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发根农杆菌转化青蒿影响因素的研究
FACTORS AFFECTING THE TRANSFORMATION OF ARTEMISIA ANNUA L. WITH AGROBACTERIUM RHIZOGENES
【摘要】 对叶盘法转化过程中各种影响因素进行研究.结果表明:发根农杆菌种类、基本培养基和青蒿株系对转化率有明显影响,而培养基pH在5.2~5.8对转化率没有明显影响;发根农杆菌ATCC15834的转化率最高;基本培养基中MS效果最好;青蒿株系025对发根农杆菌最敏感;处于对数生长期的发根农杆菌(D660nm=0.75)稀释5倍后用来转化效果最好;幼嫩的叶片的转化率比成熟叶片高;向共培养培养基中加入乙酰丁香酮等酚类物质对转化没有促进作用.在最适的转化条件下,发根农杆菌ATCC15834对青蒿株系025的转化率可达到100%.PCR检测证明,青蒿发根基因组中含有发根农杆菌Ri质粒TDNA上的rolC片段.
【Abstract】 Factors affecting Artemisia annua L. transformation with Agrobacterium rhizogenes using leaf disc method were studied. The results indicated that the effects of A. rhizogenes strains, basal media and A. annua L. strains on hairy root formation were notable, while that of medium pH ranging between 5.2~5.8 was not. Among the 5 A. rhizogenes strains used, ATCC15834 was the best; MS medium gave the best results and A. annua L. strain 025 was the most sensitive one among the 3 strains tested. After diluted for 5 times, the A. rhizogenes suspension at logarithmic phase was the most suitable preparations for the transformation. The hairy root formation frequency of immatural leaves was higher than that of matural leaves. The addition of acetosyringone to the co culture media had no obvious effects on transformation frequency. A 100% hairy root formation frequency was achieved with A. rhizogenes ATCC15834 at the optimal transformation conditions. The results of PCR amplification with the primers designed for rol C gene indicated that rol C fragment of T DNA of Ri plasmid was keen transferred into the genome of hairy root of A. annua L.
【Key words】 Artemisia annua L. Agrobacterium rhizogenes; hairy root formation frequency; Ri plasmid; hairy root;
- 【文献出处】 应用与环境生物学报 ,CHINESE JOURNAL OF APPLIED AND ENVIRONMENTAL BIOLOGY , 编辑部邮箱 ,1998年04期
- 【分类号】Q182,
- 【被引频次】26
- 【下载频次】207