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Comparison of two PVS2-based procedures for cryopreservation of commercial sugarcane (Saccharum spp.) germplasm and confirmation of genetic stability after cryopreservation using ISSR markers

  • Cryopreservation
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Abstract

Conservation of Saccharum spp. germplasm as ex situ collections of plants has a high cost, and in natural conditions, the plants remain exposed to pests, pathogens, and natural disasters. Long-term preservation of plant germplasm is important for agricultural biodiversity and food safety, so the aim of this study was to develop a cryogenic procedure for cryopreservation of sugarcane germplasm. The first study compared droplet vitrification and encapsulation-vitrification techniques for cryopreservation of in vitro shoot tips of Saccharum spp. variety Halaii. The best regeneration rate (70.9%) was obtained from 45-min PVS2 vitrification solution-treated shoot tips via the droplet vitrification technique. This technique was tested on two other Saccharum sp. varieties, and the best regeneration rates for varieties NG 57-024 and H 83-6179 were 63.3 and 76.3%, respectively. Shoots derived from cryopreserved shoot tip buds developed well-formed roots, and were easily acclimated to greenhouse conditions. The second study evaluated genetic stability of the cryopreserved varieties using ten inter-simple sequence repeat primers. A total of 211 (Halaii), 198 (H83-6179), and 201 (NG 57-024) reproducible bands, ranging from 125 to 5500 bp, were scored with this technique. One hundred genetic stability was detected from Halaii and H 83-6179 whereas 98.5% genetic stability was detected from varieties of NG 57-024. The PCR reactions showed that there was no crucial variation on genetic stability for all cryopreserved varieties.

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Acknowledgements

This work was supported by Mugla Sitki Kocman University, Scientific Research Projects Coordination Unit (Mugla, Turkey, MSKU-BAP 16/021).

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Authors and Affiliations

  1. Faculty of Science, Molecular Biology and Genetics Department, Mugla Sitki Kocman University, 48000, Kotekli, Mugla, Turkey

    Ergun Kaya

  2. Embrapa Cassava & Fruits, Caixa Postal 007, Cruz das Almas, BA, 44380-000, Brazil

    Fernanda Vidigal Duarte Souza

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  1. Ergun Kaya
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  2. Fernanda Vidigal Duarte Souza
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Correspondence to Ergun Kaya.

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Editor: Barbara Reed

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Kaya, E., Souza, F.V.D. Comparison of two PVS2-based procedures for cryopreservation of commercial sugarcane (Saccharum spp.) germplasm and confirmation of genetic stability after cryopreservation using ISSR markers. In Vitro Cell.Dev.Biol.-Plant 53, 410–417 (2017). https://doi.org/10.1007/s11627-017-9837-2

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