpful comments on the preliminary version of this manuscript. This research was financially supported by the Ministry of Science, Research and Technology of Iran for University of Kurdistan.

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Authors and Affiliations

  1. Forest Biology Laboratory, Department of Forestry, Faculty of Natural Resources, University of Kurdistan, Khanagah Campus, 416, Sanandaj, Iran

    Fatemeh Zarinjoei & Mohammad Shafie Rahmani

  2. Department of Forestry, Faculty of Natural Resources, University of Kurdistan, Khanagah Campus, 416, Sanandaj, Iran

    Naghi Shabanian

Authors
  1. Fatemeh Zarinjoei
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  2. Mohammad Shafie Rahmani
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  3. Naghi Shabanian
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Corresponding author

Correspondence to Naghi Shabanian.

Additional information

Fatemeh Zarinjoei and Mohammad Shafie Rahmani contributed equally to this study and are considered co-first authors.

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Zarinjoei, F., Rahmani, M.S. & Shabanian, N. In vitro plant regeneration from cotyledon-derived callus cultures of leguminous tree Gleditsia caspica Desf.. New Forests 45, 829–841 (2014). https://doi.org/10.1007/s11056-014-9440-x

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