Pollen morphology of Aletris L. (Nartheciaceae) and its systematic significance

In this paper, we studied pollen morphologies of seven species in genus Aletris in detail by light microscopy and scanning electron microscopy. Of these, six species were reported for the first time. The palynological characteristics do not support the infrageneric classification into two clades. The results indicated that pollen grains of Aletris are small or medium with the P/E ratio of 0.36–0.59. They are elliptic or long‐elliptic in the polar view with blunt, round or acute ends and bilateral symmetric with a monosulcate, narrow or wide, deep colpus that has length extending to the ends of pollen grains, obvious or absent colpus membranes. The pollen ornamentation is gemmate, perforated, or reticulate. The sexine is slightly or quite thicker than the nexine.     

Pollen morphology of subfamily Caryophylloideae (Caryophyllaceae) and its taxonomic significance

The Pollen morphology of subfamily Caryophylloideae (Caryophyllaceae) from different phytogeographical region of Pakistan has been evaluated. In this research, 16 species belong to 6 genera of subfamily Caryophylloideae have been studied using light (LM) and scanning electron microscopy (SEM) for both qualitative and quantitative characters. Different palyno‐morphological features were observed including; pollen ornamentation, pore ornamentation, echini arrangement, echinidensity, number of pori, size of polar and equatorial diameter, P/E ratio, exine thickness, and size of pore were studied. The palyno‐morphological characters of subfamily Caryophylloideae have taxonomically significant in identification and delimitation of species. Two pollen types, i.e., subspheroidal (15 species) and prolate (one species Vaccaria hispanica) were observed. Microechinate‐punctate and microechinate‐perforate pollen ornamentation were examined. Two pollen types of pori, i.e., prominent (5 species) and sunken (11 species) ornamentation have been reported. Three types of echini arrangement have been reported irregular, regular and rather regular, while three type of echinidensity (i.e., medium, dense and sparse) were observed. Pori numbers were found different in different species range from 5 in Silene apetala to 19‐35 in Silene vulgaris. Based on different palyno‐morphological characters, taxonomic key was develop for quick and accurate identification. The quantitative data were processed using SPSS software for average, minimum, maximum, and standard error.     

Pollen morphology of the genus Sophora (Fabaceae) and its taxonomic implications

The genus Sophora (Fabaceae) is one of the taxonomically challenging genera with high economic and medical values. In this study, the pollen morphology of 43 samples of 27 species, 4 subspecies, and 4 varieties of the genus Sophora and 3 closely related genera was examined using scanning electron microscopy to evaluate the pollen diversity of the genus and its taxonomic significance. Pollen grains of the studied species were tricolporate (rarely six-aperture), and pollen shape varied from suboblate, spheroidal, subprolate to prolate. Echinate external ornamentation was reported for the first time in some species of the genus. Aperture membrane ornamentation and outline in a polar/equatorial view were described for the first time in the genus. Principal component analysis was used to understand the relationship and discrimination between the species and the genera, with six components accounting for 79.92% of the total variance. Taxonomic keys based on pollen morphology were also constructed to easily identify the taxa of the genus through palynological characteristics. Results showed that pollen morphology alone is not sufficient to elucidate or reconstruct taxonomic relationships within the genus Sophora, but palynological assessments can provide some useful information for identifying taxonomically problematic taxa.     

Pollen morphological variation of Berberis L. from Pakistan and its systematic importance

Due to overlapping and diverse morphological characters, Berberis is among the most taxonomically complex genera. Palynology is one of the taxonomic tools for delimitation and identification of complex species. In this study, pollens of 10 Berberis species were analyzed through light and scanning electron microscopy. Qualitative as well as quantitative features (pollen shape, size, presence or absence of colpi, colpi length and width, exine thickness, ornamentation, pollen class, aperture, and polar–equatorial ratio) were measured. Five species were observed to have colpate (pantocolpate) with elongated ends, radially symmetrical, isopolar, monads, and psilate‐regulate pollens. In polar view, six pollen were spheroidal, two were ovoid, one spherical, and one oblate. Similarly, variation in pollen length was prominent and the largest pollen on polar view was recorded for B. psodoumbellata 60–65 μm (62.4 ± 0.9), while the smallest one was observed for B. lycium 29–35 μm (32.2 ± 1). The observed variation in both quantitative and qualitative features were important in taxonomic identification. This shows that palynological characters are helpful in identification of Berberis genus at the species level.     

Taxonomic importance of pollen morphology in Veratrum L. (Melanthiaceae) using microscopic techniques

Previous studies reported that intrageneric relationships of genus Veratrum of family Melanthiaceae are controversial and hard to delimit. Therefore, we observed the pollen morphological features of six species in the genus Veratrum in detail using both light microscopy and scanning electron microscopy and investigated their significance for Veratrum taxonomy. Among them, five were studied for the first time. The results demonstrated that pollen grains of Veratrum are medium in size with P/E being 0.31–0.60. Three types of shape in polar view have been observed elliptic, long‐elliptic, or wide‐elliptic with blunt or rounded at both ends. Two types of width of colpus reported narrow or wide, two types of depth of colpus reported deep or flat, and two types of length of colpus reported extend almost or do not extend to the ends, whereas two types of colpus membranes reported absent or obvious. One type of surface ornamentation was noted as reticulate. These results support species Veratrum album and Veratrum lobelianum as well as Veratrum grandiflorum and Veratrum oxysepalum as two independent species, respectively, rather than classifying Veratrum into two sections. Overall, we demonstrated that the ratio of polar axis length to equatorial axial length, pollen characteristics at the polar view, the colpus morphology, and the surface ornamentation of pollen grains of genus Veratrum have important systematic significance in identification and delimitation of species.     

Pollen diversity and its implications to the systematics of mimosaceous species by LM and SEM

In this study we have discussed pollen morphology of 14 mimosaceous species belonging to five genera by using light microscopy (LM) and scanning electron microscopy (SEM). The palynological features of species were determined by both qualitative and quantitative characters. The qualitative characters include pollen shape, colpi arrangement, and exine sculpturing. Oblate spheroidal shaped pollens were noted in Albizia procera, Albizia lebbeck, Acacia tortilis, Acacia ampliceps, and Acacia modesta, subprolate shaped pollen in Leucaena leucocephala, Prosopis julifera, and Acacia nilotica, prolate pollen in Acacia farnesiana and Prosopis glandulosa while spheroidal, sub spheroidal and sub oblate pollen grains were observed in Acacia catechu, Mimosa himalayana, Prosopis cineraria, respectively. Sparsely foveolate, reticulate, scabrate, and scrobiclate exine sculpturing were observed. Colpi with sunken ornamentation were seen in P. cineraria, P. glandulosa, P. juliflora, and L. leucocephala while colpi were absent in rest of the studied species. The quantitative characters were statistically studied using SPSS software. The current study gives important morpho‐palynological characters for identifying and validation of close related and similar taxa which will aid to the phylogenetic analysis of Mimosaceae family.     

Seed micromorphology and its taxonomic evidence in subfamily Alsinoideae (Caryophyllaceae)

Seed micromorphology of 13 species, belonging to four genera of subfamily Alsinoideae (Caryophyllaceae) were investigated with scanning electron microscopy (SEM), in order to assess their diagnostic significance at generic level and provide additional evidence on species delimitation, as well as correct identification and phylogenetic position. Genera and species of subfamily Alsinoideae exhibit great variation in ultrastructure and a high diversity of novel micromorphological characters were observed. Variation in seed shape, color, hilum, anticlinal wall, epidermal cell, cell surface, margins, and quantitative characters as length and width were studied in detail, compared, illustrated, and their taxonomic significant were discussed. Seed shapes of the species were classified as reniform, round, angular, subcircular, subreniform, and elliptical pyriform, with sub‐central, central, basal, and nearly basal hilum. Wavy, irregular, tetragonal, and elongated epidermal cells structure has been observed as an exomorphological character. The present findings show that the micromorphology of subfamily Alsinoideae provides taxonomic information and is helpful to distinguish different species. The results also explained that SEM morphology of seeds provide important data about affinity among taxa and give potential characters in delimitation of members of subfamily Alsinoideae at generic and species level. A principal component analysis allowed to highlight the most outsiders among seed micromorphology with a possible explanation. Taxonomic keys were developed based on micromorphological characters to delimit the species and useful for their quick identification within subfamily Alsinoideae.     

Pollen morphological investigations of family Cactaceae and its taxonomic implication by light microscopy and scanning electron microscopy

The family Cactaceae is the diversified group of angiosperm plants whose pollen statistics has been used for taxonomic identification. In this article, we present the pollen morphology of eight species belong to seven taxonomically complex genera of Cactaceae including Astrophytum, Cylindropuntia, Echinocereus, Echinopsis, Mammillaria, Opuntia, and Thelocactus using light and scanning electron microscopy. The pollen grains were acetolyzed, measured, described, and electron photomicrographs were taken. Cactaceae can be characterized by presenting different palynomorphological features including pollen type, sculpturing, polar and equatorial diameter, aperture orientation, exine thickness, P/E ratio, and echini features. Four types of pollen shapes, that is, prolate spheroidal (three species), subprolate (two species), prolate (two species), and oblate spheroidal in Echinocereus reichenbachii were observed. The polar and equatorial diameter observed maximum in O. ficus indica 116.95 and 112.27 μm while minimum in M. compressa 38.42 and 21.05 μm. Pollen of two types, tricolpate in members of subfamily Cactioideae and pantoporate in the Opuntioideae were examined. The fertility percentage has been observed maximum in Opuntia macrocentra (83.84%) and minimum in Opuntia ficus‐indica (57.89%). Exine sculpturing showing great variations such as granulate, reticulate, granulate perforate and micro‐echinate foveolate ornamentation was examined only in Echinopsis eyriesii. A key to species, based on pollen micromorphological attributes, has been constructed for correct identification of complex cactus species.     

Pollen morphology and its taxonomic potential in some selected taxa of Caesalpiniaceae observed under light microscopy and scanning electron microscopy

Pollen micromorphology is not only used to check the functional and structural evolution in plants but also to solve the taxonomic problem related to the classification of plants. Therefore, keeping in view the significance of pollen traits, selected taxa of the subfamily Caesalpiniaceae was collected from different geographical regions of Pakistan. The species were then analyzed under both light microscopy and scanning electron microscopy techniques to investigate the importance of micromorphological characters of pollen in the identification and classification of species. Great variation was recorded in equatorial shape, surface ornamentation, tectum, polar diameter, equatorial diameter, and exine thickness. However, little variation was observed in pollen type, polar shape, and fertility of pollen. The equatorial shape of five types was observed: prolate, prolate-spheroidal, spheroidal-subprolate, subspheroidal-prolate, and subspheroidal. Four types of surface ornamentation, psilate, granulate, clavate, and perforate, were recorded. Tectum of five types, intactate, reticulate regulate, medium reticulate, tactate, and striate, was observed. Sexine was thicker than nexine in all studied species. The largest polar diameter was observed in Caesalpinia pulcherrima 64.1 μm while the smallest in Parkinsonia aculeata 26.1 μm. The largest equatorial diameter was found in C. pulcherrima of 70.25 μm whereas the smallest in P. aculeata 27.57 μm. All the pollens analyzed were tricolporate. All studied species have a fertility ratio of more than 90%. A taxonomic key was developed to show the variation in pollen features and delimit species for the correct identification. In conclusion, the pollen traits were found useful to define species boundaries at various taxonomic ranks and will strengthen the taxonomy of this subfamily. Besides, this study also explored the palynological traits and their implication in the taxonomy of the subfamily Caesalpiniaceae.     

Systematic approach to the correct identification of Asplenium dalhousiae (Aspleniaceae) with their medicinal uses

In present study, multiple microscope techniques were used for the systematics identification of the species Asplenium dalhousiae. The plant was collected from different phytogeographical and its natural habitat of Pakistan, where it shows higher diversity. Morphology, foliar epidermal anatomy, and spore morphological characters of the species were studied in detailed using multiple microscopic techniques through light microscopy (LM) and scanning electron microscopy (SEM). LM and SEM were used for the systematics identification of the species. Traditionally, the species is used in the ailment of many diseases, so the spore morphology, anatomical features, and morphological characters are relevant to describe the species taxonomy. The importance of multiple methods of taxonomic study (e.g., documentation and morphological characteristics) for characterizing herbs are important step in systematic certification to maintain the efficacy of herbal medicines. The aim of the present study is to examine the morphological, anatomical, and spore morphology of the species A. dalhousiae in more detailed for the correct taxonomic identification and their medicinal validation from Pakistan.     

Characterization of Albanian water frog,Pelophylax shqipericus,sperm traits and morphology,by using phase contrast microscopy

Mature spermatozoa traits and morphology of endangered Albanian water frog, Pelophylax shqipericus, have been characterized for the first time through phase contrast microscopy, as part of successful implementation of in vitro fertilization technique for this species. The basic morphology of P. shqipericus spermatozoa consists of an elongated, thick, smooth‐edged, and solid‐staining head, continuing with a thin and long tail which usually extends 2.48 times the head length. The acrosome was not clearly discernible so the measurements were done on the head as a whole, while the middle section was better visible. Average length of head, including the acrosome and midsection was estimated to be 11.78 μm ± 0.32, while the tail length resulted 29.24 ± 1.75 μm. The average thickness of the head was shown to be 3.45 μm. The total sperm length resulted to be 41.02 ± 1.83 μm. The average sperm concentration was estimated of 25.5 × 10⁶/ml. Sperm amount, survival rate and motility were also measured. The sperm survival rate was maximal immediately after preparation of the suspension and tended to decrease over time of storage, reaching 50% after 72 hr. Decreased sperm motility seemed to follow the same trend as sperm viability. Sperm traits resulted to be very similar both in size and in shape with those of “Lessonae” frog group, one of the lineages of Western Palearctic species complex, suggesting a strong phylogenetic relationship among these species.     

Pollen morphology and its implication in the taxonomy of some selected taxa of the bi and tri-ovulate Euphorbiaceae of the Hainan Island by using multiple microscopic techniques

The family Euphorbiaceae is one of the most taxonomically complex and diverse family among the angiosperm. Therefore, this study aims to document the pollen morphology of the Euphorbiaceae taxa collected from different geographical regions of the Hainan Island South China, to strengthen the taxonomy and support the subfamilial classification. The Euphorbiaceae includes 23 species and 10 genera of the bi-ovulate subfamily Phyllanthoideae and tri-ovulate Crotonoideae, Euphorbioideae, and Acalyphoideae were examined under light and scanning electron microscopy. The principal component analysis was performed by using six variables to show variation and relationships between the species. A significant variation was found in the pollen morphology (exine and colpus sculpturing). The croton-type pattern of the pollen was found the diagnostic trait of the Crotonoideae. The Euhphorboideae taxa were mostly homogenous however, variation was found in the muri pattern. The Acalyphoideae, have fossulate scabrate and verrucate exine sculpturing that can delimit from the rest of sub-families. The Phyllanthoideae, showed reticulate exine, tri to tetra-colporate having various colpus morphology. The maximum pollen size was found in Croton hirtus (59.49 × 60.86 μm) and minimum in Acalypha indica (10.86 × 73 μm). Similarly, the maximum colpus width was observed in Euphorbia pulcherrima (8.3 μm). Hence pollen traits have relevance in the taxonomy and phylogenetic of the family to define species boundaries at various taxonomic levels. This study can support and strengthen the taxonomy of subfamilies and fill the palynological gap of these taxa in Hainan Island and will contribute to future systematic studies of the Euphorbiaceae.     

Systematic significance of pollen morphology and foliar epidermal anatomy of medicinal plants using SEM and LM techniques

Alternative medicinal source is a hot issue within medicine community now a days due to various side effects of allotropic drugs; most importantly the multidrug resistance phenomena. Medicinal plants provide an attractive and effective alternative to modern synthetic drugs as they contain different types of bioactive compounds, having great potential in treatment of various diseases. The present study deals with identification of micromorphological characteristics of 10 ethnomedicinally important plants of Northern Pakistan, that is, Aster himalaicus C. B. Clarke, Artemisia maritima (Berg.) Wellkomm, Isodon rugosus (Wall. ex Benth.), Myricaria squamosal Desv, Pedicularis groenlandica Retz. Polygonum aviculare L, Salvia himmelbauer E. Peter, Swertia kingie Hook. f., Tanacetum dolichophyllum (Kitam) Kitam, Verbascum epixanthinum Boiss. & Heldr. as potential source for pharmaceutical industry. The characters studied were shape and size of pollen, polar diameter and equatorial diameter, epidermal cell shape and size, stomata width and length, trichome shapes, and type. Most of the species had tricolporate pollen type but hexacolporate pollen was also observed in Isodon rugosus, Polygonum aviculare, Salvia himmelbauer. Different types of exine sculpturing observed were foveolate, scabrate, lophate, regular, steriate, and alveolate. Minimum equatorial diameter was noted in Tanacetum dolichophyllum (12.4 μm) and maximum in Myricaria squamosal (20.9 μm). Shape of epidermis cell ranged from rectangular and irregular, polygonal and pentagonal. Different types of stomata like anisocytic, actinocytic, paracytic, and diacytic were noticed. Variations in microscopic characters observed in the current study have great potential to be used as identification tools by the taxonomists for further studies.     

Morphological redescription and phylogenetic assessment of Spauligodon aspiculus (Oxyuroidea: Pharyngodonidae) infecting the white‐spotted gecko,Tarentola annularis (Squamata: Gekkonidae)

Thirty white‐spotted geckos, Tarentola annularis, from the South Sinai desert in Egypt, were examined for helminth parasites. Spauligodon aspiculus was observed to infect 19 geckos with 63.33% as a prevalence of parasitic infection. The present nematode species is separated from congeners by morphological and metrical characteristics such as lateral alae, aspinose filamentous tail, and no spicule, and three pairs of caudal papillae with posterior pair excluded from envelopment by the caudal alae in the male worms, and knobbed eggs, and postbulbar vulva in females. It compared morphometrically with other Spauligodon species described previously and showed few differences in measurements. Molecular characterization based on the partial 28S rRNA nuclear ribosomal gene sequence showed that there was a close identity, up to 72%, with other sequences retrieved from GenBank. Phylogenetic analysis showed that the parasite sequence in conjunction with existing data facilitates the investigation of the placement of this pharyngodonid species within Oxyuridae. The present species is deeply embedded in the genus Spauligodon with close relationships to previously described Spauligodon nicolauensis (gb| JN619349.1, and JF829243.1) as more related sister taxa. This study highlights the importance of combining genetic and morphological data with taxonomy in pharyngodonid species.