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.     

Microscopic techniques for characterization and authentication of oil-yielding seeds

Investigation of alternative energy sources is need of current time due to growing power crisis and associated environmental issues. Biodiesel is considered as sustainable power source and promising alternative to fossil fuels. Therefore, our current investigation aimed to identify micromorphological characters of 10 novel nonedible oil-yielding seeds through scanning electron microscopy. It was revealed from light microscopic study that there is variation in seed size from 3 to 15 mm in length and 2 to 11 mm in width. Likewise, a huge variation in color was observed such as light green, greenish yellow, blackish brown, and various shades of brown. Presence and absence of Hilum was observed, and compression of seeds varied from depressed, lateral, and dorsoventral. Seed's shape differs from ovate, clavate, triangular ovate, cuneiform, ovoid, and elliptical shape. Seed oil content fall in range of 18–58% (wt/wt). Free fatty acid content of the seeds varies from 0.3 to 3.1 mg KOH/g. Ultrastructure of seeds exhibited huge variation in shape, size, periclinal wall, anticlinal wall, and surface ornamentation. Nonedible seeds varied in wall structure from angular, wavy, dentate entire, irregular, puzzled, elongated, even, and polygonal. The periclinal wall arrangements show alteration from flat, looped, raised, depressed, lofty, even, pentagonal, polygonal, and undulate seed margins. Outcomes of this investigation recommended that scanning electron microscopy could act as a helpful tool in disclosing the hidden micromorphological characters among nonedible oil-yielding seeds and subsequently helping in correct, authentic seed identification and classification as potential feedstock for biodiesel.     

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.     

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.     

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.     

Biodiesel production process optimization from Spirulina maxima microalgae and performance investigation in a diesel engine

Biodiesel is a renewable, easily biodegradable, eco-friendly and sustainable alternative energy source. In this investigation, crude oil was extracted from Spirulina maxima microalgae through biochemical conversion method with the help of soxhlet apparatus. Biodiesel production process parameters were optimized through base transesterification. Maximum biodiesel yield achieved was 87.75 % at optimal reaction condition after transesterification, when methanol to oil ratio was 6:1, catalyst loading was 1 % KOH (wt.%), temperature was 65 °C, and stirring speed was 600 rpm for a reaction time of 70 minutes. All the physicochemical properties of the produced biodiesel were determined and compared with the ASTM D6751 specification. Finally, performance and emission of an unmodified diesel engine was evaluated with 20 % and 40 % (v/v) biodiesel blends and compared the results with ordinary Diesel fuel (DF). Using biodiesel blends improves Hydrocarbon (HC) emission by 10-15 % and Carbon monoxide (CO) emission by 9.3-13.9 %. However, Brake specific fuel consumption (BSFC), Oxides of nitrogen (NOX), Carbon dioxide (CO₂) and smoke opacity were found to be slightly higher for biodiesel blends, and Brake thermal efficiency (BTE) was found slightly lower than DF. Thus, Spirulina maxima serves as a potential feedstock for biodiesel production and prospective fuel in diesel engine application.

     

Combustion characteristics of an agricultural diesel engine using biodiesel fuel

Biodiesel has great potential as an alternative fuel for diesel engines that would reduce air pollution. It is a domestically produced, renewable fuel that can be manufactured from fresh or used vegetable oils, or from animal fats. In this study, a biodiesel fuel derived from rice bran oil was tested as an alternative fuel for agricultural diesel engines. The emissions were characterized for both neat and blended biodiesel fuels, and for conventional diesel fuel. Since this biodiesel fuel contained 11% oxygen, it strongly influenced the combustion process. The use of biodiesel fuel resulted in lower carbon monoxide, carbon dioxide, and smoke emissions, without any increase in nitrous oxide emissions. The study demonstrated that biodiesel fuel could be effectively used as a renewable and environmentally innocuous fuel for agricultural diesel engines.     

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.     

Effect of phyllanthus emblica biodiesel based lubricant on cylinder liner and piston ring

In the present time, need of biogenic lubricants is the focusing area which will be biodegradable, avirulent and eco-friendly. Current experimental tests depict the effect of ‘Phyllanthus emblica’ a non-edible feedstock through pin on disc tribo tester. Tests were conducted to evaluate the impact of 0, 10, 20 and 30 % blending of Phyllanthus emblica with SAE20W40 lubricating oil on cylinder liner and piston ring. Promising results have been manifested with 10 % blending of biodiesel (BD) with lubricating oil in terms of coefficient of friction and specific wear rate in comparison with other examined feedstock. For analysis of wear debris in the used oil analytical ferrography was also done. The effects of temperature on wear and friction characteristics have also been discussed.

     

Quality assurance and authentication of herbal drug (Argyrolobium roseum and Viola stocksii) through comparative light and scanning electron microscopy

The quality assurance and authentication of crude herbal drugs play important role in the effective therapeutic effect of herbal drug and their products. There are many reported problems in quality assurance of herbal crude drugs concerning to their correct identification. The present study was designed with the aim to document the authentication and quality assurance of the herbal crude drugs (Argyrolobium roseum and Viola stocksii) thorough light microscopy (LM) and scanning electron microscopy (SEM). The detailed foliar anatomical studies showed polygonal epidermal cells having anticlinal walls in Argyrolobium roseum while rounded epidermal cells were observed in Viola stocksii. The anomocytic stomata type was observed in Argrolobium roseum while actinocytic was noticed in Viola stocksii. The pollen of studied species appeared as tricolporate showing reticulate exine sculpturing in Argrolobium roseum while fine perforations were recorded in Viola stocksii. Furthermore, quantitative analysis of phenols, flavonoids, and antioxidant activity showed high flavonoid and phenol content in Argyrolobium roseum as compared with Viola stocksii. It was observed that Argyrolobium roseum was discriminated from the Viola stocksii based on the leaf and pollen micromorphological traits by using LM and SEM techniques. It was concluded that LM and SEM techniques were found useful for the quality assurance of botanicals and their authentication.     

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.     

Pollen micromorphological analysis of tribe Acacieae (Mimosaceae) with LM and SEM techniques

The morphology of pollen grains of the 06 species of tribe Acacieae (Mimosaceae) belonging to one genus were investigated using scanning electron microscopic (SEM) and light microscopic (LM) approach. Lactic acid acetolysis method was used for (LM) while non‐acetolysis for (SEM). The species were examined for the both qualitative and quantitative palynological features. Qualitatively the pollen was studied for the shape, exine sculpturing, pollen shape, and pollen type. Oblate spheroidal shape of pollen was observed in Accacia modesta. Subprolate shape in Accaca nilotica. In Accacia farnesiana, prolate pollen was examined while prolate spheroidal in Acacia tortilis and spheroidal pollen grains were studied in Acacia catechu, respectively. Pollen exine sculpturing of the studied tribe possesses novel variation. Foveolate, reticulate, foveolate‐psilate, faint reticulate, fossulate, perforate, and scrabate exine sculpturing were noticed. Colpi were absent in all species. Excessive variation was observed in both qualitative and quantitative character of pollen. SPSS software was use for the quantitative measurement of the pollen grains. The current study will be helpful for the identification of the problematic species and phylogenetic studies of family Mimosaceae.     

废动植物油生产转化生物柴油技术的研究

生物柴油是一种绿色环保、可再生的生物质燃料.利用废动植物油在微酸催化剂环境下制取生物柴油的酯交换工艺进行了系统研究,探索了反应温度、催化剂用量等对反应的影响,并讨论了最佳反应条件.     

Pretreatment methods to improve the kinematic viscosity of biodiesel for use in power tiller engines

Biodiesel is an environmentally friendly fuel that can replace diesel in compression ignition engines without changing the engine structure. Biodiesel is typically manufactured from vegetable oils and animal fats, which give the fuel its oxidation stability and cold-flow properties, respectively. However, the kinematic viscosity of biodiesel can cause engine performance problems such as incomplete combustion and sludge formation due to insufficient fuel atomization. To address these problems, in this study, a pretreatment technology that lowers the kinematic viscosity of biodiesel made from blended animal fat and vegetable oil was developed. The results of application of the pretreated fuel to a single-cylinder power tiller engine indicated that it produced 88.3–99.8 % of the brake power produced by conventional diesel. In addition, although the pretreated biodiesel exhaust included increased amounts of nitrogen oxides and carbon dioxide emissions, the proposed fuel also decreased the amounts of hydrocarbon and carbon monoxide emissions compared with conventional diesel emissions.

     

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.     

Palyno‐anatomical studies of monocot taxa and its taxonomic implications using light and scanning electron microscopy

Palyno‐anatomical study of monocots taxa using Light and Scanning Electron Microscopy (SEM) was first time conducted with a view to evaluating their taxonomic significance. Studied plants were collected from different eco‐climatic zones of Pakistan ranges from tropical, sub‐tropical, and moist habitats. The aim of this study is to use palyno‐anatomical features for the correct identification, systematic comparison, and investigation to elucidate the taxonomic significance of these features, which are useful to taxonomists for identifying monocot taxa. A signification variation was observed in quantitative and qualitative characters by using the standard protocol of light microscopy (LM) and SEM. Epidermal cell length varied from maximum in Allium griffthianum (480 ± 35.9) μm at the adaxial surface to minimum in Canna indica (33.6 ± 8.53) μm on abaxial surface. Maximum exine thickness was observed in Canna indica (4.46) μm and minimum in Allium grifthianum (0.8) μm. Variation was observed in shape and exine ornamentation of the pollen, shape of the epidermal cell, number, size, and type of stomata, guard cell shape, and anticlinal wall pattern. Based on these palyno‐anatomical features a taxonomic key was developed, which help in the discrimination of studied taxa. In conclusion, LM and SEM pollen and epidermal morphology is explanatory, significant, and can be of special interest for the plant taxonomist in the correct identification of monocots taxa.     

Sustainable cultivation location optimization of the Jatropha curcas L. under uncertainty: A unified fuzzy data envelopment analysis approach

Second-generation biodiesel production from non-edible high oil content feedstocks such as Jutropha curcas L. (JCL) has been found as a suitable alternative for the fossil diesel which is mostly consumed in transportation sector. Second-generation biodiesel eliminates the drawbacks of the first generation such as food-energy challenge and provides opportunity for rural development. Location optimization of JCL plays a vital role in the success and prosperity of JCL projects. In this study, sustainability and ecological indicators are defined to evaluate the performance and efficiency of the candidate locations for JCL cultivation under uncertainty. The values of defined indicators are highly tainted with uncertainty in real-life situation. To optimize the candidate locations for JCL cultivation under uncertainty, an efficient Unified Fuzzy Data Envelopment Analysis (UFDEA) model is developed. The proposed UFDEA model is verified and validated through investigating a real case study in Iran and the associated results are compared to those obtained by the crisp Unified Data Envelopment Analysis model under different levels of uncertainty. The obtained results show the applicability of the proposed UFDEA model in selecting suitable areas for JCL cultivation under uncertainty.