摘要
Rice cultivar Norin 8 and its mutant Norin 8m harbour bentazon resistance trait and bentazon susceptibility trait respectively. A total of 360 arbitrary 10-mer oligonucleotide primers were screened on the genomic DNA of Norin 8 and Norin 8m with RAPD technique. Among which, five primers produced seven polymorphic RAPD bands between Norin 8 and Norin 8m. Amplified RAPD polymorphic products were cloned and sequenced. The sequences were used to design primers for PCR. Five SCAR markers, SCAR/G18/883, SCAR/G18/890, SCAR/G18/919/948, SCAR/D10/1237 and SCAR/F03/1186, were developed from OPG18/943, OPG18/972, OPD10/1248 and OPF03/1198. F-2 progeny of 320 individuals was analyzed to map SCAR markers in relationship to ben or Ben genes. SCAR markers of SCAR/G18/883, SCAR/G18/890, SCAR/G18/919/948 were shown to cosegregate with ben or Ben genes, and SCAR/D10/1237 to be linked of Ben gene with a distance of (14.8 +/- 2.1) cM. The genetic linkage to ben gene and SCAR markers was identified by a pair of near isogenic lines H121 and Hben121. Southern blotting analysis and segregation ratio of F-2 progeny revealed that OPG18/943 and OPG18/972 were single-copy in genome, and locus of OPG18/943 and OPG18/972 were allelic and sequence tagged sites. It is the first report on molecular markers linked to ben or Ben genes. The markers are useful to marker-assisted selection for the breeding and tag ben gene with map-based cloning.
利用RAPD技术对水稻品种农林 8号 (含苯达松抗性基因Ben)和其突变体农林 8号m (含苯达松敏感致死基因ben)进行标记 ,从 36 0个 10bp寡核苷酸随机引物中筛选出 5个引物产生的 7个RAPD标记。经对多态性标记的克隆和序列分析 ,再设计PCR引物 ,将其中 4个RAPD标记OPG18/ 94 3、OPG18/ 972、OPD10 / 12 4 8和OPF0 3/ 1198转化成SCAR标记SCAR/G18/ 883、SCAR/G18/ 890、SCAR/G18/ 919/ 94 8、SCAR/D10 / 12 37、SCAR/F0 3/ 1186。通过对农林 8号×农林 8号mF2 分离群体 32 0个单株的连锁分析及在 1对含ben基因的近等基因系H12 1和Hben12 1中验证 ,标记SCAR/G18/ 883、SCAR/G18/ 890、SCAR/G18/ 919/ 94 8与Ben或ben基因共分离 ,SCAR/D10 / 12 37与Ben基因的遗传距离为 (14 .8± 2 .1)cM。经Southernblotting分析并结合F2 代分离比例表明 ,标记OPG18/ 94 3、OPG18/ 972及其转化的SCAR标记在基因组中为单拷贝序列 ,且OPG18/ 94 3和OPG18/ 972为一对等位STS位点。这是首次报道与ben或Ben基因相连锁的分子标记。本研究为利用分子标记辅助ben基因的转育及利用图位克隆技术分离ben基因提供了有用的分子标记。
基金
国家自然科学基金 (19890 3 0 0 )
安徽省自然科学基金 (0 0 0 41110 )~~
