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5-Azacytidine as a tool to induce somaclonal variants with useful traits in sugarcane (Saccharum spp.)

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Abstract

A possible strategy to produce variant sugarcane plants with beneficial traits was tested by promoting somaclonal variation in vitro through the action of the hypomethylation and mutagenic agent 5-Azacytidine (Azac). Treatment of calli in liquid medium caused high levels of necrosis. Consequently, 6- to 8-week-old calli of cultivar NCo376 were exposed to 50 and 100 μM Azac in semi-solid callus induction medium (CIM) (MS salts and vitamins, sucrose, casein hydrolysate, agar, with or without 3 mg l−1 2,4-D) for 1 week. They were then transferred to fresh CIM with 2,4-D and to CIM without 2,4-D, for 2 and 8–10 weeks, respectively. The highest callus necrosis (>60 %) and reduced recovery (<40 %) were recorded for calli treated with 100 μM Azac without 2,4-D, which also resulted in lower plant yield (12 plantlets/0.2 g calli) than the control (18 plantlets/0.2 g calli). From methylation-sensitive amplified fragment length polymorphism analyses, the highest polymorphisms (4.2 %) were also obtained from plants derived from the 100 μM Azac treatment without 2,4-D. After 9 months of field growth, Azac-derived plants exhibited phenotypic differences compared with the controls. Ex vitro screening resulted in the identification of one plant from the 100 μM Azac with 2,4-D treatment putatively tolerant to smut, and three plants from the 100 μM Azac with 2,4-D and one from the 50 μM Azac with 2,4-D treatments, potentially tolerant to the herbicide imazapyr.

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Acknowledgments

The authors thank the University of KwaZulu-Natal, South Africa Sugar Research Institute (SASRI) and National Research Foundation (NRF) for support. A Munsamy is grateful to the NRF for a postgraduate scholarship. E Albertse (SASRI) is thanked for his assistance with AFLP analysis.

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Authors and Affiliations

  1. School of Life Sciences, University of KwaZulu-Natal, Westville Campus, P. Bag X54001, Durban, Kwa-Zulu Natal, 4000, South Africa

    Ashlin Munsamy, R. Stuart Rutherford, Sandy J. Snyman & M. Paula Watt

  2. South African Sugarcane Research Institute, P. Bag X02, Mount Edgecombe, KwaZulu-Natal, 4300, South Africa

    R. Stuart Rutherford & Sandy J. Snyman

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  1. Ashlin Munsamy
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  2. R. Stuart Rutherford
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Correspondence to Sandy J. Snyman.

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Munsamy, A., Rutherford, R.S., Snyman, S.J. et al. 5-Azacytidine as a tool to induce somaclonal variants with useful traits in sugarcane (Saccharum spp.). Plant Biotechnol Rep 7, 489–502 (2013). https://doi.org/10.1007/s11816-013-0287-y

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