Production of nematode free plantlets in Polianthes tuberosa using in vitro culture techniques
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RESEARCH REPORT
Production of nematode free plantlets in Polianthes tuberosa using in vitro culture techniques Kanchan B. M. Singh3 · Jayanthi Madhavan1 · Raghunath Sadhukhan2 · Shivani Chandra3 · Uma Rao1 · Pranab Kumar Mandal4 Received: 27 April 2019 / Revised: 13 May 2020 / Accepted: 19 May 2020 / Published online: 25 August 2020 © Korean Society for Horticultural Science 2020
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. Keywords Callus · Meloidogyne incognita · Shoot tip culture · Nematode · Polianthes tuberosa · Regeneration Abbreviations MS Murashige and Skoog BAP 6-Benzylaminopurine NAA α-Naphthaleneacetic acid 2,4-D 2,4-Dichlorophenoxyacetic acid KIN Kinetin Communicated by Tae-Ho Han, Ph.D. Kanchan B. M. Singh and Jayanthi Madhavan are First authors. * Jayanthi Madhavan [email protected] 1
Division of Nematology, Indian Agriculture Research Institute (IARI), Pusa, New Delhi 110012, India
2
Bidhan Chandra Krishi Viswavidyalaya, Mohanpur, West Bengal 741252, India
3
Amity Institute of Biotechnology, Amity University, Noida 201313, Uttar Pradesh, India
4
National Institute of Plant Biotechnology, New Delhi 110012, India
IBA Indole-3-butyric acid DPI Days post inoculation
1 Introduction Ornamental crops are of immense economic importance since long time. International demands for cut flowers are rapidly expanding in the industries (Lawson 1996). Among the ornamental crops tuberose (Polianthes tuberosa: Polianthes meaning “many flowers” in Greek and tuberosa meaning “swollen or tuberou
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