不同因素对农杆菌介导的杉木转化效率的影响.pdfVIP

43 3 Vol.43, No.3 20 0 7 3 SCIENTIA SILVAE SINICAE Mar., 2 0 0 7 ＊ 席梦利　施季森 (　210037) 　:　, 6 , : 1～3 d, OD600 nm 0.1～0.4 10 ～ 15 min, 3 ～ 5 d, -1 (AS)80 μmol ·L , 3 d 。 186 (Km), 39 Km 。 CR 2 Km , (GAF )2 Km DNA 。 :　;; :S718.46;Q943.2 　　　:A　　　:1001-7488(2007)03-0046-05 An Assessment of Factors Influencing the Efficiency of Transformation of Cunninghamia lanceolata Mediated by Agrobacterium tumef aciens Xi Mengli 　Shi Jisen (Key Laboratory of Forest Tree Genetics and GeneEngineering, S tateForestry Administration 　Nanjing Forestry University 　Nanj ing 210037) Abstract:　Using plantlet shoot sections nearthe shoot tip as explants, after studying the six factors to affect transformation, we have preliminarily determined an effective system for genetic transformation.The protocol was:1 ～ 3 days for pre-culture, OD600 nm=0.1～0.4, 10 ～ 15 minutes for infection, 3 ～5 days for co-culture, and acetosyringone (AS)80 μmol ·L-1 as co- culture medium, and then lasting 3 days before selecting.And then 186 regenerated kanamycin (Km )-resistant buds were obtained from the first selection medium.39 Km -resistant budswere obtained from the second selection medium.CR analysis on 39 regenerated Km-resistant buds showed that 2 budswere CR positive.The result indicated that gastrodia antifungal protein (GAF )gene was integrated into the genome of the two Km-resistant buds. Key words:　Cunninghamia lanceolata;Agrobacterium tumef aciens mediated transformation;gastrodia antifungal protein gene 、, , (, 2001), , 。 , , 。, , , , 。 。。, (Lari decidua ) (Huang et al ., 1991;Shin et al ., 1994)、(Lari kaempferi ×L .decidua )(Levee et al., 1997)、(Picea abies )(Wenck et al ., 1999)、(Pinus taeda)(Tang et a