Identification of Prosopis juliflora and Prosopis pallida Accessions Using Molecular Markers

There has been much taxonomic confusion over the identification of Prosopis species, especially where introduced. Prosopis juliflora is the most widespread species in the arid and semi-arid tropics, although it has been confused with other species, particularly the closely related Prosopis pallida. In this study, RAPDs markers were used for the first time to distinguish between these species. Eighteen primers were used in amplification reactions, which yielded an average of 120 bands per accession. A dendrogram showing genetic similarities among accessions was constructed using UPGMA cluster analysis and the Nei and Li similarity coefficient. The genetic similarity observed between P. juliflora and P. pallida is similar to the value in sympatric Prosopis species in North America, and reconsideration of the series rank in section Algarobia is suggested. Species-specific markers confirmed that material in Burkina Faso is P. juliflora, but suggested that material collected in Brazil, Cape Verde and Senegal is P. pallida, whereas this has previously been identified as P. juliflora.     

Isolation and characterization of two antigenic glycoproteins from the pollen of Prosopis juliflora

Two antigenically active glycoprotein fractions were isolated from crude extract of the pollen of Prosopis juliflora using DEAE-cellulose ion exchange chromatography. The glycoproteins gave single band on polyacrylamide gel electrophoresis. The molecular weight of these two glycoprotein was 20,000 and 10,000 as determined by gel filtration on Sephadex G-75. With the help of crossed immunoelectrophoresis and gel diffusion crude extract exhibited twelve and three precipitating antigens suggesting its heterogeneous nature; and the purified glycoprotein fractions however formed single precipitin band on gel diffusion test and immunoelectrophoresis. As tested by ELISA the polyclonal antisera raised in rabbit showed strong binding affinity with glycoprotein of MW 20,000. These result indicates that the two glycoprotein fractions are not antigenically identical.     

Fractionation and immunochemical characterization of Prosopis juliflora pollen allergen

Prosopis juliflora pollen grain crude extract gave six different molecular weight fractions varied from 81,000 to 13,000 dalton on Sephadex G-100 gel filtration. The purity of fractions of Prosopis juliflora pollen extract were checked by polyacrylamide gel electrophoresis. The fraction had an molecular weight 20,000 dalton showed four absorption maxima whereas other fractions had single absorption maxima. Allergenic activity and nature of allergens were evaluated by in vitro Radioallergosorbent test and in vivo Passive Cutaneous Anaphylaxis test. All these tests indicated that most allergenic fractions were in the 20,000 molecular weight.     

Diazotrophic diversity in the rhizosphere of two exotic weed plants, Prosopis juliflora and Parthenium hysterophorus

This study is aimed at assessing culturable diazotrophic bacterial diversity in the rhizosphere of Prosopis juliflora and Parthenium hysterophorus, which grow profusely in nutritionally-poor soils and environmentally-stress conditions so as to identify some novel strains for bioinoculant technology. Diazotrophic isolates from Prosopis and Parthenium rhizosphere were characterized for nitrogenase activity by Acetylene Reduction Assay (ARA) and 16S rRNA gene sequencing. Further, the culture-independent quantitative PCR (qPCR) was performed to compare the abundance of diazotrophs in rhizosphere with bulk soils. The proportion of diazotrophs in total heterotrophs was higher in rhizosphere than bulk soils and 32 putative diazotrophs from rhizosphere of two plants were identified by nifH gene amplification. The ARA activity of the isolates ranged from 40 to 95 nmol ethylene h⁻¹ mg protein⁻¹. The 16S rRNA gene analysis identified the isolates to be members of alpha, beta and gamma Proteobacteria and firmicutes. The qPCR assay also confirmed that abundance of nif gene in rhizosphere of these two plants was 10-fold higher than bulk soil.     

Isolation and characterization of allergens of Prosopis juliflora pollen grains

Proteins and glycoproteins from Prosopis juliflora (Pj) pollen grains were separated by gel filtration, electrophoresis, DEAE cellulose chromatography and their molecular weight was determined by gel filtration and SDS-Polyacrylamide gel electrophoresis. The allergenic activity of different fractions were evaluated by in vivo skin prick test and in vitro gel diffusion test. It was found that fraction E of gel filtration and fraction III and IV of DEAE cellulose chromatography were most allergenic. This fraction E of gel filtration showed positive reaction with periodic acid Schiff's reagent as determined by SDS-gel electrophoresis.     

Ellagic acid 4-O-rutinoside from pods of Prosopis juliflora

From the pods of Prosopis juliflora a new glycoside, ellagic acid 4-O- rutinoside, has been characterized.     

Isolation of allergenically active glycoprotein from Prosopis juliflora pollen

An allergenically active glycoprotein was homogeneously isolated from the aqueous extract of Prosopis juliflora pollen by ConA-Sepharose affinity chromatography. The molecular weight of this glycoprotein was 20,000 dalton, determined by gel filtration and SDS-PAGE. This fraction showed a total carbohydrate concentration of 25%. The purified glycoprotein revealed immunochemically most antigenic or allergenic and demonstrated homogeneous after reaction with P. juliflora pollen antiserum, characterized by gel diffusion, Immunoelectrophoresis and Radioallergosorbent test.     

The human allergens of mesquite (Prosopis juliflora)

BACKGROUND: A computerized statistical analysis of allergy skin test results correlating patient reactivities initiated our interest in the cross-reactive allergens of mesquite tree pollen. In-vitro testing with mesquite-sensitized rabbits and a variety of deciduous tree pollens revealed so many cross-reactivities that it became apparent there could be more allergens in mesquite than previously described in the world literature. Our purpose was to examine the allergens of mesquite tree pollen (Prosopis juliflora) which elicit an IgE response in allergic humans so that future research could determine if these human allergens cross-react with various tree pollens in the same manner as did the mesquite antiserum from sensitized rabbits. METHODS: Proteins from commercial mesquite tree pollen were separated by polyacrylamide gel electrophoresis in the presence of sodium-dodecyl-sulphate. These mesquite proteins were subjected to Western blotting using pooled sera from ten mesquite-sensitive patients and goat anti-human IgE. The allergens were detected using an Amplified Opti-4-CN kit, scanned, and then interpreted by Gel-Pro software. RESULTS: Thirteen human allergens of mesquite pollen were detected in this study. CONCLUSION: The number of allergens in this study of mesquite exceeded the number identified previously in the literature. With the increased exposure to mesquite through its use in "greening the desert", increased travel to desert areas and exposure to mesquite in cooking smoke, the possible clinical significance of these allergens and their suggested cross-reactivity with other tree pollens merit further study.     

Two flavonoid glycosides from the bark of Prosopis juliflora

Two new glycosides, kaempferol 4′-methyl ether 3-O-β-d-galactopyranoside and retusin 7-O-neohesperidoside, have been characterized from the stem bark of Prosopis juliflora.     

Community Impacts of Prosopis juliflora Invasion: Biogeographic and Congeneric Comparisons

We coordinated biogeographical comparisons of the impacts of an exotic invasive tree in its native and non-native ranges with a congeneric comparison in the non-native range. Prosopis juliflora is taxonomically complicated and with P. pallida forms the P. juliflora complex. Thus we sampled P. juliflora in its native Venezuela, and also located two field sites in Peru, the native range of Prosopis pallida. Canopies of Prosopis juliflora, a native of the New World but an invader in many other regions, had facilitative effects on the diversity of other species in its native Venezuela, and P. pallida had both negative and positive effects depending on the year, (overall neutral effects) in its native Peru. However, in India and Hawaii, USA, where P. juliflora is an aggressive invader, canopy effects were consistently and strongly negative on species richness. Prosopis cineraria, a native to India, had much weaker effects on species richness in India than P. juliflora. We carried out multiple congeneric comparisons between P. juliflora and P. cineraria, and found that soil from the rhizosphere of P. juliflora had higher extractable phosphorus, soluble salts and total phenolics than P. cineraria rhizosphere soils. Experimentally applied P. juliflora litter caused far greater mortality of native Indian species than litter from P. cineraria. Prosopis juliflora leaf leachate had neutral to negative effects on root growth of three common crop species of north-west India whereas P. cineraria leaf leachate had positive effects. Prosopis juliflora leaf leachate also had higher concentrations of total phenolics and L-tryptophan than P. cineraria, suggesting a potential allelopathic mechanism for the congeneric differences. Our results also suggest the possibility of regional evolutionary trajectories among competitors and that recent mixing of species from different trajectories has the potential to disrupt evolved interactions among native species.     

Purification and characterization of the glycoprotein allergen from Prosopis juliflora pollen

Highly active glycoprotein allergens have been isolated from pollen of Prosopis juliflora by a combination of Sephadex G-100 gel filtration and Sodium dodecyl sulphate-Poly-acrylamide gel electrophoresis. The glycoprotein fraction was homogeneous, and had molecular weight 20,000. The purified glycoprotein allergen contained 20% carbohydrate, mainly arabinose and galactose. Enzymatic digestion of glycoprotein with protease released glycopeptides of molecular weight ranging from less than 1,000 to more than 5,000 on Sephadex G-25 gel filtration. Antigenicity or allergenicity testing of these glycopeptides by immunodiffusion, immunoelectrophoresis, and radioallergosorbent test indicated complete loss of allergenic activity after digestion with protease whereas incubation with beta-D-galactosidase and periodate oxidation had little affect on the allergenic activity of the glycoprotein fraction. But incubation with alpha-D-glucosidase did not affect the allergenic activity significantly. All these tests indicated that protein played significant role in allergenicity of P. juliflora pollen.     

Prosopis juliflora pollen allergen induced hypersensitivity and anaphylaxis studies in guinea pigs

In vivo and in vitro allergenic activities of Prosopis juliflora pollen allergens were measured in guinea pigs. Intracutaneous skin test showed an early wheal flare response and a late erythema-redness, sensitized with various concentrations (100, 50, 25, 5 and 1.5 micrograms/ml) of Prosopis juliflora pollen extract after administration of a challenging dose. A 50 micrograms/ml sensitizing dose of Prosopis juliflora pollen allergen gave optimum skin response as both early and late effects. The nature of immunochemical reactivity between pollen allergens and reaginic antibodies were further characterized by histamine release test, gel diffusion test, radioallergosorbent test and passive cutaneous anaphylaxis test. These tests confirm allergenicity caused by Prosopis juliflora pollen allergens and showed the binding of allergens with reaginic antibody and its regulation in guinea pigs.     

An ellagic acid glycoside from the pods of Prosopis juliflora

From the fresh pods of Prosopis juliflora, a new glycoside has been isolated and characterized as ellagic acid 4-O-α-L-rhamnosylgentiobioside.     

Bioaccumulation and effect of cadmium in the photosynthetic apparatus of Prosopis juliflora

In the present study Prosopis juliflora plants grown in hydroponics solution were exposed to 50,100 and 1000 μM CdCl₂. The cadmium uptake, transport and toxicity on the photosynthetic activities in the plants were measured at 48 h after starting cadmium treatments. The results showed that the concentration of Cd²⁺ in P. juliflora tended to increase with addition of Cd²⁺ to hydroponics solution. However, the increase of Cd²⁺ in roots and leaves varied largely. In this sense, the accumulation of Cd²⁺ in P. juliflora roots increased significantly in proportion with the addition of this metal. In contrast a relatively low level of Cd²⁺ transportation index, and bioaccumulation factor were found in P. juliflora at 48 h after of treatments. On the other hand the maximum photochemical efficiency of photosystem II (Fv/Fm) and the activity of photosystem II (Fv/Fo) ratios in P. juliflora leaf treated with Cd²⁺ not showed significantly changes during the experiment. These results suggested that the photosynthetic apparatus of P. juliflora was not the primary target of the Cd²⁺ action. Further studies will be focused in understanding the participation of the root system in Prosopis plants with the rhizosphere activation and root adsorption to soil Cd²⁺ under natural conditions.     

Classification of Prosopis juliflora invasion in the Lake Baringo basin and environmental correlations

The spread of Prosopis juliflora in the Baringo basin, Kenya, has led to severe changes in the ecosystem with negative socio‐economic impacts. The drivers that foster the invasiveness of Prosopis are not fully understood. Thus, a method to quantify the degree of infestation will support the determination of environmental preferences and the risk assessment of future Prosopis invasion. We developed a methodology for characterising and classifying degrees of Prosopis infestation in vegetation stands and propose its application in environmental correlation models. The relative cover was identified as the most suited attribute for assessing and monitoring the invasion of Prosopis. The distance of invaded stands from original plantations and environmental attributes related to water availability (ground water table, rainfall and soil water–holding capacity) have potential to predict potential or future invasion risks.     

Diversity of bacteria that nodulate Prosopis juliflora in the eastern area of Morocco

A total of 274 bacterial strains were isolated from the root nodules of Prosopis juliflora, growing in two arid soils of the eastern area of Morocco. A physiological plate screening allowed the selection of 15 strains that could tolerate NaCl concentrations between 175 and 500 mM. These were compared with 15 strains chosen from among the ones which did not tolerate high salinity. The diversity of strains was first assessed by rep-PCR amplification fingerprinting using BOXA1R and ERIC primers. An analysis of the PCR-amplified 16S rDNA gene digestion profiles using five endonucleases indicated the presence of different lineages among the taxa associated with P. juliflora nodules in the soils studied. Nucleotide sequencing of the small subunit rRNA gene and BLAST analysis showed that P. juliflora could host at least six bacterial species in this region and that the identity of those associated with high salt tolerance was clearly distinct from that of the salt-sensitive ones. Among the former, the first type displayed 99% similarity with different members of the genus Sinorhizobium, the second 97% similarity with species within the genus Rhizobium, while the third ribosomal type had 100% homology to Achromobacter xylosoxidans. Within the salt-sensitive isolates the prevailing type observed showed 98% similarity with Rhizobium multihospitium and R. tropici, a second type had 98% similarity to R. giardinii, and a further case displayed 97% colinearity with the Ensifer group including E. maghrebium and E. xericitae. All of the thirty strains encompassing these types re-nodulated P. juliflora in microbiologically controlled conditions and all of them were shown to possess a copy of the nodC gene. This is the first report detecting the betaproteobacterial genus Achromobacter as nodule-forming species for legumes. The observed variability in symbiont species and the abundance of nodulation-proficient strains is in line with the observation that the plant always appears to be nodulated and efficiently fixing nitrogen in spite of a wide range of soil and environmental conditions.     

Diurnal and Seasonal Modulation of Sucrose Phosphate Synthase Activity in Leaves of Prosopis juliflora

Diurnal changes of sucrose-phosphate synthase (SPS) activity in different seasons were measured in Prosopis juliflora (Swartz) DC. leaves. SPS activity showed large variations with two distinct peaks, one around 09:00 and another at 21:00. The diurnal pattern was apparently not due to circadian rhythm since the activities were directly related to the changes in environmental parameters (irradiance, temperature, and leaf to air vapour pressure deficit) in different seasons. During the day, the enzyme showed changes in kinetic properties, differential sensitivity to allosteric modulators, differential response to ATP concentration, to concentration of endogenous sucrose, and to protein phosphatase inhibitors. These results taken together indicate the modulation of SPS in synchrony with photosynthesis and suggest the existence of multiple levels of modulation, presumably as an adaptive response to changing environmental extremes. This revised version was published online in July 2006 with corrections to the Cover Date.     

Characterization of high temperature-tolerant rhizobia isolated from Prosopis juliflora grown in alkaline soil

A method was developed for the fast screening and selection of high-temperature tolerant rhizobial strains from root nodules of Prosopis juliflora growing in alkaline soils. The high-temperature tolerant rhizobia were selected from 2,500 Rhizobium isolates with similar growth patterns on yeast mannitol agar plates after 72 h incubation at 30 and 45 degrees C, followed by a second screening at 47.5 degrees C. Seventeen high-temperature tolerant rhizobial strains having distinguishable protein band patterns were finally selected for further screening by subjecting them to temperature stress up to 60 degrees C in yeast mannitol broth for 6 h. The high-temperature tolerant strains were NBRI12, NBRI329, NBRI330, NBRI332, and NBRI133. Using this procedure, a large number of rhizobia from root nodules of P. juliflora were screened for high-temperature tolerance. The assimilation of several carbon sources, tolerance to high pH and salt stress, and ability to nodulate P. juliflora growing in a glasshouse and nursery of the strains were studied. All five isolates had higher plant dry weight in the range of 29.9 to 88.6% in comparison with uninoculated nursery-grown plants. It was demonstrated that it is possible to screen in nature for superior rhizobia exemplified by the isolation of temperature-tolerant strains, which established effective symbiosis with nursery-grown P. juliflora. These findings indicate a correlation between strain performance under in vitro stress in pure culture and strain behavior under symbiotic conditions. Pure culture evaluation may be a useful tool in search for Rhizobium strains better suited for soil environments where high temperature, pH, and salt stress constitutes a limitation for symbiotic biological nitrogen fixation.     

Mapping Current and Potential Distribution of Non-Native Prosopis juliflora in the Afar Region of Ethiopia

We used correlative models with species occurrence points, Moderate Resolution Imaging Spectroradiometer (MODIS) vegetation indices, and topo-climatic predictors to map the current distribution and potential habitat of invasive Prosopis juliflora in Afar, Ethiopia. Time-series of MODIS Enhanced Vegetation Indices (EVI) and Normalized Difference Vegetation Indices (NDVI) with 250 m² spatial resolution were selected as remote sensing predictors for mapping distributions, while WorldClim bioclimatic products and generated topographic variables from the Shuttle Radar Topography Mission product (SRTM) were used to predict potential infestations. We ran Maxent models using non-correlated variables and the 143 species- occurrence points. Maxent generated probability surfaces were converted into binary maps using the 10-percentile logistic threshold values. Performances of models were evaluated using area under the receiver-operating characteristic (ROC) curve (AUC). Our results indicate that the extent of P. juliflora invasion is approximately 3,605 km² in the Afar region (AUC  = 0.94), while the potential habitat for future infestations is 5,024 km² (AUC  = 0.95). Our analyses demonstrate that time-series of MODIS vegetation indices and species occurrence points can be used with Maxent modeling software to map the current distribution of P. juliflora, while topo-climatic variables are good predictors of potential habitat in Ethiopia. Our results can quantify current and future infestations, and inform management and policy decisions for containing P. juliflora. Our methods can also be replicated for managing invasive species in other East African countries.     

Molecular profiling of rhizosphere bacterial communities associated with Prosopis juliflora and Parthenium hysterophorus

Prosopis juliflora and Parthenium hysterophorus are the two arid, exotic weeds of India that are characterized by distinct, profuse growth even in nutritionally poor soils and environmentally stressed conditions. Owing to the exceptional growth nature of these two plants, they are believed to harbor some novel bacterial communities with wide adaptability in their rhizosphere. Hence, in the present study, the bacterial communities associated with the rhizosphere of Prosopis and Parthenium were characterized by clonal 16S rRNA gene sequence analysis. The culturable microbial counts in the rhizosphere of these two plants were higher than bulk soils, possibly influenced by the root exudates of these two plants. The phylogenetic analysis of V1_V2 domains of the 16S rRNA gene indicated a wider range of bacterial communities present in the rhizosphere of these two plants than in bulk soils and the predominant genera included Acidobacteria, Gammaproteobacteria, and Bacteriodetes in the rhizosphere of Prosopis, and Acidobacteria, Betaproteobacteria, and Nitrospirae in the Parthenium rhizosphere. The diversity of bacterial communities was more pronounced in the Parthenium rhizosphere than in the Prosopis rhizosphere. This culture-independent bacterial analysis offered extensive possibilities of unraveling novel microbes in the rhizospheres of Prosopis and Parthenium with genes for diverse functions, which could be exploited for nutrient transformation and stress tolerance in cultivated crops.