Selection and Sequence Alignment of PCR Primers for Identifying Zizhi Strain

  Objective  The molecular markers of the cultivated strain of Ganoderma sinense, Zizhi S2 (aka Wu-Zhi No. 2), recently popularized in Fujian and surrounding provinces were studied to facilitate the authentication of the medicinal fungus.

  Method   Relevant primers of Zizhi S2 showing clear and stable bands and polymorphism were screened using PCR. Phylogenetic tree of UPMGA clustering analysis on the verified authentic cultivars was constructed to determine their relationship by genetic distance as well as antagonistic reaction. Subsequently, sequences of the selected primers were blasted on the genome of Zizhi S2 to validate the methodology.

  Result  There were 2 RAPD-PCR and 3 ISSR-PCR primers found to clearly and stably amplify the specific or polymorphic bands. However, the sites and numbers on the scaffolds of the Zizhi S2 genome that matched the sequences of the 5 primers were not same.

  Conclusion   It was confirmed that three primers(ISSR13, S1326, and S1506)could be effectively used for identification of Zizhi cultivated strains based on sequences alignment with the genome of G. sinense strain Zizhi S2.