【作者】 穆胜玉；

【导师】 白志川；

【作者基本信息】 西南大学 ， 植物学， 2007， 硕士

【摘要】 中药青蒿即黄花蒿（Artemisia annua L．），与分类学上的青蒿（Artemisia apiacea）同属菊科（Asteraeeae D，Compositae）蒿属（Artemisia）。从青蒿中分离出的青蒿素是目前所有抗疟药中起效最快、疗效最好、毒性最低的药物。然而，目前青蒿中的青蒿素含量普遍偏低，为了提高青蒿药材的产量和质量，我们利用组织培养技术进行了青蒿组织培养体系的建立及其同源四倍体育种的研究，旨在选育出有效成分含量高，叶部和花部药材产量高的优良品种，从而使多倍体所具有的根、茎、叶的巨型性在药材生产中得以充分利用。1建立并优化了青蒿组织培养下快速繁殖体系进行了青蒿愈伤组织的诱导，愈伤组织的分化，试管苗的扩大繁殖及生根试验，结果显示青蒿花序和幼叶在MS＋BA0.5mgL^-1＋NAA1.0mgL^-1培养基上能较快地诱导出愈伤组织，诱导率可达80％以上，且诱导出的愈伤组织在MS+BA8.0mgL^-1+IAA1.0mgL^-1的培养基上能够分化出健壮的绿苗，分化率为20％，青蒿丛生芽在MS+BA2.0mgL^-1+KT2.0mgL^-1的培养基上能有效地进行快繁，在生根培养基1／2NIS+NAA1.0mgL^-1上，试管苗生根率可达100％。2优化了青蒿染色体压片技术对青蒿试管苗根尖染色体显微鉴定的制片方法进行了优化试验，确立了较好的染色体显微鉴定技术和程序，并有效地应用于四倍体株系的检出。3在组织培养条件下进行了青蒿同源四倍体诱导技术的研究在培养基中添加一定浓度的秋水仙素，或者把青蒿丛生芽经一定浓度的秋水仙素溶液浸泡一定时间后再进行培养，结果表明，均可诱发青蒿四倍体的产生，其中以先用0.05％秋水仙碱浸泡，然后在含50mgL^-1秋水仙碱的培养基上培养，这一组合条件虽然青蒿试管苗存活率最低，但四倍体诱导率最高，达到69.2％。4通过优化的根尖染色体显微鉴定条件，鉴定出了10个同源四倍体株系从中选择了6个根茎粗大、枝叶繁茂、生长旺盛的四倍体株系作为进一步育种的材料加速快繁用于田间对比试验。5对青蒿四倍体各株系及原二倍体植株的试管苗进行了移栽试验青蒿试管苗在适宜的温湿条件下移栽效果较好，结果表明，成活率可达85.8％以上。6对各株系生长情况、农艺性状进行了初步观察记载结果显示，四倍体植株和原青蒿二倍体植株相比，各种农艺性状均发生了明显的变化：四倍体植株的叶子明显比二倍体亲本大而厚；而且生长势旺，叶、花序药材比二倍体粗大，显示了一般多倍体植株所表现的典型性状——巨型性。7对青蒿四倍体和二倍体进行了青蒿素含量的比较比较了两者青蒿素的含量，结果表明，青蒿四倍体和二倍体之间存在着比较大的差异，这为进一步选育高产优质的青蒿新品种提供了一定参考依据。8对四倍体株系试管苗生长率、农艺学性状和青蒿素含量综合评价通过综合评价，初步选育出5个株系为作为田间进一步选育的材料，其药用成分含量和生长率的综合指标均超过了对照品二倍体。更多还原

【Abstract】 Chinese medicine "Qinghao" is Artemisia annua L.. With Artemisia apiacea in taxology, it belongs to Artemisia of Compositae. Artemisin distilled from Artemisia annua L. is the best medicine with the highest efficency, the best effect and the lowest toxicity in treating ague nowadays. But, at present, the content of artemisin in Artemisia annua L.is very low rifely. So, a study on the breeding of autotetraploid of Artemisia annua L. by tissue culture was done with a view to develop superior varieties ofArtemisia annua L. with high yield and good quality. 1 The establishment and optimization of the technique and methods of tissue culture of Artemisia annua L..The experiments on the induction and differentiation of Callus, the subculture of cultured materials and rooting of plantlets were done. The results indicated that callus can easily be induced from anthotaxy and leaves of Artemisia annua L. plantlets on MS BA0.5mgL^-1+NAA1.0mgL^-1 medium with the inducement ratio 80%, and the callus can differentiate into strong green plantlets on MS+BA8.0mgL^-1+ IAA1.0mgL^-1 medium. The medium containing MS+BA2.0mgL^-1 +KT2.0mgL^-1 was proved to be proper in crowd buds propagation. The rooting ratio of the plantlets on 1/2MS+NAA 1.0mgL^-1 medium was 100%. 2 The method in chromosome identification of cultured material of Artemisia annua L. was optimized.A good procedure was developed after doing a series of experiments. By using the method, autotetraploids were identified from cultured plantlets of Artemisia annua L. 3 The induction of autotetraploid plant of Artemisia annua L. in the process of tissue culture.The obtained results indicated that autotetraploid can be induced by adding suitable concentration of colchicine in medium in tissue culture or by immersing the crowd buds in colchicine solution for a period of time prior to culture. The best method was proved to be by this way: First immersed in 0.05% colchicine solution,then placed in the medium contained 50mgL^-1 colchicine. Although the death ratio is the highest, the inducing ratio reached as high as 69.2%.4 On the condition of the opitimised chromosome identification, we have identified 10 autotetraploid individual plant system.We have chosen 6 individual plant system that showed wide and rough leaf, strong stem and big root for further study. At the same time, we accerate quick propagation for further comparation study in the field.5 The transplant ofplantlets.The results of transplant experiments showed that under suitable weather condition about 85.8% plantlets survived when transplanted from nutrient medium to soil.6 The identification and observation of agronomic characteristics of autotetraploid lines.The obtained results demonstrated that autotetraploid plants grew vigorously and showed wide and rough leaves,strong stems and big roots. These agronomic characteristics demonstrated the typical characters of polyploidy plants.7 The artemisin content measurement of the transplants.There is big difference in artemisin between diploid and autotetraploid transplants which provides some references for further choosing higher production plantlets.8 The general estimate of the growth ratio, agronomic characteristics and artemisin content of the tetraploid.By the general estimate, 5 transplants system were chosen for further study in the field. Their artemisin content and growth ratio surpassed the comparative diploid更多还原