【作者】 高喆；

【导师】 王迎春；

【作者基本信息】 内蒙古大学 ， 植物学， 2014， 博士

【摘要】 白刺属植物在内蒙古分布有4个种：唐古特白刺(Nitraria tangutorum Bobr.)、西伯利亚白刺(Nitraria sibilica Pall.)、齿叶白刺(Nitraria roborowskii Kom.)和泡泡刺(Nitraria sphaerocarpa Maxim.)。其中唐古特白刺为中国特有种。上述4种白刺均具有极强的抗逆能力,作为多年生落叶灌木,是防治土地荒漠化的先锋植物,亦可用于盐碱化土壤的改良。白刺属植物兼具生态价值和经济价值,对唐古特白刺果实的研究表明,其果实中富含多种花青素成分,是少有的天然抗氧化剂来源,作为食品添加剂和保健食品的开发正在展开。除泡泡刺为膜质干果外,其余3种白刺果实均为荒漠半荒漠地区少见的核果状浆果。目前对分布面积广阔的西伯利亚白刺果实成分的研究尚不充分,而对于唐古特白刺除果实以外其他部位的黄酮成分鉴定也并不系统,且分布区重叠的多个白刺属植物间缺乏化学成分的比较研究,这都限制了内蒙古地区野生白刺资源的深入开发和利用。本研究对西伯利亚白刺果实中的花青素进行了成分鉴定和含量测定,分析其果实在发育过程中花青素成分的动态变化；对唐古特白刺果实、叶片、种子、茎组织黄酮成分进行了系统分析,建立了唐古特白刺叶黄酮成分的指纹图谱并与西伯利亚白刺、齿叶白刺、泡泡刺和一种过渡类型白刺叶提取物进行了综合比较。本研究的开展对于明确西伯利亚白刺果实花青素成分、深入了解唐古特白刺不同部位黄酮成分以及开发利用白刺属下各种质资源,用于品种选育和产业开发具有重要的意义。主要研究结果如下：1.建立了高效液相色谱串联质谱(MSn)方法,从西伯利亚白刺果实甲醇提取物中鉴定了12种花青素组分,分别为：矢车菊素3-槐糖苷、矢车菊素3-戊糖苷、矮牵牛素3-芸香糖-葡萄糖苷、芍药素3-槐糖苷、矮牵牛素3-鼠李糖苷、锦葵素3-阿拉伯糖苷、芍药素3-己糖苷、矢车菊素3-顺式-香豆酰二葡萄糖苷、矢车菊素3-反式-香豆酰二葡萄糖苷、天竺葵素3-香豆酰二葡萄糖苷、芍药素3-丙二酰葡萄糖苷、矮牵牛素3-香豆酰葡萄糖苷。利用所建立的半定量方法方法,分析并比较了3个不同发育阶段和2个采集点样品中花青素组成和含量的差异,其中矢车菊素3-反式-香豆酰二葡萄糖苷含量为最高,达到27.88mg/100g。2.采用高效液相色谱串联质谱(MSn)方法对产自内蒙古西部地区的唐古特白刺果实、种子、叶片、茎组织中甲醇提取物进行了定性分析,识别出14种黄酮成分,包括槲皮素衍生物3种、山奈素衍生物3种、异鼠李素衍生物8种,均为二糖苷以上糖苷衍生物。建立了对上述黄酮成分的超高效液相色谱分析体系,通过标准品进行半定量分析,得到并比较各组织中各黄酮成分含量,结果显示唐古特白刺叶中黄酮含量达到42.43mg/g,是已报道沙棘果实中的4倍。3.建立了分析唐古特白刺叶黄酮成分的RP-HPLC方法,以黄酮成分为评价指标,分析采自4个地点、不同生长期的唐古特白刺叶片样品,通过《中药色谱指纹图谱相似度评价系统》分析所得数据,建立了分离度和重现性较好的唐古特白刺指纹图谱。使用上述分析体系同时对其他3种白刺和1种过渡类型白刺叶样品进行了分析,比较了各白刺种内和种间相似度,其中对泡泡刺的叶成分分析尚属首次。结果显示相同的遗传背景使得种内黄酮组分较种间更为一致,不同白刺种间黄酮类成分的组成和含量差异较大,揭示了白刺属下各种在化学成分层面上的丰富性,对于白刺属植物种质资源的保护和开发提供了佐证。更多还原

【Abstract】 There are four Nitraria L. species distributing in Inner Mongolia: N. tangutorum Bobr., N. sibilica Pall., N. roborowskii Kom. and N. sphaerocarpa Maxim., of which N. tangutorum is the endemic specie in China. All of the four Nitraria L. species have strong resilience. As a perennial deciduous shrub, Nitraria L. is the pioneer plant for the prevention and control of desertification, which also can be used for improving soil salinization. Nitraria L. species have ecological and economic value. The research on N. tangutorum fruit shows that the fruit of N. tangutorum is rich in a variety of anthocyanin composition, and it is a rare source of natural antioxidants. To date, it is used for developing as food additives and health food. In addition to N.sphaerocarpa, the fruits of other two Nitraria L. species are also rare drupelike berries in the desert and semi-desert areas. To date, there is not sufficient research on the fruit components of N.sibilica distributing in a widely area. Except for the fruit, the identification of flavonoids in other parts of N. tangutorum is not systematic. And the different Nitraria L. species overlapping in distribution areas have been lack of the comparative study on chemical composition. These limited the further development and utilization of wild Nitraria L. resources in Inner Mongolia. In the present study, I identified the constituents and determined the content of anthocyanins in N. sibilica fruit, and analyzed the dynamic process of anthocyanin composition during the fruit ripening; systematicly analyzed the flavonoid constituents in fruits, leaves, seeds and stem tissues of N. tangutorum; establish the fingerprint of flavonoids constituents of N. tangutorum leaves, and comprehensively compared the leaves extracts with the other three Nitraria L. species. The study has important implications for determining anthocyanin constituents of N. sibilica fruit, in-depth understanding the flavonoid constituents of N. tangutorum in different parts, and developing and utilizing various Nitraria L. germplasm resoures for breeding and industrial development. The main research results are as follow:1. Established a high-performance liquid chromatography tandem mass spectrometry (MSn) method, and identified12anthocyanin components from N sibilica fruit methanol extracts, respectively: Cyanidin3-O-sophoroside, Cyanidin3-O-hexose, Cyanidin3-O-(cis-p-coumaroyl)-diglucoside, Cyanidin3-O-(trans-p-coumaroyl)-diglucoside, Petunidin3-O-rutinoside-glucose, Petunidin3-O-rhamnoside, Petunidin3-O-(p-coumaroyl)-glucoside, Peonidin3-O-hexose, Peonidin3-O-sophoroside, Peonidin3-O-malonyl-glucoside, Pelargonidin3-O-(p-coumaroyl)-diglucoside and Malvidin3-O-arabinose. Using the established semi-quantitative method, I analyzed and compared the differences of anthocyanin composition and content of the samples between three different maturity stages and two collection locations, and found that the composition Cyanidin3-O-(trans-p-coumaroyl)-diglucoside showed the highest content, up to27.88mg/100g.2. Using a high-performance liquid chromatography tandem mass spectrometry (MSn) method, the qualitative analysis was conducted for methanol extracts of N. tangutorum fruits, seeds, leaves, stems derived from western Inner Mongolia, and identified14flavonoids constituents, including three quercetin derivatives, three kaempferol derivatives, and eight isorhamnetin derivatives, all of which are glycoside derivatives with more than two glycoside. The ultra performance liquid chromatography analysis system was established for the analysis of flavonoids composition. According to the semi-quantitative analysis for standard, I determined and compared the content of each flavonoid component in different tissues, and the results showed that the flavonoids content in N. tangutorum leaves was up to42.43mg/g, which was the four times of the reported sea buckthorn fruit.3. Established a RP-HPLC method for analyzing flavonoids constituents of N. tangutorum leaves. Using the flavonoid composition as evaluation criteria, I analyzed N. tangutorum leaf samples, which were collected from four locations and different growth stages. The N. tangutorum fingerprints with a better resolution and reproducibility were established, using the data from the analysis system of "Similarity Evaluation System for Fingerprint of Traditional Chinese Medicine". The analysis system above were used to analyze the leaf samples of three Nitraria L. species and one transition Nitraria L. specie, and compared interspecific and intraspecific similarity of the four Nitraria L. species. It is the first time to analysis the flavonoid composition of Nitraria sphaerocarpa in leaves. The results showed that the flavonoid components in interspecific were more consistent than that in intraspecific because of the same genetic background, and there were significant differences of flavonoid composition and content in different Nitraria L. species. It reveals that the richness in the chemical composition of different Nitraria L. species, which provides the evidence for the protection and development of Nitraria L. germplasm resources.更多还原