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。
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.
基金
国家自然科学基金!39660050
关键词
河套蜜瓜
子叶
再生小植株
番茄ACC合酶
遗传转化
Cucumis melo, Cotyledon, Regenerated plantlet, Tomato ACC synthase, Transformation
