【作者】 刘威生；

【导师】 李绍华； 张爱民；

【作者基本信息】 中国农业大学 ， 果树学， 2005， 博士

【摘要】 本研究采用RAPD、ISSR和SSR三种分子标记技术,分别对100余份李及其近缘种材料进行了遗传多样性检测和种间亲缘关系分析,以企为李种质资源的收集、保存、利用和主要种的系统学研究提供依据。供试的材料涵盖了7个李种(中国李(P. salicina)、乌苏里李(P. ussuriensis)、杏李(P. simonii)、樱桃李(P. cerasifera)、欧洲李(P. domestica)、黑刺李(P. spinosa)和加拿大李(P. nigra))、2个李的近缘种(普通杏(P. armeniaca)和毛樱桃(P. tomentosa))、野生欧洲李类型以及杏和李的自然或人工杂交类型。 试验结果表明,47个RAPD引物用于108份材料的扩增,共扩增出243条带,平均5.17条带,其中多态带占95.5%;12个ISSR引物用于104份材料的扩增,供扩增出103条带,平均8.6条,多态带占96.1%;47对SSR引物对96份材料进行扩增,共扩增出654个等位变异,平均每位点13.9个,100%的位点均为多态位点,观察杂合度值(H_O)为0.06-0.95,平均为0.47。样品间的两两相似系数,用RAPD检测为0.11-0.98,ISSR为0.19-1.00,SSR为0.042-0.98。三种分子标记的多态性均较高,对李种质资源的鉴定效率次序为SSR>RAPD>ISSR。 三种标记均可较好地用于李种质资源的重复收集品的剔除、同名异物或同物异名的区分、标签错误样品的更正、亲缘关系分析和多样性检测。特别是RAPD、ISSR标记,因其具有简易、快速、经济的特点,在严格控制反应条件的前提下,适合于种质圃或基层育种单位在资源管理或育种实践中采用。 从聚类分析和主坐标分析可以看出,三种分子标记均较好地把供试材料分为三大组,即“杏与杏和李杂种组”、“中国李系统品种组”和“欧洲李品种组”。如果不考虑欧洲李的极端类型,中国李系统品种的多样性高于欧洲李品种的多样性。供试材料的排序结果较好地反应了主要种的生物地理学分布,但除SSR标记外RAPD和ISSR分析结果并不反映品种的地理分布特点。 试验结果还表明,杏和李的杂种类型与杏比与李近,改良的杂种中国李品种与中国李亲缘关系近,奈李作为中国李的变种有其合理性,杏李应为中国李变种,乌苏里李应为一个独立种。杏李的种内多样性低,在其种的形成过程中,可能“建立者效应”起了重要作用。 欧洲李与黑刺李比与樱桃李亲缘关系近,试验结果不能确认欧洲李是黑刺李和樱桃李的种间杂种的假说。欧洲李的起源可能是由黑刺李演化而来,而黑刺李融合了樱桃李和其它李的种质。新疆的野生欧洲李内多样性较低,可能是通过栽培欧洲李的种子传播从欧洲传到新疆并自然化的结果,而并非是栽培欧洲李的祖先。 中国李品种可以划分为三个品种群,即“东北品种群”、“北方品种群”以及“南方和国外品种群”。日本和美国的品种与我国南方品种群亲缘关系较近,说明日本是从南方引进中国李的种质,并进而传到美国等世界各地。原产于东北的乌苏里李与中国李的东北品种群亲缘关系近,原产于华北的杏李与中国李的北方品种群亲缘关系近,说明原产于长江流域的中国李与乌苏里李渐渗杂交形成了东北品种群,与杏李渐渗杂交形成了北方品种群,从而提高了其抗寒性,扩大了适栽范围。更多还原

【Abstract】 Three molecular markers such as RAPD, ISSR and SSR were employed to detect the genetic diversity in above 100 accessions of plum and its related species sampling from Chinese National Germplasm Repository for Plums and Tianshan Germplasm Repository for Wild Fruits, with the hope of assisting the management of plum germpalsm resources and elucidating the inter-species phylogenetic relationships. The accessions tested consisted of 7 plum species, including P. salicirta, P. ussuriensis, P. simonii, P. cerasifera, P. domestica, P. spinosa and P. nigra, 2 related species including P. armeniaca and P. tomentosa, wild European plums and natural / artifical hybrids between apricot and plum.A total number of 243 bands were generated using 47 RAPD primers for 108 accessions, with a average number of 5.17 per primer, among which 95.5% of the total bands was polymorphic and a total number of 103 bands using 12 ISSR primers for 104 accessions, with an average of 8.6, among which 96.1% was polymorphic. Totally, 654 alleles were found using 47 SSR primer pairs for 96 accessions, averaging 13.9 alleles per locus, and all of 47 loci were phylymorphic. The observed heterozygosity varied from 0.06 to 0.95 for 47 loci, with an average value of 0.47. The Jaccard’s coefficients of similarity between accessions tested ranged from 0.11 to 0.98 for RAPD data, and from 0.19 to 1.00 for ISSR, from 0.042 to 0.98 for SSR. The efficiencies of three markers for identication of plum germplasm were SSR > RAPD > ISSR.Three markers involved in this study could be used for the purposes of discarding the duplicated collection, discriminating synonyms or homonyms, identificating the mislabelled accessions, analysing the relationships and detecting the diversity of plum accessions. Specially, RAPD and ISSR were suitable for utilization in germpalsm repositories or remote field breeding stations, as both were reliable, quick, uncomplicated and cheap methods if much care were exercised to control occasional contamination and the amplification conditions were identical for all reactions.The clustering and principle coordinate analysis showed that each of three markers could grouped the accessions surveyed into three main groups such as "apricot and hybrids of apricot and plum", Chinese-type plums" and "European plums". If the distinct (out of group) accessions were not considered, the higher diversity existed in Chinese plums than in European plums. The grouping results based three markers reflected the biogeographic distributions of main species but only the results based on SSR data was correlated with geographic origins of accessions.The results suggesting (1) the hybrids of apricot and plum were closer to apricot than to plum, (2) it was reasonable that ’Neili’ was classed as a variant of P. salicina, (3) P simonii should be a variant of P. salicina, (4) P. ussuriensis was an independent species, (5) founder effect had possibly played an important role on the formation of P. simonii, (6) The introduced improved Chinese plum cultivars were clustered as same group with P. salicina despite most of which were hybrids of P. salicina and other diploidy species.European plums were closer to P. spinosa than to P. cerasifera and the result could not present direct evidence to support Crane & Lawrence assumption that European plum were originated from thehybrids of both species. Our results suggested that another possible evolutional way for P. domestica was that European plum derived from P. spinosa while the later containing two or more different genomes. A low level of diversity was revealed in wild European plums collected from Xinjiang of China, implying that this wild species was brought recently from Europe to Xinjiang and had be naturalized there rather than the ancestor of cultivated European plums.Chinese-type plum cultivars could be classed into three main groups such as "northeast China cultivars", "north China cultivars" and "south China and exotic cultivars". The improved Chinese-type plum cultivars introduced from Japan and US were closely related to更多还原