关键词:
alkaloid production
black henbane
haploid
medicinal plant
transgenic plants
摘要:
Leaf, root, stem, petiole, hypocotyl, and zygotic embryo explants, as well as pollen embryoids, and re-differentiated 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 beta-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, beta-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.