Microscopic investigations of some selected species of Papilionaceae through SEM and LM from Skardu valley,northern Pakistan

Pollen morphology of 10 species and foliar epidermal anatomy of eight species of Papilionaceae from Skardu valley, northern Pakistan has been estimated for the first time. The present study was commenced with an aim to provide a detailed account of the pollen morphology by scanning electron microscopy and foliar epidermal anatomy by light microscopy. The pollen aperture was tricolporate with reticulate exine in the selected species. Stomata types are actinocytic, paracytic, and anomocytic. Irregular or polygonal with undulate or straight walls, epidermal cells were reported. A unique diversity was observed in the foliar trichomes that show the taxonomic significance of the discrimination of taxa. Non‐glandular trichomes were observed in the selected species which are unicellular with thin, long and pointed apical cells. Pollen and foliar micro morphological characters proved to be helpful for the identification of taxa at a specific level.     

Palynological investigation of some selected species of family Fabaceae from Pakistan,using light and scanning electron microscopy techniques

Pollen morphology of 11 species of family Fabaceae that is, Trifolium alexandrinum, Trifolium resupinatum, Arachis hypogaea, Lathyrus aphaca, Medicago lupulina, Vicia sativa, Lathyrus odoratus, Pongamia pinnata, Melilotus indicus, Medicago polymorpha, Medicago sativa from Pakistan has been investigated by light and scanning electron microscopy. Pollen were generally tricolporate, radially symmetrical, isopolar, elliptic in equatorial view and triangular in polar view under LM. Results showed that pollens were per‐prolate (T. alexandrinum), prolate (T. resupinatum, V. sativa, L. odoratus, Melilotus indicus, M. polymorpha, M. sativa) and sub‐prolate (A. hypogaea, L. aphaca, M. lupulina, P. pinnata). The larger polar/equatorial (P/E) ratio was found in T. alexandrinum (2.26 μm) and the smallest was found in M. lupulina (1.21 μm). The exine of T. resupinatum was 3.00 μm in thickness while others posses smaller exine thickness. The larger pore diameter was found in P. pinnata (16.01 μm) while others have smaller. The length of colpi was larger in Arachis hypogaea (32.24) while others posses smaller. Eight types of surface ornamentation (Psilate, faintly rugulate). Perforate and rugulate to verrucate have been observed under SEM. The pollens were europalynous type. Pollen morphology proved to be useful for the specific delimitation and serve as a tool for the identification and classification of taxa at specific and generic levels and can also be used as a key for the taxonomic features. Diversity in exine sculpture is helpful indicative characters for the isolation of closely related species. Hence, it is clear that both qualitative and quantitative characters of pollen can be useful for differentiating between taxa at specific level.     

Seed morphology using SEM techniques for identification of useful grasses in Dera Ghazi Khan,Pakistan

In recent study, 15 taxa of family Poaceae from Dera Ghazi Khan were investigated for morphological characterization of seeds. Scanning electron microscopy (SEM) was utilized to study morphological features of grass seeds. Moreover, economic importance of studied taxa has been discussed in present research. Grass seeds or caryopsis were collected in their mature state from different areas of Dera Ghazi Khan. Collected caryopses were subjected to SEM to investigate surface sculpture, epidermal cell shape, and cell wall patterns. Caryopsis shape observed was elliptic to oval. Also, the morphology of plant is being investigated. Cell wall pattern observed was straight and wavy. Surface sculpturing varies from smooth to reticulate. Epidermal cell shape was either wavy or irregular. Hilum position is terminal in all species. Epidermal cell structure and cell wall pattern was not observable in some species. Major variations among studied taxa were observed in terms of seed surface pattern. Five types of caryopsis surface pattern were observed namely, striate, smooth, reticulate, papillate, and granulate. Investigated taxa illustrated variations in terms of studied morphological features. Utilization of SEM has been proved very helpful in exploring seed morphological features. These observed features can aid in delimitation and identification of various grass taxa. Recent research recommends the utilization of SEM for caryopsis morphology to solve the identification issue of problematic grass genera.     

Palynological characteristics of some selected species of family Cucurbitaceae (Juss.) using light and scanning electron microscopy techniques

The pollen morphology of 11 species distributed in seven genera belonging to family Cucurbitaceae has been studied using light microscopy (LM) as well as scanning electron microscopy. Pollen morphology of this family is diverse and is almost europalynous. Nearly all types of pollen belonging to different size class and shape class with a variety of exine sculpturing are found in the family. Pollen grains are usually triporate to tricolporate, prolate, oblate‐spheroidal, spheroidal, to suboblate, radially symmetrical, and isopolar. The common exine pattern is reticulate of various forms ranging from finely reticulate to coarsely reticulate with some species having echinate type of exine ornamentation, for example, Cucurbita pepo and Praecitrullus fistulosus. Most of the studied species are somewhat triangular with respect to equatorial outlook and circular to elliptic in polar outlook. The palynological characteristics of the members of the family are useful in identification of taxa at generic level mostly. Based on the present study it is not recommended to wholly solely depend upon the pollen morphology for the identification of taxa especially at specific levels because very minor differences regarding pollen characters exist among the species of the same genera.     

Microscopic investigation of palyno‐morphological features of melliferous flora of Lakki Marwat district,Khyber Pakhtunkhwa,Pakistan

The present study was carried out to investigate the polliniferous bee flora of Lakki Marwat district, Khyber Pakhtunkhwa, Pakistan. A total of 18 species, belonging to 13 different families were identified for the foraging activities of honeybees, namely, Cenchrus biflorus, Cestrum nocturnum, Citrus limon, Combretum indicum, Datura innoxia, Duranta erecta, Hamelia patens, Helianthus annuus, Ipomoea cairica, Luffa aegyptiaca, Nerium oleander, Ocimum basilicum, Parthenium hysterophorus, Pennisetum typhoides, Prosopis cineraria, Prosopis juliflora, Saccharum spontaneum, and Ziziphus jujuba. The identified melliferous flora was collected for the morpho‐palynological investigation. Pollen were acetolyzed, measured, and described qualitatively using light microscopy. The pollen types varied from psilate to echinate and colpi from tricolpate to hexacolpate. Quantitative characteristics including polar diameter, equatorial diameter, polar to equatorial ratio, colpi length, colpi width, spine length, spine width, and exine thickness of the pollen were determined and analyzed statistically using software IBM SPSS Statistics 20. The results of the present study highlighted the significance of pollen morphology of the bee flora of the area which may lead to the identification of the potential and useful botanical sources for beekeepers and to check the honey quality marketed from the study area.     

Palynological investigation of lactiferous flora (Apocynaceae) of District Rawalpindi,Pakistan, using light and scanning electron microscopy

This was the first comprehensive pollen micromorphological investigation of lactiferous flora (Apocynaceae) of District Rawalpindi, Pakistan. The pollen morphology of 10 species of the family Apocynaceae was observed and documented using light microscopy and scanning electron microscopy. Pollen was found subspheroidal in shape in most of the species, however peroblate shape was noted in Vinca major. Exine sculpturing patterns (psilate, rugulate, scabrate, and microreticulate) were observed. The result indicated that exine ornamentation of Apocynaceae taxa is systematically informative at generic and species levels. Most of the species have tricolporate type pollen but tetraporate pollen was also observed in Trachelospermum jasminoides and tetracolpate in V. major. Minimum equatorial diameter was noted in Carissa edulis (27.13 μm) and maximum in V. major (108.25 μm). Similarly, maximum exine thickness was found in Cascabela thevetia (9.5 μm). In the present findings, the pollen morphological data are compared with available other pollen studies to evaluate the taxonomic value of pollen traits in Apocynaceae taxa by using multiple microscopic techniques. Furthermore, molecular and phylogenetic studies were recommended to strengthen the systematics of Apocynaceae taxa.     

Palyno-anatomical characters and their systematic significance in the family Apiaceae from Chitral,eastern Hindu Kush,Pakistan

The present study was performed to provide a detailed explanation of leaf epidermal anatomy and pollen micromorphological features of selected species of family Apiaceae from Chitral, eastern Hindu Kush region as the basis of forthcoming studies. In the present article pollen morphology of eight species and foliar epidermal of seven species of family Apiaceae have been examined through microscopic techniques. In results two types of pollen prolate (five species) and perprolate (three species) with three colpi have been recorded. The exine ornamentation was found to be regulate, striate, and cerebroid. Largest pollen was found in Heracleum leucocarpum with the polar diameter of 43.25 μm and equatorial diameter of 21.6 μm. Smallest pollen was observed in Elaeosticta chitralica with the polar diameter of 18.4 μm. The P/E ratio varied from 1.59 to 2.16. Regarding to foliar epidermal anatomy, three types of epidermal cells including rectangular, irregular, and polygonal with variation in anticlinal wall pattern were determined. In the selected species three kinds of stomata comprising anisocytic, anomocytic, and paracytic type were reported in the current research. The size of epidermal cells ranged from 106 × 42.50 μm in Bupleurum falcatum subsp. cernuum and 77.25 × 26.35 μm in Prangos pabularia in adaxial surface. Largest stomatal complex was found in Prangos pabularia both in adaxial 33.55 × 20.05 μm and abaxial 50.25 × 39.40 μm. All the observed quantitative and qualitative features of the species were proved to be useful in the delimitation of species at generic and species level.     

Morpho‐palynological investigations of natural resources: A case study of Surghar mountain district Mianwali Punjab,Pakistan

Surghar mountain belt has comparatively less natural resources of floral diversity because it is composed of minerals of different kinds making it less favorable for the growth of different vegetation. The pollen morphology of some selected plants from Surghar belt Mainwali has been evaluated. The pollen grains were measured and demonstrated using scanning electron microscopy (SEM). The examined plant specimens have a difference in size, shape, polarity, and their exine ornamentation. The pollen taxa show a huge variation in size and sculpture. Pollen fertility has also been estimated, shows that the selected plants are well‐known in the Surghar belt. The need of the hour is to conserve these plants having a higher fertility rate to cope with the deforestation in an area. The conclusion does not favor theories in which deforestation results in fast growth in population. It shows that the irrational management and unlawful cutting down of woods neglected by the forest department are the main causes of deforestation in the mountain belt of Mianwali. The findings show the importance of morphological characteristics in the identification of natural resource species in the area.     

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.     

Comparative pollen and foliar micromorphological studies using light microscopy and scanning electron microscopy of some selected species of Lamiaceae from Alpine Zone of Deosai Plateau,Western Himalayas

The study was conducted to highlight a detailed account of morphology of pollen chosen species of Lamiaceae through scanning electron microscopy, and the anatomical characteristics of leaf epidermis of seven species using simple light microscopy. In results, Anisomeles indica and Otostegia aucheri belong to subfamily Lamioideae because it has tricolpate pollen while the rest eight species belong to subfamily Nepetoideae (hexacolpate pollen). The exine sculpturing of pollen of studied species was found to be reticulate. In the family Lamiaceae, four kinds of stomata were found anomocytic, anisocytic, diacytic, and actinocytic, respectively. The cell wall patterns of epidermal cells were irregular or polygonal with straight or undulate walls. It was noted that the variety of the epidermal trichomes seems of taxonomically important for the identification of species of Lamiaceae. Both nonglandular and glandular trichomes were analyzed. The nonglandular trichomes were characterized with long, thin, and pointed apical unicellular cells. The nonglandular trichomes were A‐shaped in Thymus linearis. In Perovskia abrotanoides, stellate glandular trichomes were observed whereas in A. indica and Mentha royleana both glandular and nonglandular trichomes were found. In A. indica, the nonglandular trichomes were sessile and peltate in M. royleana. For the first time in this study, pollen and foliar micromorphological features of selected species of this area are carried out. These taxonomic characters were found to be important in discrimination of species from each other. In future, the detailed study with comprehensive morphology coupled with other important characters is required for delimitation of taxa at various levels.     

Light and scanning electron microscopy-based foliar morpho-anatomical comparison of selected family Rosaceae members distributed in District Lahore,Punjab, Pakistan

In the present study morpho-anatomical characterization of selected Rosaceae members distributed in District Lahore was performed. Light and scanning electron microscopy was used for systematic characterization of the selected 19 species. Distinguished morpho-anatomical features such as size and shape of epidermal cells, size and type of stomatal cells, size and shape of trichomes, oil droplets, and silica bodies were contrasted. Results reported remarkable variations which could be taxonomically useful in identification of these members. Polygonal epidermal cells were observed in Eriyobotraya japonica, Potentilla bifurca, Potentilla supina, and Prunus amygdalus. However, Prunus cerasus possessed irregular-shaped epidermal cells that can be distinguished from hexagonal epidermal cells of Prunus persica. Similarly, stomatal type varied among some members. Paracytic or perisocytic stomata were observed in E. japonica whereas P. bifurca observed paracytic and anisocytic stomata. Lengths of guard cells were also of variable sizes. The average length of guard cells ranged from 53 (52–54) μm to 74 (73–75) μm in abaxial view. Potentilla supina had biggest, while Rosmarinus officinalis had tiny guard cells. Trichomes were tubular, stellate, cylindrical, ribbon-like, glandular, and nonglandular. Silica bodies in the present investigation were bilobed, rounded, and oval-shaped. It is inferred that diverse anatomical features proved to be valuable taxonomic tools that could be fruitfully helpful in identification of plants at specific as well as generic level.     

Taxonomic investigation of selected rust fungi using scanning electron microscopy from Khyber Pakhtunkhwa,Pakistan

Rusts comprises the largest natural group of plant pathogens including approximately 8% of all described Fungi. Rust fungi are extremely plant pathogens responsible for great losses to agriculture productivity. Rust species belong to several genera among which more than half are Puccinia species. In Pakistan, rust causes severe damage to agriculture crops. Current study was carried out to identify and characterize different rust species common in the research area through microscopy and Scanning electron microscopy (SEM) in Khyber Pakhtunkhwa, Pakistan. Morpho-anatomical investigation of each collected rust species was carried out using different standard protocols. The dimensions of spores were measured and snapped under a stereomicroscope. SEM was used to examine the shape, size, and ornamentation of the spores of each rust fungus. Results revealed documentation of seven rust fungi, that is, Melampsora euphorbiae, Phragmidium barclayi, Puccinia nepalensis, P. exhausta, P. menthae, Uromyces capitatus, and Uromyces decorates belong to four different genera, were recorded. SEM revealed that spermogonia and Aecia were missing in most of the rust fungus studied. Uredinia was found in a scattered, irregular, lengthy, and epidermis-enclosed form. Urediniospores were found to be ovulating, elongated, echinulate, globose to sub-globose, ellipsoid to ovoid, and globose to sub-globose. Telia was found as sub-epidermal, amphigenous, dispersed, minute, and spherical cells. Teliospores ranged in form from cylindrical to oblong. The germ pores were detected in both apical (top cell) and basal (bottom cell) idiosyncratic and pedicel-attached cells. The techniques used in the current investigation will aid mycologists in rust identification and microscopic characterization.     

Spores morphology of the family Pteridaceae from Shandong Province,China

In this article, we explored systematically the spore morphology of Pteridaceae by observation of the species distributed in Shandong Province using scanning electron microscopy (SEM). The results showed that the spore morphology of all the species in the family is tetrahedral and trilete. The characters of spore ornamentation are intraspecies stable, but significantly different among species and genera. Spore morphology is significant in exploring the phylogenetic relationships of Pteridaceae as well as in generic and specific delimitations.     

Palyno‐morphological investigations of halophytic taxa of Amaranthaceae through SEM from Salt range of Northern Punjab,Pakistan

The pollen morphology of 11 salt tolerant plant species of family Amaranthaceae from the salt range of Northern Punjab, Pakistan has been studied. The palyno‐morphological characters were examined using light and scanning electron microscope. The examined all salt tolerant species have a slight difference in size but have similarity in shape, pore ornamentation, and polarity. The observed morphological characters of pollen grains were pollen symmetry, size, shape, pore ornamentation, pore size, number of pores, exine thickness, polar and equatorial diameter and, P/E ratio. Apolar type of pollens has been observed in all species. Shape of pollens was spheroidal. Exine sculpturing of pollen grains was scabrate (six spp), microechinate (four spp), and microechinate‐scabrate (one spp). Different pori numbers were observed in different species. The pantoporate aperturate and sunken pore ornamentation have been reported in all species. A pollen taxonomic key was developed using examined morphological characters for the accurate identification of halophytic taxa. The high fertility and low sterility of pollens confirmed that the selected halophytes are well‐established in the salt region. The findings highlight the taxonomic significance of pollen morphology in correct identification and differentiation of salt tolerant plant species.     

A new species of Cassida Linnaeus, 1758, from Turkey (Chrysomelidae: cassidinae)

A new species, Cassida alidagiense sp. nov., has been described from Kayseri province in Turkey. For the time being, the species is endemic to Turkey. Cassida alidagiense sp. nov., is related to Cassida linnavuorii Borowiec, 1986; Cassida brevis Weise, 1884; and Cassida bella Faldermann, 1837, from which it differs in the shape of the apex of cornu of the spermethaca, and it can be distinctively differentiated from these species based on color of under body and spermathecal characters especially. In addition, the paper presents ultrastructures observed by SEM of spermatheca of Cassida alidagiense sp. nov. from Turkey.     

Morpho-anatomical characterization of some members of euglenophycota (algae) of north-east Punjab,Pakistan based on light microscopy (LM) and scanning electron microscopy (SEM)

The study was carried out in four districts, that is, Gujranwala, Gujarat, Narowal and Sialkot of Punjab, Pakistan. The sampling was carried out randomly in different seasons from the water bodies especially from wastewater. Twenty-one species belonging to Euglenophycota were identified using light microscopy and scanning electron microscopy from which 04 species belong to genus Phacus, 02 species belonging to Trachelomonas and Euglena based on light microscopy. It was observed that Euglena was the most diverse genus and it is supposed to be the indicator species of the polluted water. It was observed that E. oblonga was found in maximum pH range, that is, 7.0–11.0. Similarly, E. brevicaudatus was found in maximum EC, that is, 169 ± 1.5 ms/cm these outcomes indicated that for internal examination along with LM, SEM was necessary for correct identification of algal sample up to specie level.     

Scanning electron microscopic studies on surface microstructural features of some tissues of epizootic ulcerative syndrome‐affected Puntius ticto (Hamilton, 1822) from Southern Assam of India

Epizootic ulcerative syndrome (EUS)‐affected Puntius ticto exhibited a number of abnormalities in scale, gill, skin, and muscle at surface microstructural level. Lepidontal damage including breakage of individual lepidonts and complete loss of lepidontal assembly was evident in the scale. Disturbances in the gill included fusion of gill–lamella, distortion of lamellar surface, damage of primary and secondary gill lamella as well as breakage of gill racker. The skin of EUS‐affected Puntius ticto showed breakage of epidermal cell boundary, distortion, and loss of alignment of cells including lesions at places. Loss of alignment of muscles along with breakage and distortion of individual fibers was also evident. The similarities of these abnormalities in EUS‐affected Puntius ticto with those of pesticide and other pollutant toxicity reported in some fish is discussed with the help of relevant literature. Microsc. Res. Tech., 2009. © 2008 Wiley‐Liss, Inc.