利用正交设计优化向日葵ISSR-PCR反应体系

向日葵产业的发展对调整黑龙江省种植业结构、利用中低产田、增加农民收入具有十分重要的作用,为了推广ISSR分子标记技术在向日葵分子育种上的应用,利用正交设计对向日葵ISSR-PCR反应体系中的5个因素(模板DNA、dNTP浓度、Mg2+浓度、引物浓度、Taq酶用量)在4个水平上进行优化,确立了适合向日葵ISSR-PCR反应的20μL体系:50ng模板DNA、2.0μmol·L-1dNTP、37.5μmol·L-1 Mg2+、8μmol·L-1引物、8UDNA Taq酶。

Optimization of ISSR-PCR System for Sunflower Based on Orthogonal Design

The development of sunflower industry plays an important role to adjust the structure of planting in-- dustry, use the middle and low yield fields and increase income of farmers. In order to promote sunflower mo- lecular marker and molecular breeding, the orthogonal design was used to optimize ISSR-PCR amplification sys- tem at four levels of five factors in sunflower （template DNA, dNTPs, Mg^2＋ , primer and Taq DNA polymer- ase）. The most suitable ISSR-PCR reaction system for sunflower was total 20 μL containing 50 ng template DNA,2.0 mmol·L^-1 dNTP,37.5 mmol· L^-1 Mg^2＋ ,8 mmol·L^-1 primer and 8 U Taq polymerase.