【作者】 陈静；

【导师】 何松林；

【作者基本信息】 河南农业大学 ， 园林植物与观赏园艺， 2012， 硕士

【摘要】 本研究以牡丹品种‘乌龙捧盛’、‘太平红’和‘凤丹白’为试验材料，定性定量分析不同品种试管苗与大田苗中的酚酸类物质，通过比较不同品种中酚酸类物质的种类和含量差异，并对主要酚酸类物质在试管苗生根过程中的动态变化加以研究分析，探讨酚酸类物质与牡丹试管苗生根的关系；同时，在牡丹试管苗生根培养基中添加不同种类、不同浓度的酚酸类物质，观察并统计生根情况，从而明确酚酸类物质对牡丹试管苗生根的影响，以期为解决牡丹试管苗生根困难等问题提供理论指导，早日实现牡丹的规模化和工厂化生产。主要研究结果如下：1.以绿原酸、4-甲基儿茶酚、肉桂酸、香豆酸、6-羟基香豆素、没食子酸、苯甲酸、4-羟基苯甲酸、3-羟基-4-甲氧基苯乙酮、莽草酸、儿茶酚、白藜芦醇、4-羟基苯甲酰肼、没食子酸甲酯、咖啡酸、2，3-二羟基-4-甲氧基苯乙酮、丹皮酚、芍药苷18种酚酸作为标准样品，采用LC-MS方法，定性定量分析3个品种牡丹试管苗生根早期与大田苗中酚酸类物质的种类及含量差异，结果如下：①这3个牡丹品种的大田苗中均检测到含有绿原酸、咖啡酸、苯甲酸、没食子酸、没食子酸甲酯、4-羟基苯甲酸、芍药苷、莽草酸、丹皮酚、3-羟基-4-甲氧基苯乙酮与2、3-二羟基-4-甲氧基苯乙酮共11种酚酸；而在这3个牡丹品种的试管苗中均只检测到绿原酸、咖啡酸、苯甲酸、没食子酸、没食子酸甲酯、4-羟基苯甲酸、芍药苷、莽草酸与3-羟基-4-甲氧基苯乙酮共9种酚酸，没有大田苗中含量较高的丹皮酚与2、3-二羟基-4-甲氧基苯乙酮这两种酚酸；且大田苗中各种酚酸含量均显著高于同品种试管苗中的酚酸含量，这可能与牡丹植株的生长时间及成熟度有关。②大田苗中检测到的11种酚酸中，绿原酸、丹皮酚、3-羟基-4-甲氧基苯乙酮、2，3-二羟基-4-甲氧基苯乙酮在3个牡丹品种中均为根中含量最高，部分品种的茎中也含有少许，叶中几乎没有；芍药苷、没食子酸、没食子酸甲酯与莽草酸在3个品种的根、茎、叶中均有分布，且没食子酸甲酯在3个品种中的含量分布均为叶中最高，茎中次子，根中最低；‘凤丹白’中的莽草酸含量在茎中最高，而在‘乌龙捧盛’与‘太平红’中均为叶中含量最高。2.通过对3个牡丹品种试管苗生根过程中主要酚酸动态变化的定性定量分析，结果表明：在试管苗生根培养早期，不同品种牡丹试管苗中的主要酚酸含量与对照相比均发生了变化。这些变化因酚酸种类、牡丹品种、生根时期不同而不同。3.以易生根的牡丹品种‘太平红’和‘凤丹白’为试验材料，在牡丹试管苗生根培养基中添加不同浓度、不同种类的酚酸类物质，以不添加酚酸类物质的生根培养基为对照，培养60天后统计生根情况。试验结果表明：以‘凤丹白’为试验材料时，在分别添加有绿原酸、咖啡酸、儿茶酚、莽草酸、没食子酸与没食子酸甲酯的生根培养基上，试管苗的生根率与生根指数均高于对照，且以添加咖啡酸与儿茶酚的处理生根率和生根系数最高。由此可以推断这6种酚酸有促进牡丹试管苗生根的作用；以‘太平红’为试验材料时，在添加有高浓度苯甲酸（浓度为1.0mg·L-1时除外）与4-羟基苯甲酸的培养基上，试管苗的生根率及生根指数均明显低于对照，当苯甲酸浓度升高到6.0mg·L-1时，‘太平红’试管苗甚至不生根，由此可以推断高浓度的苯甲酸与4-羟基苯甲酸具有抑制牡丹试管苗生根的作用，而芍药苷与牡丹试管苗生根的关系还有待进一步的研究验证。更多还原

【Abstract】 Three cultivars of tree peony (Paeonia suffruticosa Andr.)‘Wu LongPeng Sheng’,‘Tai Ping Hong’ and ‘Feng Dan Bai’ were used as testmaterials to analyse phenolic acids material in three cultivars of test tubeplantlet and field plants qualitatively and quantitatively. By comparingvarieties and contents of phenol acid material in the different cultivars oftree peony, and researching the dynamic changes of the main phenolicacids in the in vitro rooting process of three cultivars of tree peony，so asto discusse the relationship between phenolic acids material and rootingof peony tube seedlings; Meanwhile, in order to clear the influence ofphenolic acids material to in vitro rooting of peony, differentconcentrations and types of phenolic acids material were added in theculture medium of rooting, and then observes and statistics the rootsituation, so as to provide theoretical guidance for solving the problem ofpeony tube seedlings root, and to realize the scale and factory productionof tree peony. The main research results were as follows:Eighteen kinds of phenolic acids were used as standard, there were chlorogenic acid,4-methylcatechol, cinnamic acid, coumalicacid,6-hydroxycoumarin, gallic acid, benzoic acid,4-hydroxybenzoicacid,4-methoxy-3-hydroxyacetophenone, shikimic acid, catechol,resveratrol,4-hydroxybenzhydrazide, methyl gallate, caffeic acid,2,3-dihydroxy-4-methoxyacetophenone, paeonolum, paeoniflorin. TheLC–MS method was used to analyse the varieties and contents differenceof some phenolic acids in three cultivars of test tube plantlet and fieldplants qualitatively and quantitatively. And the results were as follows:Eleven phenolic acids were all detected in three different cultivars ofthe field plants, and it is respectively chlorogenic acid, caffeic acid,benzoic acid, gallic acid, methyl gallate,4-hydroxybenzoic acid,paeoniflorin, shikimic acid, paeonolum,4-methoxy-3-hydroxyacetophenone and2,3-dihydroxy-4-methoxyacetophenone. While only nine kinds ofphenolic acids were all detected in three different cultivars of test tubeplantlet, and it is respectively chlorogenic acid, caffeic acid, benzoic acid,gallic acid, methyl gallate,4-hydroxybenzoic acid, paeoniflorin, shikimic acid and4-methoxy-3-hydroxyacetophenone. There were notpaeonolum and2,3-dihydroxy-4-methoxyacetophenone,but this two kinds of phenolic acids content wassignificantly higher in three cultivars of field plants; and all kinds ofphenolic acids content in the field plants was significantly higher than invarieties of test tube plantlet, this could relate to growth time andmaturity of peony plant.During eleven kinds of detected phenolic acids in the field plants,four kinds of phenolic acids were all the highest content in root, it alsocontains a few in stem of part of the cultivars, and leaves almost no. It isrespectively chlorogenic acid, paeonolum,4-methoxy-3-hydroxyacetophenone and2,3-dihydroxy-4-methoxyaceto-phenone.While paeoniflorin, gallic acid, methyl gallate and shikimicacid distribute in roots, stems and leaves of three varieties, and thecontent distribution of methyl gallate in three cultivars are highest in theleaf, higher in the stem, and minimum in the root.But shikimic acid ishighest in stem of cultivars of tree peony ‘Feng Dan Bai’, while it is highest in leaves of cultivars of ‘Wu Long Peng Sheng’ and ‘Tai PingHong’.By qualitative and quantitative analysis for dynamic change ofmain phenolic acids in the in vitro rooting process of three cultivars oftree peony, the results showed that: At the early stage of rootingcultivation of test tube plantlet, the main phenolic acids content were allchanged compare with contrast in different cultivars of tissue cultureplantlet. These changes due to phenolic acids type, peony species androot period.With cultivars of tree peony (Paeonia suffruticosa Andr.)‘Tai PingHong’ and ‘Feng Dan Bai’ as experimental materials which is easy toroot,different concentration and different kinds of phenolic acidsubstances were added in rooting substrate, and rooting substrate which isnot added phenolic acid was used for comparison, then making rootstatistics after60days. The test results show that: When‘Feng Dan Bai’was used as experimental materials, which grows in rooting substrateadded chlorogenic acid, caffeic acid, catechol, shikimic acid, gallic acid and methyl gallaterespectively, the rooting rate and rooting index were significantly higherthan those of control, and caffeic acid and catechol were significantlyhigher than other phenolic acids processing, so we concluded that the sixphenolic acids have the role of promoting rooting of test tube plantlet;When ‘Tai Ping Hong’ was used as experimental materials, which growsin rooting substrate added benzoic acid (except concentration of1.0mg· L-1) and4-hydroxybenzoic acid respectively, the rooting rate androoting index were significantly lower than contrast, and whenconcentration of benzoic acid rise to6.0mg· L-1, test tube plantlet of‘Tai Ping Hong’ even don’t take root. So we can concluded that the highlevels of benzoic acid and4-hydroxy benzoic acid can inhibit peony testtube plantlet from taking root.But the relationship between paeoniflorinand taking root of peony test tube plantlet remained to be further researchand verified.更多还原