【作者】 金香兰；

【导师】 曲柏宏； 曹后男；

【作者基本信息】 延边大学 ， 果树学， 2002， 硕士

【摘要】 本试验以RAPD为研究方法，对梨属种质资源的秋于梨、白梨、砂梨和西洋梨四个系统的40个品种进行了分类研究，并对苹果梨的分类地位进行了探讨.结果表明： 在试验中，采用对SDS法可快速提取较高质量梨基因组总DNA用于扩增.并从100个引物中筛选出12个10碱基随机引物，它们的多态性百分级大多在90%以上。确定了最适RAPD反应体系：总体积为25 μL，其中模板DNA 100ng，MsCl₂ 2.0mmol／L，dNTP 200 μ mol／L,TaqDNA聚合酶1U，primer 8pmol，剩下的用重蒸节水来填充.扩增程序为：94℃预变性2分钟；94℃变性l分钟，37℃ 退火1.5分钟，72℃延伸2分钟，5个循环；94℃变性0.5分钟；37℃退火1分钟，72℃延伸1.5分钟，40个循环；最后72℃延伸10分钟。同时建立了40个品种的RAPD指纹图谱。并确定用引物OPH09和OPG01、OPH09和TG14、OPG09和OPI16以及三个以上的引物结合可区分开全部供试材料。 对梨属40个品种及类型进行了RAPD扩增，根据相似系数进行了聚类分析，能将起源于欧洲的西洋梨与起源于中国的3个种明显区分开，起源于中国的3个种虽未完全各自聚类，但各材料间都有不同的遗传距离，证明RAP D技术可作为梨种质分类鉴定的依据。 通过聚类分析树状图及苹果梨和其他品种的相似系数表明：苹果梨和日梨系统的品种亲缘关系较近，并和白梨系统的鸭梨、汤山酥梨、栖霞大香水聚在一起，认为苹果梨在遗传关系上和白梨系统有很大的相似性.同时用15个代表性品种来进行RAPD分析，用引物 0PA08和OPA18可以鉴别供试的 15个品种，聚类分析结果苹果梨仍然和白梨系统聚在一起。这证明了苹果梨应归属于白梨系统而不应归属于砂梨系统。 RAPD分析发现谢花甜与砂梨系统和白梨系统的部分品种聚合在一起，说明这花甜属于秋子梨系统的分类地位有待于进一步研究。更多还原

【Abstract】 In the experiment, the classification of pryrus germ plasma resource’s pvrus ussuriensis Maxim. Pyrus bretschneidcri Rehd. pyrus pyrifolia Nakai and pvrus communis Linn’s 40 varieties were researched. apple-pear’s classification position was made classified by RAPD. The result indicate: SDS can extract high quality genetic DNA template immediately in order to clone select twelve 10-mer primer random primer from 100 primers, the most suitable RAPD reaction system: template DNA 10ng， MgCl₂ 2.Ommol/L.dNTP 200 μmol/L, Taq DNA ploymerase 1U, primer 8pmol. the left is filled with redistilled water. The augmentation procedure is: predenature two minutes at 94℃;predenature one minutes at 94℃.reneal 1.5 minutes at 37℃,extent 2 minutes at 72℃,fine cycles; denature 0.5 minutes at 94℃. reneal 1 minute at 37℃.extent 1.5 minutes at 72℃.forty cycle:extent 10 minutes 72℃. Build forty varieties RAPD fingerprint, and determine OPRO9 and OPGOI,OPHO9 and OPGI4.OPHO9 and 0P116.and more than three primer combinations can distincuish all of the material.Random amplified polymorphic DNA （RAPD） expanding was done on 40 Pyrus cultivars. Based on the similar coefficient, the clustering analysis was done.The results showed that: Pyrus communis L. originated in Europe could be distinctly distinguished from the other three species which originated in China by RAPD technique. Three Chinese species were not clustered into different groups respectively, but there were different distance between any two accessions, It showed that RAPD technique could be used for classifying and identifying at the level of Pvrus species.Through tree diagram of the clustering analysis and apple-pear and other varieties’ index value indicate: the genetic relationship of apple-pear and pvrus bretschneideri Rehd’s variety is very near, and get together with pvrus bretsc hneideri Rehd of YaIh. Dangshansulh Xixiadaxiangshui. The hereditary relationship of apple-pear has great similarity with pYrus hretschneideri Rehd’s. Analyze fifteen representative varieties bN’ RAPD, distinguish these varieties with primer OPAOS and OPA 18, apple-pear still gets together with pyrus bretschneideri Rehd’s All of these indicate that apple-pear should belong to pyrus bretschneideri Rehd. Anyway it should not belong to pyrus.pyrzfolia Nakai.RAPD analysis indicate Xiehuatian got together with pyrus.pyrifolia Nakai and rants varieties of pvrus.bretschneideri Rehd,so whether Xiehuatian belong to pyrus.ussuriensis Maxim system’s classification position or not still need further research.更多还原