摘要
利用基因枪介导的方法 ,以LEAFY基因为目的基因 ,NPTⅡ为选择 /标记基因进行共转化。对影响基因枪法转化甘蔗的影响因素进行了探讨 ,结果表明转化前预培养 2d获得的转化效率最高 ,转化前后进行高渗处理可以提高转化效率。大量转化后 ,将LEAFY基因导入到 3个甘蔗品种Badila、ROC2 2和ROC16中去 ,获得了转LEAFY基因的甘蔗植株。其中抗性愈伤组织的获得频率最高达 11.2 % ,抗性植株的获得频率最高达 1.2 2 %。转基因植株的PCR及PCR Southern杂交检测结果表明 ,外源基因LEAFY已整合进了转基因植株基因组中。
LEAFY and NPT Ⅱ genes were co transferred into sugarcane in this study. Sugarcane calli as target tissues were bombarded by Biolistic PDS 1000/He Particle Delivery System (BIO RAD)with the plasmid DNA coated to GOLD Particle .The result demonstrated that pre culture or mannitol treatment before bombardment could improved the regeneration ability significantly. The highest transformation efficiency was 11.2% according to the estimation of the resistant calli produced on the medium with kan. The highest regeneration frequency from resistant cultures was 1.22%.The transformed plants were confirmed by PCR analysis and PCR Southern blotting.
出处
《作物学报》
CAS
CSCD
北大核心
2003年第4期541-544,共4页
Acta Agronomica Sinica
基金
农业部"948"项目资助 ( 9710 43
9810 16)
