葡萄无核基因定位与作图的研究

Mapping of Seedlessness Gene in Grapes Using SCAR Markers

以UBC 269484和GSLP1569的序列为支点,设计合成了包括UBC 269和GSLP1在内的9条引物,以葡萄 有核亲本红地球和无核亲本红光无核的DNA为模板,对这9个引物进行筛选,结果GSLP1、39970524 5号引物和 39970524 6号引物在无核亲本红光无核上扩增出了特异标记GSLP1569、39970524 5 564、39970524 6 1538和 39970524 6 1200。用这3个特异引物在红地球、红光无核、无核白和红地球×红光无核杂交组合F1代163株杂 种的DNA样上进行PCR扩增,结果4个特异标记在F1群体中与无核主效基因共分离。4个特异标记也出现于所 用组合中无核基因原始供给者无核白上。这些标记和葡萄无核主效基因相连锁。用QTXb17遗传作图软件,对葡 萄无核主效基因S定位与作图,当P=0.01时,LOD值在32.7～46.4之间,置信界限在0.2～9.9之间。这4个 特异标记和无核主效基因S处于在同一连锁群,位于无核主效基因S的两侧,覆盖基因组12.3cM。特异标记 39970524 5 564、GSLP1569、39970524 6 1538、39970524 6 1200距S基因的遗传距离分别为0.6cM、1.2cM、 4.9cM和11.1cM。

Nine primers (including UBC-269 and GSLP1) were designed and synthesized based on DNA sequences of UBC-269 (484) and GSLP1 (569).The template DNA from Red Globe (seeded paternal parent) and Flame Seedless (seedless maternal parent) were screened using these primers.For Flame Seedless,GSLP1 yielded specific marker GSLP1 (569);No.39970524-5 primer yielded specific marker 39970524-5-564; and No.6 primer yielded specific marker 39970524-6-1538 and 39970524-6-1200.GSLP1,No.39970524-5,and No.39970524-6 primers were used specifically to screen template DNA from the experimental plant materials.The results showed that the specific markers GSLP1 (569),39970524-5-564,39970524-6-1538 and 39970524-6-1200 were co-segregating with the major seedlessness gene.All these specific loci were also present in Thompson Seedless which was the initial donor of the seedlessness gene.It suggests that these SCAR markers are linked to a major grape seedlessness gene S.Markers order and map distance were estimated using the software ‘QTXb17’.This showed that GSLP1 (569),39970524-5-564,39970524-6-1538 and 39970524-6-1200 were tightly linked to gene S.When P=0.01,confidence limits for map distance ranged from 0.2 to 9.9; standard errors of map distance were from 0.6 to 1.9;LOD for linkage were from 32.7 to 46.4.These markers and the gene S were found to be in the same group.The markers were located on either side of gene S,covering 12.3 cM of the grape genome.The genetic distances between gene S and 39970524-5-564,GSLP1 (569),39970524-6-1538 and 39970524-6-1200 were 0.6 cM,1.2 cM,4.9 cM and 11.1 cM respectively.