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.     

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.

     

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.     

Dietary modulation of α-amylase activity in eight geographical strains of Sitophilus oryzae and Sitophilus zeamais

Rank transformation of specific activity values of -amylase across four strains of Sitophilus oryzae (L.) and four strains of S. zeamais Motschulsky indicates that levels of these predominant enzymes are highest in adults feeding on hulled barley or long-grain brown rice. Intermediate activity levels are found in weevils feeding on yellow corn (maize) and lowest levels are found in wheat-fed weevils. Although extracts prepared from barley contain inhibitory activity against two purified isoamylases from S. oryzae, levels of the naturally-occurring -amylase inhibitors against these two enzymes are about 2.2-fold and 6.1-fold, respectively, more concentrated in wheat. Ingestion of these amylase inhibitors and formation of an inactive enzyme:inhibitor complex with previously secreted amylase may account for the lower activity of amylase in weevils of both species feeding on wheat. Amylase levels across all strains feeding on a given diet are about 2-fold higher in S. oryzae than in S. zeamais. Significant differences in activity levels were also found between strains in both species. Since -amylase is a predominant digestive hydrolase in these species, the degree to which cereal diets affect amylase levels may indicate their suitability as potential hosts.

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Résumé La transformation de rang des valeur d'activité spécifique de l'-amylase de 4 souches de S. oryzae et de 4 souches de S. zeamais montre que les niveaux les plus élevés de ces enzymes prédominantes s'observent chez les adultes nourris d'orge mondé ou de riz brun á grains longs. Des niveaux intermédiaires d'activité ont été obtenus chez les insectes élevés sur maïs jaune, et les niveaux les plus faibles chez ceux élevés sur blé. Bien que les extraits préparés à partir d'orge présentent une activité inhibitrice de deux isoamylases purifiées de S. oryzae, les niveaux des inhibiteurs naturels -amylase de ces deux enzymes sont environ respectivement 2,2 et 6,1 fois plus concentrés dans le blé. L'ingestion de ces inhibiteurs d'amylase et la formation d'un complexe enzyme inactive/inhibiteur avec l'amylase secrétée antérieurement, peut rendre compte de la plus faible activité de l'amylase chez les charançons consommant du blé. Le niveau d'amylase de S. oryzae est 2 fois plus élevé que celui de S. zeamais pour toutes les souches élevées sur un régime donné. Des niveaux d'activité significativement différents ont été trouvés suivant les souches pour chacune des deux espèces. Puisque l'amylase est la principale hydrolase digestive de ces espèces, l'intensité de la modification des teneurs en amylase par la consommation de céréales peut indiquer leur adéquation comme hôtes potentiels.

     

Channel formation in phospholipid bilayer membranes by the toxin ofHeminthosporium maydis,race T

Summary Southern Corn Leaf Blight is caused by a toxin produced by a virulent form ofHelminthosporium maydis (Race T). The toxin has been shown to uncouple oxidative phosphorylation and dissipate Ca²⁺ gradients in mitochondria isolated from susceptible, but not resistant, corn. The possibility that the toxin acted by increasing the permeability of membranes to ions was tested using a planar bilayer membrane system. Addition of the toxin to the bilayer system, under voltage-clamp conditions, resulted in stepwise increases in current across the phospholipid bilayer, a response characteristic for channel formers. Single-channel conductance in 1m KCl is 27 pS which corresponds to 1.7×10⁷ ions sec^–1 channel^–1 at 100 mV applied potential. The toxin channels are: (i) fairly uniform in conductance, (ii) ideally selective for K⁺ over Cl^–, and (iii) most conductive to H⁺. The channel showed the following selectivity for alkali metal cations: Rb⁺>K⁺>Cs⁺>Na⁺>Li⁺ (169731) based on the most frequently observed conductance in 1m chloride salts. The toxin showed no voltage dependence over the range of –100 to +100 mV. Channel formation was also a property of a synthetic analog (Cmpd IV) of the toxin. The ability of the native toxin to form channels may be a mode of toxin action on mitochondrial membranes from susceptible corn.     

The fate of immunoglobulin G fed to larvae of Ostrinia nubilalis

Since plants can be transformed genetically to produce functional antibodies, an immunological approach may be developed for controlling their arthropod pests. Specific antibodies would protect plants from arthropods if they could gain access to the pest antigen in sufficient amounts such that the normal function of the antigen is disrupted. In order to study the fate of ingested antibodies in the body of the European corn borer (ECB), Ostrinia nubilalis (Hübner), (Lepidoptera: Pyralidae), we fed the larvae on serum-containing diet. When larvae were fed on the serum-containing diet for various lengths of time between 12 and 96 h, no significant differences were noted in the immunoglobulin G (IgG) concentration in their body. Immediately after the larvae stopped feeding, the concentrations of the IgG in their midgut was about one half that of the diet itself, but it decreased significantly after 6 h and again after 18 h (about 3 and 10 fold, respectively). Immediately after the larvae stopped feeding, the concentration of the IgG in their hemolymph was about 1/500 that in the diet. The concentration of IgG in the hemolymph of ECB larvae was influenced directly by the titer of antibodies in their diet. During the first 6 h after the larvae stop feeding the concentration of IgG in their hemolymph did not decrease significantly; however, it did so after 18 h (about 6 fold). The possibility that specific antibodies will gain access to antigens in the ECB body is discussed.     

The effects of cropping corn on the extractability of zinc added to a calcareous soil

The effects of cropping corn on the decrease in the extractability of Zn added to a calcareous soil were studied by a pot experiment and chemical extractions. The results show that the concentrations of Zn exchangeable with MgCl₂ (EXC-Zn) and extractable with DTPA (DTPA-Zn) in the soils with added Zn decreased with time. The processes associated with the decrease in extractability in DTPA of Zn added to soil can be described aptly by a diffusion equation which gives the proportion of added Zn in the non-DTPA fraction as a function of the square root of incubation time. This result suggests that the diffusion of Zn cations into microporous solids is a rate-limiting reaction. The relative diffusion rate coefficients (D/r²) were found to be 1.95×10^-10 and 3.34×10^-10 sec^-1 in the soils with added Zn of 20 and 60 mg kg^-1, respectively. Compared with uncropped soil, the concentrations of DTPA-Zn in the soils with added Zn were decreased by cropping. The decrease of DTPA-Zn in the soils in the presence of corn can be attributed to both its acquisition by corn and other processes associated with the growing of corn. The activity of plant roots would appear to enhance the process of decrease in the extractability in DTPA of Zn added to the soil. The source of Zn uptake by corn was affected by the loading or activity of Zn in soil. In the soil with low available Zn, the DTPA non-extractable Zn (non-DTPA-Zn) was mobilized and taken up by corn. In the soils with high available Zn, e.g. the recently added Zn, only EXC-Zn and a part of the DTPA-Zn were taken up by corn.     

Deoxynivalenol and contaminant mycoflora in freshly harvested Argentinian wheat in 1993

A mycological survey was carried out on wheat heads from the main production area of Argentina. The isolation frequency and relative density of species from genus Fusarium and dematiaceous fungi were calculated. F graminearum was the predominant Fusarium species; similar to that observed in the USA and Canada. An analysis of deoxynivalenol (DON) natural contamination also was performed on a limited number (44) of samples. DON contamination levels in positive samples ranged from 0.2 to 30 ppm. A stepwise regression procedure showed that, among the species analysed, F. graminearum relative density was related to the DON contamination level and that other prevalent fungi did not influence or modify that relationship.     

Changes in carbon storage in temperate humic loamy soils after forest clearing and continuous corn cropping in France

Soil samples from forest and agricultural sites in three areas of southwest France were collected to determine the effect of forest conversion to continuous intensive corn cropping with no organic matter management on soil organic carbon (C) content. Soils were humic loamy soils and site characteristics that may affect soil C were as uniform as possible (slope, elevation, texture, soil type, vegetation). Three areas were selected, with adjacent sites of various ages of cultivation (3 to 35 yr), and paired control forest sites. The ploughed horizon (0-Dt cm) and the Dt-50 cm layer were collected at each agricultural site. In forest sites, each 10 cm layer was collected systematically down to 1 meter depth. Carbon concentrations were converted to total content to a given depth as the product of concentration, depth of sample and bulk density, and expressed in units of kg m^-2. For each site and each sampled layer, the mineral mass of soil was calculated, in order to base comparisons on the same soil mass rather than the same depth. The pattern of C accumulation in forest soils showed an exponential decrease with depth. Results suggested that soil organic carbon declined rapidly during the first years of cultivation, and at a slower rate thereafter. This pattern of decrease can be fitted by a bi-exponential model assuming that initial soil organic carbon can be separated into two parts, a very labile pool reduced during the first rapid decline and more refractory fractions oxidizing at a slower rate. Sampling to shallow depths (0-Dt cm) resulted in over-estimation of the rate of carbon release in proportion to the initial amount of C, and in under-estimation of the total loss of C with age. The results for the 0–50 cm horizon indicated that losses of total carbon average about 50% in these soils, ranging in initial carbon content from 19 to 32.5 kg m^-2. Carbon release to the atmosphere averaged 0.8 kg m^-2 yr^-1 to 50 cm depth during the first 10 years of cultivation. The results demonstrate that temperate soils may also be an important source of atmospheric carbon, when they are initially high in carbon content and then cultivated intensively with no organic matter management.