Identification of novel nonedible oil seeds via scanning electron microscopy for biodiesel production

Exploration of substitute energy feed‐stocks is the much‐debated topic in the scientific society due to increasing power crises and related ecological concerns. As a source of sustainable energy, biodiesel turns out to be the best alternative to petro fuels. In this context, nonedible oil‐producing seeds might be a potential source for biodiesel production owing to their environment‐friendly nature and cost‐effectiveness. The current study, consequently, deals with the investigation and identification of micro‐morphological characters between six novel nonedible oil‐bearing seeds employing scanning electron microscopy as possible biodiesel feed‐stocks. Light microscopic examinations show that seed size varies from 0.3 to 1.3 cm in width and 0.5 to 1.5 cm in. Additionally, a large difference in seed color ranges from dark brown, black, and various shades of light brown was also witnessed. The FFA content of the seeds ranges in 0.3–4.1 mg KOH/g, and the seed oil content fall in 30–65% (w/w) range. SEM‐mediated seed ultrastructure investigations displays greater variation in seed size, shape, color, periclinal wall shape, and sculpturing and so on. All the seeds differ from rounded, ovoid, ovate, oblong, flattened, and elliptical shape. Greater variation in seed wall structure has been seen from angular, entire, irregular, straight, elongated, smooth, and polygonal. The periclinal wall arrangements show alteration from flat, depressed, elevated, smooth, pentagonal, bullate, and coarse seed margins. The results obtained from the current study suggest that scanning electron microscopy could be a beneficial tool in vitalizing the hidden micromorphological characters among various nonedible oil producing seeds, which eventually helps in exploration, correct identification, seed classification, and authentication in future.     

Implication of scanning electron microscopy as a tool for identification of novel,nonedible oil seeds for biodiesel production

Biodiesel is a promising, bio-based, renewable, nontoxic, environment friendly, and alternative fuel for petroleum derived fuels which helps to reduce dependency on conventional fossil fuels. In this study, six novel, nonedible seed oil producing feedstock were explored for their potential for sustainable production of biodiesel. It is very important to correctly identify oil yielding plant species. Scanning electron microscopy (SEM) was used as reliable tool for authentic identification of oil yielding seeds. Macromorphological characters of seeds were studied with light microscopy (LM). Outcomes of LM of seeds exposed distinctive variation in seed size from 16.3 to 3.2 mm in length and 12.4 to 0.9 mm in width, shape varied from oval to triangular, and color from black to light brown. Oil content of nonedible seed ranged from 25 to 30% (w/w). Free fatty acid content of seed oil varied from 0.32 to 2.5 mg KOH/g. Moreover, ultra structural study of seeds via SEM showed variation in surface sculpturing, cell arrangement, cell shape, periclinal wall shape, margins, protuberances, and anticlinal wall shape. Surface sculpturing varied from rugged, reticulate, varrucose, papillate, and striate. Periclinal wall arrangements confirmed variation from rough, wavy, raised, depressed, smooth, and elevated whereas, anticlinal walls pattern showed variation from profuse undulating, smooth, raised, grooved, deep, curved, and depressed. It was concluded that SEM could be a latent and advanced technique in unveiling hidden micromorphological characters of nonedible oil yielding seeds which delivers valuable information to researchers and indigenous people for precise and authentic identification and recognition.     

Scanning electron microscopy leads to identification of novel nonedible oil seeds as energy crops

Currently, exploration of alternative energy resources is hotly debated among the scientific community owing to rising energy crises and environmental issues. Biodiesel, as renewable energy source proves to be a better option and substitute to petro diesel. In this regard, nonedible seeds could be a better feedstock for synthesizing biodiesel due to their cost effectiveness and environmental friendly attributes. The present study, therefore, deals with the exploration and identification of micromorphologic features among eight novel nonedible oil yielding seeds via scanning electron microscopy (SEM) as potential feedstock for biodiesel industry. Light microscopic studies revealed that seeds size vary from 0.1–2.9 cm in length to 0.1–3 cm in width. Moreover, a great variation in seed color from black, green, and different shades of brown was also observed. Seeds ultra‐structure examination by SEM exhibit great variation in seed shape, size, color, sculpturing and periclinal wall shape and arrangement and so on. All the understudy seeds vary from rounded, irregular, subspherical, ellipsoidal, reniform, flattened, polygonal, ovate, pyriform, oblong, and globose shape. Seeds wall structure exhibits great variation from entire, angular, straight, irregular, polygonal, smooth, and elongated. The periclinal wall pattern exhibits variation from flat to slightly concave‐convex with straight, angular, undulate, or dentate seeds margin. Among the studied species only Argemone ochroleuca Linn. (Papaveraceae) possess micropylar peak, ridged raphe, and basal helium. The obtained results from the present study would therefore, suggest that SEM could be a useful tool in refreshing the veiled micromorphological features among different oil yielding seeds which in turn helps the researchers for their correct identification, exploration, authentication, and seeds classification in future.     

Identification of novel,non-edible oil seeds via scanning electron microscopy as potential feedstock for green synthesis of biodiesel

In the present era, environmental glitches associated with extensive emission of greenhouse gases (GHG) and energy crises caused by exhausting fossil fuel reservoirs has diverted researcher's interest toward alternative and renewable energy sources. Biodiesel is a renewable, biodegradable, and sustainable alternative to petro-diesel. Biodiesel synthesized from non-edible seed oils is preferred due its cost effectiveness and eco-friendly nature. Hence, our present study focused on investigation and identification of micromorphological characters of six novel, non-edible seed oil feedstock for biodiesel production via scanning electron microscopy (SEM). Results of light microscopy of seeds revealed distinct variation in seed size (15.8–1.8 mm in length and 9.4–1.1 in width), shape (round to Cuneiform), and color (from black to yellowish green). Non-edible seed oil content fall in range of 28–38% (wt/wt). Free fatty acid (FFA) content varied from 0.56 to 2.06 mg KOH/g. Multivariate analysis was performed to investigate correlation between three significant variables of seed oil yielding feedstock such as potential for biodiesel synthesis, oil content, and FFA content via principal component analysis. Ultra morphological investigation of seeds surfaces via SEM exhibited distinctive variation in surface sculpturing, cell arrangement, cell shape, periclinal wall shape, margins, protuberances, and anticlinal wall shape. Seed surface sculpturing varied from reticulate, semitectate, wrinkled, rugose, papillate, perforate, and striate. Periclinal wall arrangements confirmed variation from glabrous, raised, depressed, elevated, smooth, pentagonal, entire, and ripple margins. Whereas, anticlinal walls pattern demonstrated variation from angular, smooth, wavy, deep, dentate, entire, irregular, puzzled, elongated, curved, and depressed. Finally, it was concluded from obtained results that SEM could be a possible useful tool in disclosing veiled micromorphological characters of non-edible oil yielding seeds, which provides useful information to researchers for their correct, authentic identification, and classification in modern synthetic system.     

Detailed macro and micromorphology of grasses caryopsis using scanning electron and light microscopy

Micromorphology of 15 grass caryopsis was investigated through scanning electron microscopy for its implications in the identification of grasses. Both macro and micromorphological characters especially caryopsis shape and surface morphology were found significant in the taxonomic diagnosis. Shape morphology of caryopsis varied among the species and lanceolate, fusiform, narrow ellipsoidal, ovate, elliptical, plano-convex, fusiform, linear ovate, broadly elliptical, broadly pllipsoidal, plano-convex, oblique ovate, and subglobose shapes were recorded. Digitaria nodosa (3.31 × 0.7 mm) and Echinochloa crus-galli (3.26 × 1.44 mm) had the largest caryopsis size, while Acrachne racemosa (0.76 × 0.54 mm) and Eragrostis minor (0.62 × 0.38 mm) had the smallest caryopsis size. Macro and micromorphological characters were useful for the diagnosis of genus and species. Utilizing both qualitative and quantitative character taxonomic key at generic and species level were made.     

Using microscopic techniques for taxonomic implications of seed and fruits of Delphinium L. (sensu lato) (Ranunculaceae)

In present research, micromorphological characters of the genus Delphinium L. (sensu lato) Ranunculaceae seeds and fruits were studied using microscopic techniques. A total of 37 species were studied using light (LM) and scanning electron microscopy (SEM) to evaluate seeds and fruits micromorphological characteristics for the taxonomic identification and species relationship in the genus. For each taxon, 33 characters were observed and studied. All characters were analyzed using MVSP software and UPGMA method by using dissimilarity index (average taxonomic distance). Considerable differences were observed in trichomes shape of fruit and seed color features. The results revealed that all studied taxa are separated into three clades. The first clade consists of D. pallidiflorum and D. semibarbatum, which have black seed and two types of glandular and crispate trichomes in fruit. The second clade consists of D. speciosum, D. szowitsianum, D. dasystachyson, D. aquilegifolium, D. ursinum, C. trigonelloides, and C. camptocarpa, which had a brown color seeds, two types of glandular and villous trichomes on fruits also have been observed. The remaining species were classified as third clade having various colors including yellow, black, and brown color seeds, the trichome shapes included one of the following forms strigose, villous, trichomes less, glandular, hooked, and crispate trichomes in fruits. The aim of the present study is to use micromorphological characters of seeds and fruits for the taxonomic identification and species delimitation in the genus Delphinium.     

Macromorphological and micromorphological seed features of selected taxa of Caesalpiniaceae using light and scanning electron microscopy

Seed micromorphological and macromorphological characteristics of 12 taxa of Caesalpiniaceae from Pakistan have been studied, using light and scanning electron microscopy, to investigate the importance of seed coat features as a taxonomic tool. Great variations have been observed in color, shape, dimension, and seed surface pattern among the different genera of the family. A taxonomic key was prepared for the studied taxa. Six types of seed shapes were observed; circular, elliptical, irregular, oblong, oval, and ovoid. All examined seeds were hard except fragile seeds of Haematoxylom campechianum. Eight types of surface ornamentation have been noticed that include levigate, lugose, papillate, reticulate, reticulate irregular, reticulate regular, rhombus, and rogues. Majority of the taxa has been observed with thick ornamentation wall but thin ornamentation wall has also been recorded in few species. Fracture line of the various patterns was present in all taxa except genus Bauhinia. Three types of texture crudeness; coarse, medium, and fine have been recorded. Both micromorphological and macromorphological characters of seed are very fruitful in identification and classification of Caesalpiniaceae.     

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.     

Scanning electron microscopy as a tool for authentication of biodiesel synthesis from Linum usitatissimum seed oil

Utilization of renewable and alternative energy feedstocks such as nonedible seeds oil to deal with the increasing energy crises and related ecological concerns have gained the attention of researchers. Biodiesel is an efficient and renewable substitute for diesel engine. This work investigates the potential of inexpensive nonedible seed oil of Linum usitatissimum to synthesize biodiesel using iron sulfate green nanocatalyst through the process of transesterification. Flax seed contains about 37.5% oil content estimated through Soxhlet apparatus. Light microscopy revealed that seed size varies from 3.0 to 6.0 cm in length, 2.0 to 3.3 cm in width, and 0.7 to 1.0 mm in diameter. Color of seed varied from yellow to brown. Characterization of biodiesel is performed through GC–MS and FTIR. Scanning electron microscopy was carried out to study the morphological features of seed coat. Catalyst was characterized by scanning electron microscopy, energy diffraction X-ray, and X-ray diffraction. The diffraction peaks of Fe₃O₄ green nanoparticles were found to be in 2θ values, 30.24°, 35.62°, 38.26°, 49.56°, 57.12°, and 62.78°. Fuel properties of biodiesel are also determined and compared with ASTM standards. Linum usitatissimum biodiesel has density 0.8722 (15°C kg/L), kinetic viscosity 5.45 (40°C cSt), flash point (90°C), pour point (−13°C), cloud point (−9°C), sulfur (0.0432% wt), and total acid number (0.245 mg KOH/g). It is concluded that L. usitatissimum seed oil is a highly potential source for biodiesel production to cope with the challenge of present energy demand.     

Intraspecific variation in spermoderm pattern of tribe Acacieae (Mimosoideae) using scanning electron microscopy techniques

Current research carried out in Pakistan is the first report on spermoderm ornamentation of eight species of tribe Acacieae (Mimosoidae) by using scanning electron microscopic techniques representing two genera, Fedherbia and Acacia were examined. Different spermoderm ornamentation were observed, described and discussed for their taxonomic importance. Seeds surfaces of the studied tribe possess novel variations in macro and micro morphology. Great variations were observed in both qualitative and quantitative characters of seeds. Seeds shape was oblong, ovate to elliptical and spermoderm ornamentation was levigate, rugose, polygonal and discoid, colliculate, and papillose type. These variations in the spermoderm ornamentation can be used as an aid in identification and classification of the members of tribe Acacieae.     

Implications of foliar epidermal micromorphology using light and scanning electron microscopy: A useful tool in taxonomy of Korean irises

The genus Iris L., comprising approximately 210 species, is one of the most species-rich genera in the family Iridaceae. In this study, the first comprehensive leaf micromorphological characters of Korean irises were studied using light and scanning electron microscopy. Our objective was to evaluate the foliar micromorphological characteristics (namely epidermal cells, stomata types, and guard cell size) of Korean Iris taxa in a systematic context. All the investigated Korean Iris taxa had amphistomatic or hypostomatic leaves with anomocytic stomatal complexes. Guard cell length varied among species, ranging from 24.8 μm (I. rossii) to 56.0 μm (I. domestica). Although the presence of papillae on the outer periclinal wall is not of taxonomic significance, leaf margin pattern, guard cell size, and sunken stomata type were useful for species-level identification of Korean Iris species. The occurrence of polymorphic stomatal types was reported here for the first time, and the correlation between genome size and epidermal guard cell length was discussed.     

Taxonomic significance of caryopsis in subfamily Panicoideae (Poaceae) using scanning electron microscopy and light microscopy

In recent study, 13 taxa of subfamily Panicoideae were investigated for morphological characterization of caryopsis. Light microscope (LM) and scanning electron microscope (SEM) were utilized to study macro‐ and micro‐morphological caryopsis features respectively. Caryopsis size in studied taxa was recorded as 1.5–10 mm long and 1–4 mm wide. Caryopsis color was brown, green, yellow, and whitish‐brown. Caryopsis shape studied was obovate, elliptic, linear oblate, and round shallowly obtriangular. Hilum position is grooved and depressed. Caryopsis compression type was lateral and dorsiventral. Major variations among studied taxa were observed in terms of caryopsis surface pattern and epicuticular projection types. Six types of caryopsis surface pattern were observed viz. scabrate, rugose, striate, reticulate, papillate, and scabridulous. Structures such as silica cells, bulges, spines, prickles, granules, and bicellular microhair were studied as epicuticular projections. Major variations were observed among Cenchrus pennisetiformis and Cenchrus ciliaris as both has entirely two different types of surface patterns and epicuticular projections. Anticlinal wall thickness and pattern as well as periclinal wall texture and level were investigated. The present research work emphasized on caryopsis characterization of subfamily Panicoideae and it is recommended to establish phylogeny within subfamily Panicoideae and with other subfamilies of Poaceae.     

Utilization of foliar cuticle morphology for the identification of weedy grasses

For the robust identification of weeds of taxonomically complex families like Poaceae, scanning electron microscopy (SEM) and light microscopy (LM) are of significant implications. In this study, SEM and LM methods were used by investigating 12 Pakistani weedy grass species as an example for foliar epidermal micromorphological characters for its role in the identification of grasses species. Qualitative and quantitative foliar micromorphological variations in stomatal type, number, guard cells, subsidiary cells, silica bodies and trichomes were studied. Significant variation was observed in both qualitative and quantitative features and based on this diversity in epidermal characters, a key was drawn to delimitate the species. Variations in stomata, silica bodies (SiO₂·nH₂O), macrohairs, microhairs, hooks, papillae, prickles, and other epidermal structures were used in establishing the taxonomic key. The data were also statistically evaluated by determining its mean, standard deviation, variance, the coefficient of variance, and standard error. Foliar epidermal characters such as silica bodies shape and trichomes were taxonomically more useful than stomatal types, subsidiary, and guard cells. SEM and LM micromorphological characters were of substantial importance in the delimitation of closely related species.     

Taxonomic significance of foliar epidermal morphology in Lamiaceae from Pakistan

Foliar micromorphological features are useful to elucidate the taxonomy and systematics of the Lamiaceae species. Leaf epidermal morphology using scanning electron microscopy and light microscopy of 22 Lamiaceae species from 15 genera have been investigated with an aim to solve its taxonomic problem in the correct identification. Various foliar micromorphological features were observed to explain their importance in resolving the correct identification of Lamiaceae taxa. Two main types of trichomes were observed; glandular trichomes (GTs) and nonglandular trichomes (NGTs). GTs were further divided into seven subtypes including the capitate, subsessile capitate, sessile capitate, sunken, barrel, peltate, and clavate. Similarly, NGTs were also divided into simple unicellular and multicellular including conical, falcate, cylindrical, dendrite, papillose, and short hook shape. Quantitative measurement includes the length and width of the trichomes, stomatal complex, epidermal cells, stomata, and trichomes index. Based on the foliar micromorphological characters, a taxonomic key was developed to delimit and correctly identify studied taxa. Further molecular, other anatomical and phylogenetic studies are recommended to strengthen the systematics of Lamiaceae.     

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.     

Investigating Schizocarp morphology as a taxonomic tool in study of Apiaceae family by utilizing LM and SEM techniques

In present study, the schizocarp morphology of 14 species belonging to Apiaceae family has been investigated. Light microscopy (LM) and scanning electron microscopy (SEM) have been utilized to highlight qualitative and quantitative features of studied species. Variations have been observed in macro‐ and micro‐morphological features such as color, shape, symmetry, length, width, apex, epicuticular projections, surface patterns, anticlinal, and periclinal wall patterns. Schizocarp shapes observed were oval, round, triangular, linear, elliptic, and globose. Fruit was either homomorphic or heteromorphic. Crystalloids, stellate hair, multicellular spines, and platelets were mostly observed epicuticular projections. Surface patterns on the fruit surface were striate, rugulate‐striate, reticulate, and striato‐knotted. Both macro‐ and micro‐morphological characters can serve as an important tool in classifying Apiaceae family at various taxonomic ranks. Substantial variations observed can assist as useful constraints at various taxonomic levels as they provide reliable and constant details. Disparities observed in schizocarp features can pave a path for Apiaceae family classification based on phylogenetic and molecular studies.     

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.     

Intraspecific variation in seed morphology of tribe vicieae (Papilionoidae) using scanning electron microscopy techniques

Seed micromorphology of 12 species of tribe Vicieae (Papilionoidae) representing five genera were examined using Scanning Electron Microscope (SEM). The different seed types were described, illustrated, compared, and their taxonomic importance is discussed. Seeds exhibit great diversity in ultrastructure and a variety of novel morphological features have been determined among and within genera of the tribe. Seeds were characterized by oval to round shape, ovate to oblong outline and striate to papillate ornamentation. Radiate and wavy cell pattern has been observed as a characteristic exomorphological feature. In Vicia sativa, testa cells superficially seem to form peltate hairs. As a result, seed micromorphology with few exceptions showed variation and its taxonomic value was significant in distinguishing taxa at species level. The disparity in shape and density of papillae or protuberances and alignment of testa cells, may possibly give further insight at intraspecific level within tribe. The findings also illustrated that the use of SEM in seed morphology can reveal hidden morphological affinities among species and showed potential in delimitation of Vicieae members generally at tribe, genus, and particularly at species level.