Twenty-seven low-copy nuclear primers for Lindera obtusiloba (Lauraceae): A Tertiary relict species in East Asia.

Premise of the study: To investigate a more detailed evolutionary history of Lindera obtusiloba (Lauraceae) and other Lindera species, polymorphic low-copy nuclear primers were developed. Methods and Results: Unigenes of the L. obtusiloba transcriptome greater than 800 bp in length were randomly chosen for initial design of 168 primers. Agarose gel electrophoresis and Sanger sequencing were used to select low-copy nuclear genes. Twenty-seven primers were obtained and were used to investigate genetic diversity in 90 individuals from 24 populations. The nucleotide diversity ranged from 2.11 x 10(-3) to 8.99 x 10(-3), and haplotype diversity ranged from 0.57 to 0.97. These primers were also cross-amplified in L. aggregata, L. chunii, L. erythrocarpa, and L. glauca; up to 15 primers were successfully amplified in these related species. Conclusions: This methodology is effective for development of low-copy nuclear primers. The 27 primers developed here will be useful for evolutionary studies of L. obtusiloba and other Lindera species.