Characterization of two Peruvian maize landraces differing in boron toxicity tolerance.

Boron (B) toxicity is a major agricultural problem that causes a considerable decrease in crop yield and quality. The soil in arid and semi-arid areas is often subjected to excessive B content. Southwestern Perú (department of Tacna) is characterized by high B levels in its agricultural land and irrigation water. This work analyzes the response of two local maize (Zea mays) landraces (Pachía and Sama) from Tacna to B toxicity. Both landraces were, therefore, grown in hydroponic media under control and B toxicity conditions, and after 10 days, seedlings were harvested and B content, B-transporter gene expressions, and several morphological and physiological parameters were determined. The leaf and root soluble B content was lower in Sama than in Pachía when both landraces were subjected to high B concentrations, which could be explained by its higher expression levels of B-efflux transporters. The capacity of Sama to maintain reduced levels of soluble B in its leaves and roots led to decreased leaf damage and higher photosynthetic and growth parameters under B toxicity conditions. These results support the proposal that Sama would perform better than Pachía under excessive B conditions, thus making it a more suitable landrace to be used in soils with toxic levels of B.

Electroporation optimizes the uptake of boron-10 by tumor for boron neutron capture therapy (BNCT) mediated by GB-10: a boron biodistribution study in the hamster cheek pouch oral cancer model.

Boron neutron capture therapy (BNCT) is a promising cancer binary therapy modality that utilizes the nuclear capture reaction of thermal neutrons by boron-10 resulting in a localized release of high- and low-linear energy transfer (LET) radiation. Electrochemotherapy (ECT) is based on electroporation (EP) that induces opening of pores in cell membranes, allowing the entry of compounds. Because EP is applied locally to a tumor, the compound is incorporated preferentially by tumor cells. Based on the knowledge that the therapeutic success of BNCT depends centrally on the boron content in tumor and normal tissues and that EP has proven to be an excellent facilitator of tumor biodistribution of an anti-tumor agent, the aim of this study was to evaluate if EP can optimize the delivery of boronated compounds. We performed biodistribution studies and qualitative microdistribution analyses of boron employing the boron compound sodium decahydrodecaborate (GB-10) + EP in the hamster cheek pouch oral cancer model. Syrian hamsters with chemically induced exophytic squamous cell carcinomas were used. A typical EP treatment was applied to each tumor, varying the moment of application with respect to the administration of GB-10 (early or late). The results of this study showed a significant increase in the absolute and relative tumor boron concentration and optimization of the qualitative microdistribution of boron by the use of early EP + GB-10 versus GB-10 without EP. This strategy could be a tool to improve the therapeutic efficacy of BNCT/GB-10 in vivo.

Pre- and postnatal environmental boron exposure and infant growth: Results from a mother-child cohort in northern Argentina.

BACKGROUND: Experimental studies show developmental toxicity of boron and we recently found impaired weight and length in newborns of mothers exposed to boron through drinking water during pregnancy. OBJECTIVES: To evaluate potential impact of pre- and postnatal boron exposure on infant anthropometry. METHODS: In our mother-child cohort (n = 177) in Argentinean Andes, where drinking water concentrations of boron, lithium and arsenic have been found to vary considerably, we collected maternal blood and urine during and after pregnancy, placenta, breast milk, as well as infant urine and blood during the first 6 months of life. In all samples, boron and other potentially toxic elements were measured by ICP-MS. Infant weight (g), length (cm) and head circumference (cm) were measured at birth, 0-3 (n = 120) and/or 3-6 months (n = 120; 79 overlap) of age. RESULTS: Boron concentrations in breast milk (range: 46-786 µg/L) correlated strongly with those in maternal serum (range: 47-624 µg/L; rs = 0.94) 0-3 months post-partum. In multivariable-adjusted linear regression, urinary boron (log2-transformed; range 60-9200 µg/L) in the youngest infants, but not maternal serum boron during pregnancy, was inversely associated with body weight at both 0-3 months (B: -141, 95% CI: -240; -42, p = 0.006) and 3-6 months (B: -200, 95% CI: -377; -23, p = 0.027). Infant urinary boron was also inversely associated with head circumference at 0-3 months (B: -0.39, 95% CI: -0.74; -0.04, p = 0.028), as well as length (B: -0.57, 95% CI: -1.1; -0.03, p = 0.040) and head circumference (B: -0.30, 95% CI: -0.64; 0.04, p = 0.083) at 3-6 months. CONCLUSIONS: The observed first evidence that elevated environmental boron exposure in early infancy may adversely affect growth supports previous findings of boron-related impaired fetal growth. More research is needed to verify the findings at older age and in other populations.

Evaporation process in histological tissue sections for neutron autoradiography.

The analysis of the distribution and density of nuclear tracks forming an autoradiography in a nuclear track detector (NTD) allows the determination of 10B atoms concentration and location in tissue samples from Boron Neutron Capture Therapy (BNCT) protocols. This knowledge is of great importance for BNCT dosimetry and treatment planning. Tissue sections studied with this technique are obtained by cryosectioning frozen tissue specimens. After the slicing procedure, the tissue section is put on the NTD and the sample starts drying. The thickness varies from its original value allowing more particles to reach the detector and, as the mass of the sample decreases, the boron concentration in the sample increases. So in order to determine the concentration present in the hydrated tissue, the application of corrective coefficients is required. Evaporation mechanisms as well as various factors that could affect the process of mass variation are outlined in this work. Mass evolution for tissue samples coming from BDIX rats was registered with a semimicro analytical scale and measurements were analyzed with software developed to that end. Ambient conditions were simultaneously recorded, obtaining reproducible evaporation curves. Mathematical models found in the literature were applied for the first time to this type of samples and the best fit of the experimental data was determined. The correlation coefficients and the variability of the parameters were evaluated, pointing to Page's model as the one that best represented the evaporation curves. These studies will contribute to a more precise assessment of boron concentration in tissue samples by the Neutron Autoradiography technique.

Boron stress response and accumulation potential of the extremely tolerant species Puccinellia frigida.

Phytoremediation is a promising technology to tackle boron toxicity, which restricts agricultural activities in many arid and semi-arid areas. Puccinellia frigida is a perennial grass that was reported to hyperaccumulate boron in extremely boron-contaminated sites. To further investigate its potential for phytoremediation, we determined its response to boron stress under controlled conditions (hydroponic culture). Also, as a first step towards understanding the mechanisms underlying its extreme tolerance, we evaluated the presence and expression of genes related with boron tolerance. We found that P. frigida grew normally even at highly toxic boron concentrations in the medium (500mg/L), and within its tissues (>5000mg/kg DW). We postulate that the strategies conferring this extreme tolerance involve both restricting boron accumulation and an internal tolerance mechanism; this is consistent with the identification of putative genes involved in both mechanisms, including the expression of a possible boron efflux transporter. We also found that P. frigida hyperaccumulated boron over a wide range of boron concentrations. We propose that P. frigida could be used for boron phytoremediation strategies in places with different soil characteristics and boron concentrations. Further studies should pave the way for the development of clean and low-cost solutions to boron toxicity problems.

Pollen Morphology and Boron Concentration in Floral Tissues as Factors Triggering Natural and GA-Induced Parthenocarpic Fruit Development in Grapevine.

Parthenocarpic fruit development (PFD) reduces fruit yield and quality in grapevine. Parthenocarpic seedless berries arise from fruit set without effective fertilization due to defective pollen germination. PFD has been associated to micronutrient deficiency but the relation of this phenomenon with pollen polymorphism has not been reported before. In this work, six grapevine cultivars with different tendency for PFD and grown under micronutrient-sufficient conditions were analyzed to determine pollen structure and germination capability as well as PFD rates. Wide variation in non-germinative abnormal pollen was detected either among cultivars as well as for the same cultivar in different growing seasons. A straight correlation with PFD rates was found (R2 = 0.9896), suggesting that natural parthenocarpy is related to defective pollen development. Such relation was not observed when PFD was analyzed in grapevine plants exposed to exogenous gibberellin (GA) or abscissic acid (ABA) applications at pre-anthesis. Increase (GA treatment) or reduction (ABA treatment) in PFD rates without significative changes in abnormal pollen was determined. Although these plants were maintained at sufficient boron (B) condition, a down-regulation of the floral genes VvBOR3 and VvBOR4 together with a reduction of floral B content in GA-treated plants was established. These results suggest that impairment in B mobility to reproductive tissues and restriction of pollen tube growth could be involved in the GA-induced parthenocarpy.

Maternal-child transfer of essential and toxic elements through breast milk in a mine-waste polluted area.

OBJECTIVE: To determine the daily intake of essential micronutrients and toxic elements through breast milk in exclusive and nonexclusive breastfed infants living in an area with major mine tailing deposition (n = 24), compared with a control area (n = 11). STUDY DESIGN: The milk volume ingested by 2 to 4 and 4 to 6 month infants was measured by a stable isotopic method. Elements in milk, maternal and infant urine, and drinking water were measured by inductively coupled plasma mass spectrometry (ICP-MS). RESULTS: Similar breast milk volume and essential micronutrients intake in groups of exclusively breastfed infants, but more cadmium, boron, and lithium through breastfeeding in experimental area was found. This exposure was even higher in the nonexclusively breastfed infants, who also ingested more arsenic, boron, and lithium than exclusive breastfed infants. CONCLUSION: The use of the deuterium and the ICP-MS methods made it possible to evaluate the exact amount of essential and toxic elements ingested by infants through breast milk demonstrating that lower amount of toxic elements are transferred to exclusive breastfed infants compared with those who additionally received nonmaternal milk.

Functional characterization of Citrus macrophylla BOR1 as a boron transporter.

Plants have evolved to develop an efficient system of boron uptake and transport using a range of efflux carriers named BOR proteins. In this work we isolated and characterized a boron transporter of citrus (Citrus macrophylla), which was named CmBOR1 for its high homology to AtBOR1. CmBOR1 has 4403 bp and 12 exons. Its coding region has 2145 bp and encodes for a protein of 714 amino acids. CmBOR1 possesses the molecular features of BORs such as an anion exchanger domain and the presence of 10 transmembrane domains. Functional analysis in yeast indicated that CmBOR1 has an efflux boron transporter activity, and transformants have increased tolerance to excess boron. CmBOR1 is expressed in leaves, stem and flowers and shows the greatest accumulation in roots. The transcript accumulation was significantly increased under boron deficiency conditions in shoots. In contrast, the accumulation of the transcript did not change in boron toxicity conditions. Finally, we observed that constitutive expression of CmBOR1 was able to increase tolerance to boron deficiency conditions in Arabidopsis thaliana, suggesting that CmBOR1 is a xylem loading boron transporter. Based on these results, it was determined that CmBOR1 encodes a boric acid/borate transporter involved in tolerance to boron deficiency in plants.

VvBOR1, the grapevine ortholog of AtBOR1, encodes an efflux boron transporter that is differentially expressed throughout reproductive development of Vitis vinifera L.

Boron (B) is an essential micronutrient for normal development of roots, shoots and reproductive tissues in plants. Due to its role in the structure of rhamnogalacturonan II, a polysaccharide required for pollen tube growth, B deficiency has been associated with the occurrence of parthenocarpic seedless grapes in some varieties of Vitis vinifera L. Despite that, it is unclear how B is mobilized and accumulated in reproductive tissues. Here we describe the characterization of an efflux B transporter, VvBOR1, homolog to AtBOR1, which is involved in B xylem loading in Arabidopsis thaliana roots. VvBOR1-green fluorescent protein (GFP) fusion protein expressed in A. thaliana localizes in the proximal plasma membrane domain in root pericycle cells, and VvBOR1 overexpression restores the wild-type phenotype in A. thaliana bor1-3 mutant plants exposed to B deficiency. Complementation of a mutant yeast strain indicates that VvBOR1 corresponds to a B efflux transporter. Transcriptional analyses during grapevine reproductive development show that the VvBOR1 gene is preferentially expressed in flowers at anthesis and a direct correlation between the expression pattern and B content in grapes was established, suggesting the involvement of this transporter in B accumulation in grapevine berries.

Histomorphometric study of alveolar bone healing in rats fed a boron-deficient diet.

Bone healing after tooth extraction in rats is a suitable experimental model to study bone formation. Thus, we performed a study to determine the effects of boron (B) deficiency on bone healing by using this model. The first lower right molar of weanling Wistar rats was extracted under anesthesia. The animals were divided into two groups: +B (adequate; 3 mg B/kg diet), and -B (boron-deficient; 0.07 mg/kg diet). The animals in both groups were killed in groups of 10 at 7 and 14 days after surgery. The guidelines of the NIH for the care and use of laboratory animals were observed. The mandibles were resected, fixed, decalcified, and embedded in paraffin. Buccolingually oriented sections were obtained at the level of the mesial alveolus and used for histometric evaluations. Total alveolar volume (TAV) and trabecular bone volume per total volume (BV/TV) in the apical third of the alveolus were determined. Percentages of osteoblast surface (ObS), eroded surface (ES), and quiescent surface (QS) were determined. No statistical significant differences in food intake and body weight were observed. Histomorphometric evaluation found -B rats had 36% and 63% reductions in BV/TV at 7 and 14 days, respectively. When compared with +B rats, -B rats had significant reductions (57% and 87%) in ObS concomitantly with increases (120% and 126%) in QS at 7 and 14 days, respectively. The findings show that boron deficiency results in altered bone healing because of a marked reduction in osteogenesis.

Calcium and boron for in vitro rooting of Nothofagus nervosa.

Several mineral compounds influence adventitious rooting, and those containing boron and calcium, play an important role. They are associated to the enzyme regulation, cell division, cellular metabolism (induction stage) and roots enlargement (expression stage). The aims of this work were to study the variation of the endogenous calcium and boron concentrations along the rooting, and determine their optimal concentrations in the rooting basal medium of Nothofagus nervosa. Significant variation in endogenous calcium concentrations was detected along the rooting phase, strongly correlated with rhizogenesis phenomenons and linked to total peroxidase variation as cited elsewhere. Calcium and boron concentrations in the basal medium significantly affected the rooting. Treatment containing 60 mg.L-1 Ca and 1 mg.L-1 B produced root formation in fewer days than other combinations. The variation of these nutrient concentrations in the medium is a useful tool to improve microcutting rooting. Roots are obtained earlier and in a shorter period of time, finally resulting in more homogeneous microplants with a better developed rooting system.

Calcium and boron for in vitro rooting of Nothofagus nervosa

Several mineral compounds influence adventitious rooting, and those containing boron and calcium, play an important role. They are associated to the enzyme regulation, cell division, cellular metabolism (induction stage) and roots enlargement (expression stage). The aims of this work were to study the variation of the endogenous calcium and boron concentrations along the rooting, and determine their optimal concentrations in the rooting basal medium of Nothofagus nervosa. Significant variation in endogenous calcium concentrations was detected along the rooting phase, strongly correlated with rhizogenesis phenomenons and linked to total peroxidase variation as cited elsewhere. Calcium and boron concentrations in the basal medium significantly affected the rooting. Treatment containing 60 mg.L-1 Ca and 1 mg.L-1 B produced root formation in fewer days than other combinations. The variation of these nutrient concentrations in the medium is a useful tool to improve microcutting rooting. Roots are obtained earlier and in a shorter period of time, finally resulting in more homogeneous microplants with a better developed rooting system.(AU)