Agrobacterium-mediated Transformation of Sunflower (Helianthus annuus L.): A Simple Protocol

Rao, Sankara K and Rohini, VK (1999) Agrobacterium-mediated Transformation of Sunflower (Helianthus annuus L.): A Simple Protocol. In: Annals of Botany Company, 83 (4). pp. 347-354.

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Abstract

A simple and reproducible Agrobacterium tumefaciens-mediated transformation system was developed for sunflower (Helianthus annuusL.) ‘KBSH-1’. The objective was to substantially eliminate the in vitroregeneration component from the transformation protocol. Two-d-old seedlings with one cotyledon detached were infected with Agrobacterium tumefaciensstrain LBA 4404/pKIWI105 harbouring genes for \beta-glucuronidase (GUS) and neomycin phosphotransferase (NPT II). Following co-cultivation, seedlings were grown aseptically for 5 d on a growth regulator-free basal medium supplemented with $250 \hspace{2mm} \mu g\hspace{2mm} ml^{-1}$ cefotaxime (a bacteriostat used to control gram negative bacteria). Seedlings were screened for transient GUS expression and the shoot portions of the putative transformants were utilized for propagation as transgenic plants. The excised shoots that initiated roots following selection were subsequently transferred to a glasshouse. Molecular analysis of transgenic plants confirmed concordance between the presence of foreign genes and enzyme activity. The transformation regime facilitated rapid generation of up to 2% phenotypically normal fertile plants containing functional transgenes. The transmission and integration of the marker genes in the progeny is demonstrated.

Item Type:	Journal Article
Publication:	Annals of Botany Company
Publisher:	Annals of Botany Company
Additional Information:	Copyright of this article belongs to Annals of Botany Company.
Keywords:	Sunflower (Helianthus annuus L.);Transformation;Agrobacterium;Simple protocol
Department/Centre:	Division of Biological Sciences > Biochemistry
Date Deposited:	16 Oct 2007
Last Modified:	19 Sep 2010 04:40
URI:	http://eprints.iisc.ac.in/id/eprint/12239

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