SA和MeJA对黄芩愈伤组织黄酮类化合物含量及其合成酶的影响

发布时间：2018-05-24 17:44

  本文选题：黄芩愈伤组织 + 水杨酸　； 参考：《吉林农业大学》2017年硕士论文

【摘要】：黄芩(Scutellaria baicalensis Georgi)为唇形科多年生草本植物,根入药,药用价值高,市场需求量大,野生资源几近枯竭,而栽培黄芩受遗传和环境因素影响,质量差异较大,不易人为控制。通过组织培养技术可以消除由遗传差异带来的影响,同时具有生长周期短,不受外界环境条件限制等优势,已成为诸多药用植物获取天然药用成分的途径之一。在众多提高药效成分的方法中,外源添加诱导子已成为最常用且最有效的方法之一。SA和MeJA作为植物体内重要的生物信息素物质,在植物信号转导和抗逆反应中发挥着重要作用,也是目前药用植物应用最多的非生物诱导子。本研究以黄芩愈伤组织为材料,通过添加不同种类浓度的诱导子,研究愈伤组织的生物量及其黄芩苷等药效成分的含量,同时对黄芩苷合成途径关键酶PAL、C4H、4CL和CHS活性及其抗氧化酶SOD、POD和CAT活性进行测定,以期从代谢途径关键酶和抗氧化酶系统两个方面初步探究诱导子对黄芩愈伤组织次生代谢物积累的机制。主要研究结果如下:1、筛选不同外植体和不同浓度植物激素配比确定黄芩愈伤组织培养的最佳外植体为其带节茎段,最佳培养条件为:MS+1.0mg/L 6-BA+1.5mg/L NAA,pH 5.8,培养温度(25±1)℃,暗培养。黄芩愈伤组织生长周期为25天。2、不同浓度SA和MeJA单独处理,愈伤组织鲜重生长受到抑制;SA浓度为10μmol/L、50μmol/L,MeJA浓度为10μmol/L、50μmol/L、100μmol/L时干重略高于对照,但差异均不显著。SA和MeJA复配作用,愈伤组织鲜重生长得到促进,干重略低于对照,差异不显著。3、SA单独处理浓度为50μmol/L时黄芩苷含量积累最多,是对照的1.63倍,与对照差异显著;MeJA浓度为50μmol/L时黄芩苷含量最大,是对照的1.83倍,与对照差异显著;SA和MeJA复配作用,A9组(SA 100μmol/L+MeJA 100μmol/L)愈伤组织黄芩苷含量最高,是对照的1.10倍,与对照差异不显著。4、SA浓度为10μmol/L时SOD活性最大,POD活性略低于对照,50μmol/L时CAT活性最大;MeJA浓度为50μmol/L时POD、CAT活性最大;SA和MeJA复配作用,A4组SOD活性最大,A9组POD活性最大,A7组CAT活性最大。5、SA浓度为10μmol/L时C4H活性最大,PAL活性在50μmol/L时略低于对照,4CL活性在50μmol/L时达到最大;MeJA浓度为200μmol/L时PAL活性达到最大,50μmol/L浓度时C4H活性最大,10μmol/L浓度4CL活性最大,100μmol/L浓度CHS活性最大;SA和MeJA复配作用整体上提高了PAL、C4H、4CL、CHS活性,其中A9组PAL活性最大,A6组CHS活性最大,A7组4CL活性最大。6、SA单独处理黄芩苷含量与POD显著负相关,与CAT显著正相关;MeJA单独处理黄芩苷含量与CAT呈显著正相关;SA和Me JA复配作用,黄芩苷含量与4CL呈极显著正相关。
[Abstract]:Scutellaria baicalensis Georgii is a perennial herb of Labiatae with high medicinal value, high market demand and close depletion of wild resources. However, the quality of cultivated Scutellaria baicalensis is influenced by genetic and environmental factors, and the quality of cultivated Scutellaria baicalensis is not easy to be controlled artificially. The technique of tissue culture can eliminate the influence of genetic difference and has the advantages of short growth cycle and no restriction of external environment. It has become one of the ways for many medicinal plants to obtain natural medicinal components. Among the many methods to improve the pharmacodynamic components, exogenous inducers have become one of the most common and effective methods. SA and MeJA are important biological pheromones in plants and play an important role in plant signal transduction and stress resistance. It is also the most widely used abiotic elicitor in medicinal plants. In this study, the callus of Scutellaria baicalensis was used as material, and the biomass of callus and the content of baicalin and other effective components were studied by adding different kinds of inducers. At the same time, the activities of PALC4Hn4CL and CHS, and the activities of antioxidant enzymes SOD- POD and CAT, were also determined, which were the key enzymes of baicalin synthesis pathway. In order to explore the mechanism of secondary metabolites accumulation of Scutellaria baicalensis callus from the two aspects of key enzymes and antioxidant oxidase system of metabolic pathway. The main results were as follows: 1. The best explants with stem segments were determined by screening different explants and different concentrations of plant hormones. The optimum culture conditions were as follows: ph 5.8 of 1.0mg/L 6-BA 1.5mg/L NAA, 25 卤1 鈩，

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