三七的组织培养与多倍体诱导初步研究及三角槭的叶绿体全基因组测序

发布时间：2018-07-29 08:06

【摘要】：三七(Panax notoginseng(Burk.)F.H.Chen)是五加科(Araliaceae)之人参属植物,是我国传统的名贵中药材,主产于云南文山,栽培利用历史悠久。研究表明,三七有止血、镇痛、降血压、调节血糖、提高免疫力等功效,具有重要的药用价值。为了实现三七种质资源的科学利用,提高三七品质,满足市场需求,需要通过包括组织培养等多种技术措施,来推动三七的基础研究以及栽培育种工作,对进一步开发利用三七这一药用植物提供参考。本研究重点对三七进行组织培养与多倍体诱导的问题开展了研究。实验结果如下:1、在三七愈伤组织诱导途径中,得出三七愈伤组织的最佳外植体为茎段,茎段的最佳消毒方法为75%酒精消毒30s、5%次氯酸钠消毒30min。最佳褐化抑制剂及浓度为PVP 0.05g/L,最佳愈伤诱导培养基为MS培养基+ 3.5mg/L 2,4-D+0.8mg/LKT。2、在三七愈伤分化诱导途径中,得出愈伤组织不定根分化的最佳培养基为MS培养基+ 1.0 mg/L 2,4-D,不定芽分化的最佳培养基为MS培养基+ 1.5 mg/L GA3+ 1.5 mg/L 6-BA。3、在三七愈伤组织多倍化的诱导中,暂未筛选出多倍体诱导的最佳方案,但实验结果显示秋水仙素会影响愈伤组织的生长,尝试低浓度的秋水仙素可能会对愈伤组织的染色体的加倍有一定效果。三角槭(Acer buergerianum Miq.)又名三角枫,槭树科(Aceraceae)槭属植物。它作为一种重要的观赏植物具有巨大的商业价值。但是,随着近几年人类的活动,三角槭的生长环境已经被严重地破坏,中国野生的三角槭自然种群正在大量且快速地减少。在过去的几年里,叶绿体基因组已广泛地应用于物种的鉴定以及系统发育分析中。而通过对三角槭进行叶绿体基因组测序得到的序列可以为以后的研究者提供有用的信息,以重建复杂的进化关系。本文采用高通量测序技术对三角槭进行了叶绿体全基因组测序。完成的叶绿体基因组长度大小为156,477bP。基因组包含134个基因,其中包括89个蛋白编码基因,40个tRNA基因和8个rRNA基因。大多数基因以单拷贝基因出现,其中有23个是双拷贝基因。通过对三角槭的叶绿体基因组的系统进化分析表明,三角枫与庙台槭(A.miaotaiense),台湾红榨槭(A.morrisonense 和青榨槭(4.davidii)有着较近的亲缘关系。三角槭的叶绿体全基因组的测序能够为生物多样性保护、分子学和生态学打下一定的基础。
[Abstract]:37 (Panax notoginseng (Burk.) F.H.Chen) is the ginseng plant of the family of five acanthopanacs (Araliaceae). It is a traditional Chinese medicinal herb, which mainly produced in Wenshan of Yunnan. It has a long history of cultivation and utilization. The study shows that 37 has the efficacy of hemostasis, analgesia, lowering blood pressure, regulating blood sugar, raising immunity and so on. It is of important medicinal value. In order to achieve 37 kinds of medical value. The scientific utilization of quality resources to improve the 37 quality and meet the demand of the market, we need to promote the 37 basic research and cultivation and breeding through various technical measures including organization and cultivation, and provide reference for the further development and utilization of 37 medicinal plants. This study focuses on the question of tissue culture and polyploid induction of 37. The results are as follows: 1, in the 37 callus induction route, the best explant of 37 callus is the stem section, the best disinfection method of the stem section is 75% alcohol disinfection 30s, 5% sodium hypochlorite disinfection 30min. best browning inhibitor and the concentration of PVP 0.05g/L, the best callus culture medium is MS medium + 3.5mg/L 2,4-D+0.8mg/LKT.2, in the 37 callus induction pathway, the best medium for the differentiation of adventitious roots of callus is MS medium + 1 mg/L 2,4-D, and the best medium for adventitious bud differentiation is MS medium + 1.5 mg/L GA3+ 1.5 mg/L 6-BA.3, and the optimal polyploid induction is not selected for the induction of polyploidy in the 37 callus. But the experimental results show that colchicine can affect the growth of callus. Trying low concentration of colchicine may have a certain effect on the doubling of the chromosomes of the callus. Acer buergerianum Miq. is also known as the triangular maple and Acer (Aceraceae) Acer plant. It has a great business as an important ornamental plant. However, with the activities of human activities in recent years, the growth environment of Acer triangulation has been seriously damaged, and the natural population of Chinese wild Acer is decreasing rapidly and rapidly. In the past few years, the chloroplast genome has been widely used in species identification and phylogenetic analysis. The sequence of green genome sequencing can provide useful information for future researchers to reconstruct complex evolutionary relationships. In this paper, high throughput sequencing technology was used to sequence the whole chloroplast genome of Acer triangulation. The length of the chloroplast genome was 134 genes, including 89 eggs, including 134 genes. White coded genes, 40 tRNA genes and 8 rRNA genes. Most of the genes appear with single copy genes, of which 23 are double copy genes. By phylogenetic analysis of the chloroplast genome of the triangulation Acer, the triangular maple (maple maple) is closely related to the maple (A.miaotaiense), the Taiwan red maple (A.morrisonense and 4.davidii). The sequence of chloroplast genome of Acer negundo can lay a foundation for biodiversity conservation, molecular biology and ecology.
【学位授予单位】：昆明理工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S567.236;S792.35

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