Vunsh Ron, Li Jihong, Hanania Uri, Edelman Marvin, Flaishman Moshe, Perl Avihai, Wisniewski Jean-Pierre, Freyssinet Georges
Department of Plant Sciences, Weizmann Institute of Science, Rehovot, Israel.
Plant Cell Rep. 2007 Sep;26(9):1511-9. doi: 10.1007/s00299-007-0361-4. Epub 2007 May 10.
The monocot family Lemnaceae (duckweed) is composed of small, edible, aquatic plants. Spirodela oligorrhiza SP is a duckweed with a biomass doubling time of about 2 days under controlled, axenic conditions. Stably transformed Spirodela plants were obtained following co-cultivation of regenerative calli with Agrobacterium tumefaciens. GFP activity was successfully monitored in different subcellular compartments of the plant and correlated with different targeting sequences. Transgenic lines were followed for a period of at least 18 months and more than 180 vegetative doublings (generations). The lines are stable in morphology, growth rate, transgene expression, and activity as measured by DNA-DNA and immunoblot hybridizations, fluorescence activity measurements, and antibiotic resistance. The level of transgene expression is a function of leader sequences rather than transgene copy number. A stable, transgenic, GFP expression level >25% of total soluble protein is demonstrated for the S. oligorrhiza system, making it among the higher expressing systems for nuclear transformation in a higher plant.
单子叶植物浮萍科（浮萍）由小型、可食用的水生植物组成。少根紫萍SP是一种浮萍，在可控的无菌条件下，其生物量加倍时间约为2天。将再生愈伤组织与根癌农杆菌共培养后，获得了稳定转化的紫萍植株。在该植物的不同亚细胞区室中成功监测到绿色荧光蛋白（GFP）活性，且其与不同的靶向序列相关。对转基因株系进行了至少18个月的跟踪，经历了180多次无性繁殖（世代）。通过DNA-DNA杂交、免疫印迹杂交、荧光活性测量和抗生素抗性检测，这些株系在形态、生长速率、转基因表达和活性方面均保持稳定。转基因表达水平是前导序列的函数，而非转基因拷贝数的函数。少根紫萍系统中稳定的转基因绿色荧光蛋白表达水平证明其占总可溶性蛋白的>25%，使其成为高等植物中核转化表达较高的系统之一。
