本文關(guān)鍵詞：多花黃精組培體系建立及甾體皂苷成分提取　出處：《安徽大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 多花黃精 組織培養(yǎng) 毛狀根 甾體皂苷 超高效液相色譜

【摘要】：多花黃精(Polygonatum cyrtonema Hua),隸屬百合科(Liliaceae)黃精屬(Polygonatum),性平、味甘,具有補(bǔ)腎益精、降血糖、降血壓、降血脂、提高人體免疫力、抗衰老、抗腫瘤等功效,是藥食同源的多年生草本植物,為中國傳統(tǒng)大宗藥材,具有很好的開發(fā)與應(yīng)用價(jià)值。多花黃精中的甾體皂苷主要有薯蕷皂苷、薯蕷皂苷元等,具有降血糖、抗腫瘤、抗HIV的生物活性。在人工栽培中受種子發(fā)芽率低下、育苗周期長等因素影響,多花黃精通過自然繁殖的方式無法滿足市場需求。較傳統(tǒng)栽培方式,組織培養(yǎng)不受季節(jié)、取材數(shù)量等因素制約,可在短期內(nèi)大規(guī)模繁殖性狀穩(wěn)定的優(yōu)良種苗。同時(shí)毛狀根具有生長速度快、合成次生代謝物質(zhì)能力強(qiáng)且穩(wěn)定等優(yōu)點(diǎn),因此,可以利用毛狀根液體培養(yǎng)技術(shù)生產(chǎn)甾體皂苷。本研究建立UPLC法同時(shí)測定多花黃精野生植株和毛狀根中薯蕷皂苷和薯蕷皂苷元的含量,為黃精資源的利用提供科學(xué)依據(jù)。研究結(jié)果如下:1.低溫沙藏、培養(yǎng)基中添加不同濃度激素處理種子,找出萌發(fā)率較高的處理方法,即0℃沙藏100d,接種到MS+2.0mg/L6-BA培養(yǎng)基中,野生多花黃精種子發(fā)芽率最高,達(dá)到43.7%。2.通過三因素三水平正交實(shí)驗(yàn),篩選出多花黃精最適快繁培養(yǎng)基配方為:MS+1.0 mg/L 6-BA +0.5 mg/L NAA +0.3 mg/L IAA,芽增殖系數(shù)達(dá)到 3.2,并且長勢良好;最適生根培養(yǎng)基為1/2MS+1.0mg/LNAA,平均生根率達(dá)到86.92%;煉苗最適基質(zhì)為泥炭土:沙子2:1。3.采用正交設(shè)計(jì)方法,探究毛狀根最佳液體培養(yǎng)條件:培養(yǎng)基為液體1/2 MS,pH為5.8,搖床轉(zhuǎn)速150 r/min,26℃暗培養(yǎng),增殖倍數(shù)達(dá)到9.4。4.優(yōu)化超聲波法提取甾體皂苷的提取方法,以75%的乙醇作為提取劑,料液比1:15,超聲2 h后,用正丁醇萃取。5.采用超高效液相色譜法(UPLC)對多花黃精野生植株和毛狀根中薯蕷皂苷和薯蕷皂苷元含量進(jìn)行統(tǒng)計(jì)分析,野生植株根莖中薯蕷皂苷和薯蕷皂苷元含量分別為106.27 μg/g、84.95μg/g,毛狀根中薯蕷皂苷和薯蕷皂苷元含量分別為 89.13 μg/g、72.24μg/g。
[Abstract]:P.cyrtonema (Polygonatum cyrtonema Hua), Liliaceae (Liliaceae) Huang Jingshu (Polygonatum), flat, sweet, with a kidney Yijing, reducing blood sugar, reducing blood pressure, reducing blood fat, improve body immunity, anti-aging, anti-tumor effect, is a perennial herb medicinal and edible plant, as the traditional China the development and Application of medicinal materials, has a very good value. Steroidal saponins p.cyrtonema consist of dioscin, diosgenin, reducing blood glucose, anti tumor, anti HIV bioactivity. The low seed germination rate in artificial cultivation, breeding effect of long cycle and other factors, p.cyrtonema through natural reproduction mode can not meet the market demand. Compared with the traditional way of cultivation, tissue culture is not the season, the number of materials and other factors, can be large-scale excellent seed and seedling traits of stable propagation in a short period of time. At the same time, hairy root with fast growth, secondary synthesis The advantages, metabolism ability and stability, therefore, can make use of hairy roots in liquid culture technology to produce steroidal saponins. The establishment of simultaneous determination of dioscin and Diosgenin p.cyrtonema wild plants and hairy roots by UPLC, to provide a scientific basis for the use of Polygonatum resources. The results are as follows: 1. at low temperature sand storage, seeds of different hormone concentration added in culture medium, find a higher rate of germination processing method, namely 0 C sand 100D were inoculated into MS+2.0mg/L6-BA medium, wild p.cyrtonema seeds germination rate is highest, reaching 43.7%.2. through three factors and three levels orthogonal experiment, screening of Polygonatum cyrtonema optimal propagation medium: MS+1.0 mg/L 6-BA +0.5 mg/L NAA +0.3 mg/L IAA, bud growth coefficient reached 3.2, and grew well; the optimum rooting medium was 1/2MS+1.0mg/LNAA, the average rooting rate reached 86.92%; the optimum seedling Substrate was Peat: sand 2:1.3. by orthogonal design method, to explore the best conditions: hairy roots cultured in liquid medium for liquid 1/2 MS, pH 5.8, rotation speed of 150 r/min, 26 C dark culture, extraction of steroidal saponins extract proliferation rate reached 9.4.4. optimization by ultrasonic wave method, using 75% ethanol as extraction agent, liquid ratio 1:15, ultrasonic 2 h, extracted with n-butanol.5. by ultra performance liquid chromatography (UPLC) to carry on the statistical analysis of Dioscorea p.cyrtonema wild plants and hairy root saponin and Diosgenin content of wild plants in potato yam rhizome saponin and Diosgenin content were 106.27 mu g/g 84.95, g/g, the hairy roots of dioscin and Diosgenin content were 89.13 g/g, 72.24 g/g.

【學(xué)位授予單位】：安徽大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S567.239;R284.2

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本文編號(hào)：1372020