In Vitro Development and Cell Allocation After Aggregation of Syngeneic Wild Type and Fluorescence-Expressing Bovine Cloned Embryos

Background: The in vitro production (IVP) of embryos by in vitro fertilization or cloning procedures has been known to cause epigenetic changes in the conceptus that in turn are associated with abnormalities in pre- and postnatal development. Handmade cloning (HMC) procedures and the culture of zona-free embryos in individual microwells provide excellent tools for studies in developmental biology, since embryo development and cell allocation patterns can be evaluated under a wide range of embryo reconstruction arrangements and in in vitro embryo culture conditions. As disturbances in embryonic cell allocation after in vitro embryo manipulations and unusual in vivo conditions during the first third of pregnancy appear to be associated with large offspring, embryo aggregation procedures may allow a compensation for epigenetic defects between aggregated embryos or even may influence more favorable cell allocation in embryonic lineages, favoring subsequent development. Thus, the aim of this study was to evaluate in vitro embryo developmental potential and the pattern of cell allocation in blastocysts developed after the aggregation of handmade cloned embryos produced using syngeneic wild type and/or transgenic somatic cells. Materials, Methods & Results: In vitro-matured bovine cumulus-oocyte complexes (COC) were manually bisected after cumulus and zona pellucida removal; then, two enucleated hemi-oocytes were paired and fused with either a wild type (WT) or a GFP-expressing (GFP) fetal skin cell at the 11th and 19th passages, respectively. Following chemical activation, reconstructed cloned embryos and zona-free parthenote embryos were in vitro-cultured in microwells, for 7 days, either individually (1 x 100%) or after the aggregation of two structures (2 x 100%) per microwell, as follows: (G1) one WT cloned embryo; (G2) two aggregated WT embryos; (G3) one GFP cloned embryo; (G4) two aggregated GFP embryos; (G5) aggregation of a WT embryo and a GFP embryo; (G6) one parthenote embryo; or (G7) two aggregated parthenote embryos. Fusion (clones), cleavage (Day 2), and blastocyst (Day 7) rates, and embryonic cell allocation were compared by the. 2 or Fisher tests. Total cell number (TCN) in blastocysts was analyzed by the Student's test (P < 0.05). Fusion and cleavage rates, and cell allocation were similar between groups. On a per WOW basis, development to the blastocyst stage was similar between groups, except for lower rates of development seen in G3. However, when based on number of embryos per group (one or two), blastocyst development was higher in G1 than all other groups, which were similar between one another. Cloned GFP embryos had lower in vitro development to the blastocyst stage than WT embryos, which had more TCN than parthenote or aggregated chimeric WT/GFP embryos. Aggregated GFP embryos had fewer cells than the other embryo groups. Discussion: The in vitro development of GFP cloned embryos was lower than WT embryos, with no effects on cell allocation in resulting blastocysts. Differences in blastocyst rate between groups were likely due to lower GFP-expressing cell viability, as GFP donor cells were at high population cell doublings when used for cloning. On a per embryo basis, embryo aggregation on Day 1 resulted in blastocyst development similar to non-aggregated embryos on Day 7, with no differences in cell proportion between groups. The use of GFP-expressing cells was proven a promising strategy for the study of cell allocation during embryo development, which may assist in the elucidation of mechanisms of abnormalities after in vitro embryo manipulations, leading to the development of improved protocols for the in vitro production (IVP) of bovine embryos.

Selenate and selenite on yield and agronomic biofortification with selenium in rice

The objective of this work was to evaluate the effect of doses of selenate and selenite on rice (Oryza sativa) biofortification with Se, as well the influence of these forms of Se in the levels of P, S, Fe, and Zn in grains. The experiment was conducted in a greenhouse, in pots with 4 dm(3) of a sandy clay loam Latosol, with medium texture, in a 5x2 factorial arrangement with five doses of Se (0, 0.75, 1.50, 3.0, and 6.0 mg dm(-3)) and two forms of Se (selenate and selenite). Selenate provided greater efficiency of root uptake of Se, plant-use efficiency, translocation from roots to shoots, and content of this element in rice grains. The application of Se during fertilization influences the levels of P, S, and Zn, but does not affect those of Fe in rice grains.

NUTRITIONAL BALANCE, ESSENTIAL OIL AND WOOD PRODUCTION FROM Corymbia citriodora Hill & Johnson WITH SEWAGE SLUDGE AND MINERAL FERTILIZER APPLICATIONS

The Corymbia citriodora is one of the most important forest species in Brazil and the reason is the diversity of its use, because it produces good quality wood and the leaves may be used for essential oil production. Although, there are not many studies about species and the handling effect in the nutritional balance. This study aimed to evaluate the biomass production and nutrient balance in the conventional production of essential oil and wood of Corymbia citriodora with sewage sludge application. The experiment design established was the randomized blocks, with four replicates and two treatments: 1 - fertilization with 10 tons ha(-1) (dry mass) of sewage sludge, supplemented with K and B, and 2 - mineral fertilization. It was evaluated the aerial biomass production, the nutrient export of the leaves, the essential oil and wood production at four years old. The trees that received application of sewage sludge produced 20 % more leaves biomass than the trees with mineral fertilization, resulting in larger oil production. Besides, the trees with sewage sludge application produced 14.2 tons ha(-1) yr(-1) of woody biomass that was 27 % higher than the treatment with mineral fertilization. For both treatments the N balance was negative, but treatment with sewage sludge application (-45 kg ha(-1)) was four times lower than the observed on mineral fertilization treatment (-185 kg ha(-1)). It may be concluded in this paper that the application of sewage sludge benefits the production of leaves biomass, essential oil and wood, besides result better nutritional balance of the Corymbia citriodora production system.

Growth of seedlings Schizolobium amazonicum (Huber ex Ducke) on substrate fertilized with nitrogen, phosphorus and potassium

The nutritional management of seedlings in the nursery is one of the most important practices that influence seedling quality. The aim of this work was to evaluate the effect of nitrogen, phosphorus and potassium on the development of Schizolobium amazonicum seedlings grown in 250 cm(3) containers with a commercial substrate in the North of Mato Grosso State, Brazil. The experimental design was completely randomized design with five treatments and five replications, each replication being represented by 24 seedlings. The treatments were: control (only commercial substrate); nitrogen fertilization (150 g m(-3) N using ammonium sulfate + 1.0 kg of ammonium sulfate dissolved in 100 L of water and applied in coverage); phosphorus fertilization (300 g P2O5 m(-3) using simple superphosphate); potassium fertilization (100 g m(-3) K2O using potassium chloride + 0.3 kg of potassium chloride dissolved in 100 L of water and applied in coverage) and; complete (a mixture of the three nutrients, 150, 300 and 100 g m(-3) N, P2O5 and K2O, respectively + 1.0 kg of ammonium sulfate + 0.3 kg of potassium chloride). The commercial substrate was composted milled pine bark plus vermiculite. Evaluations of the seedlings were performed at 90 days after sowing. The complete treatment (NPK) gave the highest values for biometric and best plant indices, which express the quality. When analyzing nutrients in isolation; potassium had the lowest effect. Based on these results it can be recommended to fertilize Schizolobium amazonicum seedlings in nurseries with 150, 300 and 100 g m(-3) of N, P2O5 and K2O, respectively, plus 1.0 kg of sulfate ammonium and 0.3 kg of potassium chloride applied in coverage.

OPTIMUM RATIO OF CALCIUM AND BORON IN THE NUTRIENT SOLUTION OR IN CASTOR BEAN SHOOT FOR FRUIT YIELD AND SEED OIL CONTENT

Calcium (Ca) and boron (B) have been reported as the major macro-and micronutrient required for castor bean plant yield. The objective of this study was to determine the Ca: B ratios (in the growth media and plant tissue) for fruit yield and shoot dry weight of the castor bean (Ricinus communis L.), grown in a nutrient solution, and to evaluate Ca and B supply on concentration and total uptake of Ca, potassium (K), magnesium (Mg), and B, as well on the seed oil content. The treatments were arranged in a 3 x 3 factorial fashion, consisting of three rates of Ca (40, 80, and 160 mg L-1) and three of B (0.32, 0.96, and 1.60 mg L-1). Calcium and B rates increased the shoot and root dry weight and fruit yield at a Ca: B ratio in the nutrient solution of 166 and 100, respectively. Symptoms of B deficiency were observed in plants supplied with 0.32 mg B L-1, regardless of the Ca concentration in the nutrient solution. Plants which showed visual symptoms of Ca deficiency cultivated with 40 mg Ca L-1 presented concentration of Ca in plant tissue up to 10 g kg(-1). The concentration and total Ca and B uptake increased with the rates of them. Notwithstanding, the shoot Ca accumulation was improved by B rates. In addition, there were no decreases in K and Mg uptake due to Ca rates. Furthermore, addition of 80 mg L-1 of Ca and 1.60 mg L-1 of B in the growth media increased the seed oil content. The Ca: B ratio in the diagnostic leaf associated with the highest plant dry weight (shoot and root) and fruit yield, was 500 (16 to 20 g kg(-1) of Ca, and for 30 to 40 mg kg(-1) of B).

Soil sulfur fractions dynamics and distribution in a tropical grass pasture amended with nitrogen and sulfur fertilizers

Soil sulfur (S) partitioning among the various pools and changes in tropical pasture ecosystems remain poorly understood. Our study aimed to investigate the dynamics and distribution of soil S fractions in an 8-year-old signal grass (Brachiaria decumbens Stapf.) pasture fertilized with nitrogen (N) and S. A factorial combination of two N rates (0 and 600?kg N ha1 y1, as NH4NO3) and two S rates (0 and 60?kg S ha1 y1, as gypsum) were applied to signal grass pastures during 2 y. Cattle grazing was controlled during the experimental period. Organic S was the major S pool found in the tropical pasture soil, and represented 97% to 99% of total S content. Among the organic S fractions, residual S was the most abundant (42% to 67% of total S), followed by ester-bonded S (19% to 42%), and C-bonded S (11% to 19%). Plant-available inorganic SO4-S concentrations were very low, even for the treatments receiving S fertilizers. Low inorganic SO4-S stocks suggest that S losses may play a major role in S dynamics of sandy tropical soils. Nitrogen and S additions affected forage yield, S plant uptake, and organic S fractions in the soil. Among the various soil fractions, residual S showed the greatest changes in response to N and S fertilization. Soil organic S increased in plots fertilized with S following the residual S fraction increment (16.6% to 34.8%). Soils cultivated without N and S fertilization showed a decrease in all soil organic S fractions.

Comparison of microsatellites and isozymes in genetic diversity studies of Oryza glumaepatula (Poaceae) populations

The study of the genetic structure of wild plant populations is essential for their management and conservation. Several DNA markers have been used in such studies, as well as isozyme markers. In order to provide a better comprehension of the results obtained and a comparison between markers which will help choose tools for future studies in natural populations of Oryza glumaepatula, a predominantly autogamous species, this study used both isozymes and microsatellites to assess the genetic diversity and genetic structure of 13 populations, pointing to similarities and divergences of each marker, and evaluating the relative importance of the results for studies of population genetics and conservation. A bulk sample for each population was obtained, by sampling two to three seeds of each plant, up to a set of 50 seeds. Amplified products of eight SSR loci were electrophoresed on non-denaturing polyacrylamide gels, and the fragments were visualized using silver staining procedure. Isozyme analyses were conducted in polyacrylamide gels, under a discontinuous system, using six enzymatic loci. SSR loci showed higher mean levels of genetic diversity (A=2.83, p=0.71, A(P)=3.17, H-o=0.081, H-e=0.351) than isozyme loci (A=1.20, p=0.20, A(P)=1.38, H-o=0.006, H-e=0.056). Interpopulation genetic differentiation detected by SSR loci (R-ST=0.631, equivalent to F-ST=0.533) was lower than that obtained with isozymes (F-ST=0.772). However, both markers showed high deviation from Hardy-Weinberg expectations (F-IS=0.744 and 0.899, respectively for SSR and isozymes). The mean apparent outcrossing rate for SSR ((t) over bar (a)=0.14) was higher than that obtained using isozymes ((t) over bar (a)=0.043), although both markers detected lower levels of outcrossing in Amazonia compared to the Pantanal. The migrant number estimation was also higher for SSR (Nm=0.219) than isozymes (Nm=0.074), although a small number for both markers was expected due to the mode of reproduction of this species, defined as mixed with predominance of self fertilization. No correlation was obtained between genetic and geographic distances with SSR, but a positive correlation was found between genetic and geographic distances with isozymes. We conclude that these markers are divergent in detecting genetic diversity parameters in O. glumaepatula and that microsatellites are powerful for detecting information at the intra-population level, while isozymes are more powerful for inter-population diversity, since clustering of populations agreed with the expectations based on the geographic distribution of the populations using this marker. Rev. Biol. Trop. 60 (4): 1463-1478. Epub 2012 December 01.

Adubação mineral com cobalto e molibdênio na cultura da soja

Altas produtividades de soja requerem grandes quantidades de nitrogênio, que podem ser obtidas principalmente, a partir da fixação simbiótica. No entanto, há possibilidade da eficiência desse processo biológico ser prejudicada pela deficiência de micronutrientes, especialmente de cobalto e molibdênio. Nesse contexto, objetivou-se com o presente trabalho avaliar a eficiência agronômica e a forma de aplicação de adubação mineral com cobalto e molibdênio na cultura da soja. Para tanto, a cultivar CD-206 foi submetida a diferentes tratamentos que consistiram da combinação entre aplicação de Co e Mo via tratamento de sementes e adubação foliar. As características agronômicas avaliadas foram o número de nódulos, massa seca da parte aérea, massa seca de raiz, número de vagens, número de grãos, massa de mil grãos e produção de grãos. A aplicação de molibdênio e cobalto via sementes e/ ou adubação foliar no estádio V4 (terceira folha trifoliolada completamente desenvolvida, quarto nó) promoveram incrementos significativos no rendimento da cultura. Respostas positivas ao cobalto e molibdênio também foram observadas no número de nódulos, vagens e grãos, com aumentos de até 240 kg ha-1 no rendimento da cultura. Os parâmetros agronômicos avaliados foram afetados positivamente pela aplicação de Co e Mo, principalmente quanto aplicado tanto via semente como foliar (TS + V4), inclusive para a produtividade de grãos. A forma de aplicação não foi significativamente distinta, ou seja, tanto a aplicação via semente como via foliar foram eficientes no fornecimento destes nutrientes para a cultura da soja.

Do changes in carbon allocation account for the growth response to potassium and sodium applications in tropical Eucalyptus plantations?

Understanding the underlying mechanisms that account for the impact of potassium (K) fertilization and its replacement by sodium (Na) on tree growth is key to improving the management of forest plantations that are expanding over weathered tropical soils with low amounts of exchangeable bases. A complete randomized block design was planted with Eucalyptus grandis (W. Hill ex Maiden) to quantify growth, carbon uptake and carbon partitioning using a carbon budget approach. A combination of approaches including the establishment of allometric relationships over the whole rotation and measurements of soil CO2 efflux and aboveground litterfall at the end of the rotation were used to estimate aboveground net production (ANPP), total belowground carbon flux and gross primary production (GPP). The stable carbon isotope (delta C-13) of stem wood alpha-cellulose produced every year was used as a proxy for stomatal limitation of photosynthesis. Potassium fertilization increased GPP and decreased the fraction of carbon allocated belowground. Aboveground net production was strongly enhanced, and because leaf lifespan increased, leaf biomass was enhanced without any change in leaf production, and wood production (P-W) was dramatically increased. Sodium application decreased the fraction of carbon allocated belowground in a similar way, and enhanced GPP, ANPP and P-W, but to a lesser extent compared with K fertilization. Neither K nor Na affected delta C-13 of stem wood alpha-cellulose, suggesting that water-use efficiency was the same among the treatments and that the inferred increase in leaf photosynthesis was not only related to a higher stomatal conductance. We concluded that the response to K fertilization and Na addition on P-W resulted from drastic changes in carbon allocation.

Fatty acids as a tool to compare cachara (Pseudoplatystoma reticulatum) (Siluriformes: Pimelodidae) and hybrid (Pseudoplatystoma corruscans x Pseudoplatystoma reticulatum) larvae during early development

In this study, we investigated the physiological alterations during ontogeny for cachara (Pseudoplatystoma reticulatum) and their hybrid larvae (Pseudoplatystoma corruscans x P. reticulatum) using lipids and fatty acids as physiological tools to elucidate the basis for differences in these groups' productivity in an industrial setting. Eggs and larvae samples were collected during January and February of 2008 in the city of Bandeirantes, MS, and were divided into three primary phases: phase I (0-16 h after fertilization); phase II (24 h after fertilization to 6 days after fertilization); and phase III (7-25 days after fertilization). The larvae of both groups showed a high degree of similarity, suggesting that the hybrid larvae showed a high level of heritability from the cachara broodstock. Analysis of the total lipid content provided evidence that there is no alteration in lipid concentration during ontogeny for both groups (i.e., the cachara and hybrids). However, the fatty acid profile showed that during the endogenous feeding period (phase II), when the larvae must use the energy reserves from the mother, the cachara larvae used mainly monounsaturated fatty acids for development. This is typical for most fish species, though notably, the hybrids preferentially used saturated fatty acids. Furthermore, certain specific changes demonstrate unique patterns of energy utilization and structural substrates, which may aid in elucidating the empirical differences reported by fish farmers (i.e., that the hybrids perform better than cacharas in captivity).

Single-cell expression analysis of BMP15 and GDF9 in mature oocytes and BMPR2 in cumulus cells of women with polycystic ovary syndrome undergoing controlled ovarian hyperstimulation

To detect expression of bone morphogenetic protein 15 (BMP15) and growth differentiation factor 9 (GDF9) in oocytes, and their receptor type 2 receptor for BMPs (BMPR2) in cumulus cells in women with polycystic ovary syndrome (PCOS) undergoing in vitro fertilization (IVF), and determine if BMPR2, BMP15, and GDF9 expression correlate with hyperandrogenism in FF of PCOS patients. Prospective case-control study. Eighteen MII-oocytes and their respective cumulus cells were obtained from 18 patients with PCOS, and 48 MII-oocytes and cumulus cells (CCs) from 35 controls, both subjected to controlled ovarian hyperstimulation (COH), and follicular fluid (FF) was collected from small (10-14 mm) and large (> 18 mm) follicles. RNeasy Micro Kit (Qiagen(A (R))) was used for RNA extraction and gene expression was quantified in each oocyte individually and in microdissected cumulus cells from cumulus-oocyte complexes retrieved from preovulatory follicles using qRT-PCR. Chemiluminescence and RIA assays were used for hormone assays. BMP15 and GDF9 expression per oocyte was higher among women with PCOS than the control group. A positive correlation was found between BMPR2 transcripts and hyperandrogenism in FF of PCOS patients. Progesterone values in FF were lower in the PCOS group. We inferred that BMP15 and GDF9 transcript levels increase in mature PCOS oocytes after COH, and might inhibit the progesterone secretion by follicular cells in PCOS follicles, preventing premature luteinization in cumulus cells. BMPR2 expression in PCOS cumulus cells might be regulated by androgens.

Patterns of cell proliferation and apoptosis by topographic region in normal Bos taurus vs. Bos indicus crossbreeds bovine placentae during pregnancy

Background: Placental and fetal growth requires high rates of cellular turnover and differentiation, which contributes to conceptus development. The trophoblast has unique properties and a wide range of metabolic, endocrine and angiogenic functions, but the proliferative profile of the bovine placenta characterized by flow cytometry analysis and its role in fetal development are currently uncharacterized. Complete understanding of placental apoptotic and proliferative rates may be relevant to development, especially if related to the pathogenesis of pregnancy losses and placental abnormalities. Methods: In this study, the proliferation activity and apoptosis in different regions of normal bovine placenta (central and boundary regions of placentomes, placentomal fusion, microplacentomes, and interplacentomal regions), from distinct gestation periods (Days 70 to 290 of pregnancy), were analyzed by flow cytometry. Results: Our results indicated that microplacentomes presented a lower number of apoptotic cells throughout pregnancy, with a higher proliferative activity by the end of gestation, suggesting that such structures do not contribute significantly to normal of placental functions and conceptus development during pregnancy. The placentome edges revealed a higher number of apoptotic cells from Day 170 on, which suggests that placentome detachment may well initiate in this region. Conclusion: Variations involving proliferation and apoptotic rates may influence placental maturation and detachment, compromising placental functions and leading to fetal stress, abnormalities in development and abortion, as frequently seen in bovine pregnancies from in vitro fertilization and cloning procedures. Our findings describing the pattern of cell proliferation and apoptosis in normal bovine pregnancies may be useful for unraveling some of the developmental deviations seen in nature and after in vitro embryo manipulations.

The double task of preventing malnutrition and overweight: a quasi-experimental community-based trial

Background: The Maternal-Child Pastoral is a volunteer-based community organization of the Dominican Republic that works with families to improve child survival and development. A program that promotes key practices of maternal and child care through meetings with pregnant women and home visits to promote child growth and development was designed and implemented. This study aims to evaluate the impact of the program on nutritional status indicators of children in the first two years of age. Methods: A quasi-experimental design was used, with groups paired according to a socioeconomic index, comparing eight geographical areas of intervention with eight control areas. The intervention was carried out by lay health volunteers. Mothers in the intervention areas received home visits each month and participated in a group activity held biweekly during pregnancy and monthly after birth. The primary outcomes were length and body mass index for age. Statistical analyses were based on linear and logistic regression models. Results: 196 children in the intervention group and 263 in the control group were evaluated. The intervention did not show statistically significant effects on length, but point estimates found were in the desired direction: mean difference 0.21 (95%CI −0.02; 0.44) for length-for-age Z-score and OR 0.50 (95%CI 0.22; 1.10) for stunting. Significant reductions of BMI-for-age Z-score (−0.31, 95%CI −0.49; -0.12) and of BMI-for-age > 85th percentile (0.43, 95%CI 0.23; 0.77) were observed. The intervention showed positive effects in some indicators of intermediary factors such as growth monitoring, health promotion activities, micronutrient supplementation, exclusive breastfeeding and complementary feeding. Conclusions: Despite finding effect measures pointing to effects in the desired direction related to malnutrition, we could only detect a reduction in the risk of overweight attributable to the intervention. The findings related to obesity prevention may be of interest in the context of the nutritional transition. Given the size of this study, the results are encouraging and we believe a larger study is warranted.

The nitrogen sufficiency index underlying estimates of nitrogen fertilization requirements of common bean

Chlorophyll determination with a portable chlorophyll meter can indicate the period of highest N demand of plants and whether sidedressing is required or not. In this sense, defining the optimal timing of N application to common bean is fundamental to increase N use efficiency, increase yields and reduce the cost of fertilization. The objectives of this study were to evaluate the efficiency of N sufficiency index (NSI) calculated based on the relative chlorophyll index (RCI) in leaves, measured with a portable chlorophyll meter, as an indicator of time of N sidedressing fertilization and to verify which NSI (90 and 95 %) value is the most appropriate to indicate the moment of N fertilization of common bean cultivar Perola. The experiment was carried out in the rainy and dry growing seasons of the agricultural year 2009/10 on a dystroferric Red Nitosol, in Botucatu, São Paulo State, Brazil. The experiment was arranged in a randomized complete block design with five treatments, consisting of N managements (M1: 200 kg ha-1 N (40 kg at sowing + 80 kg 15 days after emergence (DAE) + 80 kg 30 DAE); M2: 100 kg ha-1 N (20 kg at sowing + 40 kg 15 DAE + 40 kg 30 DAE); M3: 20 kg ha-1 N at sowing + 30 kg ha-1 when chlorophyll meter readings indicated NSI < 95 %; M4: 20 kg ha-1 N at sowing + 30 kg ha-1 N when chlorophyll meter readings indicated NSI < 90 % and, M5: control (without N application)) and four replications. The variables RCI, aboveground dry matter, total leaf N concentration, production components, grain yield, relative yield, and N use efficiency were evaluated. The RCI correlated with leaf N concentrations. By monitoring the RCI with the chlorophyll meter, the period of N sidedressing of common bean could be defined, improving N use efficiency and avoiding unnecessary N supply to common bean. The NSI 90 % of the reference area was more efficient to define the moment of N sidedressing of common bean, to increase N use efficiency.

Deficiency symptoms and uptake of micronutrients by castor bean grown in nutrient solution

Castor bean is a nutrient-demanding species, but there is still little information on its micronutrient requirements. The objectives of this study were to evaluate the effects of levels of B (2.5, 12.5 and 25.0 µmol L-1), Cu (0.05, 0.25 and 0.50 µmol L-1), Mn (0.2, 1.0 and 2.0 µmol L-1) and Zn (0.2, 1.0 and 2.0 µmol L-1) in a nutrient solution on plant B, Cu, Mn and Zn concentrations and uptake, vegetative growth and fruit yield of castor bean "Iris", grown in greenhouse. The experiment was arranged in a completely randomized block design with three replicates. The first deficiency symptoms were observed for B, followed by Zn, Cu and Mn. The main changes in the cell ultrastructure due to lack of B were thickening of the cell walls and middle lamellae, distorted chloroplasts and tightly stacked thylakoids, besides the absence of starch grains. The Mn, Zn and Cu deficiencies led to disruption of chloroplasts, disintegration of thylakoids and absence of amyloplasts. The concentration and uptake of B, Cu, Mn, and Zn in castor bean plants increased with micronutrient supply in the solution. Fruit yield was drastically reduced by B and Mn deficiencies. On the other hand, the dry matter yield of the shoot and root of castor bean plants was not. In the treatment with full nutrient solution, the leaves accumulated 56 and 48 % of the total B and Mn taken up by the plants, respectively, and the seeds and roots 85 and 61 % of the total Cu and Zn taken up, respectively. This shows the high demand of castor bean Iris for B and Mn for fruit yield.