Phytotaxa 351 (4): 273–280 http://www.mapress.com/j/pt/ Copyright © 2018 Magnolia Press Article PHYTOTAXA ISSN 1179-3155 (print edition) ISSN 1179-3163 (online edition) Accepted by Joel Nitta: 22 May 2018; published: 7 Jun. 2018 https://doi.org/10.11646/phytotaxa.351.4.3 273 Ophioglossum gujaratense, a new species from Gujarat State, India SACHIN M. PATIL, RONAK N. KACHHIYAPATEL, RAVI S. PATEL & KISHORE S. RAJPUT* Laboratory of Plant Anatomy and Pteridology, Department of Botany, The Maharaja Sayajirao University of Baroda, Vadodara, Gujarat State - 390 002 (India). Author for correspondence: ks.rajput15@yahoo.com Abstract A new species, Ophioglossum gujaratense, is described from Gujarat state (India). It resembles O. polyphyllum in the presence of sheathing rhizomorph and 1–4 (rarely 5) trophophylls. On the other hand, rhizomorph morphology, common stalk, trophophyll arrangement, leaf lamina and leaf base make it distinct from O. polyphyllum. Stoloniferous roots, trophophyll number and their arrangement of the new species also resemble O. parvifolium and O. nudicaule. However, both these species lack a sheath around the leaf-stem base. A comparative account of morphologically similar species, viz. O. gujaratense, O. polyphyllum, O. parvifolium and O. nudicaule is provided. The distinctness of the new taxon has been confirmed using molecular data from chloroplast genome markers viz rbcL, trnH-psbA, trnF-trnE and trnL-trnF. Keywords: DNA barcode, leaf sheath, stoloniferous roots, pteridophytes Introduction The genus Ophioglossum (Linnaeus 1753: 1062; Ophioglossaceae Martinov) comprises nearly 45 species worldwide (Hassler & Swale 2002, Yadav & Goswami 2010), of which 13 species are reported from India (Patil & Dongare 2014, Fraser-Jenkins et al. 2017). The genus has been studied in Gujarat by Phatak et al. (1953), Chavan & Mehta (1956), Inamdar & Shah (1967), Inamdar (1970), Dixit (1984), Kachhiyapatel et al. (2015) and Rajput et al. (2016a, b). As of now four species of Ophioglossum are known from Gujarat (Kachhiyapatel et al. 2015). In a recent botanical excursion, the authors collected interesting specimens of Ophioglossum having sub-globose-tuberous rhizomes with stoloniferous roots, horizontal trophophylls in contact with the soil at a 90º angle to the fertile stalk, and a sheathing base from the slopes and base of the hills at Dhanpuri and Zand (Jambughoda Wild Life Sanctuary), Shivrajpur Gujarat (western India). The morphology of the specimens was therefore compared with similar Indian species, viz. O. polyphyllum (Braun ex. Seubert 1837: 17), O. parvifolium (Greville & Hooker 1833: 218), and O. nudicaule (Linnaeus f. 1781: 443) and molecular analysis of the plastid genome (rbcL, trnH-psbA, trnF-trnE and trnL-trnF) carried out to confirm the distinctness of the taxon. Materials and methods Collection of Plant Material:—A field survey to document pteridophyte diversity was carried out during June– October from 2014 to 2017 in forest areas of different regions of the state to collect species of Ophioglossum. One of the specimens collected from Dhanpuri, Jambughoda (Panchmahal Dist.) in the month of July 2017 was found to be different from previously reported species (Panigrahi & Dixit 1969, Manickam & Irudayaraj 1992, Patil 2014, Fraser- Jenkins et al. 2017). Subsequently the same species was also collected from Zand and Shivrajpur. Voucher specimens were deposited in BARO. DNA Isolation and PCR:—Ten individuals were packed in sterile polyethylene bags for the isolation of DNA from each collection site. Fresh leaf samples weighing 50 mg were crushed in liquid nitrogen and a fine powder obtained using a sterile motor and pestle. DNA extraction was carried out using Plant/Fungi DNA isolation kit (Sigma Aldrich Cat# E5038) or CTAB method (Doyle & Doyle 1990). Subsequently, PCR reactions were carried out using