【作者】 王庆彪；

【导师】 方智远； 张扬勇；

【作者基本信息】 中国农业科学院 ， 蔬菜学， 2013， 博士

【摘要】 结球甘蓝简称甘蓝，是世界上许多国家的主要蔬菜作物。中国自20世纪50-60年代开展甘蓝新品种选育以来，先后有200多个不同类型新品种通过审（认、鉴）定。但许多甘蓝品种的系谱尚不清楚，对甘蓝育成品种的系谱研究与我国主要农作物相比相差甚远。通过系谱分析不仅能够发现育成品种更替演变规律，总结出在育种过程中亲本选择和组合配制上的规律，而且能够发现在育种中重要的骨干亲本。在此基础上解析骨干亲本的遗传构成，挖掘其与重要农艺性状相关的特异染色体位点和区段并揭示其遗传效应，将有利于提高甘蓝育种工作中亲本选配的准确性和效率，为创造新的骨干亲本提供理论指导。本研究在对甘蓝育成品种系谱分析的基础上，利用甘蓝骨干亲本01-20及其姊妹系、衍生系以及由01-20和自交系96-100构建的DH群体，发掘骨干亲本01-20特异的染色体位点和区段并利用QTL揭示其遗传效应，研究获得以下结果：1、系统总结分析了中国甘蓝育成品种的系谱。1982～2012年中国共报导育成甘蓝品种219个，其中杂交种183个，所占比例为83.56%，尤其是近10年来育成品种中杂交种占96.74%；扁球型品种最多，为108个。对目前亲本来源清楚的176个杂交种的亲本组成进一步分析表明，176个杂交种来自261个直接亲本，可以追溯到67个我国地方品种和104个国外引进品种。其中黑叶小平头衍生品种最多（38个），其次是北京早熟（27个）。系谱分析发现亲本选配时不同地理来源或者植物学性状差异较大的两个亲本组配能够表现较强的杂种优势，并结合国内育成的几个重要甘蓝主栽品种的亲本组配特点，探讨在甘蓝育种过程中亲本选择、组合配制上的规律和特点。绘制了‘黑叶小平头21-3’、‘北京早熟01-20’两个甘蓝骨干亲本的系谱图。2、利用978对EST-SSR引物，2173对Scaffold-SSR引物和707对Indel引物在DH群体的双亲01-20和96-100间进行筛选，获得273对SSR和261对Indel多态性引物，将这些标记全部用于遗传图谱构建，获得了一张包含406个标记的甘蓝遗传图谱，包含9个连锁群，总长度为934.06cM，平均间距2.3cM。根据共有标记实现连锁群与甘蓝9条染色体的对应。3、在此图谱的基础上，根据2011-2012两年三季田间调查结果对甘蓝株型、叶片、叶球、种子、品质和抗性性状进行了QTL定位。共检测到与32个数量性状相关的QTL171个，其中与株型、叶片性状相关的QTL有58个，其中贡献率大于10%的38个；与叶球性状相关的QTL57个，贡献率大于10%的QTL共38个；与叶色、球色相关的QTL38个，贡献率大于10%的QTL有26个；与抗性和叶球品质相关的QTL11个，贡献率大于10%的有10个；与角果和种子相关的QTL7个，贡献率大于10%的QTL有5个；并且有15个性状的26个QTL在至少两季调查结果中能够重复检测到。这些QTL的定位为下一步骨干亲本01-20特异染色体位点的性状关联和遗传效应分析奠定了基础。4、在多条染色体上出现了不同性状QTL成簇聚集现象。其中Chr.1上55.9～60.9cM处聚集了与株型和叶片、叶球性状相关的QTL分别为9个、7个；Chr.3上88～92cM处聚集了与株型和叶片、叶球、球色相关性状QTL分别为5个、7个、1个；Chr.7上27.8～30.9cM处聚集了与球色、叶球相关性状QTL分别为2个、4个；Chr.9上58.6～61.5cM处聚集了与叶色、叶球相关性状QTL各3个，与品质、抗性相关性状QTL3个。这些染色体区段为将来进行QTL精细定位和克隆提供了重要的参考依据，也为甘蓝育种过程中相关联性状的辅助选择提供分子依据。5、通过对骨干亲本01-20及其姊妹系和亲本材料的表型性状研究发现，骨干亲本01-20的表型特征在于综合性状优良，与其他姊妹系相比没有明显的缺陷，既继承了其原始亲本北京早熟的早熟性特征，又在株型、叶色、球色和品质性状方面有所提高。对6个姊妹系基因组SSR/Indel位点多态性比较分析发现，01-20具有1个特异位点，01-07-258具有1个特异位点，01-20和01-07-258共同区别于其他姊妹系的特异位点有6个。利用QTL定位结果，其中5个特异染色体位点与农艺性状关联并揭示其遗传效应，表现为骨干亲本材料外叶色绿，株型紧凑、叶球圆正，中心柱较细，球叶脆嫩。其中Chr.3上Indel64位点和Chr.9上Indel353位点具有明显的不同性状聚集和染色体区段效应，是骨干亲本01-20在育种中发挥重要作用的染色体位点和区段。更多还原

【Abstract】 Cabbage (Brassica oleracea var. capitata) is a major vegetable crop widely cultivated in the world.More than200cabbage varieties of different types have been developed and released since the1950s and1960s in china. But the pedigree analysis of cabbage varieties was far behind many major crops, andmany cabbage varieties’ pedigree were unclear. The analysis of pedigree could not only find the rule ofcultivar succession, predict breeding objectives, but also reveal the rule of parental selection andcombination, provide guidance for the breeding of new cabbage varieties. The analysis of pedigree wouldbe conducted to clarify the founder parent phenomenon in cabbage. Several specific chromosomal lociand regions associated with important agronomic traits could be detected based on the revealing ofgenetic composition and breeding effects of founder parents. This study will improve the accuracy andefficiency of parental selection in cabbage breeding, and provide theoretical guide for the the creation ofnew founder parents.Based on the pedigree analysis of cabbage released,01-20(founder parent), other four sister lines(non-founder parents) and their derived lines, DH population from founder parent01-20and96-100wereused as materials for discovering specific chromosome loci and regions of founder parent01-20. Theconstructed high density linkage map and QTL analysis could clarify the relations between specifical lociof founder parent and important agronomic traits and elucidate the breeding effect of specific loci orregions.1. There were219cabbage varieties released during1982－2012in china, which included183hybrids, accounting for83.56%. The hybrid proportion reached96.74%in recent10years. The analysisof176varieties with known parental origination indicated that all the varieties were combined with261parental lines, which could be traced to171original resources, i.e.67domestic landraces and104alienaccessions. In all the original resources, the number of Heiyexiaopingtou derived varieties was the most(38), followed by Beijingzaoshu (27). The pedigree analysis showed that the combinations with twoparents from different geographical area or having great botanical characteristic difference always hadstrong heterosis. The rule of parental selection and combination was revealed based on the parentalcharacteristic of several major varieties in China. Pedigree of foundation parents Beijingzaoshu01-20and Heiyexiaopingtou21-3were drawn in this study.2. Polymorphism between parent01-20and96-100was screened with978EST-SSR,2173Scaffold-SSR,707Indel primer pairs. The polymorphic markers of273SSRs and261Indels weredetected. The linkage map has placed406SSR and Indel loci into nine groups. The map covered934.06cM, and average marker interval was2.3cM. All nine linkage group was assigned to nine chromosomeswith anchored markers.3. QTL analysis was performed to determine the markers associated with important agronomicaltraits of plant, leaf, head and seed, quality and disease resistance, according to the results of three seasonsin2011-2012. A total of171QTLs were detected for33agronomical traits. There were58QTLs for plant and leaf traits, and the number of QTLs (R2>10%) was38;57QTLs for head traits,38QTLs(R2>10%) and11QTLs were repeatedly detected in different seasons;38QTLs for leaf color and headcolor traits,26QTLs (R2>10%);11QTLs for quality and disease resistance traits,10QTLs (R2>10%);seven QTLs for silique and seed traits, five QTLs (R2>10%). These QTLs could be used for clarifing therelations between specific loci of founder parent01-20and important agronomic traits, and elucidatingthe breeding effect of specific loci.4. Several QTL clusters had been found on regions of chromosome1,3,7and9. The region in55.9～60.9cM on Chr.1contained nine plant and leaf trait QTLs and seven head trait QTLs. The regionof88～92cM on Chr.3contained five plant and leaf trait QTLs, seven head trait QTLs and one headcolor trait QTL. The region of27.8～30.9cM on Chr.7contained four head trait QTLs and two headcolor trait QTLs. The region of58.6～61.5cM on Chr.9contained three head traits QTLs, three leafcolor trait QTL and three quality and disease resistance traits. These regions provided importantreferences for the QTL fine mapping and cloning.5. According the phenotype comparison of founder parent01-20with other5sister lines, thefounder parent01-20had good comprehensive traits and no obvious defects. It not only inherited theearly maturity, but also improved in leaf color, head color and quality traits. The polymorphic patternsamong six sister lines showed that01-20had one unique allele,01-07-258had one unique allele,01-20and01-07-258had five same alleles which were however different from the other four sister lines.Based on the QTL analysis, there are five specific loci of founder parent associated with importantagronomic traits, which could elucidate the breeding effect of specific loci. Founder parent01-20possessed the greener outer leaves, smaller plant expansion, rounder head shape, less core length andbetter quality. The loci of Indel64on Chr.3and Indel353on Chr.9have significant cluster and geneticeffect.更多还原