【作者】 韩国辉； 汪卫星； 向素琼； 贾志刚； 郭启高； 秦国新； 何桥； 李晓林； 梁国鲁；

【Author】 HAN Guo-hui,WANG Wei-xing,XIANG Su-qiong,JIA zhi-gang,GUO Qi-gao,QIN Guo-xin, HE Qiao,LI Xiao-lin,LIANG Guo-lu~* (Key Laboratory of Horticulture Science for Southern Mountainous Regions,Ministry of Education/College of Horticulture and Landscape Architecture.Southwest University,Chongqing 400715 China)

【机构】 南方山地园艺学教育部重点实验室/西南大学园艺园林学院；

【摘要】 以‘软条白沙’枇杷三倍体株系为试材,采用L25（5⁶）正交设计方法,对影响枇杷SCoT-PCR反应的Mg²⁺、dNTPs、引物、Taq DNA聚合酶及模板DNA用量等因素进行优化筛选,并对最适退火温度进行了探讨。建立了适于不同倍性枇杷分析的SCoT-PCR扩增体系,即20μL的优化体系中含有:Mg²⁺1.5 mmol·L^-1,dNTPs0.35 mmol·L^-1,引物0.5μmol·L^-1,Taq酶0.5 U,模板DNA50ng。引物最适退火温度为51.8℃。应用优化的反应体系,对三个枇杷品种（‘泸州六号’:2x,3x,4x,5x;‘龙泉1号’:2x,3x,4x;‘早红3号’:2x,3x,4x）的10个不同倍性株系进行扩增,结果表明:9个引物共扩增出128条带,片段大小为100-2000 bp,其中多态性条带84条,多态率为65.6%,与相应的二倍体比较,多倍材料均发生了不同程度的遗传变异。更多还原

【Abstract】 Triploid‘ruantiaobasha’ loquat was used as materials,orthogonal design method was applied to optimize SCoT-PCR amplification system of loquat in five factors such as Mg²⁺, dNTPs,primers,Taq DNA polymerases and template DNA at five levels,and the annealing temperatureand was also optimized.An optimal SCoT-PCR amplification system（20μL） of polyploidy loquat was established,i.e.dNTPs 0.35 mmol?L^-1,Mg²⁺ 1.5 mmol ? L^-1,primer 0.5 umol?L^-1,Taq polymerase 0.5 U,template DNA 50 ng.51.8℃was the most suitable annealing temperature.10 different ploidy cultivars of‘luzhouliuhao’（2x,3x,4x,5x）,‘longquanyihao’（2x, 3x,4x） and‘zaohongsanhao’（2x,3x,4x,） were analyzed with the optimized reaction system,128 bands were obtained by nine polymorphic SCoT primers,the size range of bands was typically between 100-2000bp,of which 84 bands were polymorphic,the polymorphism rate is 65.6%,and the ploidy had greater degree genetic variation compared with the corresponding diploid.更多还原