【作者】 王艳；

【导师】 姜长阳；

【作者基本信息】 辽宁师范大学 ， 植物学， 2012， 硕士

【摘要】 艾蒿(Artemisia argyi),属菊科蒿属多年生草本植物。它是一种集药用价值及食用价值于一身的天然中草药,另外它与中国的传统节日“端午”联系紧密。由于长期大量采集艾蒿药用以及每年端午前的过度采拔,导致野生艾蒿迅速减少,乃至濒危灭绝。为保护野生资源、满足人们的需求,本研究对艾蒿的组织培养及无性系建立展开了探索,研究了不同因素对艾蒿组织培养各阶段的影响,确定了愈伤组织诱导、愈伤组织分化、生长点分化、生根的理想培养基,最终成功建立了艾蒿植株的无性再生系,为实现对艾蒿野生资源的保护、栽培对种苗的需求,以及基因库的建立奠定了技术基础。本研究的主要内容及结果如下：1.愈伤组织的诱导及增殖继代培养研究结果表明,以艾蒿的无菌嫩茎段为外植体最易诱导愈伤组织,愈伤组织诱导的理想培养基为MS+2,4-D2.0mg·L-1+蔗糖30g·L-1,同时,此种培养基还适用于艾蒿愈伤组织的增殖继代培养。2.愈伤组织的分化培养艾蒿愈伤组织分化的理想培养基是MS+6-BA1.5mg·L-1+IAA0.3mg·L-1+蔗糖30g·L-1,分化率为51.6%,平均每块愈伤组织能分化出3.2个不定芽。3.生长点的分化培养艾蒿生长点分化培养的理想培养基是MS+NAA1.0mg·L-1+GA30.5mg·L-1+6-BA3.0mg·L-1+蔗糖30g·L-1,分化率为93.3%,平均每个生长点能分化出5.3个分化芽。4.生根及继代培养生根培养适宜在恒温光照条件下进行,诱导愈伤组织分化出的不定芽生根的理想培养基是1/4MS+IAA0.2mg·L-1+蔗糖20g·L-1；诱导生长点分化芽生根的理想培养基是N6+IAA0.2mg·L-1+蔗糖20g·L-1；生根继代培养的理想培养基是1/4MS+IAA0.2mg·L-1+蔗糖20g·L-1。接种时采取将试管苗的植物形态学下端插入培养基的正插方式,生根效果最好。继代培养时每经25d就能培养出1代试管苗,平均每代的繁殖系数为3.4。5.移栽与定植移栽基质为园土,试管苗移栽成活率高达100%,定植的试管苗长势旺盛,并且保持了野生植株所具有的所有生物学性状。更多还原

【Abstract】 Artemisia argyi is the perennial herb of the genus Artemisia in the famliy Compositae. It is a kind of natural herbal medicine which combines medical value and food nutritive value together. Moreover, it has a close connection with Dragon Boat Festival which is a traditional festival of China. Because of people collecting plenty of Artemisia argyi for long term as medicine, as well as excessive picking before Dragon Boat Festival, in the recent years, the wild resources of Artemisia argyi has been reduced rapidly, even in imminent danger exterminates. In order to protect wild resources and meet the need of people, the study focused on the tissue culture and clone establishment of Artemisia argyi. Systematic studies were done on different factors which affecting the process of tissue culture of Artemisia argyi. According to the studies, the optimum media for callus induction, callus differentiation, growing point differentiation and rooting were identified and finally the asexual regeneration system of Artemisia argyi was established successfully. The study laid the foundation of the technology to realize the protection of wild resources, the need of seedlings and the establishment of the gene pool of Artemisia argyi.The main contents and results of the study were shown as follows:1. The cultivation of callus induction and subcultureThe results showed that the most suitable explant for callus induction was the aseptic tender stem; the ideal medium for callus induction was MS+2,4-D2.0mg·L-1sucrose30g-L"1, and this kind of medium applied to subculture of induced callus as well.2. The cultivation of callus differentiationThe results demonstrated that MS+6-BA1.5mg-L-1+IAA0.3mg·L-1+sucrose30g·L-1was the optimum medium for callus differentiation, the differentiation rate was51.6%, and one callus could differentiate3.2buds on average.3. The cultivation of growing point differentiationThe results manifested that the optimum medium for growing point differentiation was MS+NAA1.0mg·L-1+GAs0.5mg·L-1-BA3.0mg·L-1+sucrose30g·L-1, in this kind of medium, the differentiation rate was93.3%and one growing point could differentiate5.3buds on average.4. The cultivation of rooting and subcultureThe results demonstrated that the optimum medium for rooting of adventitious buds was1/4MS+IAA0.2mg·L-1+sucrose20g·L-1;the optimum medium for rooting of shoot buds was N6+IAA0.2mg·L-!+sucrose20g·L-1;1/4MS+IAA0.2mg·L-1+sucrose20g·L-1was the ideal medium for rooting sbuculture. The cultivation of rooting and subculture was suitable to cultivate under homothermal illumination. In the process of cultivation of rooting subculture, insert the plant morphology bottom into medium could achieve the best result. Subculture for25days could get a generation of plantlets and the average propagation coefficient of per-generation was3.4.5. Plantlet transplanting colonizationGarden soil was chosen as transplanting substrate. The survival rate of transplanted tube seedlings could be up to100%. Colonization of plantlets grew vigorously and maintained all biological traits which wild plant had.更多还原