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PubMed2005

Resolution of the structure of the allergenic and antifungal banana fruit thaumatin-like protein at 1.7-A.

The structure of a thaumatin-like protein from banana (Musa acuminata) fruit, an allergen with antifungal properties, was solved at 1.7-A-resolution, by X-ray crystallography. Though the banana protein exhibits a very similar overall fold as thaumatin it markedly differs from the sweet-tasting protein by the presence of a surface exposed electronegative cleft. Due to the presence of this electronegative cleft, the banana thaumatin-like protein (Ban-TLP) acquires a strong (local) electronegative character that eventually explains the observed antifungal activity. Our structural analysis also revealed the presence of conserved residues of exposed epitopic determinants that are presumably responsible for the allergenic properties of banana fruit towards susceptible individuals, and provided evidence that the Ban-TLP shares some structurally highly conserved IgE-binding epitopes with thaumatin-like proteins from fruits or pollen from other plants. In addition, some overlap was detected between the predicted IgE-binding epitopes of the Ban-TLP and IgE-binding epitopes previously identified in the mountain cedar Jun a 3 TLP aeroallergen. The presence of these common epitopes offers a molecular basis for the cross-reactivity between aeroallergens and fruit allergens.

Allergens↗
PubMed2005

Community environmental health assessment strengthens environmental public health services in the Peruvian Amazon.

In December 1999, the Centers for Disease Control and Prevention (CDC) and the Cooperative for Assistance and Relief Everywhere, Peru Country Office (CARE Peru), initiated the Urban Environmental Health Project (SAU, in Spanish) to strengthen environmental public health services in urban and periurban settlements in Peru. The project received funding from the Woodruff Foundation as part of the CARE-CDC Health Initiative (CCHI). The "Protocol for Assessing Community Excellence in Environmental Health" (PACE EH) guided the development of a community environmental public health assessment (CEHA) process in Cardozo, a settlement in Iquitos, Peru. The project developed a three-phase process that merged scientific understanding and community perception about local environmental health problems. In phase 1, local environmental health technicians assisted the community in understanding environmental health conditions in Cardozo and selecting priorities. During phase 2, local technicians assessed the community-selected priorities: water and sanitation. Results from recent water quality assessments revealed that 82% (9 of 11) of samples from shallow dug wells, 18% (2 of 11) from deeper drilled wells, and 61% (11/18) from household drinking containers were positive for thermotolerant coliforms. Phase 3 activities produced an action plan and an intervention to mitigate health problems associated with inadequate water and sanitation services in the Cardozo community. As a result of the CEHA process, CARE Peru obtained funding from the United States Agency for International Development (USAID) to develop and implement an environmental health risk monitoring system and the proposed water and sewage intervention in the settlement. CDC continues to provide technical assistance to the local environmental health services groups in Iquitos through an agreement with CARE Peru as part of the USAID-funded Urban Environmental Health Models Project (MUSA). Technical assistance activities and the development of the environmental health risk monitoring system have helped to strengthen the local environmental public health services delivery system.

Community-Institutional Relations↗
PubMed1998

The abundant 31-kilodalton banana pulp protein is homologous to class-III acidic chitinases.

We have identified and characterized the abundant protein from the pulp of banana fruit (Musa acuminata cv. Grand Nain), and have isolated a cDNA clone encoding this protein. Comparison of the amino terminal sequence of the purified 31 kDa protein (P31) suggests that it is related to plant chitinases. Western analyses utilizing rabbit anti-P31 antiserum demonstrate that this protein is pulp-specific in banana. A full-length cDNA clone homologous to class III acidic chitinase genes has been isolated from a pulp cDNA library by differential screening. The identity of this clone as encoding P31 was verified by comparisons between the amino-terminal peptide sequence and the cDNA sequence and cross-hybridization of the translation product of the cDNA clone with P31 antiserum. Northern and western blot analyses of RNA and protein isolated from banana pulp at different stages of ripening indicate that the cDNA and protein are expressed at high levels in the pulp of unripe fruit, and that their abundance decreases as the fruit ripens. Based on its expression pattern and deduced amino acid sequence and composition, we hypothesize that the physiological role of P31 is not for plant protection, but as a storage protein in banana pulp.

Amino Acid Sequence↗
PubMed1998

Multiple forms of polygalacturonase from banana fruits.

Three multiple forms of polygalacturonase (PG) in ripe and two in unripe banana (Musa acuminata) fruits were separated by DEAE-cellulose and further purified using Sephadex G-150 chromatography. The multiple forms can be differentiated from each other on the basis of their properties. PG1 and PG3 were identified as endo-PG and PG2 as exo-PG on the basis of decrease in viscosity, increase in reducing sugar and the reaction product. PG2 and PG3 increased with the ripening of fruits. PG1, PG2 and PG3 exhibited optimum activity at pH 3.3, 3.7 and 4.3, respectively. Complete loss of PG2 and PG1 activities occurred at 60 and 70 degrees, but PG3 retained 60 and 50% activity respectively. The three forms showed a different response towards divalent metal ions. Ca2+ activated PG1 activity only. Teepol 0.1%, inhibited PG1 activity by 25%, but PG2 and PG3 activities were completely inhibited. CTAB, 0.1%, had no effect on PG1 and PG2 activities, but inhibited PG3 activity by 40%. 2-ME stimulated PG2 and PG3 activities but had no effect on PG1 activity. Gel filtration through Sephacryl indicated M(r) of 23,200, 58,000 and 130,000, respectively, for PG1, PG2 and PG3. The substrate saturation curve for PG1 and PG2 were Michaelian, while PG3 showed biphasic curve. The Km values of PG1 and PG2 were 0.22% and 0.14%, respectively.

Cations↗
PubMed2000

Development of a multiplex immunocapture PCR with colourimetric detection for viruses of banana.

A multiplex, immunocapture PCR (M-IC-PCR) was developed for the simultaneous detection of three viruses from crude sap extracts of banana and plantain (Musa spp.). A reverse transcription step was required for Banana bract mosaic virus and Cucumber mosaic virus, which have ssRNA genomes. The detection of Banana bunchy top virus (ssDNA genome) was not adversely affected by inclusion in this step. All the three viruses could be detected simultaneously from a mixed infection. Identification and detection of individual viruses was achieved through the visualisation of discretely sized PCR amplicons by gel electrophoresis. Alternatively, a colourimetric microplate detection system utilising digoxigenin-labelled virus-specific probes was used. The latter assay was up to five times more sensitive than detection by gel electrophoresis and between 25 and 625 times more sensitive than ELISA for the various viruses. Careful selection of PCR primers was necessary to ensure the detection of a wide range of virus isolates and to avoid detrimental interactions between heterologous templates and primers.

Badnavirus↗
PubMed1999

Detection of episomal banana streak badnavirus by IC-PCR.

A polymerase chain reaction (PCR) based strategy to detect episomal banana streak badnavirus (BSV) in banana and plantain plants that carry integrated BSV sequences was developed. Antisera used in immuno-capture polymerase chain reaction (IC-PCR) are capable of binding a large number of BSV serotypes. The primers used for PCR are capable of annealing to and amplifying across the aspartic protease-reverse transcriptase domain boundaries of both episomal and integrated BSV sequences and result in similar or identical sequence size fragments from either template. However, we show that under the conditions selected for IC-PCR, nuclear, mitochondrial or chloroplast genomic sequences are not amplified and thus only captured episomal BSV is amplified. IC-PCR is suitable for the large-scale screening of Musa for episomal BSV which is necessary for germplasm movement.

Badnavirus↗
PubMed2000

Delayed ripening of banana fruit by salicylic acid.

Salicylic acid treatment has been found to delay the ripening of banana fruits (Musa acuminata). Fruit softening, pulp:peel ratio, reducing sugar content, invertase and respiration rate have been found to decrease in salicylic acid treated fruits as compared with control ones. The activities of major cell wall degrading enzymes, viz. cellulase, polygalacturonase and xylanase were found to be decreased in presence of salicylic acid. The major enzymatic antioxidants namely, catalase and peroxidase, were also found to be decreased in presence of salicylic acid during banana fruit ripening.

Journal Article↗
PubMed2001

Detection of DNA methylation changes in micropropagated banana plants using methylation-sensitive amplification polymorphism (MSAP).

The extent of DNA methylation polymorphisms was evaluated in micropropagated banana (Musa AAA cv. 'Grand Naine') derived from either the vegetative apex of the sucker or the floral apex of the male inflorescence using the methylation-sensitive amplification polymorphism (MSAP) technique. In all, 465 fragments, each representing a recognition site cleaved by either or both of the isoschizomers were amplified using eight combinations of primers. A total of 107 sites (23%) were found to be methylated at cytosine in the genome of micropropagated banana plants. In plants micropropagated from the male inflorescence explant 14 (3%) DNA methylation events were polymorphic, while plants micropropagated from the sucker explant produced 8 (1.7%) polymorphisms. No DNA methylation polymorphisms were detected in conventionally propagated banana plants. These results demonstrated the usefulness of MSAP to detect DNA methylation events in micropropagated banana plants and indicate that DNA methylation polymorphisms are associated with micropropagation.

Journal Article↗
PubMed1998

Study of the anti-hyperglycemic effect of plants used as antidiabetics.

The purpose of this research was to study the anti-hyperglycemic effect of 28 medicinal plants used in the treatment of diabetes mellitus. Each plant was processed in the traditional way and intragastrically administered to temporarily hyperglycemic rabbits. The results showed that eight out of the 28 studied plants significantly decrease the hyperglycemic peak and/or the area under the glucose tolerance curve. These plants were: Guazuma ulmifolia, Tournefortia hirsutissima, Lepechinia caulescens, Rhizophora mangle, Musa sapientum, Trigonella foenum graceum, Turnera diffusa, and Euphorbia prostrata. The results suggest the validity of their clinical use in diabetes mellitus control, after their toxicological investigation.

Animals↗
PubMed1999

Traditional healers and the treatment of sexually transmitted illnesses in rural Zambia.

Lately there has been increasing interest regarding the practice of traditional healers and their use of indigenous plants to treat illnesses. Twenty-three local healers (n'ganga) in Chiawa, rural Zambia, were interviewed about knowledge, practices, and their use of indigenous plants in the diagnosis and treatment of sexually transmitted illnesses (STIs) among male clients. They were also asked about their perceptions of modern medicine. The study revealed that all the n'ganga diagnosed and treated three main types of STIs. They named them as: songeya, doroba and bola-bola. They treated the illnesses with Strychnos cocculoides; Musa species; Solanum delegoense; Ximenia caffra; Diplorynchus condylocarpon; and Croton megalobotrys. Ten of the n'ganga perceived modern medicine to be effective against STIs and five of them sometimes referred some of the clients to the local health centre. It is being argued that a scheme to incorporate the n'ganga into STD control activities in which they can be utilised to refer clients to modern medical facilities can be beneficial. Given the necessary health information and support, the n'ganga may effectively execute this scheme.

Adult↗
PubMed2000

High content of dopamine, a strong antioxidant, in Cavendish banana.

A strong water-soluble antioxidant was identified in the popular commercial banana Musa cavendishii. It is dopamine, one of the catecholamines. For suppressing the oxygen uptake of linoleic acid in an emulsion and scavenging a diphenylpicrylhydrazyl radical, dopamine had greater antioxidative potency than glutathione, food additives such as butylated hydroxyanisole and hydroxytoluene, flavone luteolin, flavonol quercetin, and catechin, and similar potency to the strongest antioxidants gallocatechin gallate and ascorbic acid. Banana contained dopamine at high levels in both the peel and pulp. Dopamine levels ranged from 80-560 mg per 100 g in peel and 2.5-10 mg in pulp, even in ripened bananas ready to eat. Banana is thus one of the antioxidative foods.

Antioxidants↗
PubMed1998

Identification of Pythium carolinianum causing 'root rot' of cotton in Egypt and its possible biological control by Pseudomonas fluorescens.

A severe root rot disease of cotton caused by Pythium carolinianum was diagnosed in a cotton field in Beni-Musa village, 20 km southwest of El-Minia city, Egypt, during the summer of 1996. This was the first reported isolation of this fungus in Egypt. In the light of the importance of the cotton industry in Egypt, research was initiated to develop a biocontrol agent against Pythium carolinianum. In vitro agar plate technique identified a Pseudomonas fluorescens strain that was highly antagonist to Pythium carolinianum. Subsequent plant growth experiments establish that substantial disease control could be obtained by applying Pseudomonas fluorescens to the soil. Optimal control was obtained by mixing the bacteria with the soil rather than by dipping the cotton roots in the bacterial suspension immediately before planting. Disease was more severe in autoclaved soil than in nonsterile soil.

Journal Article↗
PubMed1998

Evaluation of ash from some tropical plants of Nigeria for the control of Sclerotium rolfsii Sacc. on wheat (Triticum aestivum L.).

Eleven ash samples, from organs of nine tropical plants, were screened for their abilities to inhibit mycelial growth and sclerotial germination of a Nigerian isolate of Sclerotium rolfsii on agar and in the soil. Ten ash samples showed some activity against mycelial growth of S. rolfsii in vitro. Ash samples from Delonix regia stem wood, Mangifera indica leaf and Vernonia amygdalina leaf were most effective as each totally inhibited mycelial growth of S. rolfsii in vitro. Ocimum gratissimum leaf ash, D. regia wood ash and Musa paradisiaca flower bract ash inhibited sclerotial germination on agar. Nine ash samples protected seeds against pre-emergence rot. Ash from M. indica leaf, V. amygdalina leaf and Azadirachta indica leaf protected seedlings against post-emergence infection. Eichornia crassipes ash, which was ineffective in vitro, offered some protection to seeds in soil against pre-emergence rot. The study demonstrates potentials of ash samples from tropical plants in control of S. rolfsii on wheat.

Journal Article↗
PubMed1997

Soluble and insoluble fiber contents of some Cameroonian foodstuffs.

As a result of the lack of reliable data on the fiber content of African foodstuffs, a study to determine the dietary fiber contents (soluble, insoluble and total) on a dry weight basis of a selected variety of major Cameroonian foods was conducted. The influence of processing and preparation methods on the fiber content was also assessed. Vegetables were found to be the richest source of total dietary fiber (57%), followed by legumes and seeds (30%) and fruits (16.5%). Okro (Hibiscus esculenta), plantain (Musa paradisiaca) and beans (Phaseolus spp) showed varietal differences in their soluble and insoluble fiber content, while methods of processing and preparation significantly influenced the fiber content of cassava (Manihot esculenta), corn (Zea mays) and beans.

Cameroon↗
PubMed2003

Isolation and characterization of a lectin from Annona muricata seeds.

A lectin with a high affinity for glucose/mannose was isolated from Annona muricata seeds (Annonaceae) by gel filtration chromatography on Sephacryl S-200, ion exchange chromatography on a DEAE SP-5 PW column, and molecular exclusion on a Protein Pak Glass 300 SW column. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (PAGE) yielded two protein bands of approximately 14 kDa and 22 kDa. However, only one band was seen in native PAGE. The Mr of the lectin estimated by fast-performance liquid chromatography-gel filtration on Superdex 75 was 22 kDa. The lectin was a glycoprotein with 8% carbohydrate (neutral sugar) and required divalent metal cations (Ca2+, Mg2+, and Mn2+) for full activity. Amino acid analysis revealed a large content of Glx, Gly, Phe, and Lys. The lectin agglutinated dog, chicken, horse, goose, and human erythrocytes and inhibited the growth of the fungi Fusarium oxysporum, Fusarium solani, and Colletotrichum musae.

Amino Acid Sequence↗
PubMed2004

Light- and singlet oxygen-mediated antifungal activity of phenylphenalenone phytoalexins.

The light-induced singlet oxygen production and antifungal activity of phenylphenalenone phytoalexins isolated from infected banana plants (Musa acuminata) are reported. Upon absorption of light energy all studied phenylphenalenones sensitise the production of singlet oxygen in polar and non-polar media. Antifungal activity of these compounds towards Fusarium oxysporum is enhanced in the presence of light. These results, together with the correlation of IC50 values under illumination with the quantum yield of singlet oxygen production and the enhancing effect of D2O on the antifungal activity, suggest the intermediacy of singlet oxygen produced by electronic excitation of the phenylphenalenone phytoalexins.

Antifungal Agents↗
PubMed1995

Purification and characterization of a novel phosphoenolpyruvate carboxylase from banana fruit.

Phosphoenolpyruvate carboxylase (PEPC) from ripened banana (Musa cavendishii L.) fruits has been purified 127-fold to apparent homogeneity and a final specific activity of 32 mumol of oxaloacetate produced/min per mg of protein. Non-denaturing PAGE of the final preparation resolved a single protein-staining band that co-migrated with PEPC activity. Polypeptides of 103 (alpha-subunit) and 100 (beta-subunit) kDa, which stain for protein with equal intensity and cross-react strongly with anti-(maize leaf PEPC) immune serum, were observed following SDS/PAGE of the final preparation. CNBr cleavage patterns of the two subunits were similar, but not identical, suggesting that these polypeptides are related, but distinct, proteins. The enzyme's native molecular mass was estimated to be about 425 kDa. These data indicate that in contrast to the homotetrameric PEPC from most other sources, the banana fruit enzyme exists as an alpha 2 beta 2 heterotetramer. Monospecific rabbit anti-(banana PEPC) immune serum effectively immunoprecipitated the activity of the purified enzyme. Immunoblotting studies established that the 100 kDa subunit did not arise via proteolysis of the 103 kDa subunit after tissue extraction, and that the subunit composition of banana PEPC remains uniform throughout the ripening process. PEPC displayed a typical pH activity profile with an alkaline optimum and activity rapidly decreasing below pH 7.0. Enzymic activity was absolutely dependent on the presence of a bivalent metal cation, with Mg2+ or Mn2+ fulfilling this requirement. The response of the PEPC activity to PEP concentration and to various effectors was greatly influenced by pH and glycerol addition to the assay. The enzyme was activated by hexose-monophosphates and potently inhibited by malate, succinate, aspartate and glutamate at pH 7.0, whereas the effect of these metabolites was considerably diminished or completely abolished at pH 8.0. The significance of metabolite regulation of PEPC is discussed in relation to possible functions of this enzyme in banana fruit metabolism.

Carbohydrate Metabolism↗
PubMed1994

Timing of fungal invasion using host's ripening hormone as a signal.

In many postharvest fruit diseases, fungi remain latent until the fruit ripens. How the fungus times its infection at ripening of the host is not known. We have found that the volatiles produced by the climacteric tomato, avocado, and banana fruits induce germination and appressorium formation in Colletotrichum gloeosporioides and Colletotrichum musae. Exposure of the spores of these fungi to ethylene, the host's ripening hormone, at </=1 microl/liter, caused germination, branching of the germ tube, and formation of up to six appressoria from a single spore. Propylene, an ethylene analog, but not the hydrocarbon gas methane was able to induce spore germination and multiple appressorium formation. The ethylene effect on the fungi appears to be a plant-like response as it was inhibited by silver ion and 2,5-norbornadiene; the inhibition by the latter could be reversed by higher ethylene concentrations. Ethylene induced germination and appressorium formation in the Colletotrichum sp. penetrating climacteric fruit but not in other Colletotrichum strains. That the ethylene induction of multiple appressorium formation could be relevant to postharvest infection was indicated by the observation that C. gloeosporioides spores formed multiple appressoria on normally ripening tomato that produces ethylene, whereas on transgenic tomato and orange, fruits incapable of producing ethylene, exogenous ethylene was required to induce multiple appressorium formation and lesion formation. These results strongly suggest that these fungi must have coevolved to develop a mechanism to use the host's ripening hormone as a signal to differentiate into multiple infection structure and thus time the infection process.

Journal Article↗