中国石蒜SSR体系的建立及性状对应分析

Establishment of an SSR Analysis Protocol and Corresponding Analysis of Characteristics in Lycoris chinensis

为开发利用石蒜属球根花卉资源和开展分子标记辅助选择育种,利用正交试验和单因素试验建立了中国石蒜的SSR分子标记体系:20μL含Taq酶0.5U、Mg2+1.5mmol·L-1、dNTP0.125mmol·L-1、Primer0.5mmol·L-1、DNA50ng。利用该体系对石蒜属的8个物种和中国石蒜种内35个无性系进行扩增,每对引物在种间平均得到7个多态性位点,在中国石蒜种内得到9.5个多态性位点,反映出该体系具有较好的通用性和稳定性。利用对应分析法对中国石蒜的花葶高度、开花期等性状和SSR扩增条带之间的对应关系进行关联分析,初步得到与迟花期‘Oct’性状相关的Ly97和Ly98条带,与中矮花葶高度‘Mid’、‘Low’性状相关的Ly84、Ly87和Ly97条带等,并提出开展进一步的杂交和遗传测定,对辅助选择标记的可靠性进行验证。

Based on an orthogonal design and one-way experiment,an SSR molecular marker analysis protocol was established in Lycoris chinensis for the development and utilization of Lycoris resources and marker-assisted selection,which had a total volume of 20 μL containing 0.5 U of Taq DNA polymerase,1.5 mmol ·L-1 Mg2＋,0.125 mmol· L-1 dNTP,0.5 mmol· L-1 primer and 50 ng of template DNA. This protocol had been applied to an analysis of 8 species of Lycoris and 35 clones of L. chinensis with 7 and 9.5 polymorphic loci per primer on average obtained,respectively,which reflects a good performance in universality and stability of this protocol. Analysis of the corresponding relation between SSR loci and such traits as scape height and flowering time had preliminarily shown that loci Ly 97 and Ly 98 were correlated with late flowering time‘Oct’as well as that loci Ly 84,Ly 87 and Ly 97 with scape heights ‘Mid’and‘Low’. It is suggested to verify the reliability of these markers to be used in assisted selection by further hybridization and genetic test.