Single pot sequential crystallization-distillation as a new purification procedure

A new purification procedure exploiting the simultaneous presence of a solid, liquid, and gas phase in a low surface area system is proposed and discussed. The assumptions of vanishingly low diffusion coefficients in the solid phase and that of the presence of a single “effective impurity” allow to plan the sequence of operations starting from the knowledge of just the melting and boiling points of the substance to be purified and of those of the “effective impurity”. Examples and results are presented.     

A new precipitation technique provides evidence for the permeability of Casparian bands to ions in young roots of corn (Zea mays L.) and rice (Oryza sativa L.)

Using an insoluble inorganic salt precipitation technique, the permeability of cell walls and especially of endodermal Casparian bands (CBs) for ions was tested in young roots of corn (Zea mays) and rice (Oryza sativa). The test was based on suction of either 100 µm CuSO₄ or 200 µm K₄[Fe(CN)₆] into the root from its medium using a pump (excised roots) or transpirational stream (intact seedlings), and subsequent perfusion of xylem of those root segments with the opposite salt component, which resulted in precipitation of insoluble brown crystals of copper ferrocyanide. Under suction, Cu²⁺ could cross the endodermis apoplastically in both plant species (although at low rates) developing brown salt precipitates in cell walls of early metaxylem and in the region between CBs and functioning metaxylem vessels. Hence, at least Cu²⁺ did cross the endodermis dragged along with the water. The results suggested that CBs were not perfect barriers to apoplastic ion fluxes. In contrast, ferrocyanide ions failed to cross the mature endodermis of both corn and rice at detectable amounts. The concentration limit of apoplastic copper was 0.8 µm at a perfusion with 200 µm K₄[Fe(CN)₆]. Asymmetric development of precipitates suggested that the cation, Cu²⁺, moved faster than the anion, [Fe(CN)₆]^4–, through cell walls including CBs. Using Chara cell wall preparations (‘ghosts’) as a model system, it was observed that, different from Cu²⁺, ferrocyanide ions remained inside wall-tubes suggesting a substantially lower permeability of the latter which agreed with the finding of an asymmetric development of precipitates. In both corn and rice roots, there was a significant apoplastic flux of ions in regions where laterals penetrated the endodermis. Overall, the results show that the permeability of CBs to ions is not zero. CBs do not represent a perfect barrier for ions, as is usually thought. The permeability of CBs may vary depending on growth conditions which are known to affect the intensity of formation of bands.     

Sodium-induced calcium deficiency in salt-stressed corn

Abstract The effect of the Na⁺/Ca²⁺ ratio in the root media on salt-stressed corn (Zea mays L. cvs DeKalb XL-75 and Pioneer 3906) was determined in greenhouse experiments. Plants grown in a complete nutrient solution salinized with 86.5 mol m^?3 NaCl exhibited severe Ca²⁺ deficiency symptoms at the four-leaf stage. The symptoms disappeared when part of the NaCl was replaced with 10 mol m^?3 CaCl₂ (Na⁺/Ca²⁺ molar ratio = 5.7). Salt stress at an iso-osmotic potential of ?0.4 MPa substantially decreased shoot growth at all solution Na⁺/Ca²⁺ ratios from 34.6 to 0.26. However, the dry weights of blades at 26 d of age were much less when plants were salinized with NaCl alone, particularly that of DeKalb XL-75 which was more susceptible to Na-induced Ca²⁺ deficiency than was Pioneer 3906. The growth of sheaths was similarity reduced by sail stress at all Na⁺/Ca²⁺ ratios. The symptoms of Ca²⁺ deficiency were correlated with low Ca²⁺ concentrations in the leaf tissue. Ca²⁺ concentrations in the developing blades of NaCl-stressed plants were much lower than in control plants. As the Na⁺/Ca²⁺ ratio in the solution was decreased, Ca²⁺ levels increased in both the blades and sheaths while Na⁺ concentrations greatly decreased. DeKalb XL-75 was much less effective than Pioneer 3906 in restricting the uptake of Na⁺. The results clearly indicate that NaCl stress may cause lesions and unique plant responses that are not manifested on agronomic plants grown on saline soils.     

Photoinactivation of δ-aminolevulinic acid dehydratase from maize by flavin mononuclotide

The -aminolevulinic acid dehydratase activity was irreversibly inactivated by irradiation of the enzyme in presence of flavin mononucleotide. The loss of enzyme activity was dependent on time of irradiation, concentration of FMN and intensity of irradiance. It required oxygen and was markedly enhanced in heavy water. The presence of levulinic acid (a competitive inhibitor of -ALAD) during irradiation prevented the inactivation considerably indicating photooxidative damage at or near the active site. Superoxide dismutase, sodium benzoate and sodium formate offered no protection, but singlet oxygen quenchers like azide and tryptophan were effective. NADH, electron donor to excited flavins, also prevented the loss of enzyme activity. These results indicate that singlet oxygen produced by light absorption of FMN was responsible for the photooxidative inhibition of the enzyme.Abbreviations ALAD -aminolevulinic acid dehydratase - FMN flavin mononucleotide - O₂ ^- superoxide - H₂O₂ hydrogen peroxide - 10₂ singlet oxygen - LA levulinic acid - PBG porphobilinogen - BSA bovine serum albumin - BME 2-mercaptoethanol - SOD superoxide dismutase - pHMB para-hydroxymercuribenzoate - DTT dithiothreitol - FAD flavin adenine dinucleotide - NADH nicotinamide adenine dinucleotide     

Anaerobic metabolism enzymes as markers of flooding stress in maize seeds

Maize (Zea mays L.) seeds differ in their relative tolerance to the anaerobic environment caused by flooding. Seed tolerance to flooding stress depends on cellular and metabolic processes since gross anatomical responses have not developed at the pre-emergence stage. The study reported here characterizes the activities of four anaerobic respiratory enzymes: pyruvate decarboxylase (PDC), alcohol dehydrogenase (ADH), lactate dehydrogenase (LDH), and malic enzyme (ME) in the flood-tolerant A632 and floodsusceptible Mo 17 inbred maize seeds during flooding at 10 and 25°C. Each inbred consisted of two seed lots possessing 95% and 75% germination levels. Flooding increased the activities of all four enzymes. However, no consistent correlation between anaerobic enzyme activity and flood tolerance was observed across genotype, seed quality and flooding temperature. The results indicate that it may not be feasible to use whole-seed anaerobic enzyme activities to predict maize seed performance under flooding stress. Contribution from the Soil Drainage Research Unit, USDA-ARS, Columbus, OH, in cooperation with the Ohio Agricultural Research and Development Center, The Ohio State University. OARDC Journal Article No. 66–86.     

The contribution of prehatch and posthatch development to protandry in the chrysomelid Diabrotica virgifera virgifera

Laboratory tests with eggs of Diabrotica virgifera virgifera LeConte showed that during a 10-day hatching period, hatch of male eggs predominated on the first and second days, eggs of mixed sex, with ca. 1:1 ratio, hatched on the third and fourth days, and eggs hatching from the fifth to the tenth days were nearly all female. Overall, female eggs hatched a mean of 2.9 days later than male eggs. Not only did female eggs hatch later, but the time for posthatch development to the adult stage was 1.8 days longer for females. The later egg hatch and longer posthatch development for females resulted in female adults emerging a mean of 4.7 days later than male adults. Total adult emergence lasted 14 days; of this, males predominated during the first 5 days, and females predominated during the last 9 days. Males of D. v. virgifera appear to have evolved protandry (the tendency for males to emerge before females) by developing both a postdiapause embryonic stage and a combined larval and pupal stage of shorter duration.

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Résumé L'observation, au laboratoire, pendant une période d'éclosion de dix jours, des oeufs de D. v. virgifera LeConte, a montré que les oeufs mâles prédominent les deux premiers jours d'éclosion, que les oeufs des deux sexes, avec des fréquences 0,5/0,5, ont éclos les troisième et quatrième jours, et que les oeufs éclos du cinquième au dixième jour étaient presque tous femelles. Globalement, les oeufs femelles ont éclos en moyenne 2,9 jours plus tard que les oeufs mâles. De plus, la durée du développement post-embryonnaire des femelles a demandé 1,8 jour en plus. Une éclosion plus tardive et un développement post-embryonnaire plus long ont entrainé une émergence des femelles en moyenne 4,7 jours après les mâles. La période d'émergence des adultes s'est étalée sur 14 jours; les mâles ayant dominé pendant les 5 premiers jours et les femelles pendant les 9 derniers. Les mâles de D. v. virgifera semblent avoir évolué vers la protandrie en acquerant, tant une diapause post-embryonnaire que des stades de développements larvaire et nymphal plus brefs.

     

Uptake by corn and chemical extractability of heavy metals from a four year compost treated soil

This paper gives the results from four-year field experiments on compost application, added at the maximum rate allowed by Italian legislation (30 t/ha/y). The purpose of the experiment was to evaluate any eventual heavy metal accumulation in soil and corn plants. Cadmium in corn plants increased particularly in the roots from 0.22 mg kg⁻¹ to 1.31 mg kg⁻¹, concentration of Zn and Cu increased in grains, from 26.8 to 35.8 and from 2.4 to 4.2 mg kg⁻¹ respectively. Relevant increase in the roots was detected for Zn from 34.6 to 146.8 mg kg⁻¹. Only in the 4th year Ni concentration increases in the root portion while the content of Pb and Cr in corn was generally unaffected by the compost application. Heavy metals in the soil determined by a sequential chemical extraction, indicated that extractability changed with time. A certain increase was found from the beginning to the end of the experiment particularly for Zn, from 23.3 mg kg⁻¹ to 45.1 mg kg⁻¹ in extractable forms. Nevertheless the extractable amounts are rather small in respect to the total heavy metal content of compost.     

Critical evaluation of thein situ nitrate reductase assay

Anin situ method, derived from anin vivo method, was used to determine nitrate reductase activity (NRA) in:i) excised barley and corn shoots and excised soybean leaves during a N-depletion experiment and; ii) roots and shoots of N-depleted barley and corn seedlings during induction of nitrate, reductase (NR). Nitrate reduction, calculated from thesein situ RNA measurements, was compared with estimates of each organ's nitrate reduction in light aerobic conditions from NO ₃ ⁻ consumption and a¹⁵N model (Gojonet al., 1986b). Thein situ RNA of roots strongly underestimated their¹⁵NO ₃ ⁻ reduction. In contrast, in barley and corn shoots and in the first trifoliolate leaves from 26-day-old, soybean, thein situ NRA assay gave a fair approximation of the true NO ₃ ⁻ reduction rate (relative differences ranging from −14 to +32%). In young soybean leaves (from 20-day-old plants), however, thein situ NRA strongly underestimated the actual NO ₃ ⁻ reduction. The physiological significance of thein situ NRA assay in shoots and roots, and its value for field studies are discussed from these results.     

Patch clamping corn protoplasts

Summary Cell wall regeneration around protoplasts from Black Mexican Sweet corn suspension cells has been observed using scanning electron microscopy. A coherent array of cellulose microfibrils can be seen around protoplasts two hours after they have been isolated. This array does not form in the presence of 15 mg/l Congo Red. The frequency and electrical resistance of seals made between patch clamp pipettes and the plasmalemma around corn protoplasts is not significantly affected by the presence or absence of these fibrils (p₀=0.75); it remains relatively low. Some single channel records from BMS corn protoplasts are shown.