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以197个橡胶树杂交群体样本为材料,通过对影响AFLP技术的多种关键因素进行研究,建立了一套适合橡胶树的AFLP技术优化体系。其中优化的关键的因素为:(1)酶切-连接时DNA模板为250ng,反应体系采用各3UEcoRI/MseI/T4连接酶。反应时间为10h;(2)预扩增模板为稀释一倍的酶切连接产物,用量为1μL;(3)选择性扩增要用稀释倍数为40倍的预扩增产物。本研究为橡胶树遗传图谱的构建及育种的辅助选择提供有力的工具。
Based on 197 rubber tree hybrid population samples, a set of AFLP technology optimization system for rubber tree was established by studying a variety of key factors affecting AFLP technology. The key factors for optimization are as follows: (1) DNA template is 250ng when ligated and ligated, and each 3UEcoRI / MseI / T4 ligase is used in the reaction system. The reaction time is 10h; (2) the pre-amplification template is a diluted product of digestion 1μL; (3) the selective amplification of the pre-amplification product with a dilution of 40 times. This study provides a powerful tool for the construction of rubber tree genetic map and the auxiliary selection of breeding.