Effect of boron on cell elongation and division in squash roots

This work establishes that cessation of root elongation of intact squash (Cucurbita pepo L.) plants is an early result of boron deficiency. Root elongation is slowed by 6 hours and is virtually stopped as early as 24 hours after boron is first withheld from the nutrient solution. As root elongation ceased, cell elongation progressed distally into the region normally occupied by the apical meristem and eventually the meristem became indistinguishable. Differentiation was determined by use of an elongation index in which cell length was compared to cell width. This index ranged from a low of 0.8 in boron-sufficient root meristems to a high of 3 in root meristems grown in a boron-deficient nutrient solution for 98 hours. It is concluded that a continuous supply of boron is not essential for cell elongation but is required for maintenance of meristematic activity. Boron may act as a regulator of cell division in this tissue.     

Substitution of germanium for boron in plant growth

The observation was confirmed that the addition of germanium dioxide (soluble form) to the nutrient solution can delay for a short time the appearance of boron deficiency symptoms on the shoots of sunflower plants (Helianthus annuus L.). This appeared to be true, however, only under growing conditions in which the plants had a low boron requirement. The delay in the appearance of boron deficiency symptoms by administering germanium was demonstrated in sunflower plants ranging in age from 7 to 20 days. This effect was noted whether the germanium was administered prior to or at the time the plants were transferred to minus-boron nutrient solution.     

Effect of light intensity and plant size on rate of development of early boron deficiency symptoms in tomato root tips

Young tomato plants (Lycopersicon esculentum Miller, cultivar Rutgers) grown in solution culture at 27° at 2 light intensities with adequate boron (0.1 mg/l) and treated with these 2 intensities in the absence of adequate boron developed root boron deficiency symptoms. The typical deficiency symptoms of decreased root elongation, increased depth of brown color and decreased RNA content of tips developed more rapidly at high than at low light intensity, and plant size influenced results. Plants supplied with adequate boron did not exhibit deficiency symptoms.     

Relationship between boron and calcium in the N2-fixing legume–rhizobia symbiosis

Because boron (B) and calcium (Ca²⁺) seem to have a strong effect on legume nodulation and nitrogen fixation, rhizobial symbiosis with leguminous plants, grown under varying concentrations of both nutrients, was investigated. The study of early pre‐infection events included the capacity of root exudates to induce nod genes, and the degree of adsorption of bacteria to the root surface. Both phenomena were inhibited by B deficiency, and increased by addition of Ca²⁺, resulting in an increase of the number of nodules. The infection and invasion steps were investigated by fluorescence microscopy in pea nodules harbouring a Rhizobium leguminosarum strain that constitutively expresses green fluorescent protein. High Ca²⁺ enhanced cell and tissue invasion by Rhizobium, which was highly inhibited after B deficiency. This was combined with an increased B concentration in nodules of plants grown on B‐free medium and supplemented with high Ca²⁺ concentrations, and that can be attributed to an increased B import to the nodules. Histological examination of indeterminate (pea) and determinate (bean) nodules showed an altered nodule anatomy at low B content of the tissue. The moderate increase in nodular B due to additional Ca²⁺ was not sufficient to prevent the abnormal cell wall structure and the aberrant distribution of pectin polysaccharides in B‐deficient treatments. Overall results indicate that the development of the symbiosis depends of the concentration of B and Ca²⁺, and that both nutrients are essential for nodule structure and function.     

Differences in cell wall components and allocation of boron to cell walls confer variations in sensitivities of Brassica napus cultivars to boron deficiency

Aims

The cell wall is the main binding site of boron (B) in plants, and the differences in B requirements among different plant species are determined by pectic polysaccharide contents in the cell walls. The aim of this research was to illustrate the relationship between cell wall properties and allocation of B to cell wall and the differential sensitivity of Brassica napus cultivars to B deficiency.

Methods

Two cultivars with opposite B efficiency were used to analyse the relationship among cell wall pectin contents and glycosyl composition, B uptake and allocation, gene expression and cell wall ultrastructure.

Results

The Brassica napus B-efficient cultivar Qingyou 10 was more tolerant to B deficiency, exhibiting a higher biomass production, milder B deficiency symptoms and less cell wall thickening compared to the Brassica napus B-inefficient cultivar Westar 10. These differences were attributed to two factors; the first was that Qingyou 10 accumulated more B and distributed significantly higher proportion of it to the cell wall pectins than did Westar 10 under low B supply. Also, the cell walls of Qingyou 10 exhibited relatively less B-binding sites than those of Westar 10, which was indicated by the lower cell wall extraction rates, less pectin and glycosyl residue contents under the B-deficient and B-sufficient conditions. A comparison of the KDOPS gene expression levels in the two conditions suggests that Westar 10 had a higher potential for biosynthesizing B-binding substances than did Qingyou 10, regardless of B levels.

Conclusions

These results suggest that both higher cell wall pectin polysaccharide content, and limited accumulation and allocation of B to the cell walls contribute to the greater sensitivity of Westar 10 to B deficiency. These two physiological aspects may determine the differences in B deficiency tolerance between Brassica napus cultivars Qingyou 10 and Westar 10. Comparably, the difference in accumulation and allocation of B to cell wall plays a much more important role than cell wall components to sensitivity difference of Brassica napus cultivars to B deficiency.     

Antioxidant responses and water status in Brassica seedlings subjected to boron stress

The effect of boron (B) on growth, water status and oxidative damage was investigated in the leaves and roots of 7-day-old seedlings of Brassica juncea var. Varuna. For this seedlings of Brassica were grown in solution culture with variable boron supply (0.033, 0.33, 3.3 and 33 mg B L^?1) under controlled conditions in green house. Photosynthetic pigments were found to be decreased more under excess (3.3 and 33 mg B L^–1) than deficient boron supply (0.033 mg B L^–1) when compared to control (0.33 mg B L^–1). Accumulation of hydrogen peroxide and thiobarbituric acid reactive substances content in both leaves and roots under deficient and excess boron supply suggested oxidative damage due to excessive production of reactive oxygen species. Increased activity of antioxidative enzymes: superoxide dismutase, catalase and peroxidase along with polyphenol oxidase was observed in leaves and roots under boron deficiency and excess than in control. Increased proline concentration, decreased total water content and water saturation deficit also indicated the water deficit condition in leaves and roots of boron-stressed Brassica seedlings.     

Characteristics of root boron nutrition confer high boron efficiency in Brassica napus cultivars

Background and aims

Brassica napus has high boron (B) demand, but significant genotype differences exist with respect to B deficiency. The aim of this research was to elucidate the relationship between the different sensitivities of Brassica napus cultivars to low B stress and the characteristics of B uptake and transport to characterise the regulation of B efficiency in Brassica napus.

Methods

B-efficient and B-inefficient Brassica napus cultivars were used to compare the uptake and transport of B using the stable isotope ¹⁰B tracer and grafting experiments, as well as expression of B transporters by RT-PCR.

Results

B-efficient cultivars have significant advantages with regard to B limitation. The B-efficient cultivar HZ showed less severe B deficiency symptoms and higher dry biomass than the B-inefficient cultivars LW and LB. Both the amount of total B and the ¹⁰B concentration and accumulation in the shoots and roots of B-efficient HZ were higher than those of B-inefficient cultivars. In B-inefficient LW, the amount of total B and the ¹⁰B that was transported into shoots was less than in the other three cultivars and the content and accumulation of total B and ¹⁰B in the roots of B-inefficient LB were the lowest among all of the cultivars. When the roots of B-efficient HZ were used as stocks, the grafted plants showed B-efficient characteristics, such as mild B deficiency symptoms, and higher dry biomass and B accumulation, regardless of whether they originated from B-efficient or B-inefficient cultivars. In contrast, the grafted plants with B-inefficient LW used as stocks were B-inefficient. The expressions of BnBOR1;1c, BnBOR1;2a and BnNIP5;1 were up-regulated in roots under low B stress compared with the normal B condition. However, there was no obvious difference in the expressions of the three genes or of four other BnBOR1s between B-efficient and B-inefficient cultivars in low or normal B environments.

Conclusions

These results indicate that the B efficiency of Brassica napus is controlled primarily by roots, which allow more uptake and accumulation of B in B-efficient cultivars than B-inefficient cultivars in a low B environment. However the molecular mechanism regulating B efficiency in Brassica napus remains to be determined.     

Nitrogen-induced boron deficiency in lucerne

Summary In a glasshouse experiment with a boron deficient soil the application of nitrogen was found to decrease the boron concentration and boron uptake by lucerne (Medicago sativa). Without added boron, nitrogen applications killed the lucerne, probably by inducing severe boron deficiency. With added boron, the lowest rate of nitrogen application increased lucerne yield but further additions depressed yields. The effect was due to a physiological interaction rather than an effect of the nitrogen on the availability of the boron in the soil.     

Nonessentiality of boron in fungi and the nature of its toxicity

An investigation was undertaken to determine whether any of the following fungi had a requirement for boron (B): Saccharomyces cerevisiae, Aspergillus niger, Neurospora crassa, and Penicillium chrysogenum. Boron was unessential, and hence a study was made of the concentrations of B that reduced the growth of S. cerevisiae and P. chrysogenum and the mode of action of the B toxicity. Fifty and 4000 mg B/liter, respectively, significantly (5% level) reduced the growth of the latter 2 species.

In both, glycolysis appeared to be inhibited by toxic levels of B, since the cells accumulated fructose-1,6-diP and ADP, but were low in glyceraldehyde-3-P and ATP. With S. cerevisiae growing on glucose, 150 mg B/liter significantly reduced CO₂ evolution. When glyceraldehyde was substituted for glucose, CO₂ evolution and O₂ consumption were unaffected by this level of B.

Aldolase was suspected of being inhibited by high B, and this was confirmed using a crude aldolase extract from S. cerevisiae and purified rabbit muscle aldolase. The inhibition of aldolase by B was uncompetitive.

With aldolase activity being reduced by toxic levels of B, the fungi were apparently unable to utilize carbohydrates at a rate sufficient to maintain the metabolic processes involved in growth and reproduction.

     

Addition of Phenylboronic Acid to Malus domestica Pollen Tubes Alters Calcium Dynamics,Disrupts Actin Filaments and Affects Cell Wall Architecture

A key role of boron in plants is to cross-link the cell wall pectic polysaccharide rhamnogalacturonan-II (RG-II) through borate diester linkages. Phenylboronic acid (PBA) can form the same reversible ester bonds but cannot cross-link two molecules, so can be used as an antagonist to study the function of boron. This study aimed to evaluate the effect of PBA on apple (Malus domestica) pollen tube growth and the underlying regulatory mechanism. We observed that PBA caused an inhibition of pollen germination, tube growth and led to pollen tube morphological abnormalities. Fluorescent labeling, coupled with a scanning ion-selective electrode technique, revealed that PBA induced an increase in extracellular Ca²⁺ influx, thereby elevating the cytosolic Ca²⁺ concentration [Ca²⁺]c and disrupting the [Ca²⁺]c gradient, which is critical for pollen tube growth. Moreover the organization of actin filaments was severely perturbed by the PBA treatment. Immunolocalization studies and fluorescent labeling, together with Fourier-transform infrared analysis (FTIR) suggested that PBA caused an increase in the abundance of callose, de-esterified pectins and arabinogalactan proteins (AGPs) at the tip. However, it had no effect on the deposition of the wall polymers cellulose. These effects are similar to those of boron deficiency in roots and other organs, indicating that PBA can induce boron deficiency symptoms. The results provide new insights into the roles of boron in pollen tube development, which likely include regulating [Ca²⁺]c and the formation of the actin cytoskeleton, in addition to the synthesis and assembly of cell wall components.     

Quantitative autoradiography in boron neutron capture therapy considering the particle ranges in the samples

PurposeBoron neutron capture therapy is a cellular-scale particle therapy exploiting boron neutron capture reactions in boron compounds distributed in tumour cells. Its therapeutic effect depends on both the accumulation of boron in tumour cells and the neutron fluence. Autoradiography is used to visualise the micro-distribution of boron compounds.MethodsHere, we present an equation for the relationship between boron concentration and pit density on the solid-state nuclear track detector, taking into consideration the particle ranges in the samples. This equation is validated using liver-tissue sections and boron standard solutions. Moreover, we present a simple co-localisation system for pit and tissue-section images that requires no special equipment.ResultsThe equation reproduces the experimentally observed trends between boron concentration and pit density. This equation provides a theoretical explanation for the widely used calibration curve between pit density and boron concentration; it also provides a method to correct for differences of tissue-section thickness in quantitative autoradiography.ConclusionsUsing the equation together with this co-localisation system could improve micro-scale quantitative estimation in tissue sections.     

Yield, transpiration and growth of tomatoes under combined excess boron and salinity stress

Boron is essential to growth at low concentrations and limits growth and yield when in excess. Little is known regarding plant response to excess boron (B) and salinity occurring simultaneously. The influences of B and salinity on tomatoes (Lycopersicon esculentum Mill. Cv `5656') were investigated in lysimeters. Salinity levels were 1, 3, 6 and 9 dSm^–1 and B levels were 0.028, 0.185, 0.37, 0.74, 1.11, 1.48 mol m^–3. Excess boron was found to decrease yield and transpiration of tomatoes. This effect was inhibited when plants were exposed to simultaneous B and salinity stresses. Both irrigation water salinity and boron concentration influenced water use of the plants in the same manner as they influenced yield. While yield was found to decrease with increased boron concentration in leaf tissue, increased salinity led to decreased boron accumulation. Yield response was found to correlate better to B concentration in irrigation water and soil solution than to plant tissue B content. A dominant-stress-factor model was assumed and validated. The model applies the principle that when a plant is submitted to conditions of stress caused by B in conjunction with salinity, the more severe stress determines yield. The results of this study have significance in modeling and management of high salinity high boron conditions. Under saline conditions, differences in crop yield and in water use may not be experienced over a significant range of boron concentrations.     

pH dependent salinity-boron interactions impact yield, biomass, evapotranspiration and boron uptake in broccoli (Brassica oleracea L.)

Aims

Soil pH is known to influence many important biochemical processes in plants and soils, however its role in salinity—boron interactions affecting plant growth and ion relations has not been examined. The purpose of this research was to evaluate the interactive effects of salinity, boron and soil solution pH on broccoli (Brassica oleracea L.) growth, yield, consumptive water use and shoot-boron accumulation.

Methods

A greenhouse experiment was conducted using a sand tank system where salinity-B-pH treatment solutions were supplemented with a complete nutrient solution. Sulfate-dominated irrigation waters, characteristic of groundwater in California’s San Joaquin valley (SJV), were tested at EC levels of 2, 5, 8, 11 and 14 dS?m^?1. Each salinity treatment consisted of two boron treatments (0.5 and 21 mg?L^?1) and each of those treatments was tested under slightly basic (pH?8.0) and slightly acidic (pH?6.0) conditions.

Results

Results included multiple salinity-boron-pH interactions affecting shoot biomass, head yield and consumptive water use. Broccoli fresh head yields were significantly reduced by salinity and boron, but the degree of yield reductions was influenced by pH. Relative head yields were substantially reduced in treatments with high pH and high B, particularly under low and high salinity where head yields were decreased by 89 % and 96 %, respectively, relative to those at low salinity and low boron. Intermediate levels of salinity were far less damaging. Increased salinity and boron reduced evapotranspiration (ET) and there were no salinity-boron associated interactions on ET. However, increased salinity and boron concentrations increased water use efficiency (shoot biomass/cumulative volume ET). Shoot B concentration increased with increased boron and was greater at pH?6 as compared to pH?8. Shoot boron concentration decreased with increasing salinity at both pH levels but particularly at the high substrate boron concentration.

Conclusions

It is likely that different mechanisms, yet unknown, are responsible for severe head-yield reductions at low and high salinity in the presence of high boron under alkaline conditions. We found that boron in the shoot did not accumulate by a simple passive process. Rather as boron increased from 0.5 to 21 mg?L^?1, there was a restrictive mechanism where total shoot boron (mg plant^?1) was reduced by 10 to 40 times the amount potentially supplied to the shoot by passive transport via mass flow perhaps involving complex interactions with membrane channels and B exporters. Total shoot boron concentration was a poor indicator of plant growth response.     

Relationship of boron to gibberellic Acid-induced proliferation in debudded tobacco plants

Stem applications of gibberellic acid (GA) to debudded tobacco plants (Nicotiona tabacum L., var. One Sucker) produce stem swellings that involve intense proliferation of, primarily, xylem tissue. Withholding boron from GA-treated debudded plants greatly reduces the GA-induced proliferation. This response offers a system for directly demonstrating the effect of boron on xylem formation and lignification that is unconfounded by the effect of boron on elongation growth.     

Species variability in boron requirement is correlated with cell wall pectin

Fourteen species of crop plants which differ in their reportedtissue boron requirements were grown in B-replete or B-deficientmedium. Leaf samples were collected and analysed for B and cellwall components. There was a significant positive correlationamong the species between B concentration in the leaf or thecell wall and uronic acid, rhamnose and galactose (indicativeof pectin) in the cell wall. The concentration of cell wallpectin was also positivety related with reported tissue-B requirementsand observed sensitivity to B deficiency. Boron deficiency didnot alter the amount of uronic acid present in cell walls, suggestingthat there was no effect of B deficiency on pectin metabolism.Under B-deficient conditions the amount of ‘soluble’B (i.e. B not associated with the cell wall) declined dramaticallywhile the proportion of cellular B that was ‘insoluble’(i.e. B associated with the cell wall) increased. The positiverelationship between pectin content, insoluble B and tissue-Brequirement of diverse species suggests that the amount of cellwall pectin may be significant in determining the relative tissue-Brequirements of the species. These results indicate that either(1) species with high cell wall pectin contents require greateramounts of B for the construction of the cell wall, or (2) pectinin cell walls forms an insoluble complex with B, thereby reducingits availability for other putative B-requiring metabolic functions.Thus, species with a high pectin content would have a highertissue-B requirement. Key words: Boron, deficiency, uronic acid, pectin, cell wall     

Effects of irradiance during growth on tolerance of geranium to sub- and supra-optimal boron supply

In our previous study on geranium, we showed that increases in growth irradiance from sub-optimal to near-optimal could delay boron deficiency effects on photosynthesis. In this study, we further investigated the effects of growth irradiance on tolerance to B stress by growing geranium (Pelargonium × hortorum cv. Maverick White) under sub- to supra-optimal B concentrations (4.5, 45, and 450 μM) and under three irradiances of 100, 300, or 500 μmol m^?2 s^?1 PAR for 30 d. In general, at low and medium irradiances, sub- and supra-optimal B availability decreased root and shoot dry masses, but at high irradiance, the B stress was prevented. Net photosynthetic rate decreased by the supra-optimal B concentration at the high irradiance only suggesting B-related photoinhibition. Tissue B content and root specific B uptake only modestly decreased by the low B treatment, but increased greatly by the high B availability, and the higher irradiance decreased the tissue B content and the root B uptake only at the low and medium B supplies. Interestingly, the increases in irradiance decreased the content and uptake of all other nutrients, except Fe uptake. Effects of the B stress on the content of other nutrients were variable, but the B stress often exacerbated decreases in nutrient content with the increasing irradiance which would be especially important under nutrient-limiting conditions. Hence, in this study, the B stress effects on growth were mitigated by the increases in growth irradiance, which offset negative effects on physiology, and the protective effects of irradiance were likely caused by its positive effects on plant carbon/energy status rather than on tissue B content or B uptake.     

Effects of boron toxicity on root and leaf anatomy in two Citrus species differing in boron tolerance

Key message

Typical toxic symptom only occurred in B-toxic C. grandis leaves. B-toxicity induced PCD of C. grandis leaf phloem tissue. The lower leaf free B might contribute to the higher B-tolerance of C. sinensis.

Abstract

Seedlings of ‘Xuegan’ (Citrus sinensis) and ‘Sour pummelo’ (Citrus grandis) differing in boron (B)-tolerance were irrigated with nutrient solution containing 10 (control) or 400 (B-toxic) μM H₃BO₃ for 15 weeks. Thereafter, the effects of B-toxicity on leaf photosynthesis, chlorophyll, plant B absorption and distribution, root and leaf anatomy were investigated to elucidate the possible B-tolerant mechanisms of Citrus plants. Typical toxic symptom only occurred in B-toxic C. grandis leaves. Similarly, B-toxicity only affected C. grandis photosynthesis and chlorophyll. Although total B concentration in B-toxic roots and leaves was similar between the two species, leaves from B-toxic C. grandis plant middle had higher free B and lower bound B as compared with those from C. sinensis. Effects of B-toxicity on leaf structure were mainly limited to the mesophyll cells and the phloem of leaf veins. Although irregular cell wall thickening was observed in leaf cortex cells and phloem tissue of B-toxic C. grandis and C. sinensis leaves, exocytosis only occurred in the companion cells and the parenchyma cells of B-toxic C. sinensis leaf phloem. Also, B-toxicity induced cell death of phloem tissue through autophagy in C. grandis leaf veins. B-toxicity caused death of root epidermal cells of the two Citrus species. B-toxicity restrained degradation of middle lamella, but did not alter ultrastructure of Golgi apparatus and mitochondria in root elongating zone cells. In conclusion, C. sinensis was more tolerant to B-toxicity than C. grandis. The lower leaf free B and higher bound B might contribute to the higher B-tolerance of C. sinensis.     

Development of a boron buffered solution culture system for controlled studies of plant boron nutrition

Chelated-buffered nutrient solutions are used for studies on micronutrient metals but so far no equivalent system exists for boron nutrition studies: the present investigation was initiated with that intention. From a literature review, it was noted that a range of substances form chelates with boron including polyhydric alcohols, sugars and phenolic compounds. However, none apart from hydrofluoric acid formed chelates with formation constants comparable to those of micronutrient metal chelates like diethylenetriaminepentaacetic acid (DTPA). Moreover, most chelating substances had deleterious side effects which reduced their possible use in water culture: many of the compounds are substrates for bacterial growth, some are harmful to handle, and others are toxic to plants or humans. Borosilicate glass; was tested in a laboratory experiment but found to release boron too slowly into solution to maintain constant boron concentration in solution even when very finely ground. Current investigations centre around the use of a boron-specific resin, which strongly complexes H₃BO₃ on its N-methyl glucamine functional groups. The boron sorption capacity of the resin varied from 2.2 to 5.0 mg B g^-1 resin. Boron saturated resin maintained an equilibrium solution boron concentration of 46 t M when added at the rate of 2 g of resin to 1 L of boron free triple deionised water. Plants grown in complete nutrient solution with boron saturated resin added at 1 g per litre of nutrient solution grew as well as plants grown in conventional nutrient solution containing 9.2 t M boron and their shoots contained adequate boron concentrations for growth. There was no evidence that the resin had effects on plant growth other than in releasing and equilibrating boron concentration in the nutrient solution.     

Biodistribution of sodium borocaptate (BSH) for boron neutron capture therapy (BNCT) in an oral cancer model

Boron neutron capture therapy (BNCT) is based on selective accumulation of ¹⁰B carriers in tumor followed by neutron irradiation. We previously proved the therapeutic success of BNCT mediated by the boron compounds boronophenylalanine and sodium decahydrodecaborate (GB-10) in the hamster cheek pouch oral cancer model. Based on the clinical relevance of the boron carrier sodium borocaptate (BSH) and the knowledge that the most effective way to optimize BNCT is to improve tumor boron targeting, the specific aim of this study was to perform biodistribution studies of BSH in the hamster cheek pouch oral cancer model and evaluate the feasibility of BNCT mediated by BSH at nuclear reactor RA-3. The general aim of these studies is to contribute to the knowledge of BNCT radiobiology and optimize BNCT for head and neck cancer. Sodium borocaptate (50 mg ¹⁰B/kg) was administered to tumor-bearing hamsters. Groups of 3–5 animals were killed humanely at nine time-points, 3–12 h post-administration. Samples of blood, tumor, precancerous pouch tissue, normal pouch tissue and other clinically relevant normal tissues were processed for boron measurement by optic emission spectroscopy. Tumor boron concentration peaked to therapeutically useful boron concentration values of 24–35 ppm. The boron concentration ratio tumor/normal pouch tissue ranged from 1.1 to 1.8. Pharmacokinetic curves showed that the optimum interval between BSH administration and neutron irradiation was 7–11 h. It is concluded that BNCT mediated by BSH at nuclear reactor RA-3 would be feasible.