【摘要】：蛇足石杉(Huperzias errata(Thunb.ex Murray)Trev.)為多年生草本植物,又名千層塔,民間用于消腫解毒,治療婦科疾病等,目前臨床主要用于阿爾茲海默癥、重癥肌無力、記憶力減退等疾病治療。隨著對蛇足石杉藥用植物的需求量逐漸增多,蛇足石杉野生資源遭到嚴(yán)重破壞,部分地區(qū)甚至瀕臨滅絕。目前,有研究學(xué)者擬用組織培養(yǎng)來擴大蛇足石杉繁殖,但在培養(yǎng)基配制和外植體內(nèi)生菌滅菌方面還存在一定的難度且生產(chǎn)配套設(shè)施等方面需要高額成本,因此采用蛇足石杉的營養(yǎng)繁殖來滿足社會需求顯得必要而迫切。本研究從蛇足石杉移栽基質(zhì)的理化性質(zhì)、插條處理工藝、扦插管理、生物堿提取檢測等方面進行研究,旨為蛇足石杉規(guī)范化、工廠化的人工繁殖栽培提供理論探索。主要研究結(jié)果如下:(1)采用三種混合基質(zhì)(Z1、Z2、Z3)對蛇足石杉野生植株進行移栽馴化,通過主成分分析(PCA)、方差分析、多重比較結(jié)果顯示,采用容重為0.291g/cm~3、總孔隙度為77.770%、大小孔隙比0.446、有機質(zhì)含量為14.245kg、全氮含量為2.228g/kg、速效磷含量為30.648mg/kg、速效鉀含量為163.011mg/kg的混合基質(zhì)Z1(珍珠巖:細(xì)陶粒:泥炭=2:1:1)其成活率在90d時達到95%,不定根生長情況良好,植株累計生長量最大(40.75mm),相比其余兩種混合基質(zhì)中蛇足石杉的生長量分別高出46.17%和14.49%;測定得到植株總的干物質(zhì)的量為35.523g,根冠比為0.488,其他生理指標(biāo)較其余混合基質(zhì)較優(yōu),試驗得出混合基質(zhì)Z1適合蛇足石杉的移栽。(2)利用封口玻璃瓶對蛇足石杉進行瓶插試驗,考察了混合基質(zhì)、生物激素、滅菌與否對蛇足石杉插條的生根情況、插條生長量等生長指標(biāo)的影響,試驗結(jié)果表明采用不滅菌混合基質(zhì)JZ3(珍珠巖:泥炭:黃沙=1:1:1),激素IBA濃度為100mg/L,浸泡時間為30min時蛇足石杉生根率為91.67%,生根條數(shù)為3.70根,不定根根長為13.52mm,蛇足石杉生長量為28.63mm,各項指標(biāo)較優(yōu)。(3)在蛇足石杉生物堿提取過程中,經(jīng)過多次試驗對比分析得出采用1.0%鹽酸冷浸24小時(料液比1:10),再超聲45min,用氨水調(diào)節(jié)PH至9,氯仿萃取三次,回收氯仿,所得生物堿提取率和純度較高。(4)以云南昆明理工大學(xué)玻璃溫室種植成株與其他3省份野生成株做對比,嘗試建立了 LC-MS法檢測不同產(chǎn)地蛇足石杉莖部位和葉部位石杉堿甲的含量,在正離子模式下,采用離子監(jiān)測模式(SIM)檢測,其定量分析離子為50-2000m/z,使用Agilent ZORBAX SB-C18(4.6×250 mm),流動相為甲醇:水(90:10),流速為 0.5μl/min,檢測波長為310 nm,柱溫為28℃,進樣量為10μL。結(jié)果表明:Hup A在50～400μg/mL濃度范圍內(nèi)線性關(guān)系良好(r=0.9994);Hup A在蛇足石杉葉中的含量是云南(0.0559%)江西(0.0433%)湖南(0.0462%)福建(0.0274%),Hup A在蛇足石杉莖中的含量是云南(0.0444%)江西(0.0426%)湖南(0.0326%)福建(0.0217%)蛇足石杉葉中Hup A含量大于莖。
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圖片說明：圖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，

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