摘要
以马铃薯品种PB04的叶片和茎段为外植体，对其再生和农杆菌介导的遗传转化体系进行了研究。分别比较不同转化条件（菌液浓度、感染时间、预培养及共培养时间）对马铃薯遗传转化效率的影响，以及选择压浓度和加入时间对抗性愈伤出芽的影响，结果表明：（1）在玉米素（ZT）2mg／L，萘乙酸（NAA）0．01mg／L条件下能有效地提高马铃薯愈伤组织分化率；（2）外植体经过2d的预培养，在OD600=0．8的农杆菌菌液中侵染10min后，抗性愈伤诱导率最高。（3）头孢霉素（Cef）比羧苄青霉素（Car）更适合用于马铃薯外植体的遗传转化。（4）农杆菌侵染后恢复培养5～7d再加入卡那霉素（Km）50mg／L能有效地提高外植体的转化率。
Leaf disks and stem segments from potato cultivar PB04 were used as receptor materials to study the effects of different explants and transformation conditions （the concentration of Agrobacterium tumefaciens,duration of infection,time of pre-culture and co-culture） on transformation efficiency, and the effects of kanamycin concetrations and media on sprout formation from calli. The results showed that the concentrations of zeatin （ZT） in 2 mg/L and naphthalene acetic acid （NAA） in 0.01 mg/L were most suitable for callus induction; Precultrue for 2 days before infection by Agrobacterium （OD600=0.8, 10 min）, yielded the highest rate of antibiotic-resistant callus formation; Cefotaxime is better for potato transformation than Carbenicillin; After Agrobacterium infection, cultruing for 5 - 7 d in a non-selectable medium before kanamycin selection （50 mg/L） could effectively increase the transformation rate. An optimized transformation system suitable for potato cultivar PB04 was established and the transgenic plants were obtained.
出处
《广西农业生物科学》
CAS
CSCD
2007年第1期1-7,共7页
Journal of Guangxi Agricultural and Biological Science
基金
国家转基因植物研究与产业化专项(J00-A-002)
广西十百千人才计划项目(2000201
2005-32)
广西科技攻关计划项目(桂科攻0228019-1)
