【作者】 廖永翠；

【导师】 宋明； 王晓武； 汤青林；

【作者基本信息】 西南大学 ， 蔬菜学， 2011， 硕士

【摘要】 白菜类作物(Brassica rapa, L. syn. Brassica campestris)是我国重要的芸薹属蔬菜作物和油料作物,品种类型丰富,营养价值高。但由于其具有复杂的遗传背景以及广泛的遗传变异,使得一些重要农艺性状的改良非常困难。随着人民生活水平的提高,消费者对蔬菜营养品质的关注和要求日益提高。硫甙的降解产物是构成十字花科蔬菜特殊辛香风味的主要来源,同时还可作为天然的抗虫、抗菌剂,也和昆虫的繁殖和预测有关；其中很多研究表明异硫代氰酸盐(isothiocyanate, ITC)有良好的抗癌效果和防止微生物繁殖和间接的抗氧化作用。作为十字花科植物的主要活性成分硫代葡萄糖甙的相关研究也越来越受到人们的重视,因此从分子水平上分析白菜类作物中硫代葡萄糖甙的遗传规律,才能实现改良白菜类蔬菜的营养品质,培育高有益硫代葡萄糖甙的白菜类作物新品种,发掘和利用重要的基因资源的目的。本研究利用硫甙含量差异较大、表型差异较大、生育期不同的欧洲油用型白菜Rapid Cycling (L144)和中国大白菜Z16为材料,构建了含120个株系的BC2DH群体。本研究采用HPLC方法,对白菜类作物叶片中硫甙组分和含量进行了评价,为进一步开展白菜类作物中硫甙的积累和遗传研究提供信息。在此BC2DH群体的基础上构建了白菜类作物遗传连锁图谱,并对白菜叶片中硫甙含量进行了QTL定位和遗传效应的分析。其主要结论如下：1.利用HPLC分析了白菜类作物13种类型的叶片中8种硫甙组分的含量,脂肪族硫甙是其中最主要的硫甙,所占的比例均在64%以上,但在大白菜中吲哚族硫甙所占的比例相对最高,约为28.5%。野生白菜的总硫甙含量最高,为17.3μmol·g-1DW。3-丁烯基硫甙(NAP)是含量最高变异幅度最大的,平均含量为2.68μmol·g-1DW,变异范围为0-53.87μmol·g-1DW。方差分析表明,不同基因型间脂肪族硫甙在品种间差异极显著,芳香族硫甙含量在不同季节间差异极显著。相关分析结果表明,脂肪族硫甙NAP与PRO呈负相关；脂肪族硫甙NAP与GBN间呈极显著正相关；芳香族硫甙NAS与脂肪族硫甙PRO极显著正相关。在大白菜的硫甙的主成分分析中,筛选出编号为32、46、45、41、19和47的6份硫甙含量较高的材料。2.利用欧洲油用型白菜Rapid Cycling (L144)和中国大白菜Z16构建了120个株系的BC2DH群体,开发了194个Indel标记,构建了包括10个连锁群的遗传连锁图谱,共覆盖基因组长度563.3 cM,平均图距为2.90 cM。3.采用区间和MQM的作图法,对白菜类作物硫甙的含量进行了QTL定位及遗传效应的分析。在BC2DH群体的8条连锁群上,共检测到15个控制硫甙合成的QTL位点,2个控制NAP的QTL位点,3个控制GBN的QTL位点,3个控制PRO的QTL位点,2个控制GBC的QTL,1个控制NEO的QTL,2个控制40H的QTL,2个控制NAS的QTL,3个控制脂肪族硫甙含量的QTL位点,1个控制总硫甙含量的QTL位点和1个控制GBN/脂肪族硫甙合成比例的QTL位点。LOD值在3.20-16.76范围之间,贡献率范围12.5%-77.0%。更多还原

【Abstract】 Chinese cabbage (Brassica rapa, L. syn. Brassica campestris) belongs to Brassica genus, with high nutritional value and is one of the most important vegetable crops in China. Complex genetic background and extensive variation of this crop make it difficult to improve some important agronomic traits. Consumers are having higher requirement and paying much attention to the quality of vegetable nutrition with the improvement of standard of living. Degradation products of glucosinolates constitute the primary source of special spicy flavors in cruciferous vegetables, and also been taken as natural insect resistance and antibacterial agent and be relevant with insect propagation and prediction; Many studies show that isothiocyanate has good anticancer effect and indirect antioxidant effect. The relative researches on glucosinolates are attaching much more importance as the main active ingredient of Brassica rapa. So genetic analysi of nutritional quality traits in B. rapa at molecular levels will achieve the purpose of the improvement of nutrition quality and beneficial types of glucosinolates, and exploitation and utilization to key genes in B. rapa.This study used BC2DH population developed by crossing of European oiled cabbage Rapid Cycling L144 and Chinese cabbage Z16 as parents, which were diversed in glucosinolate content, morphological traits and growth period comprising of 120 lines. Evaluaion to glucosinolate composition and content in B. rapa. leaves was conducted by HPLC, which would be useful to exploit glucosinolate accumulation and genetics in B. rapa.. Genetic linkage map of B. rapa. wasconstructed on this BC2DH population, and quantitative locus (QTL) analysis was performed for glucosinolate content. Main conclusions are listed as follows:1. Eight main glucosinolates were determined by HPLC among 13 Brassica rapa types, and aliphatic glucosinolates were the major glucosinolates, taken the ratio of above 64%; While in Chinese cabbage, indolic glucosinolates taken the highest ratio relatively about 28.5%.The total glucosinolate content of wild cabbage was the highest among all 13 different types, about 17.3μmol·g-1 DW. Gluconapin (NAP) content was at most and with the widest variation, varied from 0 to 53.87μmol·g-1 DW, and the mean content was 2.68μmol·g-1 DW. Analysis of variance indicated, for aliphatic glucosinolates, there was a significant difference between different varieties and for aromatic glucosinolates also significantly different between different seasons. Aliphatic glucosinolate NAP showed negatively correlation with PRO, and positively correlation with GBN; Aromatic glucosinolate NAS was significantly positive correlative with aliphatic glucosinolate GBN by correlation analysis. Six materials with high glucosinolate content were selected in Chinese cabbage by principal component analysis, numbered 19,32,41,45,46 and 47.2. Genetic linkage maps were constructed based on BC2DH population developed by crossing an European oiled cabbage Rapid Cycling (L144) and a Chinese cabbage variety Z16. One hundred and ninety four markers were developed to construct a 10 linkage groups genetic map covering a total distance of 563.3 cM. The average interval distance was 2.90 cM.3. Interval mapping and multiple-QTL model mapping (MQM) methods were employed in mapping and analysis of QTL controlling glucosinolate contents.15 putative QTL controlling glucosinolate,2 for NAP,3 for GBN,3 for PRO,2 for GBC,1 for NEO,2 for 4OH,2 for NAS,3 for aliphatic glucosinolates content,1 for total glucosinolate and 1 for GBN/Ali were detected in 8 linkage groups. The LOD value was from 3.20 to 16.76, and the percentages of variation explained were varied from 12.5% to 77.0%.更多还原