【作者】 黄江；

【导师】 李杨瑞； 杨丽涛；

【作者基本信息】 广西大学 ， 作物栽培学与耕作学， 2004， 硕士

【摘要】 罗汉果[Siraitia grosvenorii（Swingle）C．Jeffrey ex Lu et Z．Y．Zhang]为葫芦科罗汉果属的植物，是广西的著名特产，具有重要的医疗和保健价值。本研究利用RAPD分子标记技术，对15份罗汉果种质材料（包括10个栽培品种和5份野生种质）进行了遗传分析和鉴定，研究结果表明： 1．用改良的SDS法能够提取浓度和纯度都较高的罗汉果基因组DNA，且经RAPD扩增后能取得比较理想的扩增效果。 2．优化了RAPD反应体系，确定了适合于罗汉果RAPD-PCR的反应体系：25μl反应体系，其中含超纯水17.8μl，10×Ex Taq Buffer 2.5μl(含20mmol／L Mg²⁺)，2mmol／L dNTP 2.5μl，10 μmol／L随机引物1μl，Ex Taq DNA聚合酶0.2μl（1U），模板DNA1μl（50ng／反应）。确定了适合的PCR扩增程序为：94℃预变性4min；94℃变性30sec，37℃退火30sec，72℃延伸1min，共35个循环；72℃延伸7min。反应产物4℃保存。 3．RAPD聚类结果表明，圆果类栽培品种间亲缘关系较近，而长果类栽培品种与圆果类栽培品种间的亲缘关系较远，却与一些野生种质的亲缘关系相对较近。在圆果类中青皮果、柑子果和马铃果亲缘关系较近，聚为一类；白毛果、红毛果、柏林二号和茶山果亲缘关系较近，又聚为一类；长果类中拉江果和冬瓜果遗传距离较近，而长滩果与这两者的遗传距离较远。更多还原

【Abstract】 Siraitia grosvenorii (Swingle) C. Jeffrey ex Lu et Z. Y. Zhang is a species of Siraitia in Cucurbitaceae, which is a special local product with important medical and health care value in Guangxi province. We made genetic analysis and identification of fifteen germplasm materials of S. grosvenorii (including ten cultivated varieties and five wild materials) by RAPD markers. The rusults showed as follows:1. The genome DNA of S. grosvenorii with high concentration and purity could be obtained with the improved method of SDS, which is good enough for RAPD amplification.2. An optimized reaction system of RAPD was summed up based on the experimental results in the present study. This system has 25 μl reaction volume, including super pure water 17.8 μl,10XEx Taq Buffer 2.5μl (containing 20mmol/L Mg2+) , 2mmol/L dNTP 2.5 μl, 10 μmol/L random primer 1 μ l, Ex Taq DNA Polymerase 0.2 μl (1U) , template DNA 1 ul (50ng/reaction) . The optimal PCR amplification procedure indues: Pre-denaturation at 94℃ for 4 min; and 35 cycles of denaturation at 94℃ for 30 sec, annealing at 37℃ for 30 sec, lengthening at 72℃ for 1 min; and then again lengthening at 72℃ for 7 min. The find products were kept at 4℃.3. The results of RAPD cluster showed that the genetic relationship among the cultivated varieties of the round-fruits was close, but that between the round-fruited and the long-fruited cultivated varieties was far away from each other, and the long-fruited cultivated varieties showed even closer genetic relationship to the wild genotypes. Among the round-fruits, Qingpiguo,Ganziguo and Malingguo showed close genetic relationship and were clustered in a group, while Baimaoguo, Hongmaoguo, Bolin No.2 and Chashanguo were clustered in another group. Among the long-fruits, Lajiangguo and Dongguaguo showed close genetic distance, but they were far away from Changtanguo in genetic distance.更多还原