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番茄ACC合酶反义基因对河套蜜瓜的转化
Transformation of Cucumis melo cv. Hetau with Tomato Antisense ACC Synthase Gene
【摘要】 河套蜜瓜(CucumismeloLcvHetau)的子叶经预培养。芽诱导和生根培养,获得再生小植株,诱导率达58%。取带有番茄ACC合酶反义基因的双元载体pMQ6/JM109与农杆菌(Agrobacteriumtumefaciens)LBA4404经三亲融合后,与在MS0上萌发5d、并在MS+1mg/LNAA培养基上预培养3d的子叶共培养48h,然后转入含50mg/L卡那霉素的MS+6mg/LZT的芽诱导培养基中,1l个月后诱导生芽,待芽长1.5-2cm时转入生根培养基中,1-2周后可诱导产生大量的根,形成完整的转基因小植株。经PCR和分子杂交检测证明,目的基因已整合入河套蜜瓜的基因组中。
【Abstract】 An efficient in vitro plant regeneration system of Cucumis melo L. cv. Hetau was established. Regenerated plantlets were obtained from cotyledons after preculture, shoot inducing culture and root inducingculture. A high regeneration rate was achieved up to 58%. Cucumis melo was transformed with the antisensetomato ACC synthase gene in binary vector pMQ6 via Agrobacterium tumefaciens mediated gene transfer.Kanamycin resistant plantlets were obtained on MS medium with 6 mg/L zeatin, 50 mg/L kanamycin and 650mg/L cefotaximine. PCR and molecular hybridization analysis showed that tomato ACC synthase antisense cDNA was integreted into the genome of C. melo.
【Key words】 Cucumis melo; Cotyledon; Regenerated plantlet; Tomato ACC synthase; Transformation;
- 【文献出处】 植物学报 ,ACTA BOTANICA SINICA , 编辑部邮箱 ,1999年02期
- 【分类号】Q789
- 【被引频次】77
- 【下载频次】120