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Efficient direct plant regeneration from immature leaf roll explants of sugarcane (Saccharum officinarum L.) using polyamines and assessment of genetic fidelity by SCoT markers

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Abstract

A rapid and efficient method for in vitro direct plant regeneration from immature leaf roll explants of Saccharum officinarum L. (sugarcane) cv. Co 86032 was developed by the application of exogenous polyamines (PA). The effect of explant source from apical meristems and pre-culture of explants in the dark on shoot regeneration was studied. Adventitious shoot regeneration occurred on the proximal regions of immature leaf roll explants when pre-incubated in the dark for 2 wk and the regeneration response was decreased from the middle to distal end. A higher number of direct shoots (130 primary shoots explant−1) and multiple shoots (657 secondary shoots explant−1), were obtained with a combination of spermidine (103.27 μM), spermine (49.42 μM), and putrescine (31.04 μM) along with plant growth regulators. Shoot induction was increased up to twofold and multiplication was increased up to threefold in the medium supplemented with PA. Profuse rooting was observed in putrescine (93.12 μM), spermidine (68.84 μM), and spermine (24.71 μM), with mean number of 57 roots. A twofold increase in the number of roots was observed in medium supplemented with PA with respect to control cultures, which facilitated the successful transplantation and acclimatization process of in vitro propagated sugarcane plants. Histology and scanning electron microscopy analyses supported adventitious direct shoot regeneration from immature leaf roll explants. The genetic stability of in vitro regenerated plants was confirmed using start codon targeted polymorphism marker system.

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Acknowledgments

The authors are grateful to the University Grants Commission (UGC), Government of India, for the financial support [F.No.41-516/2012 (SR) and No.F.3-27/2011 (SAP-II)].

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Authors and Affiliations

  1. Department of Biotechnology, Bharathidasan University, Tiruchirappalli, Tamil Nadu, 620024, India

    Dorairaj Sathish, Venkatachalam Vasudevan, Dhandapani Elayaraja & Markandan Manickavasagam

  2. Department of Biotechnology, Bishop Heber College, Tiruchirappalli, Tamil Nadu, 620017, India

    Jeevaraj Theboral

  3. Division of Crop Improvement, Indian Council of Agricultural Research-Sugarcane Breeding Institute, Coimbatore, Tamil Nadu, 641007, India

    Chinnaswamy Appunu

  4. Department of Biotechnology, Vellore Institute of Technology, Vellore, Tamil Nadu, 632014, India

    Ramamoorthy Siva

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  1. Dorairaj Sathish
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Correspondence to Markandan Manickavasagam.

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Editor: Jayasankar Subramanian

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Figure s1

Banding pattern obtained from Start Codon Targeted (SCoT) polymorphism marker analysis a) S4, b) S5, c) S6, d) S7, e) S8, f) S9, g) S10, h) S17, and i) S25 in donor mother plant, and in vitro regenerated plants of Saccharum officinarum L. (M - mother plant; Lane 1 to 8 – in vitro regenerated plants). (JPG 6775 kb)

Figure s2

Banding pattern obtained from Start Codon Targeted (SCoT) polymorphism marker analysis a) S26, b) S32, c) S33, and d) S34 in donor mother plant, and in vitro regenerated plants of Saccharum officinarum L. (M - mother plant; Lane 1 to 8 – in vitro regenerated plants). (JPG 3092 kb)

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Sathish, D., Vasudevan, V., Theboral, J. et al. Efficient direct plant regeneration from immature leaf roll explants of sugarcane (Saccharum officinarum L.) using polyamines and assessment of genetic fidelity by SCoT markers. In Vitro Cell.Dev.Biol.-Plant 54, 399–412 (2018). https://doi.org/10.1007/s11627-018-9910-5

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