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1、 摘 要 I 摘摘 要要 我国是世界上第一个利用水稻杂种优势的国家。由于机械混杂、生物学混杂和 不法经营者的人为掺假，迫切需要对杂交水稻种子真实性和纯度进行鉴定。传统的 田间形态学鉴定是目前最为普遍的方法，但受环境和基因显隐性的影响很大，鉴定 周期较长，特别是难以区分种子形态及遗传背景相近的材料。杂种优势主要来源于 杂合基因间的相互作用，其遗传学基础在于双亲间的遗传差异。随着分子生物学的 发展， DNA 分子标记技术为品种真实性和纯度鉴定、 杂种优势预测提供了新的手段。 SSR 标记因其众多的优点而成为杂种优势预测的重要方法之一。 SSR 标记是建立在 PCR 基础上的 DNA 标记，与其他
2、DNA 标记相比，具有遗 传方式简单、共显性、多态性好、DNA 用量少、且操作简便、重复性好、稳定可靠 等特点，因而成为目前使用最广的 DNA 分子标记技术。 本研究以籼型杂交水稻为研究材料， 利用 SSR 标记构建籼型杂交水稻骨干亲本 的指纹图谱。 并在此基础上， 利用 SSR 标记研究亲本间遗传差异与杂种优势的关系。 论文工作取得的主要研究结果如下： 1 建立了籼型杂交建立了籼型杂交水稻水稻骨干骨干亲本亲本 SSR 指纹图谱指纹图谱 以120个籼型杂交水稻骨干亲本为研究材料， 从国标中公布的48对SSR引物中筛 选出12个多态性高、重复性好、杂带少且在水稻12条染色体上较均匀分布的引物作
3、为核心引物。12对SSR引物在120份材料中共扩增出42个多态性片段，平均每对引物 可以检测3.50个等位基因。其中RM590检测到2个等位基因，RM8209、RM598、 RM586、 RM346、 RM201和RM7102分别检测到3个等位基因， RM1、 RM241、 RM5473 和RM224分别检测到4个等位基因，而RM8277则检测到 6个，表明12对SSR引物在 120份材料中具有较好的多态性。 利用这12对核心引物成功地构建了籼型杂交水稻骨 干亲本的SSR指纹图谱，为水稻品种的种质鉴定奠定了基础。 2 利用利用 SSR 标记研究籼型杂交水稻亲本间的遗传差异与杂种标记研究籼型杂交
4、水稻亲本间的遗传差异与杂种 优势的关系优势的关系 基于 SSR 指纹图谱的亲本遗传差异与籼稻杂种优势关系的研究 II ( 1 ) 以协优和汕优两组材料作为研究对象，两组材料中都是由同一母本不同父 本组成，共 17 份材料。首先利用筛选出的 47 对 SSR 标记构建 17 份材料的指纹图 谱，47 对 SSR 引物在 9 份协优材料中共扩增出 130 个多态性片段，平均每对引物 可以检测 2.77 个位点，而同样的 47 对 SSR 引物在 8 份汕优材料中扩增出了 116 个 多态性片段， 平均每对 SSR 引物可以检测 2.47 个位点， 然后对扩增结果进行 “0” 、 “1” 数字化，
5、通过统计分析软件分别计算出两组材料中各父本和母本的遗传距离。 在 17 份材料中，不育系和恢复系之间的遗传距离变幅在 0.5040.875，平均遗传距 离为 0.669，其中遗传距离最大的组合是协 A 和 7954，遗传距离为 0.875；遗传距离 最小的组合是协 A 和 2DZ057，遗传距离为 0.504。 ( 2 ) 与此同时，在田间种植两组材料，成熟时期对两组材料进行考种，考察每 份材料的产量相关性状，包括：株高、穗数、穗长、每株总粒、每株实粒、结实率、 千粒重、理论产量及实际产量。通过对杂交组合亲本间的遗传差异与杂种优势的关 系分析得知不同材料、不同遗传距离范围之间，遗传距离与产量性
6、状之间的相关性 有很大差别，在一定范围内，亲本间遗传差异与杂种优势成正相关。 关键词：关键词：杂交水稻；微卫星；指纹图谱；遗传距离；杂种优势 ABSTRACT III Abstract China is the first country which makes use of rice heterosis. Because of machinal, biological admixture and superadd man-made adulteration of some illegal operators, the identification of genuinenss and vari
7、etal purity of Hybrid Rice are urgently concerned by people. Although the traditional morphological identification of the field is still the most common method at present, it is significantly affected by environment and dominant or recessive gene, a longer cycle of identification, especially it is v
8、ery difficult for the material having similar morphology and genetic background to provide accurate results identified. Heterosis in hybrid mainly comes from the interaction of gene; its genetic base is based on genetic distance of parents. With the development of molecular biology, DNA molecular ma
9、rkers provide a new means for identification of genuinens and purity of variety and heterosis prediction. SSR markers have become one of important methods predicting heterosis because of its many advantages. SSR is a kind of DNA marker based on PCR, comparing with other DNA markers, it has many adva
10、ntages such as a simple mode、codominance、good polymorphism、a few of DNA used、simple operation、good reproducibility、stable、 reliable and so on, thus it become the most widely used DNA molecular markers. Establish the SSR fingerprint map of 120 backbone parental lines in indica hybrid rice. Basing on
11、this, analyze the correlation between genetic distance and rice parents with SSR molecular marker. The results are as follow: 1 Establishment of SSR fingerprint map of 120 backbone parental lines in indica hybrid rice 120 backbone parental lines in indica hybrid rice were analyzed by using 12 SSR (s
12、imple sequence repeats) primers selected from 48 SSR of National Standard that dispersed on 12 chromosomes in rice. A total of 42 polymorphic bands were detected among 120 lines, and each primer pair detected 3.50 alleles on average. RM590 detected two alleles, RM8209, RM598, RM586, RM346, RM201 and
13、 RM7102 detected three alleles, RM1, RM241, RM5473 and RM224 detected four alleles, and RM8277 detected six alleles, it showed that 12 SSR primers had a good polymorphism. Then establish the 基于 SSR 指纹图谱的亲本遗传差异与籼稻杂种优势关系的研究 IV fingerprint map of 120 backbone parental lines in indica hybrid rice, the f
14、ingerprint could provide a foundation for germplasm identification of rice varieties. 2 Correlation between genetic distance of rice parents and heterosis with SSR molecular marker (1) 17 materials from same female parent and different male parent were studied, which was consisted of Xieyou and Shan
15、you. At first, establish the SSR fingerprint map of 17 parental lines in hybrid rice, a total of 130 polymorphic bands were detected by 47 SSR among 9 Xieyou varieties, and each primer pair detected 2.77 alleles on average. Likewise, a total of 116 polymorphic bands were detected by 47 SSR among 8 S
16、hanyou varieties, and each primer pair detected 2.47 alleles on average, and transfered the amplified results into “0” or “1”, finally calculated the genetic distance (GD) of male and female parent of the two hybrid combinations respectivly. The range of genetic distance of the 17 parental SSR markers was from 0.504 to 0.875 (0.669 on average), in all the combination, Xie A and 7954 had the longest genetic distance (0.875), and Xie A and 2DZ057 had the shortest gene
