Ascorbic acid metabolism in ageing recalcitrant sal (Shorea robusta Gaertn. f.) seeds

Changes in ascorbate content and its enzymatic utilization pattern were studied in embryonic axes and cotyledons of sal seeds undergoing rapid loss of viability, at ambient conditions. Ascorbate levels were significantly higher initially in the embryonic axes (0.32 mg/g fresh weight) and cotyledons (0.21 mg/g fresh weight) of freshly mature, relatively hydrated (42.2% moisture content) and 100% viable sal seeds. It declined sharply as the tissues; embryonic axes and cotyledons, desiccated with absolutely no detectable amount in non-viable seeds (21% moisture content). Significantly strong correlation was obtained between desiccation of embryonic axes (r = 0.96) and cotyledon (r = 0.97) with loss of ascorbate levels and loss of germinability. Higher rates of ascorbic acid utilization (AAU) recorded in the embryonic axes of 100% viable seed declined sharply as the seed viability reduced due to desiccation below 36.8% moisture content. AAU was not detected in the cotyledons.     

Protein modification by Amadori and Maillard reactions during seed storage: roles of sugar hydrolysis and lipid peroxidation

The non-enzymatic modifications of proteins through Amadori and Maillard reactions play an important role in the loss of seed viability during storage. In the present study, the contribution of sugar hydrolysis and lipid peroxidation to Amadori and Maillard reactions, and to seed deterioration was investigated in mung-bean (Vigna radiata Wilczek). The contents of glucose and lipid peroxidation products in seed axes increased significantly during storage. The accumulation of Amadori products in seed axes was correlated to the lipid peroxidation, whereas the accumulation of Maillard products was closely correlated to sugar hydrolysis. The rate of accumulation of Maillard products was not well correlated to the content of Amadori products in both seed axes and protein/glucose model system, reflecting the complex nature of Amadori and Maillard reactions. The content of Amadori products in seed axes increased during the early stages of seed ageing, whereas the content of Maillard products increased steadily during the entire period of storage. The accumulation of Maillard products in seed axes was associated with the decline of seed vigour. These data suggest that, during seed ageing, sugar hydrolysis and lipid peroxidation are coupled with non-enzymatic protein modification through Amadori and Maillard reactions.     

Ageing-induced changes in glutathione system of sunflower seeds

The glutathione system is thought to be involved in defence mechanisms present in plant tissues. The efficacy of this system was evaluated in large seeds of sunflower ( Helianthus annuus L. cv. Peredovik) in response to accelerated ageing (43°C/75% relative humidity from 1 to 11 days). Differences between the embryo axis and cotyledons in relation to the glutathione system were also investigated. Additionally, lipid peroxidation was determined by measuring the malondialdehyde (MDA) content. All assays were performed using dry seeds and seeds subsequently hydrated by imbibition in distilled water for 12 h at 25°C. Accelerated ageing caused a marked decrease in seed viability, accompanied by an increase in mean germination time. There were no changes in total glutathione in dry seeds. However, the distribution in its reduced (GSH) and oxidized (GSSG) forms revealed that ageing produced a slow conversion from GSH to GSSG. As the ageing period increased, this effect was accompanied by a decrease in glutathione reductase (GR, EC 1.6.4.2) activity. The results also indicated that the GSH system exerts a different response in the embryo axis as compared with the cotyledon: (1) the GSH levels decreased less in the cotyledons than in axes of aged seeds, and (2) the GSSG level in cotyledons was independent of ageing, while its amount increased in aged embryo axes. These different responses, in conjunction with the lower MDA levels in large as compared with small seeds, indicate a possible protective role of the reserve lipids. The efficacy of the GSH system in aged seeds was associated with seed viability, as revealed by multiple regression analysis. Upon imbibition, aged seeds were able to restore their GSH levels, reaching values approximating those of unaged seeds.     

The influence of plants on atmospheric methane in an agriculture-dominated landscape

The primary objective of this study was to clarify the influence of crop plants on atmospheric methane (CH₄) in an agriculture-dominated landscape in the Upper Midwest of the United States. Measurements were carried out at two contrasting scales. At the plant scale, CH₄ fluxes from soybean and corn plants were measured with a laser-based plant chamber system. At the landscape scale, the land surface flux was estimated with a modified Bowen ratio technique using measurements made on a tall tower. The chamber data revealed a diurnal pattern for the plant CH₄ flux: it was positive (an emission rate of 0.4?±?0.1 nmol m^?2 s^?1, average of soybean and corn, in reference to the unit ground area) during the day, and negative (an uptake rate of ?0.8?±?0.8 nmol m^?2 s^?1) during the night. At the landscape scale, the flux was estimated to be 14.8 nmol m^?2 s^?1 at night and highly uncertain during the day, but the available references and the flux estimates from the equilibrium methods suggested that the CH₄ flux during the entire observation period was similar to the estimated nighttime flux. Thus, soybean and corn plants have a negligible role in the landscape-scale CH₄ budget.     

Trace elements influenced by environmental changes in Lake Biwa: (II) Chemical variations in the hypolimnion over the last half-century

The history of the deep north basin of Lake Biwa extends over 430,000 years. Although it has probably been oxic and oligotrophic since its formation, human impacts have been changing lake conditions. In this paper, we discuss long-term changes in the chemistry of bottom water by compiling literature and through our own data over the last half-century. Long-term records show an increase in temperature, decrease in dissolved oxygen (DO), and increase in nutrients in bottom water. The stoichiometry among oxygen and nutrients indicates that changes are basically consistent with aerobic decomposition of organic matter. These changes are most likely the result of global warming and local eutrophication. Of particular note, yearly minimum DO concentrations <50 µmol kg^?1 have started to occur frequently at ~90 m depth since 1999. Manganese (Mn) concentrations in bottom water are at their minimum during the turnover period and at a maximum during the late stratification period each year. Yearly minimum Mn concentration has been within a narrow range over the last 30 years (0.25 ± 0.07 µmol kg^?1, n = 12). However, abnormally high Mn concentrations (up to 9.3 µmol kg^?1) were observed in 2007, caused by reductive release of a substantial amount of Mn from suboxic sediments and subsequent oxidation in bottom water. The concentration of arsenic (As) has gradually increased over the last 20 years in a similar manner, with a homologous element of phosphorus (P), resulting in an observed range of 17–29 nmol kg^?1 in 2010. The accumulation rate was ~0.8 nmol kg^?1 year^?1 for As and ~6 nmol kg^?1 year^?1 for P.     

Relevance of amadori and maillard products to seed deterioration

The possible role of Amadori and Maillard reactions in the deterioration of dry seeds was investigated using model systems and whole soybean seeds, Glycine max cv Hodgson. In model systems of glucose plus an enzyme (lysozyme), the production of Amadori products was accelerated by higher temperature and relative humidity. The reaction between glucose and lysozyme at 50°C, 75% relative humidity, leads to a progressive decline in enzymatic activity. During accelerated aging of soybean seeds (40°C, 100% relative humidity), a sequence is observed in which the Amadori products increase with time and then decline under conditions in which the Maillard products increase in the axes. Loss of germinability occurs at the time when the Maillard products increase in the soybean axes. These results are suggestive of a role for nonenzymic glycation in soybean seed deterioration during accelerated aging.     

Effects of ageing on peroxidase activity and localization in radish (Raphanus sativus L.) seeds

Peroxidase activity was assayed in crude extracts of integument, cotyledons and embryo axis of radish seeds, deteriorated under accelerated ageing conditions. Over five days of ageing, in which germination decreased from 100 to 52%, the enzyme activity in integument was higher than that in other seed parts, increasing in the first days of ageing and then decreasing sharply in extremely aged seeds. Polyacrylamide gel electrophoresis analysis showed four peroxidase isoenzymes with MM of 98, 52.5, 32.8 and 29.5 kDa in the embryo axis of unaged seeds, and only the 32.8 and 29.5 kDa MM isoforms in the integument and cotyledons. In these parts of the seed, only the 29.5 kDa MM isoenzyme increased in activity in early days of ageing and decreased there-after. In the embryo axis, the 29.5 kDa MM isoenzyme activity increased slowly in the first day of ageing, while the 98 and 52.5 kDa MM isoenzyme activities disappeared. A cytochemical localization of peroxidase activity in the various tissues showed that main differences between unaged and extremely aged seeds occurred in the embryo axis.     

Carbon Dioxide and Methane Fluxes From Tree Stems,Coarse Woody Debris,and Soils in an Upland Temperate Forest

Forest soils and canopies are major components of ecosystem CO₂ and CH₄ fluxes. In contrast, less is known about coarse woody debris and living tree stems, both of which function as active surfaces for CO₂ and CH₄ fluxes. We measured CO₂ and CH₄ fluxes from soils, coarse woody debris, and tree stems over the growing season in an upland temperate forest. Soils were CO₂ sources (4.58 ± 2.46 µmol m^?2 s^?1, mean ± 1 SD) and net sinks of CH₄ (?2.17 ± 1.60 nmol m^?2 s^?1). Coarse woody debris was a CO₂ source (4.23 ± 3.42 µmol m^?2 s^?1) and net CH₄ sink, but with large uncertainty (?0.27 ± 1.04 nmol m^?2 s^?1) and with substantial differences depending on wood decay status. Stems were CO₂ sources (1.93 ± 1.63 µmol m^?2 s^?1), but also net CH₄ sources (up to 0.98 nmol m^?2 s^?1), with a mean of 0.11 ± 0.21 nmol m^?2 s^?1 and significant differences depending on tree species. Stems of N. sylvatica, F. grandifolia, and L. tulipifera consistently emitted CH₄, whereas stems of A. rubrum, B. lenta, and Q. spp. were intermittent sources. Coarse woody debris and stems accounted for 35% of total measured CO₂ fluxes, whereas CH₄ emissions from living stems offset net soil and CWD CH₄ uptake by 3.5%. Our results demonstrate the importance of CH₄ emissions from living stems in upland forests and the need to consider multiple forest components to understand and interpret ecosystem CO₂ and CH₄ dynamics.     

Transport of magnesium ions in the phloem of Ricinus communis L. seedlings

During growth of Ricinus communis seedlings, magnesium ions are mobilized in the endosperm, taken up by and accumulated to very high levels (150 μmol·g FW^?1) in the cotyledons, and translocated to hypocotyl and roots. The magnesium gain from days 6 to 7 in the cotyledons and the seedling axis necessitates a total up-take rate of 600 nmol·h^?1-seedling^?1 and the phloem translocation rate must amount to 200 nmol·h^?1. seedling^?1. The phloem loading of magnesium and the regulatory properties of this process were investigated, making specific use of the ability to collect pure phloem sap from the cut hypocotyl of 6-d-old Ricinus seedlings. The concentration of magnesium in sieve-tube sap (5 mM) was fairly constant under many incubation conditions, e.g. incubation in magnesium-free buffer, incubation with different cations (K⁺, Na⁺, NH ₄ ⁺ ) or anions (Cl^?, NO ₄ ^- , SO ₄ ^2- ), or incubation with sucrose and amino acids. Even addition of magnesium chloride to the cotyledons did not enhance phloem loading of magnesium ions. Therefore the high magnesium content of the cotyledons was sufficient for continuous phloem loading of magnesium, irrespective of external ionic conditions. Also, the flow rate of sieve-tube sap did not influence the magnesium concentration in the sap. Only the incubation with sulfate and phosphate ions increased the magnesium-ion concentration in the phloem. Magnesium sulfate offered to the cotyledons caused a threefold increase of magnesium ions in the sieve-tube sap, which was inhibited by Na⁺, NH ₄ ⁺ and Ca²⁺ in rising order, but not by K⁺. Incubation with phosphate for a prolonged period (8 h) led to an increased mobilization of intra-cotyle-donary magnesium and an enhanced phloem loading of mobilized magnesium. It is concluded that phosphate availability is a decisive factor for mobilization and translocation of magnesium ions within the plant.     

Endogenous ABA content in relation to maturation of somatic embryos in <Emphasis Type="Italic">Tulip</Emphasis>a (L.) ‘Apeldoorn’ cultures

The aim of the present study was to estimate the endogenous abscisic acid (ABA) content in tulip ‘Apeldoorn’ torpedo and mature somatic embryos. Moreover, the effect of exogenous ABA and/or its inhibitor fluridone on somatic embryo maturation and conversion into plantlets was investigated. Torpedo-stage somatic embryos were subcultured on media containing 5 μM of picloram and 1 μM of 6-benzyl-aminopurine (BAP)—control, and combinations of ABA (0 or 10 μM) and/or fluridone (0 or 30 μM) for 1 week. Then, the torpedo embryos were transferred to a maturation medium containing 0.25 μM of α-naphthaleneacetic acid (NAA) and 2.5 μM of BAP, without ABA and fluridone treatment, and cultivated under darkness or light for ten weeks. Endogenous ABA content (first time measured in tulip somatic embryos) was evaluated by ELISA test. The obtained results revealed that the highest level of endogenous ABA, at 17.45 nmol g^?1 dry weight (DW), was recorded in torpedo-stage of tulip embryo development, only after 1 week of ABA treatment, and was nearly 10 times higher in comparison with the control. Simultaneous addition of ABA and fluridone to the medium resulted in the lowering of the ABA concentration to 9.58 nmol g^?1 DW. During ten weeks of maturation of the embryos, the endogenous ABA content in mature tissue of tulip somatic embryo considerably decreased to an amount 0.87–1.33 nmol g^?1 DW (irrespective of ABA and fluridone treatment) and did not differ significantly from control (0.59 nmol g^?1 DW). Exogenous ABA and fluridone significantly decreased the growth value of fresh weight (FW) of the tulip torpedo-shaped and mature embryos under light conditions. Percentage of the DW of the torpedo embryos treated with exogenous ABA was significantly higher (15.43–17.02) in comparison with the control (10.87). Three to three and a half times more malformed mature embryos were noted under light conditions than in darkness, irrespective of ABA and fluridone treatment. The highest percentage of mature embryos forming shoots (conversion) was observed under light conditions in the control and after fluridone treatment (26 and 20%, respectively).     

Isocitrate lyase in germinating seeds of Prunus dulcis

Isocitrate lyase activity increased in cotyledons of germinating seeds of Prunus dulcis (almond) which had been induced to germinate by either stratification or treatment with gibberellic acid (GA). Germination of embryos, growth of the embryonic axis and activity of isocitrate lyase increased with increasing concentrations of GA from 10^?7 to 10^?3 M.     

Optimization of protease production by Streptomyces sp. A6 using statistical approach for reclamation of shellfish waste

Protease producing Streptomyces sp. A6 was isolated from intertidal zone of the coast of Diu (Gujarat, India). Plackett–Burman method was applied to identify important factors (shrimp waste, FeCl₃, ZnSO₄ and pH) influencing protease production by Streptomyces sp. A6. Further optimization was done by response surface methodology using central composite design. The concentrations of medium components for higher protease production as optimized using the above approach were (g l^?1): Shrimp waste, 14; FeCl₃, 0.035; ZnSO₄, 0.065 and pH, 8.0. This statistical optimization approach led to production of 129.02 ± 2.03 U ml^?1 of protease which was 4.96 fold higher compared to that obtained using the unoptimized medium. The protease production was scaled to 3 l in a 5-l bench fermenter using optimized medium which further increased the production by 63.4%. Deproteinization and chitin recovery obtained at the end of fermentation was 85.12 ± 4.7 and 70.58 ± 1.33%, respectively. The present study is the first report on statistical optimization of medium components for production of protease by Streptomyces species using cheaper raw material such as shrimp waste. The study also explored the possibility Streptomyces sp. A6 for reclamation of shrimp wastes.     

β-Estradiol Protects Embryo Growth from Heavy-Metal Toxicity in Germinating Lentil Seeds

Mammalian sex hormones spread in the environment from both natural and anthropogenic origin. In the present study, we found that treatment with β-estradiol (E) could improve embryo growth and alleviate unsuitable availability of nutrients imposed by cadmium and copper toxicity during germination of lentil (Lens culinaris Medik.) seeds. The length of embryonic axes decreased in the presence of 100 μM CdCl₂ or 200 μM CuCl₂. Addition of 10^?6 M E in the germination media could greatly reverse the inhibitory effect of heavy-metal (HM) stress on post-germination events. The cotyledons of E-treated seeds also tended to (1) retain higher protease and amylase activities, (2) breakdown more storage compounds (albumin, globulin, and starch), and (3) have higher contents of free amino acids and glucose than controls without added E (Cd or Cu applied individually). Further investigations showed that exposure to HM dramatically provoked the solute leakage in imbibition medium, whereas the combination of HM with E significantly reduced the loss of nutrients. Moreover, the seed Cd and Cu contents were not significantly different between the cotreatment of Cd or Cu with E and no treatment, meaning that E was not responsible for preventing HM accumulation in seed tissues.     

Meristematic nodule culture: a new pathway for in vitro propagation of<Emphasis Type="Italic"> Eucalyptus globulus</Emphasis>

Morphogenesis was induced in Eucalyptus globulus seeds, cotyledons, hypocotyls and leaves from in vitro clonal plantlets. Globular structures were observed after 2 weeks induction on B5 culture medium supplemented with 10% coconut water, 0.05–0.5 mg l^?1 6-benzylaminopurine (BAP) and 0.5 mg l^?1 indole-3-butyric acid (IBA). These continued to proliferate under dark conditions until the 2nd to 3rd subculture. Following transfer to a photoperiod of 16 h light, shoots evolved from these globular structures and developed further to plantlets. The influence of several factors, including culture medium composition, sucrose concentration, the type, concentration and combination of growth regulators and the presence of coconut water was studied. The percentage of explants showing globular structure formation and the number of globular structures per explant were evaluated. Macroscopic, histological and scanning electron microscopic studies revealed that the morphogenic process involved mainly organogenic nodules with fewer globular somatic embryos. The nodules gave rise to shoots and subsequently complete plants following incubation on B5 Gamborg medium containing 0.5 mg l^?1 IBA and 30 g l^?1 sucrose, which promoted root formation.     

Optimization of heterologous expression of the phytase (PPHY) of Pichia anomala in P. pastoris and its applicability in fractionating allergenic glycinin from soy protein

The phytase (PPHY) of Pichia anomala has the requisite properties of thermostability and acidstability, broad substrate spectrum, and protease insensitivity, which make it a suitable candidate as a feed and food additive. The 1,389-bp PPHY gene was amplified from P. anomala genomic DNA, cloned in pPICZαA, and expressed extracellularly in P. pastoris X33. Three copies of PPHY have been detected integrated into the chromosomal DNA of the recombinant P. pastoris. The size exclusion chromatography followed by electrophoresis of the pure rPPHY confirmed that this is a homohexameric glycoprotein of ~420 kDa with a 24.3 % portion as N-linked glycans. The temperature and pH optima of rPPHY are 60 °C and 4.0, similar to the endogenous enzyme. The kinetic characteristics K _m, V _max, K _cat, and K _cat/K _m of rPPHY are 0.2 ± 0.03 mM, 78.2 ± 1.43 nmol mg^?1 s^?1, 65,655 ± 10.92 s^?1, and 328.3 ± 3.12 μM^?1 s^?1, respectively. The optimization of medium components led to a 21.8-fold improvement in rPPHY production over the endogenous yeast. The rPPHY titer attained in shake flasks could also be sustained in the laboratory fermenter. The rPPHY accounts for 57.1 % of the total secreted protein into the medium. The enzyme has been found useful in fractionating allergenic protein glycinin from soya protein besides dephytinization.     

Productivity and Temperature as Drivers of Seasonal and Spatial Variations of Dissolved Methane in the Southern Bight of the North Sea

Dissolved CH₄ concentrations in the Belgian coastal zone (North Sea) ranged between 670 nmol l^?1 nearshore and 4 nmol l^?1 offshore. Spatial variations of CH₄ were related to sediment organic matter (OM) content and gassy sediments. In nearshore stations with fine sand or muddy sediments, the CH₄ seasonal cycle followed water temperature, suggesting methanogenesis control by temperature in these OM-rich sediments. In offshore stations with permeable sediments, the CH₄ seasonal cycle showed a yearly peak following the chlorophyll-a spring peak, suggesting that in these OM-poor sediments, methanogenesis depended on freshly produced OM delivery. This does not exclude the possibility that some CH₄ might originate from dimethylsulfide (DMS) or dimethylsulfoniopropionate (DMSP) or methylphosphonate transformations in the most offshore stations. Yet, the average seasonal CH₄ cycle was unrelated to those of DMS(P), very abundant during the Phaeocystis bloom. The annual average CH₄ emission was 126 mmol m^?2 y^?1 in the most nearshore stations (~4 km from the coast) and 28 mmol m^?2 y^?1 in the most offshore stations (~23 km from the coast), 1260–280 times higher than the open ocean average value (0.1 mmol m^?2 y^?1). The strong control of CH₄ by sediment OM content and by temperature suggests that marine coastal CH₄ emissions, in particular in shallow areas, should respond to future eutrophication and warming of climate. This is supported by the comparison of CH₄ concentrations at five stations obtained in March 1990 and 2016, showing a decreasing trend consistent with alleviation of eutrophication in the area.     

Estimation of carbon biomass and community structure of planktonic bacteria in Lake Biwa using respiratory quinone analysis

The relationship between bacterial respiratory quinone (RQ) concentration and biomass was assessed for Lake Biwa bacterial assemblages to evaluate the utility of bacterial RQ concentration as an indicator of bacterial carbon. The biomass estimated from the RQ concentration correlated well with that from cell volume, indicating that RQ concentration is an appropriate indicator of bacterial biomass. The estimated carbon content per unit of RQ (carbon conversion factor) of bacteria was 0.67 mg C nmol RQ^?1. Bacterial carbon biomass, which was estimated from the RQ concentration using the conversion factor, ranged between 0.008 and 0.054 mg C L^?1 (average 0.025 mg C L^?1) at 5 m depth and between 0.010 and 0.024 mg C L^?1 (average 0.015 mg C L^?1) at 70 m depth. Ubiquinone-8-containing bacteria dominated the epilimnion and hypolimnion. Compared to conventional image analysis, bacterial RQ analysis is a less laborious method of simultaneously determining bacterial biomass and community.     

Sunflower seed deterioration as related to moisture content during ageing, energy metabolism and active oxygen species scavenging

The objectives of the present work were to investigate whether loss of sunflower (Helianthus annuus L.) seed viability was affected by the embryo moisture content (MC) during seed pretreatment at 35°C, and was related to changes in energy metabolism and in the antioxidant defence system. Non‐dormant seeds were equilibrated at MC of the embryonic axis ranging from 0.037 to 0.605 g H₂O g^?1 dry matter (DM) for 1 day at 15°C, and they were then placed at 35°C for various durations up to 14 days before the germination assays at 15°C. As expected, the higher the MC, the faster was seed deterioration. There existed a negative linear relationship between the time taken for germination to drop to 50% (P₅₀) and the embryonic axis MC ranging from 0.108 and 0.438 g H₂O g^?1 DM. In dry seeds, adenosine triphosphate (ATP), adenosine diphosphate (ADP) and adenosine monophosphate represented 6.3, 14.8 and 70.9% of the adenylate pool, respectively, and the energy charge (EC) was very low (0.14). ATP and ADP levels and EC increased sharply during the first day of equilibrium of seeds at a MC above 0.158 g H₂O g^?1 DM. Subsequent controlled deterioration at 35°C resulted in a decrease in the adenylate pool, and consequently in ATP level. The higher the energy metabolism during ageing, the lower was seed viability. Loss of seed viability was associated with an accumulation of H₂O₂, and then of malondialdehyde (MDA) suggesting that lipid peroxidation was not the only cause of seed deterioration. When there was a sublinear relationship between H₂O₂ content in the embryonic axis and seed viability, MDA accumulation only occurred when 50% of the seed population died within 7 days, i.e. when MC was higher than 0.248 g H₂O g^?1 DM. Ageing was associated with a decrease in the activity of superoxide dismutase, catalase and glutathione reductase, the main enzymes involved in cell detoxification. The involvement of seed MC, as key factor of ageing is discussed with regards to energy metabolism and the regulation of active oxygen species accumulation.     

Plant growth promoting rhizobacteria enhanced leaf organic acids,FC-R activity and Fe nutrition of apple under lime soil conditions

Iron chlorosis in the calcareous soils is one most important stress factors worldwide that limits photosynthesis and decreases fruit yield and quality. Certain soil rhizobacteria produce organic compounds such as plant acids and they may reduce the soil rhizosphere pH and affect ferric chelate reductase (FC-R) activity in root. However, there is no knowledge regarding changes in organic acids content and FC-R activities of leaf due to rhizobacterial root inoculation. Therefore, the efficiency of six plant growth promoting rhizobacteria (PGPR) were tested on apple cv. Braeburn on M9 and MM106 rootstocks. The results of the experiment showed leaf organic acid contents, iron quantity of soil, root and leaf and root and leaf FC-R activity were significantly affected via rhizobacteria applications in apple plants. In MM106 and M9, there was a remarkable increase in Fe in M3 inoculated soil by 95 and 89%, respectively, compared to control. Average increases in citric, malic, malonic, butyric and lactic acid in the leaf were obtained from rhizobacterial root inoculations of 25.1, 21.8, 29.6, 18.0 and 18.2% in Braeburn/MM106, respectively. In Braeburn/M9, MFDCa1 application increased all organic acid concentrations compared to the control. MFDCa2 treatment caused the maximum leaf FC-R activity in Braeburn on M9 and MM106 (60.9 and 50.3 nmol Fe⁺² g^?1 FW h^?1, respectively) while the least values were determined in the control (33.5 and 29.9 nmol Fe⁺² g^?1 FW h^?1, respectively). This study showed the bacterial strains tested in our study may be used as a biofertilizer instead of Fe fertilizers.