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Development of Efficient Regeneration and Transformation Systems in Alstroemeria

出版	Wageningen Universiteit, 2005*
ISBN	90850428019789085042808
URL	http://books.google.com.hk/books?id=ltUUAAAACAAJ&hl=&source=gbs_api

註釋. Both compact embryogenic callus (CEC) and friable embryogenic callus (FEC) were induced from leaves with axil tissues. Alstroemeria plants were regenerated from somatic embryos induced from CEC and FEC within six to seven months. Protoplasts were successfully and efficiently isolated from FEC culture. After culture of protoplasts in liquid culture and micro-colony formation in solid medium, embryogenic callus cultures were obtained with high efficiency from isolated protoplasts. Plants with healthy roots were then produced and established in the greenhouse after 8 months of protoplast isolation. FEC cultures were transformed using the Alstroemeria mosaic virus (span class=SpellEAlMV ) coat protein gene and a 3'-untranslated region sequence generated by particle bombardment. The presence of transgenes was confirmed by the expression of the GUS gene, luciferase, as well as by PCR. Over 20 independent transgenic virus-resistant lines were obtained by particle bombardment protocol developed. Agrobacterium -mediated transformation was made to transform FEC of Alstroemeria by using a high efficient protocol. Finally, the FEC culture system was subjected to particle bombardment and Agrobacterium -mediated transformation in Alstroemeria.