VALIDATION OF SINGLE NUCLEOTIDE POLYMORPHISMS IN RIBOSOMAL DNA INTERNAL TRANSCRIBED SPACER SEQUENCES OF WOLFFIA MICROSCOPICA (GRIFF). KURZ

Duckweeds are aquatic plants with 35 species and 2 hybrid species belonging to five genera (Spirodela, Landoltia, Lemna, Wolffiella, and Wolffia) within the Lemnaceae family. The only surviving Wolffia microscopica (Griff.) Kurz (accession number 9276) was lost from all collections in 2009. Fortunately, some accessions of this species were rediscovered in Sughad, India, and Bangladesh in 2014. The purpose of this study is to identify single nucleotide polymorphisms (SNPs) in the internal transcribed spacer sequence (ITS) region of 10 Wolffia microscopica accessions. In all, 31 plasmids from 8 accessions were investigated, resulting in 15 plasmid variants and 24 SNPs. However, based on the outcome of the SNP evaluation techniques (restriction enzyme, polymerase chain reaction analysis using certain SNPs, and web-based allele specific primer design primers), these SNPs were not Wolffia microscopica subspecies-specific. Therefore, for the 10 investigated Wolffia microscopica accessions, the ITS region works well for species-level classification but is not appropriate for subspecies level classification.