BIOSCIENCES BIOTECHNOLOGY RESEARCH ASIA, November 2014. Vol. 11(Spl. Edn.), p. 17-25 A Comparative Study of Spore Morphology of Some Pteridoideae Subfamily Genera Alexander Alexandrovich Kuznetsov1, Alexey Vladimirovich Vaganov2, Mikhail Viktorovich Skapcov3, Andrey Sergeevich Erst4 1National Research Tomsk State University, Laboratory of Biogeochemical and of Remote Methods of Monitoring Environmental, prospekt Lenina, 36; 634050, Tomsk, Russia. 2,3Altai State University, South-Siberian Botanical Garden, Lenina st. 61; 656049, Barnaul, Russia. 4National Research Tomsk State University, Laboratory of Biodiversity and Ecology, prospekt Lenina, 36; 634050, Tomsk, Russia; Central Siberian Botanical Garden of the Siberian Branch of Russian Academy of Sciences, Zolotodolinskaya str., 101; 630090, Novosibirsk, Russia. doi: http://dx.doi.org/10.13005/bbra/1435 (Received: 27 September 2014; accepted: 10 October 2014) Using the method of scanning electronic microscopy (SEM), a comparative study of ten representatives of subfamily Pteridoideae C.Chr. ex Crabbe, Jermy & Mickel family Pteridaceae E.D.M.Kirchn. was carried out. A comparative study of morphological characters of investigated spores has revealed characters that allow considering the relatedness of the studied species to one subfamily – Pteridoideae. Those characters include spore form – triangular-roundish, laesura rays are straight, merged into sporoderm (laesura lips), tubercles on the spore surface, and in some cases “cerebriform” folds, exosporium surface without excrescences. For species of the genera Afropteris and Taenitis, we described characters that allow considering them apart from the complex of characters belonging to the species of genera Actiniopteris, Anopteris, Onychium, Jamesonia and Pteris. Key words: subfamily Pteridoideae, family Pteridaceae, Actiniopteris, Jamesonia, Onychium, Pteris, Anopteris, Afropteris, Taenitis, morphology of the spores, scanning electronic microscopy (SEM). Family Pteridaceae E.D.M. Kirchn. is except Antarctica. The maximum species diversity considered one of the most complex families is observed in tropical and subtropical latitudes of according to the amount of unresolved issues in the Old and New Worlds. the systematics of its taxonomic ranks. Partly due The system of the family Pteridaceae was to insufficient knowledge of this family, as well as repeatedly reprocessed based on the ferns systems the lack of evidential techniques, the final volume being developed. In the modern period, the use of of the family is not determined. Members of the molecular genetics to solve systematic problems family Pteridaceae are spread across all continents in a number of major works1-3 allowed to prove relatedness of five subfamilies to a family Pteridaceae based on in-depth analysis of previously proposed systems: Cryptogrammoideae S. Linds.; Pteridoideae * To whom all correspondence should be addressed. C.Chr. ex Crabbe, Jermy a. Mickel; E-mail: [email protected] Ceratopteridoideae (J. Sm.) R.M. Tryon; 18 TSOI & PESHKOV, Biosci., Biotech. Res. Asia, Vol. 11(Nov. Spl. Edn.), pgs 17-25 (2014) Vittarioideae (C. Presl) Crabbe, Jermy a. Mickel; members of the subfamily Pteridoideae. Cheilanthoideae W.C. Shieh. To construct a Photographs of the ferns in the works of Belling system of the family, the information on individual and Heusserl, as well as in Nayar with Devi’s can genes of the chloroplast and nuclear DNA were only be used to describe the form and carry out used to recreate the intergenetic family connections very “rough” measurements for biometric research. in the system Pteridaceae family by constructing It is impossible to identify and describe the detailed phylogenetic trees. morphological structures, laesura structures, Subfamilies Ceratopteridoideae and equatorial ridges, tubercles on the sides of a spore Cheilanthoideae, being brought by a number of and describe the surface of exosporium based on authors to the rank of families are considered the the photographs given in those papers. The exact most stable system of ferns in the world. In determination of the size of all spore structures particular, due to the presence of stable characters and their description is required to identify the of external morphology. In subfamilies common characters on intergeneric, specific and Vittarioideae, Cryptogrammoideae and intersectional levels. Thus, in the works dated last Pteridoideae, a reverse situation is observed – so century, the description of spores, as the far, there is no definitive evidence to prove the photographs themselves do not contain enough inclusion into specified subfamilies of the number information about the spore morphology of this of ferns. In the course of studying ferns, for complex group of ferns and require modern high- historical reasons, genera, which could not be tech equipment and new additional data. reasonably related to a particular taxonomic However, survey works on the category for various reasons, were included in the morphology of fern spores of Pteridaceae family subfamily Pteridoideae. allow yet to receive general information on a As a result, in the work on fern spores of number of stable characters common for subfamily Pteridaceae family from the territory of Formosa Pteridoideae: tetrahedral spores with thiradial (Taiwan)4 due to the use of “raw” and incomplete laesura, triangular-roundish or roundish-triangular, system of ferns of Pteridaceae family, as well as a have a clear exosporium with a distinct surface misunderstanding of its volume, an erroneous ornamentation and total absence of perisporium, spore morphology classification was offered. the surface of exosporium varies from rough to Consequently, for the family Pteridaceae was having large tubercles and roller-like thickenings. proposed a classification with two types of spores – bilateral and trilete-tetrahedral spores. In the light MATERIALS AND METHODS of the above, it is necessary to correlate the results of one’s anatomical and morphological and Spores for the study were selected from molecular genetic studies with new information on herbarium materials stored in the herbarium of the the taxonomy of any given taxonomic group. National Museum of Natural History (R), the Apart from the information obtained by Botanical Garden and Botanical Museum Berlin- methods of molecular genetics, in order to determine Dahlem (B), the V.L. Komarov Botanical Institute the phylogenetic boundaries of genera and of the Russian Academy of Sciences (LE) and the establish a clear distinction between them, the National Research Tomsk State University (TC). results of microphotography scanning microscope Spores were investigated using scanning have been often used over the last years. electron microscope “Philips SEM 525-M”, and Micrographs of spores obtained with a scanning electron-ion scanning microscope “Quanta 200 3D” electron microscope are more informative; provide situated in Tomsk Material Testing Center for more information to resolve disputes in the collective use of the National Research Tomsk State systematics of complex fern groups. University, a scanning electron microscope Spore morphology of individual members “Hitachi – S 3400 N” (Tomsk) in the laboratory of of the subfamily Pteridoideae at some point were the Institute of water and Ecological Problems paid enough attention in large reports devoted to (Barnaul) and JEOL JSM-6390LA Analytical morphology of fern spores4-8. However, these Scanning Electron Microscope of Center for works provide the descriptions only for some collective use of the V.L. Komarov Botanical TSOI & PESHKOV, Biosci., Biotech. Res. Asia, Vol. 11(Nov. Spl. Edn.), pgs 17-25 (2014) 19 Institute of the Russian Academy of Sciences is rugose, with folds (3.0) 5.4 (7.8) µm broad. The (Saint-Petersburg). Spores were fixed using a proximal side has tuberculum-like ornamentation, carbon adhesive tape; to reduce the influence of tubercles are small, (1.1) 2.75 (4.4) µm in diam. The the charge a method of thermal spraying by folds and tubercles have a distinct outline: the chromium, carbon or gold-palladium mixture in a tubercles are roundish in outline, dense beside the vacuum sputtering plant. Spore surface was pole and sparse on the margins. The surface of the scanned in high vacuum at an accelerating voltage exosporium is coarse-grainulate, with sparse of 2 kV and a magnification of 1000 to 7000 times, roundish excrescences (0.2)1.25(2.3) µm in diam. at and 10,000 to 16,000 times. The measurements of the distal side, and (0.2) 0.7 (1.2) µm in diam. at the spores were carried out using “SIAMS proximal side. Investigated specimen: Nilghiris, MesoPlant” and “Photometer” programs. Mardas. Legit. Boord of Reuenue (P). An analysis was performed on the Actiniopteris dimorpha Pichi-Serm., 1962, following spore morphological characters: 1 – Webbia 17, 1: 18, f. 2. equatorial diameter, µm; 2 – polar axis; 3 – laesura SEM-description. Spores in a polar length; 4 – laesura width; 5 – the width of the position are triangular-roundish or irregular- equatorial ridges; 6 – roller-like thickening width ; triangular-roundish with straight sides (non 7 – roller-like fold thickness on the distal side of lociniate). The equatorial diameter is the spore; 8 – roller-like fold thickness on the (46.5)54.5(62.5) µm. The distal side in equatorial proximal side of the spore; 9 – roller-like thickening position is hemispherical, the proximal side is width around laesura; 10 – diameter of tubercles convex, with an acute pole. Equatorial ridge is on