Acacia Farnesiana Research Articles

Tolerance, arsenic uptake, and oxidative stress in Acacia farnesiana under arsenate-stress

ABSTRACTAcacia farnesiana is a shrub widely distributed in soils heavily polluted with arsenic in Mexico. However, the mechanisms by which this species tolerates the phytotoxic effects of arsenic are unknown. This study aimed to investigate the tolerance and bioaccumulation of As by A. farnesiana seedlings exposed to high doses of arsenate (AsV) and the role of peroxidases (POX) and glutathione S-transferases (GST) in alleviating As-stress. For that, long-period tests were performed in vitro under different AsV treatments. A. farnesiana showed a remarkable tolerance to AsV, achieving a half-inhibitory concentration (IC50) of about 2.8 mM. Bioaccumulation reached about 940 and 4380 mg As·kg−1 of dry weight in shoots and roots, respectively, exposed for 60 days to 0.58 mM AsV. Seedlings exposed to such conditions registered a growth delay during the first 15 days, when the fastest As uptake rate (117 mg kg−1 day−1) occurred, coinciding with both the highest rate of lipid peroxidation and the strongest up-regulation of enzyme activities. GST activity showed a strong correlation with the As bioaccumulated, suggesting its role in imparting AsV tolerance. This study demonstrated that besides tolerance to AsV, A. farnesiana bioaccumulates considerable amounts of As, suggesting that it may be useful for phytostabilization purposes.

Parasaissetia nigra(Hemiptera: Coccidae) and Its Parasitoids from the GenusCoccophagus(Hymenoptera: Aphelinidae), with Description of a New Species from Tamaulipas, México

Abstract A list of parasitoids of the genus Coccophagus Westwood that parasitize the soft scale Parasaissetia nigra (Nietner), in the world, is given. Data on the biology of P. nigra in Mexico are presented. A key to species of Coccophagus associated to P. nigra in Mexico, including possible species of parasitoids, was prepared. A new species, Coccophagus minor Myartseva sp. nov., reared from P. nigra on mistletoe, Phoradendron quadrangulare (Kunth) Griseb., growing over leaves and shoots of huisache, Acacia farnesiana (L.) Willd., in Tamaulipas, Mexico, is described.

DESARROLLO DE PLÁNTULAS DE HUIZACHE (Acacia farnesiana) EN SUSTRATOS CON VERMICOMPOST

Se estudió el efecto del vermicompost, como parte del sustrato de crecimiento, sobre el desarrollo de plántulas de huizache (Acacia farnesiana) colocadas bajo un sombreador rústico construido a base de carrizo, durante un periodo de 240 días. Se evaluaron cinco mezclas de vermicompostarena con proporciones: 10:90, 20:80, 30:70, 40:60 y 50:50 (% volumen) y como testigo se utilizó suelo natural de la zona donde los huizaches se desarrollan. Las semillas para producir las plántulas se obtuvieron de la zona donde se desarrollan los huizaches. Los tratamientos se repitieron 30 veces y de cada uno se cosecharon cinco plántulas para evaluar la altura, diámetro de tallo, número de ramificaciones, peso fresco y peso seco. El efecto de los tratamientos se determinó mediante un diseño completamente al azar y se aplicó la prueba de comparación de medias DMS(5 %). El desarrollo del huizache en los diferentes sustratos no fue uniforme. Las variables presentaron las respuestas más sobresalientes con la mezcla vermicompost-arena 10:90, por lo que se puede concluir que el vermicompost, en baja concentración, favoreció el desarrollo de A. farnesiana.

Genetic Description of Acacia Species Based on Different Markers

Five Acacia species: Acacia Tortilis ssp. radiana, Acacia farnesiana, Acaciastenophylla, Acacia sclerospermaand Acacia saligna were used in the current research.Morphological, biochemical and molecular markers used to describe the genetic variations amongAcacia species. Data showed highly significant differences among the five species concerning themorphological parameters. Acacia Tortilis ssp. Radiana collected from (Siwa Oasis and Borg Al-Arab city) showed the highest values comparing with other species. The highest values for spinelength (mm) were 28.75 and 19.25 mm, in respect. The lowest mean value 6.50 mm was recordedin Acacia farnesianafor Leaf length (cm) data showed that Acacia sclerosperma and Acacia salignahave the highest mean values (19.25 and 26.08 cm). While the lowest leaf length (2.85 cm) wasrecorded in Acacia Tortilis ssp. Radiana collected from Siwa.One cathode common band (Pex.C1) was found for all the species. While, five anodal(Pex.A1; Pex.A2; Pex.A3, Pex.A4 and Pex.A5) bands were recorded for all species in differentmolecular weights. (Pex.A2, Pex.A3 and Pex.A5) was recorded in Acacia tortilisssp. radiana(Siwa), (Pex.A2,andA4) were recorded in Acacia tortilis ssp. radiana (Borg Al-Arab).In molecular markers measured, out of 156 fragments, 5 fragments were produced for theprimer OPA-18 in the six samples of Acacia species with molecular weights ranging from 251 to 832bp. and polymorphism (40%). While, 18 fragments were observed with OPB-03 primer (11 unique)and 3 polymorphic with polymorphism (93.3%). Ten fragments with molecular weights from 326 to1503 bp were recorded for OPC-02 primer and (70 %) polymorphism. Eleven fragments with widemolecular weight range extended from 299 to 3178 bp recorded to OPD-03. This primer revealed(72.7%) polymorphism. Finally, primer OPE-12 gave nine fragment bands with 77.8 %polymorphism.

Examination of seed characters of Vachellia farnesiana (L.) Wight & Arn. as potentially applicable species in Serbia under climate change conditions

Seeds of sweet acacia (Vachellia farnesiana (L.) Wight & Arn.) were mechanically scarified and treated with concentrated sulfuric acid in order to determine the permeability degree of the seed coat. The obtained results suggest a stronger form of seed coat dormancy that prevents potential invasiveness after introduction. The species is recommended for limited cultivation in outdoor conditions and unlimited use in interior spaces as full size or bonsai trees.

Plants Used in the Treatment of Piles by the Scheduled Caste Community of Andro Village in Imphal East District, Manipur (India)

The present study was carried out at Andro Village in Imphal East district (Manipur) in the remote North-Eastern corner of India. The village is inhabited by a scheduled caste community known as ‘Lois’. The scheduled caste people of Andro Village have a very good knowledge about the treatment of various diseases and ailments with plants. Hemorrhoids’ (Piles) is commonly occurring ailment in Andro village and the villagers treat Piles successfully with the help of different plant species found within their village and the adjoining Baruni (Nongmaiching) Hills. Every elderly people of Andro have common knowledge and easy cure for many common aliments and most of the elderly people uses and prepare different types of medicines from different plant parts. A total of 29 medicinal plant species belonging to 27 genera and distributed over 19 families were found to be used by the schedule caste community of Andro in the treatment of rectum disease-piles. Some of the species used are Acacia farnesiana (Linn.) Willd., Azadirachta indica A.Juss., .Chamaesvce hirta (L.) Millsp, Elsholtzia blanda (Benth.) Benth., Hedychium coronarium Koenig ex Retz., Lantana camara Linn., Leucaena leucocephala (Lamk.) de Wit, Nicotiana plumbaginifolia Viv., Xylosma longifolium Clos etc.

Pharmacodynamic studies on the isolated active fraction of Acacia farnesiana (L.) willd.

Background:Acacia farnesiana is a medicinal plant that grows throughout tropical parts of Indian subcontinent, particularly in sandy soils of river beds in Northern India. The objective of the present study was to evaluate the anti-hyperglycemic activity of the extracts using glucose tolerance test. Isolation of an active fraction (AF) from the active extract (water extract) using alcohol precipitation and to get insight to the mechanism of action of the AF of A. farnesiana.Materials and Methods:Glucose uptake by isolated rat diaphragm of the AF was performed. Further the effect of release of Insulin from isolated and cultured pancreatic β-cell was determined. Besides, effect of oral administration of the AF was compared with that of intraperitonial administration. The effect of AF on serum glucose levels in orally glucose loaded rats was compared with that of intraperitoneal glucose loaded rats.Results:The water extract significantly lowered the blood glucose level. When precipitated with alcohol, the activity was found in the soluble fraction. Glucose uptake in the isolated rat hemidiaphragm, was increased by the AF at 40 μg/ml concentration, the AF did not significantly influence insulin release from cultured islets. The AF was found to be effective in orally glucose loaded in contrast to intraperitonial route.Conclusion:Our findings suggest that this plant is promising for further studies leading to the development of valuable medicine for diabetes.

English

Pollen micromorphological characters such as pollen size, shape, number of associated monads, colpi and ornamentation of the tectum surface, of seven species of Acacia belonging to the family Mimosaceae were studied, using light microscope (LM) and scanning electron microscope (SEM). The study showed that the pollen size range from 42.1 to 69.4 µm, pollen shape is round to semi- round, the number of associated monads is 16 or 32, and the colpus is Y - H shaped. The tectum surface ornamentation among the species is variable, it is foveolate in Acacia ehrenbergiana; psilate-foveolate in Acacia nilotica, Acacia laeta and Acacia negrii; and micro-reticulate in Acacia farnesiana, Acacia oerfota and Acacia tortilis ssp. raddiana. An artificial key based on micromorphological characters is provided. Key words: Mimosaceae, Acacia, pollen morphological characters, Saudi Arabia.

Isolation, via 454 sequencing, and characterization of microsatellites for Vachellia farnesiana (Fabaceae: Mimosoideae).

• Premise of the study: We isolated 15 polymorphic microsatellite markers from Vachellia farnesiana for use in population genetic studies to determine the native range of the species.• Methods and Results: Initially, 454 shotgun sequencing was used to identify and design primers for 68 microsatellite loci. Of these, we trialed 47 loci in the target species, and 42 (89%) amplified a product of expected size. Fifteen of the 47 loci were screened for variation in 21 individuals from the native range of V. farnesiana in southern Mexico and 20 from northwestern Australia. Fourteen loci were polymorphic, with observed heterozygosity ranging from 0.026 to 1.00 (mean = 0.515) and two to 12 alleles per locus (average = 5.2). Cross-amplification was successful in four to 11 loci in three other Vachellia species.• Conclusions: The new microsatellite loci will be useful in understanding genetic variation and investigating the role of human-mediated dispersal in the current distribution of V. farnesiana.

Medicinal and non-medicinal uses of some plants found in the middle region of Saudi Arabia

Kingdom of Saudi Arabia is gifted with a wide range of flora, consisting of different species of trees, shrubs and herbs. It contains hydrocarbon, edible and medicinal plants. Eighty three species (belonging to thirty four plant families) have been investigated for their traditional and medicinal potentiality. The ethnopharmacy of Saudi Arabia represents a good example of a strongly interconnected integration between straight forward familiar medicinal remedies, health, diet and traditional healing practices characterized by cultural specific symbolism. Some of these species have uses other than being medicinal; examples include Adiantum capillus-veneris as refreshing summer drink, Beta vulgaris as source of sugar, Chenopodium murale for making dye, Eleusine indica for paper manufacture,Acacia farnesiana for making perfumes and dyes. Sixty four species are used for treating ailments of different digestive system organs (stomach and intestinal pain, appetizer, emetic). Five species are used as a remedy for diabetes; 18 species are used as diuretic plants; 14 are used for liver and spleen disorders; 17 for rheumatoid and joint inflammation pain; 9 plants for treating cancer and tumors; 24 are used for different skin diseases; 9plants are used as antimicrobial, fungicide and insecticide and 19 are used for sexual diseases. Some of these plants have been used in medicinal remedy, as food and source of non-edible oil. Key words: Medicinal plants, economic uses, Saudi Arabia.

In vitro Ruminal Gas Production Kinetics of Four Fodder Trees Ensiled With or Without Molasses and Urea

This study investigated if the addition of urea (U), molasses (M) or their 1:1 (v/v) mixture during ensiling increases the nutritional value of forage from four fodder trees (Prunus persica, Leucaena esculenta, Acacia farnesiana, and Prunus domestica). Forage samples of fodder trees were collected in triplicate (three individual samples of each species) and subjected to an in vitro gas production (GP) procedure. Fermentation at 24 h (GP24), short-chain volatile fatty acids (SCFA), and microbial crude protein production (MCP), in vitro organic matter digestibility (OMD), metabolizable energy (ME) and dry matter degradability (DMD) were estimated. Forage samples were incubated for 72 h in an incubator at 39°C and the volume of GP was recorded at 2, 4, 6, 8, 10, 12, 24, 48, and 72 h of incubation using the reading pressure technique. The rumen fermentation profiles were highest for P. persica, which showed the highest (P<0.0001) DMD, ME, OMD, SCFA, GP24 and MCP. On the other hand L. esculenta had the lowest (P<0.0001) DMD, SCFA, MCP; P. domestica had the lowest (P<0.0001) OMD. The addition of M to silage increased (P<0.0001) ME and OMD, as well as GP. However, the addition of U and the mixture of U and M reduced (P<0.0001) DMD, ME, OMD, SCFA, GY24 and MCP. These results show that P. persica has the highest nutritive value and L. esculenta the lowest for ruminants. Additionally, the addition of M to forage from fodder trees increases rumen GP and fermentation, which may improve nutrient utilization in ruminants.

Green Synthesis of Silver Nanoparticles Using Acacia farnesiana (Sweet Acacia) Seed Extract Under Microwave Irradiation and Their Biological Assessment

In the present study, Acacia farnesiana (Sweet acacia) seed extract is used to reduce Ag+ → Ag0 under microwave irradiation. The formation of silver nanoparticles (AgNPs) is monitored by recording the UV–Vis absorption spectra for surface plasmon resonance (SPR) peak at ~450 nm. The absorbance of SPR increases linearly with increasing temperature of the reaction mixture. Rapid reduction of silver ions occurred to form AgNPs, 80–90 % yield in about 150 s. A marginal decrease in pH and increase in solution potential (E) of the reaction mixture during the formation of AgNPs are in agreement with the proposed mechanism. XRD pattern of the AgNPs agree with the fcc structure of Ag metal, and the calculated crystallite size is ~17 nm. FT-IR and solid-state 13C NMR spectra indicate the functional groups of flavonones and terpenoids (biomolecules from plant extract) which are adsorbed on AgNPs, thereby the present method led to in situ biofunctionalization/bio-capping of AgNPs. TG analysis shows the thermal decomposition of these plant residues present on AgNPs at about 250 °C. The spherical shape of the particles with a diameter (ϕ) in the range of ~15–20 nm is evident from FE-SEM image. Elemental analysis by EDX analysis confirms the presence of Ag as the only major element. The in vitro antibacterial screening of AgNPs shows that these bio-capped AgNPs have higher inhibitory action for E. coli and S. aureus followed by B. subtilis and P. aeruginosa. In addition, AgNPs show very good antioxidant property.

Contribution à l’amélioration de la germination chez une endémique forestière du Maroc, le cyprès de l’Atlas

La cupressacée à Cupressus atlantica, qui abrite une diversité biologique originale, joue un rôle socio-économique et écologique important. Cependant, ces formations, souvent présteppiques, sont soumises à des pressions croissantes d’origine principalement antropique. En outre, d’autres facteurs tels que la sécheresse, l’absence de régénération ou encore les ravageurs des cônes et des graines, menacent à terme l’avenir du cyprès de l’Atlas. Actuellement, les services forestiers marocains adoptent des orientations qui consistent à utiliser de plus en plus des espèces autochtones, notamment le Cyprès atlasique, dans les programmes de reboisement. Ainsi, il est très important de maîtriser l’itinéraire technique de production des plants de qualité. Cela nécessite une intensification des efforts en matière de recherche pour améliorer les connaissances des différentes étapes de ce processus, depuis la collecte des semences jusqu’à l’acclimatation des plants en passant par le stockage et la germination des graines. La présente étude vise à apporter une contribution concernant le comportement germinatif de ce conifère. En premier lieu, plusieurs traitements de prégermination (traitement chimique, traitement par l’eau à différentes températures, traitement mécanique qui consiste à frotter les graines avec du papier à sable) ont été expérimentés afin d’améliorer la germination par la levée de la dormance des graines. Les résultats relatifs à ces prétraitements ont montré que ces derniers jouent un rôle primordial dans la levée de la dormance et améliorent par conséquence la germination. En effet, les taux de germination les plus élevés 67,77 % et 75,55 % ont été enregistrés respectivement au niveau des graines traitées avec de l’eau chaude (60 oC) et par la scarification mécanique. Le traitement avec de l’acide sulfurique concentré a conduit à des taux de germination très faibles voire nuls, contrairement à ce qui a été observé chez d’autres espèces forestières et à ce qui avait été précédemment reporté chez le Cyprès atlasique. C. atlantica possèdent une dormance physique attribuée au tégument dur, qui peut entraver la régénération naturelle. La scarification mécanique et l’eau chaude (60 oC) sont les traitements les plus efficaces pour améliorer la germination des graines du C. atlantica. Le stress hydrique et le stress salin ont un effet inhibiteur sur la germination des graines du cyprès de l’Atlas. Ces résultats, concernant la germination, premier maillon de l’opération de reboisement, peuvent contribuer à la restauration des espaces dégradés et la réhabilitation des écosystèmes aride et semi-aride.

Extraction of some secondary metabolites &amp;Thin layer chromatography from different parts of Acacia farnesiana L.

Acacia farnesiana L. is an Indian plant used in Ayurvedic treatment for many diseases. A qualitative analysis by thin layer chromatography & a quantitative analysis by standard chemical protocol of secondary metabolites in the pod wall (pericarp) and seeds of the Acacia farnesiana L. has been studied. Using thin layer chromatography (TLC) different components like Alkaloids, Saponin, Flavonoids, and Terpenoides are isolated & identified. The Rf values of the developed spots in the different solvent systems are noted. In the quantitative analysis, alkaloids, saponins, terpenoids & flavonoids are extracted by using the standard chemical protocol. These results may be helpful for rationale use of this plant in the modern system of health care.

Seed bank dynamics of Acacia farnesiana (L.) Willd. and its encroachment potential in sub-humid grasslands of eastern Australia

Shrub encroachment involves abiotic and biotic factors that regulate demographic factors influencing seed production, storage, germination, and subsequent recruitment. In the rangelands of semi-arid and arid Australia, the thorny acacia, Acacia farnesiana (L.) Willd., is apparently encroaching into grasslands and changing the structure, composition, and functioning of native grasslands. The potential for A. farnesiana to expand rapidly in response to changes in land use was examined by quantifying the reproductive output, dispersal, seed-bank density, and germination of seeds of A. farnesiana. Even in the absence of high grazing pressure, low numbers of seeds were produced and these were mainly locally dispersed, although long-distance dispersal via floods could occur. Pre-dispersal seed predation is likely to reduce the seed output, and despite strong physical seed dormancy, soil-stored seed banks were not large. Seed dormancy is broken by scarification but not strongly synergistically with fire or by fire alone, and fire may have a small effect on recruitment. Hence, pulse recruitment seems unlikely given the match between seedling densities and seed-bank density. These patterns suggest that A. farnesiana is unlikely to encroach rapidly into grasslands unless there is widespread landscape disturbance that triggers broad-scale dispersal, such as floods, and or physical scarification of seed.

Qualitative Analysis of Legume Pericarp (Pod Wall) and Seeds of Acacia Farnesiana L.

Present study deals with the qualitative analysis of ethanolic extract of Legume pericarp (pod wall) and seeds of Acacia farnesiana (L). In which we analyze 22 Phytochemical, which are use full for controlling the diseases in Human beings. In India, Acacia farnesiana L. is known as Mulla tumma, Kampu tumma in local area and it is commonly known as Aroma and sweet acacia also. The aim of the present study is to investigate the presence or absence of phytochemicals such as Flavonoids, Alkaloids, Steroids, Proteins, Carbohydrates, Tannin, Amides, Terpenoides, Amines, Phenol, Test for Unsaturation, Carboxylic acid, Test for NH2, Nitrogen, Sulphur, Halogen, Starch, Saponin, Ascorbic acid, Glycosides, Reducing Sugar and Triterpenoids contents of the selected medicinal plants. The ethanolic extract of legume pericarp indicates the presence of major bioactive compound compare to seeds.

Análisis de la restauración pasiva post-pecuaria en el matorral espinoso tamaulipeco del noreste de México

La superficie con actividad ganadera en México es alrededor de 56 % del territorio nacional. El matorral es el ecosistema más distribuido e históricamente más utilizado para fines pecuarios en las zonas áridas y semiáridas de México. Para analizar la restauración pasiva postpecuaria en el matorral espinoso tamaulipeco del noreste de México se establecieron cuatro sitios de muestreo de 40 m x 40 m (1600 m2). En los sitios de muestreo se midieron todos los árboles y arbustos ≥3 cm de diámetro a 10 cm. Se utilizaron los indicadores ecológicos de abundancia, dominancia, frecuencia, índice de valor de importancia, índice de Margalef e índice de Shannon &amp; Wiener. Se registraron dos especies dominantes (Vachellia farnesiana y Prosopis glandulosa) con el 74 % de la dominancia absoluta, mientras el 26% está dividido entre las 10 especies restantes. Se registró una abundancia absoluta de 3050 individuos, donde Vachellia farnesiana representa el 60 % del total. De acuerdo a la distribución diamétrica, se observó que de todos los individuos de Vachellia farnesiana, el 57 % (168 individuos) se encuentran en el rango de 0 cm a 10 cm de diámetro, lo cual indica que se está regenerado activamente.

Desert species adapted for dispersal and germination during floods: Experimental evidence in two Astrophytum species (Cactaceae)

Dispersal of organisms by water is known as hydrochory and is an important means of seed transportation. Here we present experimental evidence of possible dispersal and germination of seeds in water for arid land cacti. Seeds from Astrophytum spp. have unusual traits for plants from arid environments, traits that are typical of water dispersed seeds. Astrophytum capricorne and A. ornatum seeds disperse by floating in the Chihuahuan Desert. Seeds of other native species (Acacia farnesiana, Opuntia streptacantha and Prosopis laevigata (mesquite) were used as control, as well as commercial beans (Phaseolus vulgaris). Seed structures of A. capricorne and A. ornatum are consistent with water dispersal (hat shape, a funicular envelope covering a prominent hilum, and air chambers throughout the tegument). Seeds of A. capricorne showed the greatest water permeability, higher than those of A. ornatum which had similar permeability to common beans and mesquite. Acacia farnesiana and Opuntia streptacantha had the lowest permeability. Seeds of A. capricorne without a funicular layer floated more than those with a funicular layer. All seeds of A. ornatum floated initially, but 10% of those with funicular layer sank after a 30 days floating period. P. laevigata seeds floated also, but less than those of Astrophytum spp., and seeds of A. farnesiana and Phaseolus vulgaris did not float. Germination of A. capricorne was high under all treatments, both when immersed in a water layer or only moistened. Germination of A. ornatum was generally low (<30%), with <5% germination for seeds immersed in water and for those placed on a water film. Seeds of A. capricorne germinated slower in a water layer than in other treatments. No difference in germination rate was found across all treatments in A. ornatum.

Soil macrofauna in relation to soil and leaf litter properties in tree plantations

Soil communities exert strong influences on the processing of organic matter and nutrients. Plantations of trees, especially of nitrogen fixing ones, may affect the soil macrofauna through litter quality and quantity. This study was conducted in a randomized block design with three blocks consisting of Populus euphratica, Eucalyptus camaldulensis, Eucalyptus microtheca, Acacia farnesiana, Acacia salicina, Acacia saligna, Acacia stenophylla and Dalbergia sissoo monoculture plantations that were established in 1992. Soils and soil macrofauna were sampled in November 2006. Leaf litterfall was collected from November 2006 to November 2007 at bi-weekly intervals. Macroinvertebrate abundance and biomass were consistently higher in A. salicina plantations than in the others, whereas they were lowest in E. camaldulensis. Tree species and nitrogen fixing trees significantly influenced the soil macrofauna richness. The results suggest that the earthworm distribution is regulated by leaf litter quality (Ca, C and N) whereas the macrofauna richness is regulated by leaf litter mass, soil organic carbon and leaf litter Mg. Totally, it was revealed that the tree species clearly affected macrofauna whereas nitrogen fixation did not. &amp;nbsp;

A study on the allergenicity and ontogeny of Acacia farnesiana pollen grains in guinea pigs

Pollen grains as the angiosperm male gametophytes transfer male genetic material during sexual reproduction. Pollen grains are among the important plant allergens, such that almost 80–90 % of plant allergens are of pollen origin. Acacia farnesiana is a plant with economical values, and due to its resistance to dry climates, it has had a widespread distribution in Southern Iran. This study was aimed at investigating the allergenicity and the ontogeny of pollen grains of A. farnesiana. Pollen grains were collected from the blossoms and flowers of A. farnesiana in the suburbs of Ahvaz-Iran. Pollen extracts (15 %) were prepared in PBS (pH 7.2). 4–6-week-old male guinea pigs (Hartly) were used for allergenicity tests. Skin tests showed a significant increase in flare diameter. Clinical tests also showed a significant change in the levels of eosinophils, neutrophils, and IgE. Histotechnical analysis was performed on male gametophytes, and photographs were taken using a camera-equipped light microscope. Pollen characteristics were identified using both light and electron microscopy. Sporoderm ultrastructure and pollen morphology were studied using Scanning Electron Microscopy. PAGE analysis of the total protein content of mature pollen grains showed seven clear bands of 10–83 kDa.