我国紫薇属植物AFLP分子标记体系的优化

Optimization of AFLP molecular marker system in Lagerstroemia plants in China

以采自广西、云南和河南等地的紫薇属Lagerstroemia植物紫薇L.indica,南紫薇L.subcostata,福建紫薇L.limii和桂林紫薇L.guilinensis叶片为材料,利用改良CTAB(十六烷基三甲基溴化铵)法提取叶片基因组DNA,获得了高质量的紫薇叶片DNA,并以其基因组DNA为模板进行了酶切连接,利用酶切连接的产物稀释一定倍数作为预扩增的模板,最后以稀释一定倍数的预扩增产物进行选择性扩增,进行紫薇和南紫薇的AFLP(扩增片段长度多态性)银染反应体系的优化。AFLP体系中每一步反应都设置了不同的反应体系,采用了160对引物作为初选引物,筛选出了10对适合紫薇基因组扩增的引物。结果表明,适宜紫薇基因组扩增的最佳酶切、预扩增和选择性扩增体系为:酶切连接体系1;预扩体系3;选扩体系3,反应体系中Mg2+质量浓度为1.2×10-6kg.L-1时扩增效果较好。

This study dealt with optimization of amplified fragment length polymorphism （AFLP） analysis based on the high quality DNA extracted by the improved cetyl trimethyl ammonium bromide （CTAB） method from the leaves of Lagerstroemia indica, L. subcostata, L. lime and L. guilinensis which were gathered in Guangxi, Yunnan and Henan Province. And their DNA were used as the template to carry on restriction-ligase reaction, then the product was diluted the certain multiple as template to take pre-amplification, finally the product was diluted and used as template to carry on selective amplification, and the best system of restrictionligase reaction, pre-amplication and silver staining suit for L. indica had been optimized. In the AFLP system each step of response had established different reacting system. The 10 pairs of primer suit for L indica were selected from 160 pairs of elementary primer. The result indicated that, the best systems suitable for the Lagerstroemia gene group were as follows： restriction-ligase reaction system 1; pre-amplification system 3 ; selective amplification system 3, in reacting system the best Mg2＋ concentration was 1.2×10-6kg.L-1. [ Ch, 1 fig. 7 tab. 20 ref. ]