Manescu C, Hamamouch N, Maios C, Harfouche A, Doulis A G, Aravanopoulos F A
Mediterranean Agronomic Institute of Chania, Chania, Greece.
Genet Mol Res. 2011 Aug 30;10(3):1891-909. doi: 10.4238/vol10-3gmr1179.
Gene mapping for a Cupressus species is presented for the first time. Two linkage maps for the Mediterranean cypress (Cupressus sempervirens) varieties, C. sempervirens var. horizontalis and C. sempervirens var. pyramidalis, were constructed following the pseudo-testcross mapping strategy and employing RAPD, SCAR and morphological markers. A total of 427 loci (425 RAPDs, two SCARs) representing parents and F(1) progeny were screened for polymorphism with 32 random decamer and two SCAR primers. A morphological marker defined as "crown form" was also included. Of 274 polymorphic loci, the 188 that presented Mendelian inheritance formed the mapping dataset. Of these loci, 30% were mapped into seven linkage groups for the horizontalis (maternal) and four linkage groups for the pyramidalis (paternal) map. The putative "crown form" locus was included in a linkage group of both maps. The horizontalis and the pyramidalis maps covered 160.1 and 144.5 cM, respectively, while genome length was estimated to be 1696 cM for the former variety and 1373 cM for the latter. The four RAPD markers most tightly linked to crown form were cloned and converted to SCARs. Each of the cloned RAPD markers yielded two to three different sequences behaving as co-migrating fragments. Two SCAR markers, SC-D05(432) and SC-D09(667), produced amplified bands of the expected sizes and maintained linkage with the appropriate phenotype, but to a lesser extent compared to their original RAPD counterparts. These linkage maps represent a first step towards the localization of QTLs and genes controlling crown form and other polygenic traits in cypress.
首次展示了一种柏木属物种的基因图谱。按照伪测交作图策略并使用RAPD、SCAR和形态学标记，构建了地中海柏木（Cupressus sempervirens）变种水平叶柏（C. sempervirens var. horizontalis）和塔形柏（C. sempervirens var. pyramidalis）的两个连锁图谱。用32个随机十聚体引物和两个SCAR引物筛选了代表亲本和F(1)后代的总共427个位点（425个RAPD、两个SCAR）的多态性。还包括一个定义为“树冠形态”的形态学标记。在274个多态性位点中，呈现孟德尔遗传的188个位点构成了作图数据集。在这些位点中，30%被定位到水平叶柏（母本）的7个连锁群和塔形柏（父本）的4个连锁群中。假定的“树冠形态”位点包含在两个图谱的一个连锁群中。水平叶柏和塔形柏的图谱分别覆盖160.1和144.5厘摩，而前者变种的基因组长度估计为1696厘摩，后者为1373厘摩。与树冠形态最紧密连锁的四个RAPD标记被克隆并转化为SCAR。每个克隆的RAPD标记产生两到三个表现为共迁移片段的不同序列。两个SCAR标记，SC-D05(432)和SC-D09(667)，产生了预期大小的扩增条带，并与适当的表型保持连锁，但与它们原来的RAPD对应物相比程度较小。这些连锁图谱是朝着定位控制柏树树冠形态和其他多基因性状的QTL和基因迈出的第一步。
