A microdroplet cell culture based high frequency somatic embryogenesis system for pigeonpea, Cajanus cajan (L.) Millsp.

Kumar, N.U. and Gnanaraj, M. and Sindhujaa, V. and et al, . (2015) A microdroplet cell culture based high frequency somatic embryogenesis system for pigeonpea, Cajanus cajan (L.) Millsp. Indian Journal of Experimental Biology, 53 (9). pp. 600-610.

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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 μI 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 10(6) 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.

Item Type: Article
Uncontrolled Keywords: Callus culture, Choline chloride, Crop improvement, Kinetin, Lithium chloride, Phytohormones, Plantlet regeneration, Polyethylene glycol, Putrescine, Spermidine
Author Affiliation: Department of Plant Morphology and Algology, School of Biological Sciences, Madurai Kamaraj University, Madurai-625 021, Tamil Nadu, India
Subjects: Plant Protection
Divisions: Pigeonpea
Depositing User: Mr B Krishnamurthy
Date Deposited: 10 Jan 2017 06:54
Last Modified: 10 Jan 2017 06:54
URI: http://eprints.icrisat.ac.in/id/eprint/14676

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