Decroocq V, Favé M G, Hagen L, Bordenave L, Decroocq S
INRA Centre de Bordeaux, U.R.E.F.V, 71 Avenue E. Bourleaux, 33883 Villenave d'Ornon France.
Theor Appl Genet. 2003 Mar;106(5):912-22. doi: 10.1007/s00122-002-1158-z. Epub 2002 Dec 3.
EST microsatellite markers were developed in apricot (Prunus armeniaca L.) and grape (Vitis vinifera L.). cDNA libraries from either apricot leaves or grape roots were used in an enrichment procedure for GA and CA repeats. The transferability of EST simple sequence repeat (SSR) markers from apricot and grapevine to other related and unrelated species was examined. Overall, grape primers amplified products in most of the Vitaceae accessions while the apricot primers amplified polymorphic alleles only in closely related species of the Rosaceae. In this taxonomic family, ten EST SSR loci were tested, and one single primer pair, PacB22, was amplified across species and sections in the Prunoideae and Maloideae. Sequencing of EST SSR loci in other species and genera confirmed a higher level of conservation in the microsatellite motif and flanking regions in the Vitaceae compared to the Rosaceae. Two distinct fragments of the PacB22 locus amplified across the Malus and Pyrus genera; however, while the coding region was highly conserved, the microsatellite repeat motif was no longer present. The banding pattern was explained by base substitution and insertion/deletion events in the intronic region of PacB22. This study includes the determination of the degree of polymorphism detected among species and genera in two unrelated taxonomic families and the evaluation of the information provided by the microsatellite repeats and the flanking regions.
在杏(Prunus armeniaca L.)和葡萄(Vitis vinifera L.)中开发了EST微卫星标记。来自杏叶或葡萄根的cDNA文库用于GA和CA重复序列的富集过程。研究了EST简单序列重复(SSR)标记从杏和葡萄向其他相关和不相关物种的转移性。总体而言,葡萄引物在大多数葡萄科种质中扩增出产物,而杏引物仅在蔷薇科的近缘物种中扩增出多态性等位基因。在这个分类科中,测试了10个EST SSR位点,并且一对单一引物PacB22在李亚科和苹果亚科的物种和组间都能扩增。对其他物种和属中EST SSR位点的测序证实,与蔷薇科相比,葡萄科微卫星基序和侧翼区域的保守程度更高。PacB22位点的两个不同片段在苹果属和梨属中扩增;然而,虽然编码区高度保守,但微卫星重复基序不再存在。条带模式是由PacB22内含子区域中的碱基替换和插入/缺失事件解释的。本研究包括确定两个不相关分类科中物种和属之间检测到的多态性程度,以及评估微卫星重复序列和侧翼区域提供的信息。