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使用20组共400条RAPD随机引物,在“薄皮”和“垂枝”两个板栗亲本无性系及其5个杂交后代进行筛选,共有187条引物获得了清晰、重复性好的多态性条带,占供试引物的46.75%。应用筛选出的多态性引物在两亲本及其60个杂交后代中进行扩增,获得的多态性条带在后代中的分离比用卡方测验(α=0.05)分析,结果共有143个标记符合1:1测交分离比,其中来源于母本“薄皮”板栗的有88个,来源于父本“垂枝”的有55个。采用作图软件Mapmaker和回交群体模型分别对符合测交分离比的标记进行亲本特异的标记分群和连锁分析,设定LOD值为3和最大重组值θ为0.50作为可信统计度与最大连锁标记数量之间的最佳组合,其中母本的标记定位了10个连锁组,父本的标记定位了7个连锁组。母本“薄皮”板栗遗传连锁图共包含88个标记,覆盖了板栗基因组总长约823.1cM(Kosambi,以下同),占基因组的66.7%;父本“垂枝”板栗遗传连锁图共包含55个标记,覆盖了板栗基因组总长约720.8cM,占基因组的41.7%。母本“薄皮”板栗遗传连锁图上标记间的遗传距离为1.6~26.5cM,平均9.4cM,连锁组的大小从15.5cM到166.3cM,平均为82.3cM;父本“垂枝”板栗遗传连锁图上标记间的遗传距离为2.1~39.9cM,平均13.1cM,连锁组的大小从53.5cM到139.6cM,平均为103.0cM。
A total of 400 random amplified polymorphic DNA (RAPD) primers were used to screen two clonal chestnut clones and their five hybrid progenies in “Thin skin” and “Weeping sticks”. A total of 187 primers obtained clear and reproducible Polymorphic bands, accounting for 46.75% of primers. Polymorphic primers were screened out in both parents and their 60 hybrids. The segregation of the polymorphic bands obtained was analyzed in the offspring using the chi-square test (α = 0.05). As a result, 143 The markers were segregated in a ratio of 1: 1, of which 88 were derived from the female parent Chestnut and 55 from the male parent. Using mapping software Mapmaker and backcross population models, the marker-specific segregation and linkage analysis were performed respectively on the markers with segregation ratio, setting LOD value as 3 and maximum recombination value as 0.50 as credible statistic and maximum linkage The best combination of the number of markers, in which the male parent’s marker locates 10 linkage groups, the male parent’s marker locates 7 linkage groups. The total length of the chestnut genome was about 823.1cM (Kosambi, the same below), accounting for 66.7% of the total genome. The genetic linkage map of the male parent “Chestnut” A total of 55 markers, covering the chestnut genome total length of about 720.8cM, accounting for 41.7% of the genome. The genetic distance between the markers on the genetic map of the chestnut was from 1.6 to 26.5cM with an average of 9.4cM and the linkage group size was from 15.5cM to 166.3cM with an average of 82.3cM. "The genetic distance between the markers on chestnut genetic linkage map was 2.1 ~ 39.9cM, with an average of 13.1cM. The linkage group size ranged from 53.5cM to 139.6cM, with an average of 103.0cM.