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Biomonitoring of organochlorines, glutathione, lipid peroxidation and cholinesterase activity among pesticide sprayers in mango orchards.

BACKGROUND: Pesticide sprayers in mango orchards of Malihabad, Lucknow (India) are generally exposed to organophosphate (OP) and pyrethroid pesticides. We determined the pesticide exposure levels along with their biochemical and clinical effects in 31 sprayers, compared with 18 controls. METHODS: Assay of acetyl and butyrylcholinesterases (AChE, BChE respectively) as an indirect measurement of OP exposure and levels of malondialdehyde (MDA) and glutathione (GSH) were estimated in blood samples to determine their impact on redox potential. Organochlorines were estimated by GLC-ECD. RESULTS: Significantly inhibited AChE, BChE activities and higher MDA level were found among sprayers compared to controls (p<0.05). Mean of total organochlorines were surprisingly higher (97.65+/-13.38 ppb) in sprayers than in those of controls (20.42+/-3.56 ppb) (p<0.05). Respiratory morbidity (32.4%), ocular problems (8.8%), gastrointestinal (17.6%) and skin problems (23.5%) were found in sprayers. There was significant correlation between AChE and GSH (r=0.29, p<0.05) and AChE with MDA (r=-0.34, p<0.05). CONCLUSION: Results indicated the significantly enhanced lipid peroxidation in sprayers correlated with cholinesterases inhibition. A small sample size limits the significance of this study. However, it paves the way for a larger Indian study with extended practical significance.

Adolescent↗

Isolation and characterization of antioxidant and antibacterial compound from mango ginger (Curcuma amada Roxb.) rhizome.

The chloroform extract of mango ginger (Curcuma amada Roxb.) rhizome was subjected to antioxidant activity-guided purification by repeated silica gel column chromatography to obtain a pure antioxidant compound. The structure was deduced by analyzing UV, IR, liquid chromatography-mass spectrometry (LC-MS) and two-dimensional heteronuclear multiple quantum coherence transfer spectroscopy (2D-HMQCT) NMR spectral data, and named it as "Amadannulen", a novel compound. It exhibited DPPH radical scavenging activity, super oxide radical scavenging activity, lipid peroxidation inhibitory activity and metal chelating activity. Amadannulen also showed antibacterial activity against both Gram-positive and Gram-negative bacteria tested. It also exhibited bactericidal activity against M. luteus, B. cereus and B. subtilis.

Anti-Bacterial Agents↗

Season effects on leaf nitrogen partitioning and photosynthetic water use efficiency in mango.

The key parameters of photosynthetic capacity (maximum carboxylation rate (V(cmax)), electron transport capacity (J(max)) and dark respiration rate (R(d))) and the slope (m) of the stomatal conductance model of Ball et al. [Progress in photosynthetic research, Martinus Nijhoff, Dordrecht, 1987] were measured for a whole growing season in fully expanded leaves of 12-year-old mango trees cv. Cogshall in La Réunion island. Leaf nitrogen partitioning into carboxylation (P(c)) and bioenergetic (P(b)) pools were computed according to the model of Niinemets and Tenhunen [Plant Cell Environ 1997;20: 845-66]. V(cmax), J(max), R(d), P(c) and P(b) remained relatively stable over the whole study period, with the exception of the period of linear fruit growth when J(max), R(d) and P(b) were slightly lower, and leaf non-structural carbohydrate content higher. During the pre-floral and floral periods, m decreased by more than 50%, indicating an increase in photosynthetic water use efficiency and m increased again during the period of linear fruit growth. Our results show that, in tropical orchard conditions characterized by mild seasonal climatic changes and non-limiting water supply, leaf nitrogen partitioning is rather stable. Our results also advocate for more studies on the effect of phenology on m and photosynthetic water use efficiency, which is of paramount importance for building coupled biochemical models of photosynthetic carbon assimilation.

Climate↗

Cloning and characterization of differentially expressed genes of internal breakdown in mango fruit (Mangifera indica).

Internal breakdown in mango fruits has become a major concern in recent years. This disorder renders the fruits unfit for human consumption. The overall loss due to this disorder is about 35-55%. Environmental and physiological factors like high temperature, humidity, respiration and low transpiration rates have been attributed to cause spongy tissue due to reduced loss of heat from fruits. Biochemical studies have shown that there is a reduction in pH, total soluble solids, ascorbic acid, total sugars and carotenoids, low reducing and non-reducing sugar contents, lower amylase and invertase activities and high acid and starch content in spongy tissue affected pulp. There are no reports on molecular studies to determine changes in gene expression in these tissues. The present study was conducted using PCR based subtractive hybridization and RNA gel blot analysis of a few selected genes. The latter showed a higher expression of catalase, ubiquitin, alcohol dehydrogenase, coproporphyrinogen oxidase and keratin associated protein. A lower expression of ribosomal gene, fructose bisphosphate aldolase and cysthathionine gamma synthase was also noticed in spongy tissue. Biochemical studies indicated a lower amylase activity and a lower content of the total and reducing sugars in spongy tissue as compared to healthy tissue. Analyses of results indicate that oxidative stress may be one of the causes for formation of spongy tissue, which affects the expression of many genes. The role of these genes in spongy tissue formation is discussed.

Fruit↗

Death and decay in the trees of Mango (Mangifera indica L.).

Death and decay of trees of Mango (Mangifera indica L.) var. Kesar due to fungal infection was studied histologically. Fungal infection in the trees was observed due to various reasons like mechanical injuries in the stem, pruning of the branches, through the inflorescences, attack of Ambrosia beetle and termites. In the initial stage, fungal spores get settled on the flowers due to presence of nectar, followed by their germination and entry of the hypha into peduncle, which gradually spreads into younger branches. The inflorescences were first attacked by Fusarium moniliformis followed by other fungi like Alternaria, Chetomium sp., Aspregillus ellipticus, Aspregillus niger, etc. Fungal mycelia gradually invade the xylem tissues from the top of the branches and spread basipetally ultimately causing death of the infected branches. During monsoon, the crevices on the surface of bark of the healthy plants supported the growth of fungi like Pleurotus, Auricularia, Xyleria, Daldinia sp., and Polyporous sp. The removal of bark from such infected trees revealed minute holes on the surface of the woody cylinder made by Ambrosia beetles. During wet season fungal mycelia makes an easy entry into the xylem through the wounded portion of the stem or pruned branches. Initial entry of the hyphae into xylem was seen through the ray cells. Then the hyphae enter into the lumen of axial elements lining the ray cells through pits and intracellular spaces. The vessel elements located in the xylem (transition zone) between healthy and infected portion were filled with tyloses while axial and ray parenchyma showed heavy accumulation of tannin contents. On the other hand, the infected xylem was also found devoid of reserve metabolites while in normal trees, axial and ray parenchyma showed heavy accumulation of starch grains.

Animals↗

Detection of unusual carotenoid esters in fresh mango (Mangifera indica L. cv. 'Kent').

The carotenoid pattern of mango cv. 'Kent' was investigated by LC-(APcI)MS analyses. In solvent extracts from the mesocarp an unusual carotenoid ester was identified as violaxanthin dibutyrate. For unequivocal identification of butyric acid by an independent method, total lipids were isolated by solvent extraction from the fruit flesh and analyzed by GC after saponification and subsequent methylation. Thus, evidence of butyric acid (1.6 area%) was provided. To the best of our knowledge, this is the first report on a xanthophyll dibutyrate in plants. Additionally, further carotenoid peaks were tentatively assigned to 9-cis-violaxanthin and neochrom or luteoxanthin, respectively, by their UV/vis and MS data of the saponified extracts.

Butyric Acid↗

Structural and rheological properties of polysaccharides from mango (Mangifera indica L.) pulp.

The structure and rheological properties of water-soluble polysaccharides from industrialized mango pulp were investigated. Soluble fraction (SF) 2 was heterogeneous on high performance size exclusion chromatography, giving two peaks as determined by multi-angle laser light scattering and refractive index detectors. The presence of starch in SF2 was demonstrated by a positive iodine reaction and by 13C nuclear magnetic resonance (NMR) spectroscopy. The presence of pectic polysaccharides was shown by a calorimetric method, 13C-NMR spectroscopy and carboxyl reduction. The main pectic polysaccharide was polygalacturonic acid; type I rhamnogalacturonan was also detected. Analysis of the rheological properties of SF2 showed a pseudoplastic behavior up to 3 g x l(-1). 'Creep and recovery' tests and analysis performed under a dynamic state revealed a weak gel character for solutions at concentrations of 15, 20 and 30 g x l(-1).

Iodine↗

Protein kinase activities in ripening mango, Mangifera indica L., fruit tissue. I: Purification and characterization of a calcium-stimulated casein kinase-I.

A Ca(2+)-stimulated protein kinase (PK-I), active with dephosphorylated casein as exogenous substrate, was purified from ripening mango fruit. The purification procedure involved 30-70% ammonium sulphate fractionation and sequential anion exchange-, affinity-, hydrophobic interaction- and gel filtration chromatography. PK-I was purified ca. 40-fold with an overall yield of < 1%. The final specific activity in the presence of 0.1 mM Ca2+ was 55 nmol min-1 mg-1. Analysis of the most highly purified preparations revealed a monomeric enzyme with an M(r) of 30.9 kDa and pI of 5.1. PK-I efficiently phosphorylated casein and phosvitin, but did not phosphorylate histone II-S, histone III-S, protamine sulphate or bovine serum albumin. PK-I activity was stimulated by micromolar concentrations of Ca2+ and was dependent on millimolar Mg2+ concentrations, which could not be substituted with Mn2+. PK-I activity was stimulated by, but was not dependent on Ca2+. Calmodulin and calmodulin inhibitors did not affect PK-I activity, but heparin and cAMP acted as inhibitors. The pH and temperature optima of the enzyme under standard reaction conditions were 6.5 and 35 degrees C, respectively. The kinetic reaction mechanism of PK-I was studied by using casein as substrate. Initial velocity and product inhibition studies with ADP as product inhibitor best fit an ordered bi-bi kinetic mechanism with the Mg(2+)-ATP complex binding first to the enzyme followed by binding of the protein substrate. The K(m)ATP and K(m)casein of PK-I were 9 microM and 0.26 mg ml-1, respectively. The KiADP of PK-I was 9 microM.

Adenosine Triphosphate↗

Protein kinase activities in ripening mango, Mangifera indica L., fruit tissue. III. Purification and characterisation of a calcium-regulated protein kinase.

A calcium-dependent protein kinase (PK-III), not requiring calmodulin for activity, was purified from extracts of ripening mango fruit tissue. Purification was achieved by ammonium sulfate fractionation and sequential anion exchange-, hydrophobic interaction-, dye ligand affinity- and gel filtration chromatography; which allowed a recovery of 1-5% of the total available kinase activity. The final specific activity in the presence of 1 mM Ca2+ was consistently 9 nmol min-1mg-1. The purified enzyme was a monomer with a Mr of 49000, but was resolved by denaturing electrophoresis into two related protein bands of 49 and 45 kDa. Enzyme activity was activated >30-fold by micromolar amounts of free calcium and was dependent upon millimolar Mg2+ or Mn2+ concentrations. Calmodulin (1 microM) had no effect on PK-III activity but the calmodulin antagonists, calmidazolium and chlorpromazine, inhibited PK-III in a dose-dependent manner over a range of 0 to 100 microM. The results suggest a regulatory domain that is similar to calmodulin. PK-III phosphorylated histone III-S and to a lesser extent casein, but did not phosphorylate histone II-S, phosvitin or protamine sulfate. The enzyme phosphorylated substrate proteins on either serine or threonine but not tyrosine. Some endogenous substrates and the ability to autophosphorylate were revealed by autoradiographic studies. PK-III displayed a broad pH optimum (pH 6.6-9.5), and the optimum reaction temperature with histone III-S as substrate was 35 degreesC. The kinetic reaction mechanism of PK-III was studied by using casein as substrate. The KmATP and Kmcasein of PK-III were determined as 10 microM and 1.0 mg ml-1, respectively.

Adenosine Triphosphate↗

Bioindication of air pollution effects near a copper smelter in Brazil using mango trees and soil microbiological properties.

A field study near the copper smelter of a large industrial complex examined air pollution effects on vegetation and soil parameters in Camaçari (northeast Brazil). Close to the smelter, soil pH-value was lower and total acidity as well as organic carbon contents were higher compared with a site far from the source and two reference sites. The acidification of top soil particularly and the drastically enhanced plant-available copper concentrations were caused by atmospheric deposition. High sulphur and copper deposition significantly reduced microbial biomass and altered functional diversity of soil microorganisms (arylsulphatase and xylanase). Large accumulations of sulphur, arsenic and copper were detected in mango leaves (Mangifera indica) growing downwind from the smelter suggesting potential food chain-mediated risk.

Air Pollutants↗

Application of FTIR spectroscopy for the quantification of sugars in mango juice as a function of ripening.

FTIR-ATR spectroscopy and multivariate analysis were used for quantification of sugars in mango juices as a function of ripening. Calibration was based on sucrose/glucose/fructose mixtures, with six concentration levels and following a triangular experimental design. PLS1 regression of the spectra first derivatives gave the best results, enabling quantification of fructose, sucrose, and glucose with 1.4, 1.4, and 4.9% prediction errors, respectively. Throughout ripening, sucrose and fructose were accurately quantified by PLS-FTIR, whereas the accuracy of glucose determination decreased at later stages, when concentrations fell to 0.6-1.5 g/L. These results enabled a correlation with fruit ripening stage to be established. This may be particularly useful to detect over-ripening in fresh fruits, a period when other indicators (pH and % soluble solids (SS)) do not change significantly; this knowledge may help in predicting fruit stability to transport and storage. Similar information obtained for nonfresh juices (in which pH and %SS may be masked by additives), may help determine whether the source fruits had suitable ripening stages.

Beverages↗

Enzymatic degradation of cell wall polysaccharides from mango (Mangifera indica L.) puree.

Ripe mango puree (Smith cultivar) was treated with fungal polysaccharidases containing pectinolytic, hemicellulolytic, and cellulolytic activities for 2 h at 50 degrees C. A loss of 30% of the cell wall material (CWM) was measured. CWM polysaccharides were hydrolyzed to varying degrees: 88, 65, and 65% of, respectively, galacturonic acid-, arabinose-, and rhamnose-containing polymers were hydrolyzed, whereas 50% of cellulose was degraded. After 30 min of treatment, the ethanol precipitation test on the serum was negative, indicating that pectic substances were rapidly hydrolyzed. Oligogalacturonic acids (degree of polymerization, 1-12) were observed in the serum. A viscosity drop of 90% was measured after 2 h, confirming the dominant role of pectic substances in puree viscosity.

Cell Wall↗

Conservation genetics of bush mango from central/west Africa: implications from random amplified polymorphic DNA analysis.

Genetic variation was assessed in the two bush mango species, Irvingia gabonensis and I. wombolu, valuable multipurpose fruit trees from central and west Africa that are currently undergoing domestication. A total of 130 individuals sampled from Cameroon, Nigeria and Gabon were analysed using 74 random amplified polymorphic DNAs (RAPDs). Significant genetic integrity was found in the two morphologically similar species (among-species analysis of molecular variance [AMOVA] variance component 25.8%, P < 0.001), with no evidence of hybridization, even between individuals from areas of sympatry where hybridization was considered probable. Results suggest that large-scale transplantation of either species into new habitats will probably not lead to genetic introgression from or into the other species. Therefore, subsequent cultivation of the two species should not be hindered by this consideration, although further studies on the potential for hybridization/introgression between these species would be prudent. Significant genetic differentiation of both species (among-countries within species, nested AMOVA variance component 9.8%, P < 0.001) was observed over the sampled regions, and genetic similarity of samples decreased significantly with increasing geographical distance, according to number of alleles in common (NAC) analysis. 'Hot spots' of genetic diversity were found clustered in southern Nigeria and southern Cameroon for I. wombolu, and in southern Nigeria, southern Cameroon and central Gabon for I. gabonensis. The possible reasons for this distribution of genetic variation are discussed, but it may reflect evolutionary history, as these populations occur in areas of postulated Pleistocene refugia. The application of these results to domestication programmes and, in the light of extensive deforestation in the region, conservation approaches, is discussed.

Africa, Central↗

Genomes of the ex-type strains of Elsino&#xeb; mangiferae and E. perseae, the causal agents of scab on mango and avocado.

Elsino&#xeb; species are slow-growing, hemibiotrophic to necrotrophic fungi that cause scab diseases on economically important fruit crops. Genome resources for many host-specific species remain limited. We report high-quality draft genome assemblies for the ex-type strains of Elsino&#xeb; mangiferae (CBS 226.50) and E. perseae (CBS 406.34), causal agents of mango and avocado scab, respectively. Among 5 approaches tested, a Nanopore-only NextDenovo assembly produced the most contiguous genomes, yielding 24.5 Mb (E. mangiferae) and 25.1 Mb (E. perseae) assemblies with 13 and 18 contigs, respectively, BUSCO completeness scores of &#x223c;94%, and multiple putative telomere-to-telomere chromosomes. Gene prediction identified 9,134 and 9,243 genes, respectively. Functional annotation revealed enrichment of metabolic and regulatory pathways, including those involved in posttranslational modification, protein transport, and secondary metabolism. Carbohydrate-active enzyme repertoires were small but conserved, consistent with stealth pathogenicity strategies and low plant cell wall degradation. Both genomes encoded large secretomes (>850 proteins), diverse protease repertoires (>300 proteins), Ecp2-like effector proteins, and multiple biosynthetic gene clusters, including clusters with similarity to those associated with elsinochrome and ACT-toxin II biosynthesis, some of which may contribute to host-pathogen interactions and disease development. A large fraction of genes lacked functional characterization, suggesting incomplete databases and/or the presence of lineage-specific genes potentially involved in virulence or host adaptation. These genome resources fill critical gaps for underrepresented Elsino&#xeb; species and provide taxonomically anchored references essential for diagnostics, comparative genomics, and research into the molecular basis of host specificity and pathogenicity in scab-causing fungi.

Persea↗

Seed germination and seedling development in the mango (Mangifera indica L.).

Mango (Mangifera indica L., cv Ruby) seeds taken from ripe fruit showed no dormancy. They germinated at temperatures between 5 and 40 degrees C, but germination was most rapid near the upper end of this range (25-40 degrees C). The fresh seeds had a high moisture content (85%, dry weight basis) and quickly died on dehydration. The optimal temperature for growth of the seedlings was close to 30 degrees C. High temperatures (40 degrees C) and temperatures below 15 degrees C were lethal. Growth of the stem occurred in successive flushes separated by rest periods. When the leaves of the preceding flush finished growing, the axis lengthened beneath the apical bud.

Journal Article↗

Inhibitory effect of flowering and early fruit growth on leaf photosynthesis in mango.

Carbohydrate and nitrogen contents, chlorophyll fluorescence and gas exchange were measured in leaves from both vegetative and reproductive terminal shoots of 12-year-old flowering mango trees. Reproductive shoot leaves were close to swelling floral buds, inflorescences or panicles bearing set fruits. Leaves close to inflorescences had lower rates of mitochondrial respiration (Rd) and net photosynthesis (Anet), and lower stomatal conductance (gs) and quantum efficiency of photosystem II under actinic light than vegetative shoot leaves. Leaf nitrogen concentration, which decreased from the beginning until the end of flowering, was lower in leaves close to inflorescences than in vegetative shoot leaves. However, these differences and changes were counterbalanced by an increase in leaf mass-to-area ratio so that leaf nitrogen per unit leaf area (Na) remained nearly constant during the whole flowering period, except in leaves close to panicles bearing set fruits. Net CO2 assimilation rate simulated by a biochemical model of leaf photosynthesis (Urban et al. 2003) was much higher than Anet measured at an ambient CO2 partial pressure (Ca) of either 36 or 70 Pa. The overestimation of Anet was more pronounced in leaves close to inflorescences, to panicles bearing set fruits and to reversing inflorescences (characterized by the appearance of leaves in terminal positions on inflorescences) than in vegetative shoot leaves. It is concluded that low Anet in leaves close to inflorescences was probably due neither to changes in Na nor to a decrease in Rubisco activity induced by low gs, but rather to a decrease in electron flow in photosystem II. This decrease was not directly associated with higher starch or soluble sugar contents.

Chlorophyll↗