Adam-Blondon A-F, Roux C, Claux D, Butterlin G, Merdinoglu D, This P
URGV, INRA, 2 rue Gaston Crémieux, 91058, Evry 5708, France.
Theor Appl Genet. 2004 Sep;109(5):1017-27. doi: 10.1007/s00122-004-1704-y. Epub 2004 Jun 4.
The aim of the present work was to develop a microsatellite marker-based map of the Vitis vinifera genome (n=19), useful for genetic studies in this perennial heterozygous species, as SSR markers are highly transferable co-dominant markers. A total of 346 primer pairs were tested on the two parents (Syrah and Grenache) of a full sib population of 96 individuals (S x G population), successfully amplifying 310 markers. Of these, 88.4% markers were heterozygous for at least one of the two parents. A total of 292 primer pairs were then tested on Riesling, the parent of the RS1 population derived from selfing (96 individuals), successfully amplifying 299 markers among which 207 (62.9%) were heterozygous. Only 6.7% of the markers were homozygous in all three genotypes, stressing the interest of such markers in grape genetics. Four maps were constructed based on the segregation of 245 SSR markers in the two populations. The Syrah map was constructed from the segregations of 177 markers that could be ordered into 19 linkage groups (total length 1,172.2 cM). The Grenache map was constructed with the segregations of 178 markers that could be ordered into 18 linkage groups (total length 1,360.6 cM). The consensus S x G map was constructed with the segregations of 220 markers that were ordered into 19 linkage groups (total length 1,406.1 cM). One hundred and eleven markers were scored on the RS1 population, among them 27 that were not mapped using the S x G map. Out of these 111 markers, 110 allowed to us to construct a map of a total length of 1,191.7 cM. Using these four maps, the genome length of V. vinifera was estimated to be around 2,200 cM. The present work allowed us to map 123 new SSR markers on the V. vinifera genome that had not been ordered in a previous SSR-based map (Riaz et al. 2004), representing an average of 6.5 new markers per linkage group. Any new SSR marker mapped is of great potential usefulness for many applications such as the transfer of well-scattered markers to other maps for QTL detection, the use of markers in specific regions for the fine mapping of genes/QTL, or for the choice of markers for MAS.
本研究的目的是开发基于微卫星标记的葡萄(Vitis vinifera,n = 19)基因组图谱,由于SSR标记是高度可转移的共显性标记,该图谱对这种多年生杂合物种的遗传研究很有用。在一个由96个个体组成的全同胞群体(S×G群体)的两个亲本(西拉和歌海娜)上测试了总共346对引物,成功扩增出310个标记。其中,88.4%的标记在两个亲本中至少有一个是杂合的。然后在雷司令(Riesling)上测试了总共292对引物,雷司令是自交产生的RS1群体(96个个体)的亲本,成功扩增出299个标记,其中207个(62.9%)是杂合的。在所有三种基因型中,只有6.7%的标记是纯合的,这突出了此类标记在葡萄遗传学中的价值。基于两个群体中245个SSR标记的分离构建了四张图谱。西拉图谱是根据177个标记的分离构建的,这些标记可排列成19个连锁群(总长度1172.2 cM)。歌海娜图谱是根据178个标记的分离构建的,这些标记可排列成18个连锁群(总长度1360.6 cM)。共识S×G图谱是根据220个标记的分离构建的,这些标记排列成19个连锁群(总长度1406.1 cM)。在RS1群体上对111个标记进行了评分,其中27个在使用S×G图谱时未被定位。在这111个标记中,110个使我们能够构建一张总长度为1191.7 cM的图谱。利用这四张图谱,估计葡萄的基因组长度约为2200 cM。本研究使我们能够在葡萄基因组上定位123个新的SSR标记,这些标记在之前基于SSR的图谱(Riaz等人,2004年)中未被排序,平均每个连锁群有6.5个新标记。任何新定位的SSR标记在许多应用中都具有很大的潜在用途,例如将分布良好的标记转移到其他图谱上进行QTL检测、在特定区域使用标记进行基因/QTL的精细定位,或用于MAS标记的选择。