Efficient transformation and regeneration of diverse cultivars of peanut (Arachis hypogaea L.) by particle bombardment into embryogenic callus produced from mature seeds

Livingstone, D.M. and Birch, R.G. (1999) Efficient transformation and regeneration of diverse cultivars of peanut (Arachis hypogaea L.) by particle bombardment into embryogenic callus produced from mature seeds. Molecular Breeding, 5 (1). pp. 43-51.

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Abstract

A method is presented to efficiently transform cultivars of both botanical types of groundnut which involves (1) particle bombardment of embryogenic callus derived from mature seeds, (2) escape-free (not stepwise) selection for hygromycin B resistance, (3) brief osmotic desiccation followed by sequential incubation on charcoal and cytokinin-containing media, resulting in efficient conversion of transformed somatic embryos into fertile, non-chimaeric, transgenic plants. The method produces three to six independent transformants per bombardment of 10 cm² embryogenic callus. Potted transgenic plant lines can be regenerated within 9 months of callus initiation, or 6 months after bombardment. Transgene copy number ranged from one to 20 with multiple integration sites. There was ca. 50% coexpression of hph and luc or uidA genes coprecipitated on separate plasmids. Luciferase (luc) reporter gene expression was confirmed in T1 progeny from each of six tested independent transformants. Insufficient seeds were produced under containment conditions to determine segregation ratios. The practically of the technique for efficient cotransformation with selected and unselected genes is demonstrated using major commercial groundnut varieties in Australia (cv. NC-7, a Virginia market type) and Indonesia (cv. Gajah, a Spanish market type)

Item Type: Article
Additional Information: SNNigam Collection
Uncontrolled Keywords: fertile transgenic plants, groundnut, hygroycin, microprojectiles, peanut, Arachis hypogaea
Author Affiliation: Department of Botany, University of Queensland, Brisbane, Qld 4072, Australia
Subjects: Plant Production
Crop Improvement
Divisions: Groundnut
Depositing User: Mr B Krishnamurthy
Date Deposited: 17 Apr 2013 11:11
Last Modified: 17 Apr 2013 11:11
Official URL: http://dx.doi.org/10.1023/A:1009658313170
URI: http://eprints.icrisat.ac.in/id/eprint/10257

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