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．
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.
出处
《应用与环境生物学报》
CAS
CSCD
1998年第4期349-353,共5页
Chinese Journal of Applied and Environmental Biology
基金
国家自然科学基金
关键词
青蒿
发根农杆菌
发根诱导率
RI质粒
发根
Artemisia annua L. Agrobacterium rhizogenes
hairy root formation frequency
Ri plasmid
hairy root
