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Effect of the NO donor “sodium nitroprusside” (SNP), the ethylene inhibitor “cobalt chloride” (CoCl2) and the antioxidant vitamin E “α-tocopherol” on in vitro shoot proliferation of Sideritis raeseri Boiss. & Heldr. subsp. raeseri

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Abstract

In the present study, the effects of the NO donor “sodium nitroprusside” (SNP) and the ethylene inhibitor “CoCl2” combined with 2.2 μM 6-benzyladenine (BA), and the antioxidant vitamin E “α-tocopherol” in combination with 2.2 μM zeatin (ZT) and 0.2 μM α-naphthaleneacetic acid (NAA) on in vitro shoot proliferation of Sideritis raeseri Boiss. & Heldr. subsp. raeseri, one of the seven endemic mountain tea species of the Greek flora were investigated. The basal nutrient culture medium used in all experiments was the MS (Murashige, Skoog, Physiol Plant 15:473–497, 1962). Taking simultaneously into consideration all shoot proliferation parameters evaluated, the explants performance was optimum and significantly better among others when treated with 2.2 μM BA + 10 μM SNP or 2.2 μM BA + 2.5 μM CoCl2, or 2.2 μM ZT + 0.2 μM NAA + 11.6 μM α-tocopherol. In particular, 10 μM SNP gave 4.6 ± 0.6 shoots/explant and 100% shoot multiplication percentage while 20 μM SNP exhibited the greatest shoot length (17.5 ± 1.8 mm). However, SNP (10–40 μM) caused hyperhydricity symptoms to the 30–100% of explants. CoCl2 applied at 2.5 μM led to the production of 5.5 ± 0.7 shoots/explant, 11.8 ± 1.1 mm shoot length and 100% shoot multiplication percentage and concurrently decreased the percentage of vitrified explants from 33.3 to 20%. 11.6 μM α-tocopherol gave 6.7 ± 0.8 shoots/explant, 14.5 ± 0.7 mm shoot length, 100% shoot multiplication percentage, diminishing vitrification percentage from 41.6 to 8.3% (33.3% decrease) and completely inhibiting the appearance of necrotic explants, indicating its antioxidant properties as a vitamin. Shoot-tip explants responded better and exhibited better shoot proliferation results when cultured in MS medium with 2.5 μM CoCl2 for 5 weeks or 11.6 μM α-tocopherol for 9 weeks rather than with SNP for 4 weeks. In this study, three new different protocols were established for the in vitro shoot proliferation stage of S. raeseri.

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Acknowledgements

This research project was funded under the Action “Research & Technology Development Innovation projects (AgroETAK)”, MIS 453350, in the framework of the Operational Program “Human Resources Development”. It was co-funded by the European Social Fund and by National Resources through the National Strategic Reference Framework 2007–2013 (NSRF 2007–2013) coordinated by the Hellenic Agricultural Organisation “DEMETER” (Institute of Genetic Breeding and Natural Resources/Scientific supervisor: Dr Eleni Maloupa).

Author contributions

VS performed all the experimental work, designed and managed the experiments, did all the statistics and wrote the first draft of the manuscript. EM contributed by scientific advices, checked the manuscript and gave her approval prior to its submission for potential publication. All the authors have read and approved the final manuscript.

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  1. Institute of Breeding and Plant Genetic Resources, Hellenic Agricultural Organization—‘Demeter’, Laboratory for the Conservation and Evalutation of Native and Floricultural Species, Thermi, Thessaloniki, Greece

    Virginia Sarropoulou & Eleni Maloupa

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Correspondence to Virginia Sarropoulou.

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Sarropoulou, V., Maloupa, E. Effect of the NO donor “sodium nitroprusside” (SNP), the ethylene inhibitor “cobalt chloride” (CoCl2) and the antioxidant vitamin E “α-tocopherol” on in vitro shoot proliferation of Sideritis raeseri Boiss. & Heldr. subsp. raeseri . Plant Cell Tiss Organ Cult 128, 619–629 (2017). https://doi.org/10.1007/s11240-016-1139-6

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