DNA-based test for detection of annual and intermediate ryegrass

摘要

We have developed a novel TaqMan quantitative PCR (Q-PCR) based DNA test for detecting annual and/or intermediate ryegrass types in perennial ryegrass. This DNA test was designed using an insertion/deletion (InDel) site in the LpVRN2_2 gene. The new DNA test is more reliable, accurate, and cost effective in detecting annual and intermediate type contamination in perennial ryegrass, having a sensitivity of 0.04% in a sample size of 5000 seeds. Use of a higher sample size (12.5-fold higher compared to the SRF test) provides additional accuracy in detecting the level of contamination A forward and reverse set of primers also identified an approximately 450 bp fragment in or near the LpVRN1 promoter, the fragment being present for all perennial, but not annual, varieties tested.