摘要
以高羊茅品种猎狗5号的胚性愈伤组织作为受体材料,通过gus基因瞬时表达检测,探讨预培养培养基、预培养时间、菌液浓度、感染时间、乙酰丁香酮浓度、共培养条件等不同因素对农杆菌介导转化的影响,结果表明,高羊茅品种猎狗5号遗传转化优化条件为胚性愈伤组织先在预培养培养基(MS+6-BA0.5mg/L+NAA0.5mg/L)预培养5d;然后用农杆菌菌液(OD600=0.5),感染时间10～20min,在23￣25℃的温度下,共培养基(MS+蔗糖30g/L+葡萄糖10g/L+CA0.5g/L+Glu0.5g/L+2,4-D1.5mg/L+AS150滋mol/L+agar8g/L,pH5.2～5.6)上共培养3d。在共培养基中添加AS150滋mol/L有助于提高gus基因瞬时表达率。
The embryogenic callus of the cultivar tall fescue (Festuca arundinace a Schreb.) 'Houndog 5' were used as explant to study the effects on different fa ctors of pre-culture media, pre-culture time, bacterial concentrations, infectio n time, acetosyringone concentrations and co-culture conditions on transient exp ression frequency of gus gene. The results showed that the condition of optimiza tion of the cultivar tall fescue 'Houndog 5' transgened by Agrobacterium-mediate d had been pre-culture for 5 days in pre-culture medium (MS+6-BA 0.5mg/L+NAA 0.5 mg/L), and then Agrobacterium tumefaciens bacterial concentrial concentration (O D600=0.5), infection time for 10~20min, and co-culture 3 days in co-culture medi um (MS+sucrose 30g/L+glucose 10g/L+CA 0.5g/L+Glu 0.5g/L+2,4-D 1.5mg/L+AS 150?滋m ol/L+agar 8g/L, pH5.2~5.6) at the temperature from 23℃ to 25℃. To be higher tr ansient expression frequency of gus gene, it is important to add AS 150?滋mol/L to co-culture medium.
出处
《分子植物育种》
CAS
CSCD
2005年第3期375-380,共6页
Molecular Plant Breeding
基金
国家自然科学基金资助项目“彩叶草坪草资源的评价与利用”(30370147)资助。
