【作者】 赵善仓；

【导师】 王宪泽； 田纪春；

【作者基本信息】 山东农业大学 ， 作物产品品质改良与质量检测控制， 2009， 博士

【摘要】 彩色小麦为特殊粒色的小麦,有灰、紫、蓝、绿等颜色,是珍贵的种质资源,是培育小麦新品种的基础。小麦籽粒颜色主要由遗传基因决定,同时还受环境如光照、温度和施肥等条件的影响。小麦不仅是人类和动物营养的主要来源,同时小麦籽粒中还含有体外抗氧化的生物活性物质,由于小麦籽粒含有的抗氧化物质活性较高,小麦已成为抵抗疾病促进健康的活性物质重要的膳食来源。小麦籽粒中的天然抗氧化活性物质主要包括黄酮类化合物、VE、胡萝卜素、生物碱、强心甙、木酚素、甾醇等。小麦籽粒中基本色素为类胡萝素、叶黄素和花色苷,其中花色苷具有较高的抗氧化活性。本研究应用超高效液相色谱配以串联质谱技术对彩色小麦籽粒中的花色苷进行分离与鉴定,同时对彩色小麦籽粒中麦黄酮含量、维生素E含量、B族维生素含量、籽粒色素含量以及麦黄酮、维生素E、B族维生素含量与籽粒色素含量之间的相关性进行系统研究,主要研究结果如下。1、小麦籽粒中花色苷类化合物发展了一种彩色小麦籽粒中花色苷分离与鉴定的系统方法,应用超高效液相色谱配以串联质谱和二极管阵列检测技术,采用全扫描、母离子扫描,子离子扫描对彩色小麦籽粒中的花色苷进行分离与鉴定。小麦籽粒中花色苷用90%甲醇水溶液(含0.5%的甲酸)超声提取,SPE C18萃取柱净化处理,液相色谱分离,质谱鉴定。彩色小麦籽粒中鉴定出14种花色苷类化合物包括:飞燕草色素-己糖苷、飞燕草色素-芦丁苷、矢车菊素-葡萄糖苷、矢车菊素-芦丁苷、牵牛花色素-芦丁苷、芍药素-己糖苷、芍药素-芦丁苷、锦葵色素-芦丁苷、矢车菊素-丙二酰葡萄糖苷、芍药素-丙二酰-葡萄糖苷、矢车菊-(未鉴定出)、芍药素-芦丁苷、锦葵色素-芦丁苷、芍药素-己糖苷。黑小麦76及其杂交后代中普遍存在的花色苷为:矢车菊素-葡萄糖苷、矢车菊素-芦丁苷、芍药素-葡萄糖苷、芍药素-丙二酰-葡萄糖苷;冬鉴系列小麦中普遍存在的花色苷为矢车菊素-葡萄糖苷、矢车菊素-芦丁苷、芍药素-己糖苷、芍药素-丙二酰-葡萄糖苷等4种花色苷;所研究的彩色小麦中普遍存在的花色苷为:矢车菊-己糖苷、矢车菊素-芦丁苷。2、小麦籽粒中麦黄酮与色素含量的关系麦黄酮(tricin)是小麦中主要的黄酮类化合物。麦黄酮从全麦粉中用甲醇超声提取。应用超高效液相色谱配以串联质谱技术,多反应检测(MRM)精确测定了小麦籽粒中麦黄酮含量。分光光度法测定小麦籽粒中色素含量。研究结果表明小麦籽粒中麦黄酮含量受基因型的影响,并存在着明显的超高亲杂种优势,因此,可通过有性杂交,培育对人体有保健作用的高麦黄酮含量的品种;籽粒色素含量与麦黄酮的含量呈现显著正相关,籽粒颜色越深小麦,其籽粒中的麦黄酮一定高。通过比较简单的籽粒色素含量测定,可以作为筛选不同品种的小麦中麦黄酮含量的一个选择指标。3、小麦籽粒中维生素E与色素含量的关系应用超高效液相色谱串联质谱法,多反应检测(MRM)模式,精确测定彩色小麦籽粒中的脂溶性维生素E的四种同系物α-VE、β-VE、γ-VE、δ-VE的含量。研究结果表明维生素E在小麦籽粒中的含量由基因控制,籽粒色素含量高的小麦,其籽粒中维生素E总量也高,籽粒色素含量与维生素E的含量呈显著正相关。4、小麦籽粒中B族维生素与花色苷含量的关系小麦籽粒含有多种B族维生素如维生素B1、维生素B2、维生素B6、烟酰胺、泛酸、生物素和叶酸等。本研究应用超高效液相色谱-串联质谱法(UPLC–MS/MS),多反应检测(MRM)模式,精确测定小麦籽粒中维生素B1、维生素B2、维生素B6的含量。研究表明小麦籽粒中B族维生素含量存在着明显的超高亲杂种优势,因此,可通过有性杂交,培育高B族维生素含量的小麦品种;籽粒色素含量与B族维生素的含量并无显著正相关,籽粒色素含量的差异不能反映B族维生素含量的差异,籽粒颜色深,其籽粒中的B族维生素不一定高。更多还原

【Abstract】 Colored grain wheat, naturally occurs with special colors such as red, blue, purple, and black, is a precious germplasm resource as well as the basis for culturing new varieties. Wheat grain color was controlled by genes, but also by the environment conditions such as light, temperature and fertilization, etc. Wheat is a major source of nutrients for humans as well as livestock, and also contains antioxidant activity of biological in vitro. Colored grain wheat is also recognized as a significant source of antioxidants which promote health and reduce the risk of disease.Wheat naturally contains numerous classes of antioxidant compounds such as flavonoids, VE, carotenoids, alkaloids, cardiac glycosides, lignans, sterols and others. Basic wheat pigments include xanthophylls, carotenoids, anthyocyanins, anthocyanins are known to exhibit good antioxidant activity. In this study determination the contents of tricin,vitamin E and B vitamins in wheat grains, and the correlations between tricin ,vitamin E and B vitamins with the total anthocyanin content in the grain were also analyzed. The main results as follows.1.The anthocyanins in Wheat grainA systematic method for anthocyanin identification using tandem mass spectrometry (MS/MS) coupled to Ultra performance liquid chromatography (UPLC) with photo-diode array detection (PDA) was developed ,including full scan, parent scan, daugther scan, to isolation and identification of anthocyanins in colored wheat grain. Wheat grains were extracted by ultrasonically assisted extraction (UAE) of anthocyanins with 90% methanol-water solution (containing 0.5% formic acid), purification on SPE C18 was then performed. Identified 14 kinds of anthocyanins: delphinindin- glucoside/galacoside, delphinindin- rutinoside, cyanidin- glucoside/galacoside, cyanidin-rutinoside, petunidin-rutinoside, peonidin- glucoside/galacoside, peonidin-rutinoside, malvidin-rutinoside, cyanidin-malonyl glucoside, peonidin-malonyl glucoside, cyanidin-(No Identified), peonidin -rutinosides, malvidin-rutinosides, peonidin-glucoside.The result showed that cyanidin- glucoside/galacoside, cyanidin-rutinoside, peonidin- glucoside/galacoside and peonidin-malonylglucoside are ubiquitous existed in Black Wheat 76 and it sib-lines of their hybridized offspring; cyanidin- glucoside/galacoside, cyanidin-rutinoside, peonidin- glucoside/galacoside, peonidin-malonylglucoside are ubiquitous existed in Dongjian series; cyanidin-rutinoside are ubiquitous existed in all colored grain wheat.2.The correlations between tricin and the total anthocyanin contentTricin (4′, 5, 7-trihydroxy-3′, 5′-dimethoxyflavone), an important natural active substance,was found to be the dominant flavone pigment in wheat. A specific Ultra performance liquid chromatography tandem mass spectrometry method was developed to accurately determine tricin content in wheat varieties with different color grains. Spectrophotography was used to determine the content of anthocyanins in wheat grain. The results showed that the tricin content in wheat was gene-regulated, Significant heterobeltiosis was achieved through sexual hybridization for breeding wheat cultivars with high tricin content. The contents of tricin and total anthocyanin in wheat grain showed a strong positive correlation. The tricin content was high in colored wheat grain, especially in deeply colored grain.3.The correlations between Vitamin E Contents and the total anthocyanin contentA specific Ultra performance liquid chromatography tandem mass spectrometry (UPLC/MS/MS) method was developed to accurately determineα-VE、β-VE+γ-VE、δ-VE and total VE contents. The correlation between vitamin E and the total anthocyanin content in the grain were analyzed. The results showed that the content of vitamin E and the content of anthocyanins in wheat grain showed a strong positive correlation. The total pigment content may be use as a selection index for high vitamin E contents in wheat breeding.4、Correlation between the Contents of B vitamin and Total anthocyanin contentThere are variety of B vitamins in wheat grain, such as thiamine, riboflavin and pyridoxine. A specific Ultra performance liquid chromatography tandem mass spectrometry (UPLC/MS/MS) method was developed to accurately determine content in wheat varieties with different color grains. The results indicated that there are essential differences for B vitamins content of wheat grains among genotypes, controlled by genetic effects. Genetic improvement can raise the B vitamins content of wheat grain. Significant heterobeltiosis was achieved through sexual hybridization for breeding wheat cultivars with high B vitamins content. The contents of B vitamins and total anthocyanin in wheat grain did not show significant correlations. Deep colored grain of wheat was not rich in B vitamins.更多还原