Volume 24, Issue 3 (9-2011)                   3 2011, 24(3): 342-354 | Back to browse issues page

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Agrobacterium–mediated transformation of Minou (Citrus sp.) and Darabi (Citrus grandis) and regeneration of transgenic explants expressing β–glucoronidase (uid A) gene.. 3. 2011; 24 (3) :342-354
URL: http://ibs.org.ir/article-1-3-en.html
Abstract:   (2342 Views)
Agrobacterium–mediated transformation and regeneration of two Iranian citrus genotypes, including Minou (Citrus sp.) and Darabi (Citrus grandis) was carried out in optimized conditions developed for most of citrus species. Additionally the effects of cocultivation period and concentration of Benzyl aminopurin (BAP), notably affecting transgenesis, were also evaluated on a set of 50 explants. Internodal stem segments of 1 cm length from 6–8 month old greenhouse grown seedlings were cocultivated with A. tumefaciens strain C58C1 (GV 3850) caring pBI121 plasmid with introduced β–glucoronidase (uid A) gene. A 72 hr cocultivation on a medium rich in BAP (5 mg/l) markedly improved transformation frequencies as it increased dividing cells competent for transformation at the cut ends of explants. In these conditions, a transformation efficiency of 34.8 % and 30.1 % was achieved for Minou and Darabi, respectively, as surveyed by GUS expression biochemical assay and confirmed by PCR amplification of uid A gene. The transgenic explants continued to growth in shoot induction medium and produced 5–10 cm shoots competent for grafting. This is the first report of transformation and regeneration of Iranian citrus genotypes.
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Type of Study: Research |
Received: 2016/12/25

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