Abstract

• In-silico trait-linked phylogenetic framework of 9 Paphiopedilum species distributed in India based on combined nrDNA and cpDNA ( ITS2 + matK + rbcL ) dataset by MP and Bayesian analysis. • Ancestral trait tracing of 47 characters viz., morphological and chromosomal using Mesquite 3.61. • Sharing of passport traits to public may augment to prevent unethical trade of Paphiopedilum species throughout the distributional range. Paphiopedilum Pfitzer is one of the largest horticulturally utilized genera of slipper orchids. Similar characters of slipper orchids within Paphiopedilum may camouflage identity and mislead a nonprofessional to species substitution. All the species of Paphiopedilum are enlisted in CITES Appendix I and barred for unethical trading from wild sources. An in-silico study on the trait-linked phylogenetic framework of the Paphiopedilum species distributed in India aimed to identify the species passport trait which may prevent unethical trade. The phylogenetic framework was reconstructed using downloaded DNA sequences of ITS2, matK, and rbcL genes from GenBank for 22 slipper orchids of the subfamily Cypripedioideae including 9 Paphiopedilum species of India. Parsimonious and Bayesian interpretations were carried out using PAUP version 4.0a and MrBayes with ancestral trait reconstruction of 47 characters by Mesquite 3.61. The combined DNA ( ITS2 + matK + rbcL ) dataset supported the monophyly of the genus Paphiopedilum with a distinct clade and section Barbata (Krzl.) V.A. Albert & Borge Pett. with a subclade. The monophyly of the conduplicate-leaved genera Paphiopedilum, Phragmipedium, and Mexipedium was congruent to recent taxonomic classifications whereas plicate-leaved Selenipedium and Cypripedium showed polytomy in parsimonious tree. Ancestral reconstruction exhibited basal leaves, conduplicate leaves and deciduous perianth as symplesiomorphic traits. Undulate-revolute, rounded synsepal, ciliate lip; pubescent dorsal sepal, glabrous, glossy lip; coriaceous rough leaves , shortly hirsute peduncle; hairy lip; glandular pubescent peduncle; and brownish dorsal sepal with greenish yellow margin can be considered as passport traits for P. fairrieanum, P. spicerianum, P. venustum, P. charlesworthii, P. druryi and P. hirsutissimum respectively.

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