Shoot Regeneration from Cultured Leaf Explants of Lycium barbarum and Agrobacterium-Mediated ransformation1
Russian Journal of Plant Physiology, Vol. 48, No.4, 2001, pp. 453-458. From Fiziologiya Rastenii, Vol. 48, No.4, 2001, pp. 529-535.，-0001，（）：
A method for fast plant regeneration via organogenesis directly from Lycium barbarum leaf explants has been developed. The key factor for shoot regeneration was the presence of benzyladenine (BA) in the medium. NAA could only induce root formation and explant callusing. Murashige and Skoog (MS) medium supplemented with 2mg/l BA and 0.5mg/l NAA is the most efficient condition for shoot formation, with up to 92.6% shoot regeneration and no callus formation. All adventitious shoots cultured on MS medium supplemented with 1mg/l IAA formed an extensive root system. Regenerated plants were morphologically normal and were also proved to be diploid (2n=24). Using the optimized regeneration system, the genetic transformation of L. barbarum was carried out mediated by Agrobacterium tumefaciens EHA101(pIG121Hm). 11.8% leaf explants produced kanamycin-resistant shoots after infection by A. tumefaciens. The putative transgenic nature of plants was confirmed by GUS assay and PCR analysis. Expression of the nptⅡ gene in the regenerated plants was also detected by observing the callus formation by leaf pieces on MS medium containing 0.2mg/l 2,4-D and 0-100mg/l kanamycin.
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