Pb hyperaccumulation and tolerance in common buckwheat (Fagopyrum esculentum Moench)

Common buckwheat grown in Pb-contaminated soil was found to accumulate a large amount of Pb in its leaves (8,000 mg/kg DW), stem (2,000 mg/kg DW), and roots (3,300 mg/kg DW), without significant damage. This indicates that buckwheat is a newly recognized Pb hyperaccumulator, which is defined as a plant containing over 1,000 mg/kg of Pb in its shoots on a dry-weight basis. Moreover, it was shown that application of the biodegradable chelator methylglycinediacetic acid trisodium salt at concentrations of up to 20 mmol/kg resulted in a more than five times higher concentration of Pb in the shoot without notable growth inhibitation at up to 10 mmol/kg. These results indicate that buckwheat is a potential phytoremediator of Pb-contaminated soils.     

Characterization of heat-induced aggregates of globulin from common buckwheat (Fagopyrum esculentum Moench)

Some physicochemical properties and the microstructure of heat-induced aggregates of globulin from common buckwheat (Fagopyrum esculentum Moench) (BWG) formed at 100 °C in 0.01 M phosphate buffer containing 1.0 M NaCl, pH 7.4 were studied. Differential scanning calorimetric (DSC) analysis shows a re-distribution of native and extensively denatured proteins in the heat-induced aggregates of BWG, particularly in the ISA fraction. Sodium dodecyl sulfate polyacrylamide gel electrophoretic (SDS-PAGE) analysis suggests the occurrence of both dissociation and association of molecules and the involvement of intermolecular disulfide linkages during thermal aggregation. Transmission electron microscopy (TEM) reveals that native BWG appeared as uniform compact globules with diameters ranging between 11.7 and 12.5 nm. TEM examination of the buffer-soluble aggregates, fractionated by sucrose density gradient ultracentrifugation, demonstrates the formation of strand-like small aggregates and large compact globular soluble macroaggregates.     

Form of aluminium for uptake and translocation in buckwheat (Fagopyrum esculentum Moench)

 The forms of Al for uptake by the roots and translocation from the root to the shoot were investigated in a buckwheat (Fagopyrum esculentum Moench, cv. Jianxi) that accumulates Al in its leaves. The Al concentration in the xylem sap was 15-fold higher in the plants exposed to AlCl₃ than in those exposed to an Al-oxalate (1:3) complex, suggesting that the roots take up Al in the ionic form. The Al concentration in the xylem sap was 4-fold higher than that in the external solution after a 1-h exposure to AlCl₃ solution and 10-fold higher after a 2-h exposure. The Al concentration in the xylem sap increased with increasing Al concentration in the external solution. The Al uptake was not affected by a respiratory inhibitor, hydroxylamine, but significantly inhibited by the addition of La. These results suggest that Al uptake by the root is a passive process, and La³⁺ competes for the binding sites for Al³⁺ on the plasma membrane. The form of Al in the xylem sap was identified by ²⁷Al-nuclear magnetic resonance analysis. The chemical shift of ²⁷Al in the xylem sap was around 10.9 ppm, which is consistent with that of the Al-citrate complex. Furthermore, the dominant organic acid in the xylem sap was citric acid, indicating that Al was translocated in the form of Al-citrate complex. Because Al is present as Al-oxalate (1:3) in the root, the present data show that ligand exchange from oxalate to citrate occurs before Al is released to xylem. Received: 10 December 1999 / Accepted: 3 February 2000     

An angiotensin-I converting enzyme inhibitor from buckwheat (Fagopyrum esculentum Moench) flour

A compound that inhibited angiotensin-I converting enzyme (ACE) activity was isolated from buckwheat powder. This compound is thought to be the hydroxy derivative of nicotianamine and its chemical structure is 2'-hydroxynicotianamine. This compound showed a very high inhibitory activity toward ACE, and the IC(50) was 0.08 microM. Only this hydroxy analog was found in buckwheat powder, at about 30 mg/100g, and no nicotianamine was detected. However, nicotianamine was detected in the buckwheat plant body. 2'-hydroxynicotianamine was also found in other polygonaceous plants.     

Enhancing Biological Control of Leafrollers (Lepidoptera: Tortricidae) by Sowing Buckwheat (Fagopyrum esculentum) in an Orchard

Two successive trials were carried out in stone-fruit and apple orchards in the summers of 1994-95 and 1995-96 respectively to determine the effect of floral resources on leafroller parasitoid numbers. Orchards were divided into eight blocks, each containing one replicate of a control and a treatment. The treatment had buckwheat (Fagopyrum esculentum Moench) drilled (as 3 m 2 m plots) in the centre of each replicate, while the control areas were kept weed free with herbicides. Catches of parasitoids in yellow pan traps were compared between the control and treatment areas. In the first trial, on average 10 times more parasitoids were captured in the buckwheat plots than in the controls. However, few of the parasitoids were natural enemies of leafrollers; the catches were dominated by Anacharis sp., a parasitoid of the brown lacewing ( Micromus tasmaniae Walker), itself a beneficial insect. Leafrollers were very rare in this orchard, which explains the absence of their parasitoids in the traps. In the second season, with the experiment conducted in an orchard with a history of leafroller populations, higher numbers of Anacharis sp. were again caught in buckwheat than in control plots. However, significantly higher (34% compared with 20%) levels (P< 0.05) of parasitism (by Dolichogenidea tasmanica (Cameron)) of released leafroller larvae were observed in buckwheat plots than in controls, despite the small plot size. Also, higher numbers of D. tasmanica were captured in yellow sticky traps 2 m above the buckwheat than in the equivalent position in the control plots.     

Light emitting diode(LED)‐based trapping of the greenhouse whitefly (Trialeurodes vaporariorum)

Visual traps like yellow sticky card traps are used for monitoring and control of the greenhouse whitefly (Trialeurodes vaporariorum). However, reflected intensity (brightness) and hence, attractiveness depend on the ambient light conditions, and the colour (wavelength) might not fit with the sensitivity of whitefly photoreceptors. The use of light emitting diodes (LEDs) is a promising approach to increase the attractiveness, specificity and adaptability of visual traps. We constructed LED‐based visual traps equipped with blue and green high‐power LEDs and ultraviolet (UV) standard LEDs according to the putative spectral sensitivities of the insects' photoreceptors. In a series of small‐scale choice and no‐choice recapture experiments, the factors time of day as well as light intensity and light quality (colour) of LED traps were studied in terms of attractiveness compared to yellow traps without LEDs. Green LED traps (517 nm peak wavelength) were comparably attractive in no‐choice experiments but clearly preferred over yellow traps in all choice experiments. The time of day had a clear effect on the flight activity of the whiteflies and thereby on the trapping success. Blue LEDs (474 nm) suppressed the attractiveness of the light traps when combined with green LEDs suggesting that a yet undetected photoreceptor, sensitive for blue light, and an inhibiting interaction with the green receptor, might exist in T. vaporariorum. In choice experiments between LED traps emitting green light only or in combination with UV (368 nm), the green‐UV combination was preferred. In no‐choice night‐time experiments, UV LEDs considerably increased whitefly flight activity and efficacy of trapping. Most likely, the reason for the modifying effect of UV is the stimulating influence on flight activity. In conclusion, it seems that the use of green LEDs alone or in combination with UV LEDs could be an innovative option for improving attractiveness of visual traps.     

Efficiency of photosynthesis in continuous and pulsed light emitting diode irradiation

The light utilization efficiency and relative photon requirement of photosynthesis in pulsed and continuous light from light emitting diodes (LEDs) has been measured. First, we chacterized the photon requirement of photosynthesis from light of LEDs that differ in spectral quality. A photon requirement of 10.3±0.4 was measured using light from a 658 nm peak wavelength (22 nm half band width) LED over the range of 0–50 mol photons m^–2 s^–1 in 2 kPa O₂ in leaves of tomato (Lycopersicon esculentum Mill., cv. VF36). Because the conversion of electrical power to photons increased with wavelength, LED lamps with peak photon output of 668 nm were most efficient for converting electricity to photosynthetically fixed carbon. The effect of pulsed irradiation on photosynthesis was then measured. When all of the light to make the equivalent of 50 mol photons m^–2 s^–1 was provided during 1.5 s pulses of 5000 mol photons m^–2 s^–1 followed by 148.5 s dark periods, photosynthesis was the same as in continuous 50 mol photons m^–2 s^–1. When the pulse light and dark periods were lengthened to 200 s and 19.8 ms, respectively, photosynthesis was reduced, although the averaged photon flux density was unchanged. Under these conditions, the light pulses delivered 10¹⁷ photons m^–2, which we calculate to be equivalent to the capacitance of PS I or PS II. Data support the theory that photons in pulses of 100 s or shorter are absorbed and stored in the reaction centers to be used in electron transport during the dark period. When light/dark pulses were lengthened to 2 ms light and 198 ms dark, net photosynthesis was reduced to half of that measured in continuous light. Pigments of the xanthophyll cycle were not affected by any of these pulsed light treatments even though zeaxanthin formation occurred when leaves were forced to dissipate an equal amount of continuous light.Abbreviations CWF cool white fluorescent - EPS xanthophyll epoxidation state - LED light emitting diode - LUE light utilization efficiency - PFD photon flux density - PR photon requirement (for CO₂ fixation) - PS II primary donor in Photosystem II - RPR relative photon requirement     

Blue organic light‐emitting diodes based on fluorene‐bridged quinazoline and quinoxaline derivatives

Two blue emitters based on fluorene‐bridged quinazoline and quinoxaline derivatives were prepared via the Suzuki reaction. Their photoluminescent properties were investigated. Furthermore, theoretical studies on these materials using the density functional theory calculation were conducted. To explore their electroluminescent properties, multilayered organic light‐emitting diodes were fabricated with the following device structure: indium–tin–oxide (180 nm)/4,4′‐bis(N‐(1‐naphthyl)‐N‐phenylamino)biphenyl (50 nm)/blue emitting materials ( 1 and 2 ) (30 nm)/bathophenanthroline (35 nm)/8‐hydroxy‐quinolinato lithium (2 nm)/Al (100 nm). Two devices showed efficient blue emission with the external quantum efficiencies of 1.58% and 1.30%, respectively, at 20 mA/cm², and Commission Internationale dÉclairage coordinates of (0.18, 0.24) and (0.19, 0.27) at 6.0 V. These results suggest that the self‐aggregation properties of emitters would have considerable effects on their photoluminescent and electroluminescent properties.     

Plant regeneration from cotyledon tissues of common buckwheat (Fagopyrum esculentum Moench)

Summary Plants were regenerated from cotyledon tissue of greenhouse grown seedlings of common buckwheat (Fagopyrum esculentum Moench.). Maximum callus regeneration was induced on Murashige and Skoog (MS) medium containing 2,4-D (2.0 mg l⁻¹) and kinetin (KIN) (0.2 mg l⁻¹) and either 3 or 6% sucrose. Friable callus was transferred to MS media containing KIN and benzylaminopurine (BAP) at varied concentrations for embryogenic callus induction. The optimum medium for embryogenic callus induction was found to be MS medium supplemented with 0.2 mg l⁻¹ KIN, 2.0 mg l⁻¹ BAP and 3% (w/v) sucrose. Variation of sucrose from 3 to 6% did not show any significant effect on callus induction or embryogenesis. Regeneration of embryonic callus varied from 13 to 32%. Whole plants were obtained at high frequencies when the embryogenic calluses with somatic embryos and organized shoot primordia were transferred to half-strength MS media with 3% sucrose. Regenerated plants after acclimation were transferred to greenhouse conditions, and both vegetative and floral characteristics were observed for variation. This regeneration system may be valuable for genetic transformation and cell selection in common buckwheat.     

Determination of riboflavin in urine by capillary electrophoresis-blue light emitting diode-induced fluorescence detection combined with a stacking technique

A simple, inexpensive and reliable method for the simultaneous, routine analysis of riboflavin in urine by capillary electrophoresis-light emitting diode (LED)-induced fluorescence detection is described. Using a blue LED as the light source, the detection limit of riboflavin was determined to be 0.48 microg/ml and was improved to 20 ng/ml when a stacking technique was applied. In the analysis of an actual sample, various concentrations of riboflavin were distributed in the urine samples over a period of 9 h after the ingestion of a vitamin B(2) tablet.     

Retinal damage induced by commercial light emitting diodes (LEDs)

Spectra of “white LEDs” are characterized by an intense emission in the blue region of the visible spectrum, absent in daylight spectra. This blue component and the high intensity of emission are the main sources of concern about the health risks of LEDs with respect to their toxicity to the eye and the retina. The aim of our study was to elucidate the role of blue light from LEDs in retinal damage. Commercially available white LEDs and four different blue LEDs (507, 473, 467, and 449 nm) were used for exposure experiments on Wistar rats. Immunohistochemical stain, transmission electron microscopy, and Western blot were used to exam the retinas. We evaluated LED-induced retinal cell damage by studying oxidative stress, stress response pathways, and the identification of cell death pathways. LED light caused a state of suffering of the retina with oxidative damage and retinal injury. We observed a loss of photoreceptors and the activation of caspase-independent apoptosis, necroptosis, and necrosis. A wavelength dependence of the effects was observed. Phototoxicity of LEDs on the retina is characterized by a strong damage of photoreceptors and by the induction of necrosis.     

Anther culture and androgenetic plant regeneration in buckwheat (Fagopyrum esculentum Moench)

Anther culture for haploid induction of buckwheat was studied over a period of five years. Approximately 24,000 anthers were isolated and cultured on different culture media. The regeneration capacity was generally very low. Data are presented for experiments that included 7278 anthers on which 99 calluses were formed and 20 buds regenerations were noted. Regeneration occurred most readily on gellan-gum solidified media, with 90 g l^-1 maltose, 2.5 mg l^-1 BA, 0.5 mg l^-1 IAA, and preferably in darkness. Haploid cells, as established by chromosome counts, were observed in eight regenerants. Several abnormalities of pollen development in vitro were detected. Starch presence in pollen as a possible sign of androgenic capacity was studied. Microspores in uninucleate and early binucleate stages contained only proplastids, while in adult pollen grains a number of amyloplasts were present.Abbreviations BA benzyladenine - IAA indole-3-acetic acid - 2,4 D-2,4-dichloro-phenoxyacetic acid - IBA indole-3-butyric acid - 2iP 6- c,c-dimethylallylaminol-purine - NAA -naphthalene acetic acid     

Isolation and properties of a metalloproteinase from buckwheat (Fagopyrum esculentum) seeds.

A homogeneous preparation of metalloproteinase, purified 1000-fold, was obtained from buckwheat (Fagopyrum esculentum) seeds. The Mr of the enzyme, determined by SDS/PAGE, was 34,000 (it was 39,000 by gel chromatography). Its pH optimum was 8.0-8.2 with 13 S globulin, from buckwheat seeds, as substrate. Atomic-absorption spectroscopy revealed the presence of one Zn2+ ion per enzyme molecule. The enzyme was completely inhibited by EDTA (1 mM), zincone (1 mM) and 1, 10-phenanthroline (1 mM). The metalloproteinase performed limited proteolysis of the following seed storage proteins: 13 S globulin from buckwheat seeds and 11 S globulin from soybean (Glycine max) seeds. It hydrolysed three peptide bonds formed by the amino groups of Leu15, Tyr16 and Phe25 in the oxidized B-chain of insulin. In its main properties the enzyme is similar to metalloproteinases of animal and bacterial origin.     

Genes outside the S supergene suppress S functions in buckwheat (Fagopyrum esculentum)

BACKGROUND AND AIMS: Common buckwheat (Fagopyrum esculentum) is a dimorphic self-incompatible plant with either pin or thrum flowers. The S supergene is thought to govern self-incompatibility, flower morphology and pollen size in buckwheat. Two major types of self-fertile lines have been reported. One is a type with long-homostyle flowers, Kyukei SC2 (KSC2), and the other is a type with short-homostyle flowers, Pennline 10. To clarify whether the locus controlling flower morphology and self-fertility of Pennline 10 is the same as that of KSC2, pollen tube tests and genetic analysis have been performed. METHODS: Pollen tube growth was assessed in the styles and flower morphology of KSC2, Pennline 10, F1 and F2 plants that were produced by the crosses between plants with pin or thrum and Pennline 10. KEY RESULTS: Pollen tubes of Pennline 10 reached ovules of all flower types. The flower morphology of F1 plants produced by the cross between thrum and Pennline 10 were thrum or pin, and when pin plants were used as maternal plants, all the F1 plants were pin. Both plants with pin or short-pin flowers, whose ratio of style length to anther height was smaller than that of pin, appeared in F2 populations of thrum x Pennline 10 as well as in those of pin x Pennline 10. CONCLUSION: The results suggest that Pennline 10 possesses the s allele as pin does, not an allele produced by the recombination in the S supergene, and that the short style length of Pennline 10 is controlled by multiple genes outside the S supergene.     

Agrobacterium-mediated transformation and plant regeneration of buckwheat (Fagopyrum esculentum Moench.)

Genetic transformation of buckwheat (Fagopyrum esculentum Moench.) and regeneration of transgenic plants were obtained by using Agrobacterium tumefaciens strains as vectors. Buckwheat cotyledons were excised from imbibed seeds, co-cultivated with A. tumefaciens and subjected to previously reported protocols for callus and shoot regeneration. The transformation with oncogenic strains was confirmed by opine and DNA analyses of tumour tissue extracts. Plants were regenerated on cotyledon fragments incubated with strain A281, harboring pGA472, which carries the neomycin phosphotransferase II gene for kanamycin resistance. The transformation of resistant shoot clones was confirmed by NPTII enzyme assay and DNA hybridization. A large number of transformed shoots were rooted and fertile plantlets were raised in the greenhouse. Transgenic plants comprised pin and thrum clones, which were allowed to cross-pollinate. In about 180 R₂ seeds tested for kanamycin resistance, the ratio of resistant to sensitive seedlings was roughly 3:1.Abbreviations BAP 6-benzylaminopurine - 2,4-D dichloro-phenoxyacetic acid - 2iP 6-(, ,-dimethylallyl-amino)-purine - IBA indole-3-butyric acid - IAA indole-3-acetic acid - Km kanamycin - NPTII neomycin phosphotransferase II     

Spatial characteristics of aluminum uptake and translocation in roots of buckwheat (Fagopyrum esculentum)

The detoxification of aluminum (Al) in root tips of the Al accumulator buckwheat by exudation of oxalate leading to reduced Al uptake (Al resistance) is difficult to reconcile with the Al accumulation (Al tolerance). The objective of this study was to analyze resistance and tolerance mechanisms at the same time evaluating particularly possible stratification of Al uptake, Al transport and oxalate exudation along the root apex. The use of a minirhizotron made it possible to differentiate between spatial responses to Al along the root apex with regard to Al uptake and organic acid anion exudation, but also to measure at the same time Al and organic acid transport in the xylem. Al accumulates particularly in the 3‐mm root apex. The study showed that Al taken up by the 10‐mm root apex is rapidly transferred to the xylem which differentiates in the 10 to 15‐mm root zone as revealed by a microscopic study. Al induces the release of oxalate from the root apex but particularly from the subapical 6–20 mm root zone even when Al was applied only to the 5‐mm root apex suggesting a basipetal signal transduction. Citrate proved to be the most likely ligand for Al in the xylem because Al and citrate transport rates were positively correlated. In conclusion, the data presented show that the Al‐induced release of oxalate, and Al uptake as well as Al accumulation are spatially not separated in the root apex.     

Luminescent properties of MAl(SO4)2Br:Eu3+ (M = Sr or Mg) red phosphors for near‐UV light‐emitting diodes

Eu³⁺‐activated MAl(SO₄)₂Br phosphors (where M = Mg or Sr) are successfully prepared using a wet chemical reaction technique. The samples are characterized by X‐ray diffraction (XRD) and photoluminescence (PL) spectroscopies. The XRD pattern revealed that both the samples are microcrystalline in nature. PL of Eu³⁺‐doped SrAl(SO₄)₂Br and MgAl(SO₄)₂Br phosphors exhibited characteristic red emission coming from the ⁵D₀ → ⁷F₂ (616 nm) electron transition, when excited by 396 nm wavelength of light. The maximum intensity of luminescence was observed at a concentration of 1 mol% Eu³⁺. The intensity of the electric dipole transition at 616 nm is greater than that of the magnetic dipole transition at 594 nm. The results showed that MAl(SO₄)₂Br:Eu³⁺, (M = Mg, Sr) phosphors have potential application in near‐UV light‐emitting diodes as efficient red‐emitting phosphor. Copyright © 2014 John Wiley & Sons, Ltd.