Sequence characteristics and phylogenetic implications of the nrDNA internal transcribed spacers (ITS) in the genus Nymphaea with focus on some Indian representatives

Abstract

Nymphaea, an aquatic perennial herb with exceptionally beautiful flowers and floating leaves, is well represented globally. Out of ten species reported from India, the internal transcribed spacers (ITS) region of nrDNA was investigated in seven species of Nymphaea viz. N. alba var. rubra, N. caerulea, N. × marliacea, N. nouchali, N. pubescens, N. rubra and N. tetragona. Barring N. pubescens, whereby double peaks detected in the sequencing chromatograms may be due to random mutations occurring in some of the ITS paralogues, the additional signals detected for N. alba var. rubra and N. rubra are probably influenced by recent hybridization and introgression. Our study on sequence characteristics of ITS 1 and ITS 2 revealed high G + C content (ITS 1, 45.5–48.4%; ITS 2, 50.2–51.5%) and sequence divergence. Percentage of sequence divergence based on substitution and substitution plus indels is 44.15 and 57.19, respectively, for the ITS 1; 29.74 and 47.96% were recorded for the ITS 2. Although highly variable, conserved motifs within the ITS 1 and ITS 2 region of Nymphaea were identified and are found to be common throughout the order Nymphaeales. Sequence analysis of the ITS 1 and ITS 2 failed to detect any variation between two morphotypes of N. nouchali, namely N. nouchali JD 06 and N. nouchali JD 07, differing in flower color and found at the same geographical location. However, on comparison with another specimen of N. nouchali found at a different location, they showed considerable variation in nucleotide composition. Complemented by sequence data retrieved from GenBank, phylogenetic tree reconstruction of the genus Nymphaea based on neighbor-joining, maximum parsimony, maximum likelihood and Bayesian inference methods is presented and discussed.