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Direct shoot organogenesis from shoot tip explants of a highly medicinal valued tree Pterocarpus marsupium Roxb.

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Abstract

An in vitro regeneration system for propagation of a valuable medicinal tree, Pterocarpus marsupium, using shoot tip (ST) explants derived from 7-d-old axenic seedlings has been successfully developed. Murashige and Skoog (MS) medium containing cytokinins (BA, mT, or Kn) and auxins (NAA, IAA) in different concentrations and combinations showed to have a marked stimulatory effect on the regeneration output. Of cytokinins, meta-topolin (mT 7.0 μM) proved optimum dose for multiple shoot induction yielding 6.50 ± 0.49 shoots per explant with mean shoot length (4.00 ± 0.16 cm) in 70% cultures after 6 wk. Whereas, supplementation of low concentration of auxin (1.0 μM NAA) with optimal cytokinin (7.0 μM mT) favored enhanced shoot induction with increased percent response. At this level, a maximum of 13.54 ± 0.34 shoots per explant with (5.30 ± 0.10 cm) mean shoot length were recorded in 80% cultures after 12 wk. Thereafter, microshoots were isolated and their basal end treated with high doses of indole-3-butyric acid (250 μM) for 5 d followed to transplant onto various potting substrates for ex vitro root formation. A maximum of 3.63 ± 0.0.08 roots per microshoot and mean root length of 3.59 ± 0.07 cm with highest rooting frequency (67.7%) was observed after 4 wk of transplantation. During ex vitro rooting-cum-acclimatization, various physiological parameters were found to be fluctuating initially for 35 d; thereafter, an increasing trend was observed in net photosynthetic rate, intercellular CO2 concentration, and chlorophyll a/b estimation. Overall, the successfully acclimatized regenerants when transferred in natural condition showed about 96.7% survival rate after 3 mo. Assessment of genetic integrity of tissue culture raised plantlets was accomplished by DNA–based ISSR primers. A total of 35 bands with an average of 4.38 bands per primer were scored in monomorphic banding pattern. Thus, it is apparent that the mode of in vitro regeneration protocol was suitable for obtaining true-to-type and disease-free plants.

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Abbreviations

BA:

6-Benzyladenine

C i :

Intercellular CO2 concentration

DAT:

Days after transfer

g s :

Stomatal conductance

IAA:

Indole acetic acid

IBA:

Indole-3-butyric acid

Kn:

Kinetin

mT:

Meta-topolin

NAA:

α-Naphthalene acetic acid

PGRs:

Plant growth regulators

P N :

Photosynthetic rate

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Acknowledgments

The authors extend their appreciation to the International Scientific Partnership Program (ISPP) at the King Saud University for providing research support through ISPP (#0082). The authors are grateful to the District Forest Officer, Forest Department of Madhya Pradesh (India), for providing the fruits of P. marsupium. Award of the UGC-BSR Faculty Fellowship (2017) to M. Anis by the University Grants Commission, New Delhi, is gratefully acknowledged.

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Authors and Affiliations

  1. Department of Botany, Aligarh Muslim University, Aligarh, 202 002, India

    Anees Ahmad, Mohammad Anis & Mehrun Nisha Khanam

  2. Department of Botany and Microbiology, College of Science, King Saud University, P.O. Box 2455, Riyadh, 11451, Saudi Arabia

    Mohammad Anis & Abdulrahman A. Alatar

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  1. Anees Ahmad
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  2. Mohammad Anis
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  3. Mehrun Nisha Khanam
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  4. Abdulrahman A. Alatar
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Correspondence to Mohammad Anis.

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Editor: Yong Eui Choi

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Ahmad, A., Anis, M., Khanam, M.N. et al. Direct shoot organogenesis from shoot tip explants of a highly medicinal valued tree Pterocarpus marsupium Roxb.. In Vitro Cell.Dev.Biol.-Plant 56, 670–681 (2020). https://doi.org/10.1007/s11627-020-10115-7

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