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应用正交设计研究了IBA、6-BA、GA3和CM等植物生长调节剂在龙眼(Dimocarpus longana Lour.)与荔枝 (Litchi chinensis Sonn.)细胞悬浮培养中的作用,并就悬浮培养细胞对Km的敏感性以及用金刚沙对胚性愈伤组织在 液体状态下进行创伤和用分别带有AP1和APBD基因的农杆菌对创伤的愈伤组织进行遗传转化进行了探讨。结果 表明:IBA、6-BA、GA3和CM均可显著地提高悬浮培养系的产量,但GA3导致细胞的快速生长和褐变。悬浮培养系 细胞在液体培养基中比在固体培养基中对Km较敏感。用金刚沙对胚性愈伤组织在液体状态下进行创伤后进行农 杆菌介导转化可以得到更多的Km抗性胚状体。PCR检测结果显示,AP1及AP-D基因均已转入龙眼的胚状体中。
The effect of plant growth regulators such as IBA, 6-BA, GA3 and CM on the cell suspension culture of Dimocarpus longana Lour. And Litchi chinensis Sonn. Was studied by orthogonal design. The effects of suspension cell culture on Km And the genetic transformation of wound callus with Agrobacterium tumefaciens in wounding of embryogenic callus in liquid state and Agrobacterium with AP1 and APBD genes respectively. The results showed that IBA, 6-BA, GA3 and CM could significantly increase the yield of suspension culture, but GA3 led to rapid cell growth and browning. Suspension-cultured cells are more sensitive to Km in liquid media than in solid media. Km-resistant embryoid bodies can be obtained by agrobacterium-mediated transformation of diamond embryogenic calluses in liquid state. PCR results showed that AP1 and AP-D genes have been transferred into embryoid bodies of longan.