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Production of nematode free plantlets in Polianthes tuberosa using in vitro culture techniques

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Abstract

Tuberose (Polianthes tuberosa) cultivation is tremendously affected due to Meloidogyne incognita infection. Histological study of in vitro nematode infected tuberose roots showed that the root infection initiated within 2 days post inoculation (DPI) and root gall formation occurred at 6 DPI indicating established infection on the roots. The life cycle of M. incognita in tuberose roots completed within 45 DPI evident by the formation of large number of eggs. Our study established in vitro methods like shoot tip culture and callus mediated regeneration of tubers collected from nematode infection fields to obtain completely nematode free plantlets. Tubers of different varieties produced multiple shoot bud on Murashige and Skoog (MS) media containing 4 mg L−1 6-benzylaminopurine (BAP) and 0.1 mg L−1 α-naphthaleneacetic acid (NAA). Maximum numbers of plantlets were obtained for Calcutta Single (14.4 ± 2.0 per plant). Embryogenic callus was induced on MS medium supplemented with altered concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D), NAA and BAP from leaf, flower, root and tuber explant. The maximum callus induction was obtained on MS containing 1 mg L−1 2,4-D, 1 mg L−1 NAA and 0.5 mg L−1 BAP (100%) and 1 mg L−1 2,4-D and 2.25 mg L−1 BAP (96.7%). Regeneration of tuber callus was achieved on MS with 0.5 mg L−1 Kinetin (KIN) within 3 weeks. Plantlets were rooted on ½ MS with 0.5 mg L−1 Indole-3-butyric acid for 25 days. The in vitro regeneration protocol developed can thus be used for producing disease free plantlets for mass propagation.

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Abbreviations

MS:

Murashige and Skoog

BAP:

6-Benzylaminopurine

NAA:

α-Naphthaleneacetic acid

2,4-D:

2,4-Dichlorophenoxyacetic acid

KIN:

Kinetin

IBA:

Indole-3-butyric acid

DPI:

Days post inoculation

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Acknowledgements

The authors wish to thank Department of Science and Technology (SERB) for funding the project to carry out the research work. We would like to acknowledge and extend our gratitude to IARI, ICAR, New Delhi, India for infrastructure facilities and Amity University, Noida for providing Ph.D. registration.

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KBMS and MJ performed the experiments, statistical analysis and wrote the manuscript and editing. UR planned the nematode infection experiments. RS has provided the tuberose tubers from the nematode infected plots and planned the tissue culture experiments along with SC. PKM has done the final editing of the manuscript and also helped in the microscopy work. The manuscript has been read and approved by all the authors.

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Correspondence to Jayanthi Madhavan.

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The authors declare that there are no conflicts of interest.

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Communicated by Tae-Ho Han, Ph.D.

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Singh, K.B.M., Madhavan, J., Sadhukhan, R. et al. Production of nematode free plantlets in Polianthes tuberosa using in vitro culture techniques. Hortic. Environ. Biotechnol. 61, 929–937 (2020). https://doi.org/10.1007/s13580-020-00263-5

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