Occidentalis Seeds Research Articles

Free Radical Scavenging-Related Antioxidant Properties and Inhibitory Activities of Telfairia occidentalis Seed Protein Hydrolysates and Peptide Fractions Against Two key Enzymes Implicated in Diabetes Mellitus

Free Radical Scavenging-Related Antioxidant Properties and Inhibitory Activities of Telfairia occidentalis Seed Protein Hydrolysates and Peptide Fractions Against Two key Enzymes Implicated in Diabetes Mellitus

Bioherbicide from Azadirachta indica Seed Waste: Exploitation, Efficient Extraction of Neem Oil and Allelopathic Effect on Senna occidentalis

Bioherbicides are an alternative to minimize the damage caused to the environment using agrochemicals. This study had the objective of extracting neem oil from ripe waste fruits that Azadirachta indica A. Juss and optimizing the process using solvents (or a mixture of solvents) with different polarities. Then, through a solid-liquid extraction system (Soxhlet), the solvents hexane, methanol/hexane (1:1), ethanol, and hexane/ethanol (1:1) were used to determine the process with the highest yield and most efficiency. The physicochemical parameters of the extracted oil (density, acidity value, iodine value, saponification value, esters value, and molecular weight) and the % of free fatty acids were determined. In addition, the allelopathic properties of the oil (0%, 2%, 3%, and 4% m/v) on septic weed Senna occidentalis seeds were evaluated, analyzing their growth and development parameters (germination, germination speed, hypocotyl, and radicle length). Hexane was the most efficient (4 h) in neem oil extraction, with the highest yield (43%). It also provided a better oleic and linoleic acid content (41.3% and 18.6%), similar to ethanol extraction (41.1% and 20.22%). Moreover, the allelopathic properties were more prominent for the oils extracted with hexane and hexane/ethanol. This optimized process provides an efficient alternative to obtain a natural herbicidal potential for strategically controlling harmful plants.

Effects of Roasting Periods on the Nutritive Value of Telfaira occidentalis (Fluted Pumpkin) Seeds

Effect of roasting on the proximate composition, anti-nutrients, and selected mineral components of fluted pumpkin seeds were obtained by extracting the seeds from the pod, sun drying, and dividing them into three portions, one untreated and the other two roasted for 30 and 60 minutes, respectively. The samples were pulverized separately using a mechanical grinder and chemically analyzed. After employing different roasting time ranging from 0 minutes to 60 minutes; moisture content reduced from 0.86-0.27% with a coefficient of variation (CV) of 53%, crude protein reduced from 30.28-21.46% with a CV of 19%, crude fiber 2.06-2.49% with a CV of 10%, ash content 4.13-4.33% with a CV of 5%, fat content 38.13-37.75% with a CV of 1.2%, and carbohydrate 24.54-32.70% with a CV 16%. Copper changed from 0.303 to 0.114% with a CV of 100%, lead from 1.070 to 0.007% with a CV of 128%, potassium from 7.54 to 7.62% with a CV of 0.68%, and calcium from 0.513-0.20% with a CV of 20.62%. The anti-nutrient, on the other hand, exhibited a steady reduction. Oxalate concentrations changed from 148.5-104.5% with a CV of 17%, cyanide concentrations from 0.5 to 0.4% with a CV of 14%, and phytate concentrations from 425.1-175.0% with a CV of 45%. On the other hand, processing times increased the amount of ash, crude fiber, carbohydrate, lead, potassium, and calcium in the product. The anti-nutrient decreased as the processing time grew. As a result of processing fluted pumpkin seeds for 60 minutes, fluted pumpkin seeds of greater protein content are produced, which can be used as a food supplement.

Hippocampal Astrogliotic Reduction in Scopolamine Hydrobromide-Induced Alzheimer's Type Cognitive Dysfunction Wistar Rats Following Administration of Aqueous Extract of Telfairia Occidentalis (pumpkin) seeds.

Astrocytes are small star-shaped glial cells that maintain normal human brain physiology including secretion of several active compounds and the formation of blood brain barrier. Reactive astrocytes support regenerating axons and also, actuate some genes responsible for the induction of synapse formation. In this study, the effect of aqueous extract of Telfairia occidentalis seeds on hippocampal astrogliosis was probed using scopolamine-induced Alzheimer's type cognitive dysfunction Wistar rats. Thirty Wistar rats weighed between 180-200g were randomly grouped into five designated A, B, C, D and E. Each group contained six rats. Alzheimer's type cognitive dysfunction was induced in groups B to E by administering intraperitoneally, 1 mg/kg body weight of scopolamine for seven days before Donepezil and the aqueous extract of Telfairia occidentalis seeds for fourteen days. Twenty-four hours after the last administration, the animals were sacrificed; their brain tissues perfused and stained with glial fibrillary acidic protein (GFAP) dye. Results revealed prominently stained astrocytes with their processes intact (group A). Some densely stained numerous astrogliosis with hypertrophied fibres were noticed in group B. Group C demonstrated prominent astrocytes with hypertrophied fibres, group D, moderately stained astrogliosis with hypertrophied fibres while group E showed numerous astrocytes with prominent nuclei and hypertrophied fibres. In conclusion, there was reduced hippocampal astrogliosis mostly in group D treated with Telfairia occidentalis which may neutralize oxidative stress and enhanced learning and memory in the Wistar rats of the present study.

Hippocampal Astrogliotic Reduction in Scopolamine Hydrobromide-Induced Alzheimer’s Type Cognitive Dysfunction Wistar Rats Following Administration of Aqueous Extract of Telfairia Occidentalis (pumpkin) seeds.

Astrocytes are small star-shaped glial cells that maintain normal human brain physiology including secretion of several active compounds and the formation of blood brain barrier. Reactive astrocytes support regenerating axons and also, actuate some genes responsible for the induction of synapse formation. In this study, the effect of aqueous extract of Telfairia occidentalis seeds on hippocampal astrogliosis was probed using scopolamine-induced Alzheimer’s type cognitive dysfunction Wistar rats. Thirty Wistar rats weighed between 180-200g were randomly grouped into five designated A, B, C, D and E. Each group contained six rats. Alzheimer’s type cognitive dysfunction was induced in groups B to E by administering intraperitoneally, 1 mg/kg body weight of scopolamine for seven days before Donepezil and the aqueous extract of Telfairia occidentalis seeds for fourteen days. Twenty-four hours after the last administration, the animals were sacrificed; their brain tissues perfused and stained with glial fibrillary acidic protein (GFAP) dye. Results revealed prominently stained astrocytes with their processes intact (group A). Some densely stained numerous astrogliosis with hypertrophied fibres were noticed in group B. Group C demonstrated prominent astrocytes with hypertrophied fibres, group D, moderately stained astrogliosis with hypertrophied fibres while group E showed numerous astrocytes with prominent nuclei and hypertrophied fibres. In conclusion, there was reduced hippocampal astrogliosis mostly in group D treated with Telfairia occidentalis which may neutralize oxidative stress and enhanced learning and memory in the Wistar rats of the present study.

Hepatic encephalopathy in swine experimentally poisoned with Senna occidentalis seeds: Effects on astrocytes

Senna occidentalis may be accidently ingested by humans and animals. In this study, the percentages of S. occidentalis seeds necessary for experimental reproduction of hepatic encephalopathy were determined in a pig model and the biochemical and microscopic pathology is described in detail, with emphasis on the astrocytes. The experimental groups (G1, G2 and G3) were fed rations containing 5%, 7.5% and 10% of S. occidentalis seeds for 7–11 days. Pigs from the three experimental groups showed incoordination, ataxia, disorientation, head pressing, anorexia, recumbency and depression. In addition, the enzymes aspartate aminotransferase, alkaline phosphatase and creatine phosphokinase increased in all treated animals, which also showed higher serum total bilirubin and ammonia levels than in the control group (C). Microscopically, all experimental animals revealed acute hepatocellular swelling, multifocal coagulative necrosis in the pancreas, necrosis in the cardiac muscle, severe spongiosis in brain white and grey matter, and Alzheimer type II astrocytes in grey matter of the cerebral cortex. These cells were glial fibrillary acidic protein (GFAP) negative in G3. In white matter, a decrease in the positive area occupied by GFAP-immunolabelling and in the number of astrocytes per immunoreactive area was observed in G3 animals (5.35 ± 1.14% and 410 ± 45 cells/mm2, respectively) compared to the C animals (13.93 ± 1.59% and 581 ± 36 cells/mm2, respectively). This loss of GFAP was accompanied by alterations in astrocyte morphology, such as shrinkage of the cell body and retraction of the extending processes. This pig model of ammonia-mediated astrocyte damage could be used to study not only poisoning by S. occidentalis, but also other medical conditions resulting in hepatoencephalopathy.

Pomade Production from the Metallic Soaps of Telfairia occidentalis Seed Oil

Mature fruits of Telfairia occidentalis were purchased from a local market in Ikono Local Government Area of Akwa Ibom State, Nigeria. The seeds were separated from the fleshy mesocarp, washed with water and sundried. The cotyledons were separated from the seed coat and then oven-dried at 105 – 110 °C for 24 hours. The dry seeds were blended into powder and the oil cold-extracted with n-hexane. The oil extracted was used to prepare copper, nickel and zinc soaps. The soaps were characterised using standard methods and then applied as additives in the production of body pomades. Several tests were performed on the control and the metallic soap pomades to establish the effectiveness of the metallic soaps in pomade production. All the prepared metallic soaps were sparingly soluble in kerosene and methanol and exhibited no foaming characteristics. The properties of the pomades imply that the metallic soaps of T. occidentalis have good potential in the cosmetics industry.

Physical Characteristics and the Effect of Boiling and Fermentation on the Nutritional Value of <i>Telfairia occidentalis</i> Seeds

This work evaluates some physical parameters (the weight, length, diameter of fruits and seeds, number of seeds per fruit) and the effect of boiling and fermentation on the nutritional value of Telfairia occidentalis (fluted pumpkin) seeds. Firstly, a survey was done in the city of Yaounde on the different treatments applied to the seeds before cooking. From the results of the survey, the seeds were divided into three groups: raw, boiled and fermented. The moisture, lipid, protein, fibre, carbohydrate and ash contents were analysed using AOAC methods and minerals by atomic absorption spectrophotometry. The results showed that T. occidentalis fruit averagely weighed 6.35 kg and contained about 90 seeds. The decorticated seeds had an ovoid shape, 3.70 cm long and weighed 8.91 g. Boiling led to an increase in lipid (16.29% - 31.44%) and carbohydrate (19.20% - 21.8%) but a decrease in protein (54.06% - 34.17%) contents. Fermentation increased the crude fibre (0.70% - 1.1%) but decreased the ash content (4.07% - 3.14% DM). Boiled seeds had higher calcium, magnesium, potassium and sodium, while fermented seeds had higher zinc levels. Boiling proved better in preserving most of the seed nutrients. These seeds could be used to prevent some mineral deficiencies and their high proteins suggest their potential for the formulation of infant foods.

Subacute Hepatotoxicity of Extracts of Senna occidentalis Seeds in Swiss Albino Mice.

Senna occidentalis is potentially toxic to humans and animals. Its seeds are crop contaminant weeds in some localities where liver disease is prevalent. This study assessed the subacute hepatotoxicity of S. occidentalis seeds in mice model. Three groups of female Swiss Albino mice (25–28 g, aged 8–10 weeks) received distilled water (control), 400, and 1000 mg/kg extract of S. occidentalis seed, respectively. At the end of the study, body weight and liver organ weight were recorded, and tissue and blood samples were collected and analyzed. The results indicated that the extract treated groups, at both doses, showed significant (p ≤ 0.001) decrease in mean body weight gain in the fourth week of the experiment. Besides, the extract treated groups showed significant (p ≤ 0.001) elevation of liver enzyme markers: alanine aminotransferase and aspartate aminotransferase. Also, histopathological examinations of liver tissue showed moderate microvesicular steatosis of hepatocytes and mild inflammation in the 400 mg/kg treated group as well as marked micro- and macrovesicular steatosis, focal area necrosis, and periportal inflammation with mononuclear cell infiltration in the 1000 mg/kg treated group. Thus, these findings show that S. occidentalis seeds exhibit hepatotoxicity in mice, characterized by changes in liver tissue architecture and liver enzyme levels.

Purification of antibacterial proteins from Coffee senna (Senna occidentalis) seeds

Senna occidentalis (L.) Link formally known as Cassia occidentalis is a popular herb in folk medicine for the treatment of a wide range of microbial infections. Crude, ammonium sulphate precipitated and dialyzed proteins of S. occidentalis seeds were evaluated for their antibacterial potential by agar well diffusion and broth dilution techniques, against ten bacterial isolates made up of five Gram positive bacteria; Staphylococcus aureus, Streptococcus pyogenes, Enterococcus Sp, Listeria monocytogene and Bacillus subtilis and five Gram negative bacteria; Escherichia coli, Klebsiella pneumonia, Pseudomonas aeruginosa, Salmonella typhi and Shigella dysentria. The proteins were isolated by gel filtration on sephadex G-75 column and tested for antibacterial activity. The crude, dialyzed and precipitated proteins were active against all Gram positive bacterial isolates tested but were inactive against all Gram negative bacterial isolates used. The Minimum Inhibitory Concentration (MIC) of the crude, precipitated and dialyzed proteins against the most significantly (p < 0.05) sensitive bacterial B. subtilis were 0.67, 0.73 and 3.34 mg/ml respectively while the Minimum Bactericidal Concentration (MBC) were 2.67, 1.46 and 6.67 mg/ml respectively. Isolation however, resulted in total loss of antibacterial activities. SDS/PAGE analysis showed that the seed contains different proteins with molecular weights ranging from 14,400 to 66,200 kDa. The results of this study suggest that S. occidentalis seed contains proteins that have narrow spectrum, synergetic antibacterial activity against important food spoilage bacterial and can therefore be exploited for the development of novel antibacterial agent(s) for food preservation.

Equilibrium, Adsorption Isotherm and Kinetic Studies on the Removal of Methylene Blue Dye from Aqueous Solution onto Senna Occidentalis Seeds Activated Carbon

This paper studied the adsorption potentials and efficiency of low cost activated carbon produced from Senna occidentalis seeds for the removal of Methylene Blue dye in aqueous solutions. Batch adsorption experiments were carried out to investigate the effects of variables such as initial dye concentration, carbon dosage, contact time, pH and temperature to the process. Similarly, adsorption isotherms consisting Langmuir, Freundlich and Temkin were employed and found Langmuir Adsorption model fitted best. However, Kinetic models consisting pseudo first-order, pseudo second-order and intra-particle diffusion were also used to investigate the kinetic nature of the process, and found that pseudo second-order model better explained the process. Keywords: Equilibrium, Adsorption Isotherm, Kinetic Studies, Activated Carbon and Methylene Blue Dye DOI : 10.7176/CMR/11-4-04 Publication date : April 30 th 2019

Effect of ultraviolet radiation the germination rate of tree seeds

Extensive use of conifers is highly advisable for urban landscaping, since they are ever-green, low exploited and long-living compared to their deciduous counterparts. The paper presents the effect of ultraviolet (UV) radiation on the germination rate of Thuja occidentalis (2014), Picea fennica (2016) and Pinus sylvestris (2016) seeds exposed to environmentally friendly UV light-emitting diodes (LEDs). We used two sources of UV: the first one consisted of 54 LEDs with power of 2 W, the second one – 81 LEDs of 3 W. UV radiation was mostly within the A range (> 90%) and had a length of 315 nm. It was found out that UV radiation improved the germination rate of Picea fennica seeds from Quality Class 2 to Quality Class 1, i.e. from 75% to 86%, thus reducing the planting rates and, subsequently, the planting costs. After being exposed to the UV dosage of 2 kJ/m2, Thuja occidentalis seeds had 12.8% better germination rate than the control sample, resulting in faster growth and earlier full-grown shoots. UV-A radiation did not have identifiable positive effects on the pine seeds.

Assessment of Liver Histomorphology and Curative Effect of Chloroform Extract of Telfairia Occidentalis Seed on Carbon Tetrachloride (CCL4) Induced Liver Toxicity in Wistar Rats

Background: Telfairia occidentalis seed is a multipurpose plant with many attributes and considerable therapeutic potentials. Carbon Tetrachloride is one of the oldest and most widely used toxins for experimental induction of liver fibrosis in laboratory animals. The Liver is usually the organ to receive the effect of this chemical thereby causing hepatotoxicity, simply because of its method of metabolism, as well as carrying out the first pass effect by metabolizing endogenous and exogenous substances hence its consideration. Aim: The aim is to evaluate the histomorphologic anatomy of the liver with Telfaira occidentalis seed upon CCL4 induced toxicity in wistar rats. Methods: Telfairia occidentalis seeds were harvested and oven dried at the temperature of 400c for 4 days with an extract made from it using chloroform while the seed was immersed in a bowl of chloroform and stirred. The Lethal dose (L50) was made using the proposed new method and at 5000 mg/kg, no mortality occurred. In this experiment, a total of 20 adult wistar rats of 4 groups with 5 Rats in each group were used; Group 1 (Control group) received food and water ad libidum, Group 2 (Negative Control) received food, water ad libidum and CCL4 alongside Olive oil at the same volume of 0.5 ml, Group 3 received food, water ad libidum, CCL4 alongside olive oil at the same volume of 0.5 ml and 400 mg/kg of the extract, Group 4 received food, water ad libidum, CCL4 alongside olive oil at the same volume of 0.5 ml and 800 mg/kg of the extract which all lasted for 14 days. At the 15th day, the rats were sacrificed and histological slides were made. Results: The histological slides showed the rats in group 1 had their normal features; rats in group 2 had several abnormal features and rats in group 3 and 4 showed hepatocurative effect by bringing the liver gradually back to normal. Conclusion: At the end, it was shown that Telfairia occidentalis seed has constituents that can improve the status of an impaired liver.

Identification of seeds based on molecular markers and secondary metabolites in Senna obtusifolia and Senna occidentalis

BackgroundSenna obtusifolia and Senna occidentalis (Leguminosae), whose seeds have similar appearance and chemical constituents, are easily confused in using their seeds. To elucidate the similarities and differences between S. obtusifolia seeds and S. occidentalis seeds, three molecular markers and high performance liquid chromatography (HPLC) were employed to evaluate the seeds characteristics of these two medicinal herbs.ResultsThe results showed that selected 3 ISSR and 7 SCoT primers could distinguish S. obtusifolia seeds from S. occidentalis seeds based on the specific band and UPGMA dendrogram. ITS2 sequence indicated that the intra-specific similarity of 20 S. obtusifolia and 16 S. occidentalis was 99.79 and 100.0%, respectively, while the inter-specific similarity between S. obtusifolia and S. occidentalis was 89.58%. Although phylogenetic analysis revealed that these two species had a close relationship, they were assigned to different branches. HPLC fingerprint results showed that seeds of S. obtusifolia and S. occidentalis shared some secondary metabolites, but aurantio-obtusin was not detected in S. occidentalis seeds which could differentiate S. obtusifolia seeds from S. occidentalis seeds.ConclusionsThe present study not only compared the seeds characters of S. obtusifolia and S. occidentalis from molecular and secondary metabolites levels, but also provided a convenient method to identify S. obtusifolia seeds and S. occidentalis seeds effectively.

PHYTOCHEMICAL SCREENING AND ANTIMICROBIAL ACTIVITY OF THUJA OCCIDENTALIS SEEDS EXTRACTS AGAINST THE ISOLATED COMMON SKIN INFECTING MICROORGANISMS

Objective: This study represents the first attempt to investigate the antimicrobial activity of Thuja occidentalis seeds extracts against the isolated skin infecting microorganisms.Methods: T. occidentalis seeds were powdered and extracted continuously by Soxhlet apparatus using 96% ethanol to obtain the total crude ethanolic extract. The extracts with increasing polarity were successively prepared with petroleum ether, chloroform, ethyl acetate, and methanol using the Soxhlet apparatus.The extracts were screened for their phytochemical constituents. Their antimicrobial activity against standards microorganism and the isolated skin infecting microorganisms was evaluated using the agar diffusion method.Results: The result revealed the presence of flavonoids, alkaloids, terpenoids, tannins and cardiac glycosides. The antimicrobial activity result showed that Escherichia coli, Pseudomonas aeroginosa, Staphylococcus aureus, Bacillus subtilis and Candida albicans were found to be sensitive to the extracts of T. occidentalis seeds with inhibition zones range between 20±0.57-12±1.25 mm. The isolated skin infecting microorganisms; Staphylococcus aureus and Staphylococcus epidermidis were found to be highly susceptible to the extracts of T. occidentalis seeds with inhibition zones of 28±1.02-22±0.45 mm and minimum inhibitory concentration (MIC) values of 6.25 mg/ml.Conclusion: From the results it concludes, T. occidentalisseeds extracts had a considerable level of antimicrobial activity. They are more effective towards gram-positive than gram-negative bacteria. Hence, the active principles present in the extracts are particularly powerful effective against the skin infected isolated bacteria. In conclusion, there is a proportional increasing in the antimicrobial activity with increase of the solvents polarity. This may attribute to the fact that the chemical constituents responsible for the antimicrobial activity are of high polarity, such as tannins.

Extraction of bio-oil from non-edible novel source Senna occidentalis seeds

ABSTRACTValuable oil within the seeds of Senna occidentalis is extracted by completely different strategy. However, ancient Soxhlet extraction was compared with microwave-assisted extraction. Soxhlet extraction was known to enhance the yield of bio-oil production. Three polar solvents specifically methanol, acetone, isopropyl alcohol and three non-polar solvents specifically chloroform, hexane, toluene were used. Soxhlet equipment has been established to be the simplest technique with the highest oil yield. Parameters like size fractions of the crushed seeds, extraction time, aging of seeds, and selection of solvents have been optimized in order to enhance oil yield. The bio-oil was characterized using (FT-IR) and GC/MS. Chemical reaction price, iodine variety, relative density, carbon residue content, flash purpose, fire point, hot price, and kinematic consistency were investigated to characterize fuel quality of the bio-oil. The maximum oil yield of 23.46% was obtained with isopropanol at 82.6°C and 3.5 h of optimum extraction time.

Toxicity of Senna occidentalis seeds in laying hens and its effects on egg production

ABSTRACTSenna occidentalis is a toxic leguminous plant found in many tropical and subtropical regions of the world and causes poisoning mainly in confined animals. The seeds are the most toxic part of the plant and may be present in animal rations. The main toxic component of the S. occidentalis seed is a dianthrone, an anthraquinone-derived compound that affects mitochondrial function. This study evaluated the effects on egg production of low-level contamination of the S. occidentalis in the laying hens’ diet. Forty-eight one-day-old pullets were randomly allocated into two treatment groups: control, birds that received no experimental treatment; and external and internal tegument (ET/IT), birds that received a diet containing 0.2% of ET/IT of S. occidentalis seeds throughout their life cycle (42 weeks). The birds were monitored for clinical signs of poisoning, and the production and quality of eggs were recorded. Necropsies were conducted at the end of the experimental period. None of the layers showed any clinical signs of poisoning, decreases in feed intake or alterations of the body weight gain. A marked reduction in egg production and, consequently, a lower feed efficiency in ET/IT group were measured. Ovaries were the most affected organ in birds from the ET/IT group, and yolk leaking and dysplasia of the inner layer of the vitelline membrane were observed. S. occidentalis was shown to be toxic for laying hens. Considering these results, it is feasible to assume that the constant presence of low concentrations of S. occidentalis seeds in rations represents a threat to the poultry industry.

Effect of roasting regime on phytochemical properties of Senna occidentalis seeds

Senna occidentalis seeds were roasted at varying temperatures of 190, 210 and 230 o C each for 10, 15 and 20 min. Phytochemicals of the roasted seeds were determined using standard methods. The phytochemicals analysed were tannins, saponins, flavonoids, alkaloids, glycosides, oxalate and phenolics. Phytochemicals are compounds hypothesized for much of the disease-protection provided by diets high in fruits, vegetables, legumes, cereals and plant-based beverages. This study has clearly shown that roasting time and temperature have significant effects on the seed parameters analyzed. There was an increase in tannin, alkaloid, saponin and phenolic contents and a decrease in the contents of flavonoids and oxalates.

Effect of roasting regime on phytochemical properties of Senna occidentalis seeds

Senna occidentalis seeds were roasted at varying temperatures of 190, 210 and 230 oC each for 10, 15 and 20 min. Phytochemicals of the roasted seeds were determined using standard methods. The phytochemicals analysed were tannins, saponins, flavonoids, alkaloids, glycosides, oxalate and phenolics. Phytochemicals are compounds hypothesized for much of the disease-protection provided by diets high in fruits, vegetables, legumes, cereals and plant-based beverages. This study has clearly shown that roasting time and temperature have significant effects on the seed parameters analyzed. There was an increase in tannin, alkaloid, saponin and phenolic contents and a decrease in the contents of flavonoids and oxalates.

Characterization of the orthodox Pinus occidentalis seed and pollen proteomes by using complementary gel-based and gel-free approaches

This work presents an analysis of Pinus occidentalis pollen and seed proteomes, in which both gel-based and gel-free approaches have been used. Proteins were extracted from P. occidentalis seeds and pollen by using the TCA/acetone/phenol precipitation protocol, and protein extracts were subjected to 1- and 2-DE coupled to MALDI-TOF-TOF as well as to shotgun (nLC-LTQ-Orbitrap) analysis. All bands (1-DE) and the most abundant spots (2-DE) were excised, trypsin digested and the resulting peptides analyzed by MALDI TOF/TOF. In order to increase the proteome coverage, a gel free approach was used. Proteins were identified from mass spectra by using three different databases, including UniProtKB, NCBI and a Pinus spp. custom database [2]. The gel-based approach resulted in 42 (seeds) and 94 (pollen) protein identifications, while the shotgun approach permitted the identification of 187 (seed) and 960 (pollen) proteins. Proteins were classified based on their corresponding functional categories. In seeds, storage proteins were the most abundant ones, and some allergens and proteases were also identified. In pollen proteins related to general metabolism were the most predominant. Data are compared and discussed from a methodological and biological point of view, taking into account the particularities of the seed and pollen organs. Biological significanceIn this work we characterized P. occidentalis proteome with seeds and pollen samples implementing two complementary approaches for the analysis. We found a high content of storage protein, stress response and metabolism related proteins in the seed proteome. Similarly, in the pollen proteome we found predominant groups of proteins related to metabolism and stress response.