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High-throughput in vitro culture system targeting genetic transformation in sugarcane

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Abstract

Conventional recombination breeding suffers setbacks for genetic improvement of sugarcane owing to its narrow gene pool, large complex genome, rare flowering, low fertility, long breeding cycle, and complex environmental interactions. However, production of transgenic plants can be a better alternative to improve quality traits and resistance to biotic and abiotic stresses. The authors report a ready in hand efficient in vitro culture protocol for genetic transformation in a popular sugarcane cv. Sabita (CoOR 03151) of Odisha. 2, 4-D at 3.0 mg/l resulted in highest callus induction frequency (87.8%) with white, friable, and nodular embryogenic calli suitable for plantlet regeneration. 2 mg/l BAP resulted in moderately higher number of shoots/responsive callus and higher percentage of plant survival. However, addition of TDZ (0.05 m/l) with 2 mg/l BAP in R medium (modified MS) may be considered optimum for regeneration of profuse healthy plantlets. A combination of PBZ (0.05 mg/l) with BAP (2 mg/l) and TDZ (0.05 mg/l) revealed profuse multiple shoots (25 microshoots/responsive callus) and higher percentage of survival during follow-up plant establishment than above hormonal supplementation. Among various hormone recipes in MS medium with 3.0 mg/l NAA resulted in excellent rhizogenesis response (88.0%) with more or less normal rooting within 2 weeks. The above efficient and highly reproducible in vitro culture system can be amenable for genetic transformation in this crop.

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Abbreviations

BAP:

6-Benzylaminopurine

2,4-D:

2,4-Dicchlorophenoxyacetic acid

GA3 :

Gibberellic acid

IAA:

Indole-3-acetic acid

IBA:

Indole-3-butyric acid

Kn:

Kinetin(6-Furfuryladenopurine)

NAA:

α-Naphthalene acetic acid

PBZ:

Paclobutrazol

PVP:

Polyvinylpyrrolidone

RH:

Relative humidity

TDZ:

Thidizuron

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Acknowledgements

We sincerely acknowledge and thank all researchers for their valuable contributions included in the text as references. Besides, special thanks to Breeder, AICRP (Sugarcane), Nayagarh, Odisha (OUAT) for providing the plant material for the study.

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This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

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Authors and Affiliations

  1. Department of Agricultural Biotechnology, College of Agriculture, Odisha University of Agriculture & Technology (OUAT), Bhubaneswar, 751003, India

    Swapan K. Tripathy & Dinesh Manohar Ithape

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  1. Swapan K. Tripathy
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  2. Dinesh Manohar Ithape
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SKT conceived the idea, performed the study and data analysis, and wrote the paper. DMI assisted in the in vitro culture work.

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Correspondence to Swapan K. Tripathy.

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Tripathy, S.K., Ithape, D.M. High-throughput in vitro culture system targeting genetic transformation in sugarcane. J. Crop Sci. Biotechnol. 23, 325–335 (2020). https://doi.org/10.1007/s12892-020-00040-x

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