The effects of 5 factors (template DNA, Mg2+, dNTPs, Taq DNA polymerase, and primer) on the polymerase chain reaction (PCR) were investigated to optimize the start codon targeted polymorphism (SCoT)-PCR system of Dactylis glomerata L., using an orthogonal design L16 (45). A suitable SCoT-PCR system for D. glomerata was established; the 20 μL reaction volume contained 3.0 mM Mg2+, 0.2 mM dNTPs, 1.0 U Taq DNA polymerase, 0.2 μM primer, 20 ng template DNA, and 2 μL 10X buffer.
Dactylis glomerata L.
Orchardgrass is a highly variable, perennial forage grass that is cultivated throughout temperate and subtropical regions of the world. Despite its economic importance, the genetic relationship and distance among and within cultivars are largely unknown but would be of great interest for breeding programs.
The accurate identification of orchardgrass (Dactylis glomerata L.) cultivars is necessary to ensure purity for consumers, the effective utilization of cultivars, and to protect the intellectual property for breeders. Therefore, this study aimed to use SSR to construct DNA fingerprinting of orchardgrass cultivars. The genetic diversity of 32 orchardgrass cultivars originated from 21 countries, but grown in China, was assessed using a set of 29 SSR markers distributed across 9 linkage groups of the orchardgrass genome.