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以普通栽培番茄(Solanum lycopersicum)99165-30为母本,野生多毛番茄(Solanum habrochaites)LA1777为父本进行杂交,通过单粒传得到了含有80个F5︰6家系的重组自交系分离群体,利用荧光AFLP分子标记技术构建番茄分子遗传连锁图谱。AFLP标记采用MseⅠ和EcoRⅠ两种内切酶及荧光标记(IRD-700或IRD-800)的E+3和非荧光标记的M+3引物组合进行选择性扩增,扩增结果经95℃预变性后在6%变性聚丙烯酰胺凝胶上电泳2.5h,运用LICOR公司的NEN Global Edition IR2 DNA Analyzer(Model 5200 LI-COR Biosciences,Lincoln,NE)荧光扫描检测DNA多态性。对RILs群体中产生分离的274个AFLP标记运用JoinMap3.0软件分析,得到一张番茄分子遗传连锁图谱,图谱总长度为662cM,共包括18个主要连锁群,125个多态性分子标记。每条连锁群上的标记数在3～22个之间,连锁群的长度在14.0～58.0cM的范围内,平均图距在2.27～13.3cM。总平均距离5.3cM,本研究中构建的番茄永久遗传图谱,为番茄分子辅助育种及重要农艺性状的定位奠定了基础。
Solanum lycopersicum 99165-30 was used as the female parent and Solanum habrochaites LA1777 as the male parent to obtain the segregation population of 80 inbred lines with 80 F5: Construction of Tomato Molecular Genetic Linkage Map Using Fluorescent AFLP Markers. The AFLP marker was selectively amplified using a combination of MseI and EcoRI endonucleases and E + 3 fluorescent markers (IRD-700 or IRD-800) and a non-fluorescent labeled M + 3 primer. The amplification results were pre-conditioned at 95 ° C After denaturation, electrophoresis was performed on a 6% denaturing polyacrylamide gel for 2.5 h and the DNA polymorphism was detected by fluorescence scanning using NEN Global Edition IR2 DNA Analyzer (Model 5200 LI-COR Biosciences, Lincoln, NE) from LICOR. A total of 272 AFLP markers isolated from RILs population were analyzed using JoinMap 3.0 software. A molecular linkage map of tomato was obtained. The total length of the map was 662 cM, including 18 major linkage groups and 125 polymorphic molecular markers. The number of markers on each linkage group ranged from 3 to 22, and the linkage group length ranged from 14.0 to 58.0 cM with an average distance of 2.27 to 13.3 cM. The total average distance was 5.3 cM. The permanent genetic map of tomato constructed in this study laid the foundation for tomato molecular assisted breeding and the location of important agronomic traits.