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Somatic embryogenesis and Agrobacterium-mediated transformation in Bixa orellana L.

Parimalan, R. and Akshatha, Venugopalan and Giridhar, P. and Ravishankar, G. A. (2011) Somatic embryogenesis and Agrobacterium-mediated transformation in Bixa orellana L. Plant Cell Tissue and Organ Culture, 105. pp. 317-328.

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Abstract

Establishment, maintenance, regeneration, and transformation of somatic embryos by both direct and indirect means (callus-mediated) was achieved for Bixa orellana, a tropical plant whose seeds produce commercially edible ‘annatto pigment,’ which mainly constitutes an apocarotenoid called bixin. Callus-mediated methodology was found to be efficient in producing a greater number of embryos in a short time. The maximum of 28 somatic embryos were produced in 16–18 weeks when immature zygotic embryonic stalks were inoculated onto Murashige and Skoog (MS) medium containing B5 vitamins supplemented with 0.44 lM benzyladenine (BA), 0.054 lM a-naphthaleneacetic acid (NAA), 2.89 lM gibberellic acid (GA3), 0.02 lM triiodobenzoic acid (TIBA), and 0.011 lM triacontanol (TRIA). Callus initiation from hypocotyl explants was obtained on MS medium supplemented with 1.07–2.14 lM NAA and 10.2 lM BA. In 3 months, somatic embryos were produced when callus was inoculated onto MS medium supplemented with 4.44 lM BA, 40 lM AgNO3, and 0.011 lM TRIA. Somatic embryos were efficiently regenerated on MS basal solid and liquid media supplemented with 0.44–4.4 lM BA, 0.54–2.69 lM NAA, 4.92 lM 2iP, 2.1 lM calcium D-pantothenate, 0.21 lM biotin, 227.7 lM cysteine HCl monohydrate, and 108.6 lM adenine sulfate. Agrobacterium tumefaciens GV 3101 harboring pCAMBIA 1305.2 binary vector-mediated stable transformation of somatic embryos exhibited a transformation frequency of 2.56%. As somatic embryogenesis in any perennial system is useful in terms of both commercial and scientific nature, this somatic embryo-based transformation protocol for the commercially important dye-yielding tropical plant B. orellana is useful for its improvement through genetic engineering.

Item Type: Article
Uncontrolled Keywords: Agrobacterium tumefaciens Annatto Regeneration Rooting Triacontanol
Subjects: 500 Natural Sciences and Mathematics > 10 Plants > 06 Trees And Shrubs
500 Natural Sciences and Mathematics > 10 Plants > 05 Tissue Culture
Divisions: Plant Cell Biotechnology
Depositing User: Food Sci. & Technol. Information Services
Date Deposited: 07 Feb 2011 05:05
Last Modified: 22 May 2012 10:55
URI: http://ir.cftri.com/id/eprint/9906

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