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枣(Ziziphus jujuba Mill.)遗传连锁图谱构建及性状的QTL定位研究

Construction of Genetic Linkage Map and Mapping QTL for Some Traits in Chinese Jujube (Ziziphus jujuba Mill.)

【作者】 申连英

【导师】 葛会波;

【作者基本信息】 河北农业大学 , 果树学, 2005, 博士

【摘要】 枣(Ziziphus jujuba Mill.)为鼠李科(Rhamnaceae)枣属(Ziziphus Mill.)植物。是我国特有的宝贵种质资源。本研究利用2个综合性状优良的鲜食枣品种冬枣和临猗梨枣杂交建立了第一个枣属植物遗传分离群体,绘制了枣属植物的第一张高密度分子遗传连锁图谱,在此基础上进行了性状的分子标记研究。 本研究采用“局部隔离法”,选用花粉发芽率低的冬枣和花粉发芽率较高的临猗梨枣进行混合栽植自然杂交授粉,对由此获得的冬枣实生子代采用AFLP银染技术进行杂种鉴定,建立了由161株冬枣×临猗梨枣杂交子代组成的枣F1代分离群体。采用筛选出的50对EcoRI/MseI引物组合,对分离群体中的150株个体进行了AFLP多态性检测,采用Joinmap3.0CP作图模型构建了包含333个AFLP多态性位点的14个连锁群组成的枣高密度遗传连锁图谱。连锁群的LOD值在4~7范围之间,覆盖基因组总长度1237.4cM,标记间平均图距3.9cM。 应用MapQTL4.0软件,选用Lander提出的区间作图法对枣性状进行了QTL检测。共检测到相关QTL110个。其中与枣树各枝干类型节间长度(包括不同发育时期)性状相关的QTL30个,与枣树叶片形态(包括不同发育时期)相关的QTL30个,与枣树叶片NPK含量有关的QTL13个,与枣树抗寒性有关的QTL16个和与枣树抗病性有关的QTL21个。在这110个QTL中,有29个主效QTL,107个QTL与AFLP标记共分离或紧密连锁(与相邻AFLP标记图距为0~5cM),87个QTL与AFLP标记共分离或紧密连锁(与相邻AFLP标记图距在0~1cM),28个与AFLP标记共分离或紧密连锁的QTL的LOD值大于3.5。根据不同性状的QTL在连锁群中的分布特征,确定出LG4为与枣树生长与形态特征密切相关的连锁群。LG9为与枣树叶片形态密切相关的连锁群。与枣树氮磷钾效率相关的QTL偏向于分散分布,与枣树抗寒性有关的QTL趋向于集中分布,并且偏向于分布在连锁群的端部。涉及枣树各类枝条节间长度、叶部形态、叶片NPK含量、抗寒性和抗病性等性状的110个QTL在枣遗传连锁图谱存在着2个或2个以上QTL共分离或紧密连锁现象。全基因组范围内共有30处由2个或2个以上QTL共分离或紧密连锁区域。 枣树高密度遗传连锁图谱的绘制和性状QTL定位为挖掘枣树有益基因打下了基础,为枣树分子遗传育种提供了依据。

【Abstract】 Chinese Jujube (Ziziphus jujuba Mill.), a precious germplasm resources originated from China, belongs to Ziziphus, Rhamnaceae family. The first high-density genetic linkage map in Ziziphus was constructed based on a F1 segregating population derived from a cross between two jujube varieties with good complex characters, Dongzao (Ziziphus jujuba Mill. cv. Dongzao) and Linyilizao(Ziziphus jujuba Mill. cv. Linyilizao). QTLs linked to some agronomic traits were located in the map.F1 segregating population of 161 individuals was identified by AFLP silver staining technique from 165 Dongzao progenies of open pollination of Dongzao and Linyilizao thanks to the few germinating pollen with Dongzao and a lot with Linyilizao. 150 F1 individuals were scored for marker polymorphisms. A high density genetic map was constructed using Joinmap3.0 setting CP (cross pollination population) type data based on the AFLP markers generated by 50 EcoRI/MseI primer pairs, 333 AFLP markers located on 14 linkage groups, with an average distance of 3.9cM between adjacent markers, covering a total distance of 1237.4cM of Chinese jujube genome. It was the first extensive genetic linkage map in Ziziphus to-date, the marker density of which could be used for mapping QTL linked to quantitative traits.110 putative QTLs was located in the genetic linkage map with MapQTL4.0 software using interval mapping method, 30 QTLs for length between nodes on different branches, 30 QTLs for some leafy traits, 13 QTLs for leaf NPK content, 16 QTLs for cold resistance and 21 QTLs for disease resistance, among them involved 87 QTLs with distance from the nearest AFLP marker between 0-1 cM, 20 QTLs with distance from the nearest AFLP marker between 1-5 cM, 29 QTLs with major effects, and 28 major QTLs co-located or closely linked with the nearest AFLP marker(with distance from the nearest AFLP marker between 05cM).Genes would cluster together on the linkage groups for their similar functions, QTLs related to length between nodes on different branches and some leafy traits closely distributed on LG4, and 9 QTLs for leafy traits located on LG9, QTLs for cold-resistance apparently clustered at top or tail of related linkage groups, contrary to 13 QTLs related to leaf NPK content fell onto 10 linkage groups. 30 regions were detected, which two or more QTLs controlling different traits co-located or closely linked each other in the genetic linkage map.These results would be useful for interest gene cloning and marker-assisted selection for genetic improvement in Chinese jujube.

【关键词】 遗传连锁图谱AFLPQTL
【Key words】 Chinese jujubeGenetic linkage mapAFLPQTL
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