蛇足石杉人工繁殖及石杉碱甲提取研究
发布时间：2019-07-09 11:59
【摘要】：蛇足石杉(Huperzias errata(Thunb.ex Murray)Trev.)为多年生草本植物,又名千层塔,民间用于消肿解毒,治疗妇科疾病等,目前临床主要用于阿尔兹海默症、重症肌无力、记忆力减退等疾病治疗。随着对蛇足石杉药用植物的需求量逐渐增多,蛇足石杉野生资源遭到严重破坏,部分地区甚至濒临灭绝。目前,有研究学者拟用组织培养来扩大蛇足石杉繁殖,但在培养基配制和外植体内生菌灭菌方面还存在一定的难度且生产配套设施等方面需要高额成本,因此采用蛇足石杉的营养繁殖来满足社会需求显得必要而迫切。本研究从蛇足石杉移栽基质的理化性质、插条处理工艺、扦插管理、生物碱提取检测等方面进行研究,旨为蛇足石杉规范化、工厂化的人工繁殖栽培提供理论探索。主要研究结果如下:(1)采用三种混合基质(Z1、Z2、Z3)对蛇足石杉野生植株进行移栽驯化,通过主成分分析(PCA)、方差分析、多重比较结果显示,采用容重为0.291g/cm~3、总孔隙度为77.770%、大小孔隙比0.446、有机质含量为14.245kg、全氮含量为2.228g/kg、速效磷含量为30.648mg/kg、速效钾含量为163.011mg/kg的混合基质Z1(珍珠岩:细陶粒:泥炭=2:1:1)其成活率在90d时达到95%,不定根生长情况良好,植株累计生长量最大(40.75mm),相比其余两种混合基质中蛇足石杉的生长量分别高出46.17%和14.49%;测定得到植株总的干物质的量为35.523g,根冠比为0.488,其他生理指标较其余混合基质较优,试验得出混合基质Z1适合蛇足石杉的移栽。(2)利用封口玻璃瓶对蛇足石杉进行瓶插试验,考察了混合基质、生物激素、灭菌与否对蛇足石杉插条的生根情况、插条生长量等生长指标的影响,试验结果表明采用不灭菌混合基质JZ3(珍珠岩:泥炭:黄沙=1:1:1),激素IBA浓度为100mg/L,浸泡时间为30min时蛇足石杉生根率为91.67%,生根条数为3.70根,不定根根长为13.52mm,蛇足石杉生长量为28.63mm,各项指标较优。(3)在蛇足石杉生物碱提取过程中,经过多次试验对比分析得出采用1.0%盐酸冷浸24小时(料液比1:10),再超声45min,用氨水调节PH至9,氯仿萃取三次,回收氯仿,所得生物碱提取率和纯度较高。(4)以云南昆明理工大学玻璃温室种植成株与其他3省份野生成株做对比,尝试建立了 LC-MS法检测不同产地蛇足石杉茎部位和叶部位石杉碱甲的含量,在正离子模式下,采用离子监测模式(SIM)检测,其定量分析离子为50-2000m/z,使用Agilent ZORBAX SB-C18(4.6×250 mm),流动相为甲醇:水(90:10),流速为 0.5μl/min,检测波长为310 nm,柱温为28℃,进样量为10μL。结果表明:Hup A在50～400μg/mL浓度范围内线性关系良好(r=0.9994);Hup A在蛇足石杉叶中的含量是云南(0.0559%)江西(0.0433%)湖南(0.0462%)福建(0.0274%),Hup A在蛇足石杉茎中的含量是云南(0.0444%)江西(0.0426%)湖南(0.0326%)福建(0.0217%)蛇足石杉叶中Hup A含量大于茎。
文内图片：
图片说明：图１．１蛇足石杉全株逡逑Ｆｉｇｕｒｅ邋１．１邋ｗｈｏｌｅ邋ｐｌａｎｔ邋ｏｆ邋Ｈｕｐｅｒｚｉａ邋ｓｅｒｒａｔａ邋（Ｔｈｕｎｂ．ｅｘ邋Ｍｕｒｒａｙ）邋Ｔｒｅｖ．逡逑
[Abstract]:Snake foot Huperia glyptostroboides (Huperzias errata (Thunb.ex Murray) Trev.) It is a perennial herbaceous plant, also known as Thousand-layer Tower, which is used to reduce swelling and detoxification, treat gynecological diseases and so on. At present, it is mainly used in the treatment of Alzheimer's disease, myasthenia gravis, memory loss and so on. With the increasing demand for medicinal plants of Taxus chinensis, the wild resources of Taxus chinensis have been seriously damaged, and some areas are even on the verge of extinction. At present, some researchers plan to use tissue culture to expand the propagation of Taxus chinensis, but there are still some difficulties in the preparation of culture medium and the sterilization of germs in explants, and the production supporting facilities need to be high. Therefore, it is necessary and urgent to use the nutritional propagation of Taxus chinensis to meet the needs of society. In this study, the physical and chemical properties, cutting treatment technology, cutting management and alkaloid extraction and detection of transplanting substrate of Taxus chinensis were studied in order to provide theoretical exploration for standardization and industrial artificial propagation and cultivation of Taxus chinensis. The main results are as follows: (1) three mixed matrices (Z1, Z2, Z3) were used to transplanting and domesticate the wild plants of Taxus chinensis. The results of principal component analysis (PCA),) showed that the bulk density was 0.291g / cm 鈮，
本文编号：2512133
文内图片：
图片说明：图１．１蛇足石杉全株逡逑Ｆｉｇｕｒｅ邋１．１邋ｗｈｏｌｅ邋ｐｌａｎｔ邋ｏｆ邋Ｈｕｐｅｒｚｉａ邋ｓｅｒｒａｔａ邋（Ｔｈｕｎｂ．ｅｘ邋Ｍｕｒｒａｙ）邋Ｔｒｅｖ．逡逑
[Abstract]:Snake foot Huperia glyptostroboides (Huperzias errata (Thunb.ex Murray) Trev.) It is a perennial herbaceous plant, also known as Thousand-layer Tower, which is used to reduce swelling and detoxification, treat gynecological diseases and so on. At present, it is mainly used in the treatment of Alzheimer's disease, myasthenia gravis, memory loss and so on. With the increasing demand for medicinal plants of Taxus chinensis, the wild resources of Taxus chinensis have been seriously damaged, and some areas are even on the verge of extinction. At present, some researchers plan to use tissue culture to expand the propagation of Taxus chinensis, but there are still some difficulties in the preparation of culture medium and the sterilization of germs in explants, and the production supporting facilities need to be high. Therefore, it is necessary and urgent to use the nutritional propagation of Taxus chinensis to meet the needs of society. In this study, the physical and chemical properties, cutting treatment technology, cutting management and alkaloid extraction and detection of transplanting substrate of Taxus chinensis were studied in order to provide theoretical exploration for standardization and industrial artificial propagation and cultivation of Taxus chinensis. The main results are as follows: (1) three mixed matrices (Z1, Z2, Z3) were used to transplanting and domesticate the wild plants of Taxus chinensis. The results of principal component analysis (PCA),) showed that the bulk density was 0.291g / cm 鈮，
本文编号：2512133
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