【作者】 鲁敏；

【导师】 汤浩茹；

【作者基本信息】 四川农业大学 ， 果树学， 2011， 博士

【摘要】 本试验采用原产于四川的白梨(Pyrus bretschneideri)代表品种‘金花’为材料,比较苹果属(Malus)、西洋梨(P. communis)、砂梨(P. pyrifolia)和白梨(P. bretschneideri)之间的亲缘关系,探讨砂梨(P. pyrifolia)与西洋梨(P.communis)种间杂种‘身不知’与西洋梨(P. communis)及砂梨(P. pyrifolia)之间的亲缘关系,并对定位于‘身不知’与‘金花’遗传连锁图谱中的苹果SSR标记进行多态性评价,选择具代表性及多态性高的SSR标记进一步用于梨属植物的亲缘归类、品种鉴别及芽变鉴别等各个领域。其主要研究结果如下①101个苹果SSR标记中,在‘身不知’ב金花’双亲及4个子代中无扩增条带的SSR引物24对,不具多态性的SSR引物17对；在双亲及62株F1代群体中不符合孟德尔遗传规律的SSR引物11对；在双亲作图时不具连锁关系的SSR引物22对；最后用于‘身不知’与‘金花’遗传连锁图谱构建的苹果SSR引物28对。②24个苹果SSR标记在‘身不知’图谱上表现出连锁关系,其中包括9条遗传连锁片段,覆盖207cM,平均距离13.8cM。15个苹果SSR标记在‘金花’图谱上表现出连锁关系,连锁图谱由5条遗传连锁片段构成,总长152.4 cM。两个遗传连锁图谱可由10个共显性SSR标记连接,分别分布于第9、11、14和17连锁群。相对于‘金花梨’而言,苹果SSR标记更趋向于转移到‘身不知’梨。③比较苹果属(Malus)、西洋梨(P. communis)、砂梨(P. pyrifolia)和白梨(P. bretschneideri)之间的亲缘关系,发现苹果SSR标记青睐西洋梨多于亚洲梨,且苹果与欧洲梨、中国梨及日本梨之间的保守区位于第9连锁群、第11连锁群和第14连锁群。第14连锁群上的SSR标记最为密集,包括CH03d08、CH04f06、CH05g07和CH05g11等。④采用比较作图与亲缘聚类两种方式探讨了砂梨(P. pyrifolia)与西洋梨(P. communis)种间杂种‘身不知’与西洋梨(P. communis)及砂梨(P. pyrifolia)之间的亲缘关系,认为‘身不知’与西洋梨(P. communis)显示了更近的亲缘关系。⑤用于构建‘身不知’ב金花’F1群体遗传连锁图谱的28对苹果SSR引物中,从等位基因数、杂合度和香农指数综合来看,多态性SSR标记排名前十位的分别为CH04a12、CH05d04、CH05g11、CH05c07、CH04d02、CH04f06、CH04h02、CH01f03b、CH03d02和CH01h01,位于第9、11、12、14和17号连锁群。其中CH04a12与CH05d04显示了最高的多态性；而CH03d02和CH05c07在现有的西洋梨,亚洲梨甚至苹果图谱中都具有普遍代表性。采用此10对多态性SSR标记进一步用于梨属植物的亲缘归类、品种鉴别及芽变鉴别等各个领域。⑥根据10对SSR引物对已知的6个西洋梨(P. communis),2个秋子梨(P.ussuriensi),9个白梨(P. bretschneideri),13个砂梨(P. pyrifolia)品种及5个没有分类信息的品种‘南宫慈’、‘红长青’、‘早七黄’、‘石宝生’和‘川霞’以及5个种间杂交种‘身不知’、‘早酥’、‘鄂梨1号’、‘鄂梨2号’和‘中华玉梨’共40个梨材料的扩增数据,将‘早七黄’、‘石宝生’和‘鄂梨1号’归为砂梨(P. pyrifolia)类群,‘南宫慈’、‘红长青’、‘鄂梨2号’和‘中华玉梨’归为白梨(P. bretschneideri)类群,‘川霞’、‘早酥’和‘身不知’归在西洋梨(P. communis)类群。苹果梨归在秋子梨(P. ussuriensi)系统。⑦任何一对单独的SSR引物都不能将‘二十世纪’梨近缘集团鉴别开来,必须两个以上的标记同时作用才能将其全部区分开来。2对或3对引物共同作用,才能对‘二十世纪’梨近缘集团进行品种鉴别。根据10对SSR引物对‘二十世纪’梨近缘集团的扩增数据,以Dice相似系数采用UPGMA聚类,得到的‘二十世纪’梨近缘集团的相似系数在0.84～0.95之间,系统树可以划分为5个组。⑧10对SSR引物对4对芽变品种及1对姊妹品种进行了SSR分析,均能产生较多、清晰的多态性指纹图谱(等位基因)。其中CH03d02能将所有4对芽变品种及1对姊妹品种全部鉴别开来,显示了很好的鉴别力。更多还原

【Abstract】 In this paper’Jinhua’(Pyrus bretschneideri)’was primary taken as representative Chinese pear of Sichuan province in China to compare genetic map of apple and pear (P. communis, P. pyrifolia, and P. bretschneideri), and to compare the genetic map of’Mishirazi’to P. pyrifolia and P. communis. The apple SSRs mapped in ’Mishirazi’and’Jinhua’were evaluated in polymorphism. The high representative and polymorphic SSR markers were chosen for further genetic classification of Pyrus, cultivar and mutant identification. The main results are as follows:①In 101 apple SSR markers, twenty-three were no amplification, and seventeen were no segregation in’Mishirazi’,’Jinhua’and their four progenies; eleven were complex segregation in’Mishirazi’,’Jinhua’and their sixty-two progenies; twenty-two were no linkage relationship and twenty-eight were linked in’Mishirazi’ and’Jinhua’.②wenty-four and fifteen apple-mapped SSR markers showed linkage relationship in’Mishirazi’and’Jinhua’respectively. Genetic linkage map segments of these 2 pears were aligned by 10 co-dominant markers that showed segregating alleles in both parents, separately distributed in linkage group segments 9,11,14 and 17. And apple SSRs were more preferable to transfer and conserve to’Mishirazi’than’Jinhua’, speculating that’Mishirazi’inherited slightly more features from its male parent, grouped it in P. communis.③he’Jinhua’was primary taken as representative Chinese pear to compare genetic map of apple and European pear (P. communis), Japanese pear (P. pyrifolia) and Chinese pear (P. bretschneideri).The alignment of the species maps based on JoinMap analysis made it possible to verify the degree of synteny between the apple and pear genomes. The apple SSR loci showed the same order and corresponding distances in the pear and apple maps, strongly suggesting the presence of highly conserved regions between the two genomes, and that apple SSRs were more preferable to transfer and conserve to European pears than Asian pears. The most conserved segments of linkage group between apple and pear were located in LG 9, LG 11 and LG 14 relying on 101 apple-mapped SSRs. The SSRs in LG 10 and 12 wouldn’t like to link each other in the cultivar of P. bretschneideri in China and P. Pyrifolia in Japan, respectively.④he relationship compared’Mishirazi’to P. pyrifolia and P. communis was investigated using comparative mapping and phylogenetic clustering. And the results were got that’Mishirazi’showed closer relationship to P. communis.⑤n the twenty-eight apple SSRs that mapped in’Mishirazi’and’Jinhua’, the former ten order of polymorphism was CH04al2, CH05d04, CH05g11, CH05c07, CH04d02, CH04f06, CH04h02, CH01f03b, CH03d02 and CHOlh01 from the allele numbers, heterozygosity and Shannon’s Information index, which located in linkage group 9,11,12,14 and 17. The ten high representative and polymorphic SSR markers were chosen for further genetic classification of P., cultivar and mutant identification.⑥’Zaoqihuang’,’Shibaosheng’and’Eli 1’were classified in P. pyrifolia; ’Nangongci’,’Hongchangqing’,’Eli 2’and’Zhonghuayuli’were classified in P. bretschneideri;’Chuanxia’,’Zaosu’and’Mishirazi’were classified in P. communis. ’Pingguoli’was classified in P. ussuriensi.⑦ny pair of individual SSR primers was not able to distinguish’Nijisseiki’ related group. It’s required that two or more than two markers had to meanwhile work. Similarity coefficient of’Nijisseiki’related group was from 0.84 to 0.95. Systematic tree could be divided in 5 groups.⑧en pairs of SSR primers were used in four pair mutant cultivars and one pair of sister cultivar, which could produce more and clear fingerprint polymorphism (allele), in which CH03d02 could identify all the ten cultivars, which showed good distinguishing ability.更多还原