Transformation of pollen embryo-derived explants by <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis> in <Emphasis Type="Italic">Hyoscyamus niger</Emphasis> |
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Authors: | Email author" target="_blank">Shanjun?TuEmail author RS?Sangwan V?Raghavan DPS?Verma BS?Sangwan-Norreel |
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Institution: | (1) Laboratoire Androgenèse et Biotechnologie, Faculté des Sciences, Université de Picardie Jules Verne, 33 rue Saint-Leu, 80039 Amiens Cedex, France;(2) Department of Plant Biology, The Ohio State University, Columbus, Ohio, USA;(3) Verma, Biotechnology Centre, The Ohio State University, Columbus, Ohio, USA;(4) Institute of Biotechnology, Zhejiang Wanli University, 8 South Qian-hu Road, Ningbo, PRC, 315100 |
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Abstract: | Leaf, root, stem, petiole, hypocotyl, and zygotic embryo explants, as well as pollen embryoids, and redifferentiated tissues from pollen embryoid-derived plantlets of Hyoscyamus niger L. (black henbane) were inoculated with Agrobacterium tumefaciens, harboring binary vectors (pGS Gluc1) and then cultured on media containing kanamycin. Transient -glucuronidase activity and kanamycin resistant callus formation were influenced by explant origin. Transgenic calluses were obtained at a frequency of up to 30% from all the explants tested. However, transgenic shoots were obtained only from the hypocotyl of plantlets derived from pollen embryoids. Transformation was confirmed by the ability of leaf segments to produce kanamycin resistant calluses, -glucuronidase histochemical and flurometric assays, polymerase chain reaction and Southern blot analysis. The results show that pollen embryoid-derived explants may be an alternative source for both efficient transformation and regeneration of transgenic plants in recalcitrant species. |
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Keywords: | alkaloid production black henbane haploid medicinal plant transgenic plants |
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