Please use this identifier to cite or link to this item: http://nopr.niscair.res.in/handle/123456789/32165
Title: A microdroplet cell culture based high frequency somatic embryogenesis system for pigeonpea, Cajanus cajan (L.) Millsp.
Authors: Kumar, Nagan Udhaya
Gnanaraj, Muniraj
Sindhujaa, Vajravel
Viji, Maluventhen
Manoharan, Kumariah
Keywords: Callus culture;Choline chloride;Crop improvement;Kinetin;Lithium chloride;Phytohormones;Plantlet regeneration;Polyethylene glycol;Putrescine;Spermidine
Issue Date: Sep-2015
Publisher: NISCAIR-CSIR, India
Abstract: A protocol for high frequency production of somatic embryos was worked out in pigeonpea, Cajanus cajan (L.) Millsp. The protocol involved sequential employment of embryogenic callus cultures, low density cell suspension cultures and a novel microdroplet cell culture system. The microdroplet cell cultures involved culture of a single cell in 10 µl of Murashige and Skoog’s medium supplemented with phytohormones, growth factors and phospholipid precursors. By employing the microdroplet cell cultures, single cells in isolation were grown into cell clones which developed somatic embryos. Further, 2,4-dichlorophenoxyacetic acid, kinetin, polyethylene glycol, putrescine, spermine, spermidine, choline chloride, ethanolamine and LiCl were supplemented to the low density cell suspension cultures and microdroplet cell cultures to screen for their cell division and somatic embryogenesis activity. Incubation of callus or the inoculum employed for low density cell suspension cultures and microdroplet cell cultures with polyethylene glycol was found critical for induction of somatic embryogenesis. Somatic embryogenesis at a frequency of 1.19, 3.16 and 6.51 per 106 cells was achieved in the callus, low density cell suspension cultures and microdroplet cell cultures, respectively. Advantages of employing microdroplet cell cultures for high frequency production of somatic embryos and its application in genetic transformation protocols are discussed.
Page(s): 600-610
URI: http://hdl.handle.net/123456789/32165
ISSN: 0975-1009 (Online); 0019-5189 (Print)
Appears in Collections: IJEB Vol.53(09) [September 2015]

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