CAJ | 학술논문

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12.3 cM.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0.6 cM、1.2 cM、4.9 cM和11.1 cM.
이UBC-269484화GSLP1569적서렬위지점,설계합성료포괄UBC-269화GSLP1재내적9조인물,이포도유핵친본홍지구화무핵친본홍광무핵적DNA위모판,대저9개인물진행사선,결과GSLP1、39970524-5호인물화39970524-6호인물재무핵친본홍광무핵상확증출료특이표기GSLP1569、39970524-5-564、39970524-6-1538화39970524-6-1200.용저3개특이인물재홍지구、홍광무핵、무핵백화홍지구×홍광무핵잡교조합F1대163주잡충적DNA양상진행PCR확증,결과4개특이표기재F1군체중여무핵주효기인공분리.4개특이표기야출현우소용조합중무핵기인원시공급자무핵백상.저사표기화포도무핵주효기인상련쇄.용QTXb17유전작도연건,대포도무핵주효기인S정위여작도,당P=0.01시,LOD치재32.7～46.4지간,치신계한재0.2～9.9지간.저4개특이표기화무핵주효기인S처우재동일련쇄군,위우무핵주효기인S적량측,복개기인조12.3 cM.특이표기39970524-5-564、GSLP1569、39970524-6-1538、39970524-6-1200거S기인적유전거리분별위0.6 cM、1.2 cM、4.9 cM화11.1 cM.
Nine primers (including UBC-269 and GSLP1) were designed and synthesized based on DNA sequences of UBC-269484 and GSLP1569.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 GSLP1569;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 GSLP1569,39970524-5-564,39970524-6-1538 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 GSLP1569,39970524-5-564,39970524-6-1538 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,GSLP1569,39970524-6-1538 and 39970524-6-1200 were 0.6 cM,1.2 cM,4.9 cM and 11.1 cM respectively.