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Iron supplementation promotes in vitro shoot induction and multiplication of Baptisia australis

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Abstract

A simple and efficient system was developed for the rapid in vitro multiplication of Baptisia australis using nodal explants derived from axenic cultures. Among the four cytokinins [benzylaminopurine (BA), kinetin, N6-(2-isopentenyl)adenine (2-iP) and zeatin] tested for axillary bud induction, BA at 4.4 µM proved to be the most effective treatment for induction of multiple shoots from nodal explants after six weeks. Supplementation of Murashige and Skoog (MS) and Driver and Kuniyuki walnut (DKW) media with a chelated iron source, ethylenediamine Bis (2-hydroxyphenylacetic acid) (FeEDDHA) and BA had a favorable effect on axillary bud proliferation from explants (apical buds and nodal segments) further improving the regeneration response as well as number of shoots induced. DKW media proved to be superior to MS for multiple shoot induction of B. australis. A combination of 4.4 µM BA and 23 µM of FeEDDHA in DKW produced a positive response resulting in 100% of nodal segments responding and regenerating 7.2 shoots per explant. The highest rooting percentage was observed on MS media containing 1 µM IBA. In vitro-rooted plants were successfully acclimated in a greenhouse with a survival rate of 97%. The micropropagation method described in this study has a potential for commercial multiplication of this horticultural plant.

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Abbreviations

BA:

6-Benzylaminopurine

DKW:

Driver and Kuniyuki walnut

FeEDDHA:

Ethylenediamine Bis (2-hydroxyphenylacetic acid) ferric-sodium salt

IBA:

Indole-3-butyric acid

MS:

Murashige and Skoog

NAA:

Naphthalene acetic acid

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Acknowledgements

The authors are grateful to the Ontario Ministry of Agriculture and Rural Affairs for funding and also wish to thank Robert Nichols for general lab support, Jeremy Boychyn for providing plant materials and Shuping Li for technical assistance with media preparation.

Author contributions

PP designed and carried out all the experiments, analyzed the data, prepared the manuscript. MS helped with designing the experiments, arranged for technical help and revised the manuscript. JAS read and revised the manuscript. PKS conceived the work, revised and finalized the manuscript. All authors read and revised the manuscript.

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Authors and Affiliations

  1. Gosling Research Institute for Plant Preservation, Department of Plant Agriculture, University of Guelph, Guelph, ON, N1G 2W1, Canada

    Priya Padmanabhan, Mukund R. Shukla, J. Alan Sullivan & Praveen K. Saxena

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  1. Priya Padmanabhan
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  2. Mukund R. Shukla
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  3. J. Alan Sullivan
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  4. Praveen K. Saxena
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Correspondence to Priya Padmanabhan.

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Padmanabhan, P., Shukla, M.R., Sullivan, J.A. et al. Iron supplementation promotes in vitro shoot induction and multiplication of Baptisia australis . Plant Cell Tiss Organ Cult 129, 145–152 (2017). https://doi.org/10.1007/s11240-016-1165-4

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  • DOI: https://doi.org/10.1007/s11240-016-1165-4

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