澳洲坚果SSR体系的建立及F_1代的鉴定

为了探索澳洲坚果SSR-PCR的可行性,采用单因素分析法对影响PCR的各个因素进行分析,建立SSR-最佳反应体系。结果表明:在25μL PCR反应体系中,当Mg~(2+)浓度、d NTPs浓度、引物浓度、Taq DNA聚合酶用量分别为2.50 mmol/L、0.10 mmol/L、0.40μmol/L、1.25 U时,PCR扩增效果最好。筛选到19对可得到扩增结果的SSR引物,其中有2对在以900为母本、294为父本的组合中PCR扩增结果存在差异,可以应用于900×294组合的F1代鉴定。

Establishment of a SSR Reaction System for Identification of F1 Generation of Macadamia

In order to explore the feasibility of SSR-PCR in Macadamia, a singular factor test was used to optimize SSR-PCR amplification system for macadamia. The results showed the optimal PCR reaction system was 2.50 mmol/L Mg2+, 0.10 mmol/L dNTPs, 0.40 μmol/L primers, and 1.25 U Taq DNA polymerase in a volume of 25 μL. Nineteen pairs of SSR primers with amplifications were obtained in SSR-PCR, but only two pairs of primers had different results in PCR amplification in the combination of 900 as female and 294 as male parents. These two pairs of primers can hence be used to identify the F1 generation of the 900 ×294 combination. Therefore the SSR-PCR could be used for identification of hybrid progeny in macadamia.