Development of SSCP markers for genes related to oil synthesis in cotton | 棉花油分合成相关基因的SSCP标记开发

صينى. 摘　　要：本研究根据棉花油份合成相关基因（lysophosphatidic acid acyltransferase,LPAAT;diacylglycerol acyltransferase,DGAT;phosphoenolpyruvate carboxylase,PEPC）设计了75对引物,在8个棉花品种中运用SSCP技术对其进行多态性检测。结果表明,在75对引物中,经PCR扩增,有55对引物在1.5%的琼脂糖凝胶电泳上出现清晰的条带。55对引物又经SSCP分析,21对引物在四个陆地棉之间产生47个多态性位点;16对引物在4个海岛棉间产生30个多态性位点;39对引物在这8个棉花品种间共产生104个多态性位点,13对引物在4个陆地棉之间和四个海岛棉之间同时出现多态性。由上可知,运用SSCP技术分析棉花油份合成相关基因得到的多态性位点较多。进一步经聚类分析后可知,3个陆地棉（中12,SG747,苏远7252）,3个海岛棉（海7124,Giza75,海岛棉3-79）分别聚在了一起。

إنجليزي. Genetic variation exists in cotton seed oil content, and genes and gene expression related to oil content and lipid synthesis are poorly understood. In this study, a total of 75 primer pairs were designed for oil synthesis-related genes that encode lysophosphatidic acid acyltransferase （LPAAT）, acyl-coa： diacylglycerol acyltransferase （DGAT） and phosphoenolpyruvate carboxylase （PEPC）. Eight genotypes were selected from the cultivated tetraploid cotton including four G. hirsutum genotypes and four G. barbadense genotypes. 55 primer pairs each amplified a clear single band from the genomic DNA when assayed by 1.5% agarose gels. Using the single strand conformation polymorphism （SSCP） analysis based on non-denaturing polyacrylamide gel electrophoresis, 21 primer pairs yielded 47 polymorphic loci among the four G. hirsutum genotypes, while 16 pairs yielded 30 polymorphic loci among the four G. barbadense genotypes. The SSCP polymorphism was much higher between the two species where a total of 39 pairs produced 104 polymorphic markers. Meanwhile,13 pairs yielded polymorphism among the four G. barbadense genotypes and among the four G. hirsutum genotypes. The polymorphic SSCP markers grouped six of the eight genotypes into two species groups, as expected. The results showed that SSCP technique can detect high polymorphism in cultivated tetraploid cotton, which can facilitate mapping of genes for seed oil production.