文档介绍：发根农杆菌Ri质粒转化甘草发状根的研究摘要甘草(6lycyrrihizag)属于豆科(Leguminosae)甘草属(61ycyrrhiza)灌木状多年生草本植物,以根和茎入药,是最常见的药用作物之一。在《神农本草经》中亦列为上品,具有补脾益气、清热解毒、润肺止咳、缓急止痛、调和诸药等之功效。甘草多年来一直是国内外研究的热点。除药用价值外,甘草在食品、饮料、卷烟、化妆品等工业中有着重要的作用,需求的日益增加仅依靠从野生药用植物中提取有用的次生代谢产物势必导致大量消耗野生药用植物资源,甚至导致物种灭绝。人工栽培,周期长,部分药用成分含量降低。而采用组织培养技术可以控制植物生长环境,加快其繁殖速度,并可以从细胞培养物直接提取有效成分。为了更深入的了解甘草及保护其野生资源,本研究进行了以下方面的工作:以乌拉尔甘草为材料,用发根农杆菌转化甘草,并诱导发状根愈伤组织,探讨了激素等对愈伤组织的影响,为植株再生和从培养物中直接提取有效成分奠定基础。其研究结果如下: l本实验利用发根农杆菌成功诱导出了甘草发状根,A4对子叶诱导率最高,预培养时间为2天,共培养时间为4天最佳,除菌浓度Cef(300mg/L)。 2MS为甘草发状根最佳培养基,最佳培养条件:交替光照,23℃,暗培养30天为一周期 3发状根细胞生长曲线为近“S”型,对数生长期在10-20天,蔗糖消耗量在对数生长期最大,NH4+的消耗主要在对数生长期前后,25天以后趋于稳定。由此断定,在接种后25—30 天是收获甘草细胞适宜时期。 4检测了不同愈伤组织的可溶性多糖、可溶性淀粉含量及多酚氧化酶、过氧化物酶的活力。绿色甘草发状根愈伤组织的可溶性多糖、可溶性淀粉含量及多酚氧化酶、过氧化物酶活性最高。 5检测了不同愈伤组织的内源激素(ABA、IAA)水平,甘草发状根愈伤组织培养中, 绿色发状根愈伤组织中多糖含量较高并表现较高的淀粉活性,内源IAA在绿色发状根愈伤组织有积累高峰,而ABA在绿色发状根愈伤组织水平较低。这些变化表明绿色甘草发状根愈伤与胚性愈伤组织有密切的关系。关键词:甘草,愈伤组织,发状根,农杆菌 Studies on the ic Transformation of61ycyrrihiza Induced by Root Inducing Plasmid(Ri)of Agrobacterium rhizogenes Abstract: The experimental materialS of thiS study were callus which were 1icorice(61yeyrrhiza uralensisf rsch)。whose roots and rhizomes are the mon Chinese herbatm edicine,a kind of perennial suffurticosa herbal plants from 61ycyrrhiza species Legume family,one ofthe traditional Chinese Medicines used to reinforce the function of the spleen and medical value, the 1icorice iS widely used in food,beverage,cigarette,cosmetics and other industries as natural resources can’t fulfill require ofmedicine and other of1iquoice has been very hot inthe and tissue culture are considered to be hopeful way to a11eviate the crisiS of natural culture technic iS a good method,it can control grow condition ofplants,accelerate pace of propagate。and distill effectual element from as material inthe for 1icorice tissue culture technology was not mature only few papers were reported,We study the cou