Stable isotope labelling and zinc distribution in grains studied by laser ablation ICP-MS in an ear culture system reveals zinc transport barriers during grain filling in wheat.

Zinc (Zn) deficiency has been recognized as a potential risk for human health in many developing regions where staple food with low micronutrient density represents a major proportion of the diet. The success of strategies to increase Zn content in the edible part of crops requires better understanding of Zn transport to, and distribution within, the grains. The transfer of Zn from the growth medium to wheat (Triticum aestivum) grains in an ear culture system was investigated by using the stable Zn isotope (70) Zn, and the spatial distribution of Zn within the grains was studied by laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS). Zinc was readily transported in the stem up to the rachis. More Zn accumulated in the stem when higher amounts of Zn were supplied to the medium. Once Zn was transported into the grain, Zn accumulated particularly in the crease vascular tissue. The gradient of (70) Zn concentration between crease vascular tissue, aleurone layer and endosperm demonstrates that Zn is distributed within grain through the crease phloem. These results suggest that two barriers of Zn transport into wheat grains may exist: between the stem tissue rachis and the grain, and the maternal and filial tissues in the grain.

Biofortification and localization of zinc in wheat grain.

Zinc (Zn) deficiency associated with low dietary intake is a well-documented public health problem, resulting in serious health and socioeconomic problems. Field experiments were conducted with wheat to test the role of both soil and foliar application of ZnSO4 in Zn concentration of whole grain and grain fractions (e.g., bran, embryo and endosperm) in 3 locations. Foliar application of ZnSO4 was realized at different growth stages (e.g., stem elongation, boot, milk, dough stages) to study the effect of timing of foliar Zn application on grain Zn concentration. The rate of foliar Zn application at each growth stage was 4 kg of ZnSO4·7H2O ha(-1). Laser ablation (LA)-ICP-MS was used to follow the localization of Zn within grain. Soil Zn application at a rate of 50 kg of ZnSO4·7H2O ha(-1) was effective in increasing grain Zn concentration in the Zn-deficient location, but not in the locations without soil Zn deficiency. In all locations, foliar application of Zn significantly increased Zn concentration in whole grain and in each grain fraction, particularly in the case of high soil N fertilization. In Zn-deficient location, grain Zn concentration increased from 11 mg kg(-1) to 22 mg kg(-1) with foliar Zn application and to 27 mg kg(-1) with a combined application of ZnSO4 to soil and foliar. In locations without soil Zn deficiency, combination of high N application with two times foliar Zn application (e.g., at the booting and milk stages) increased grain Zn concentration, on average, from 28 mg kg(-1) to 58 mg kg(-1). Both ICP-OES and LA-ICP-MS data showed that the increase in Zn concentration of whole grain and grain fractions was pronounced when Zn was sprayed at the late growth stage (e.g., milk and dough). LA-ICP-MS data also indicated that Zn was transported into endosperm through the crease phloem. To our knowledge, this is the first study to show that the timing of foliar Zn application is of great importance in increasing grain Zn in wheat, especially in the endosperm part that is the predominant grain fraction consumed in many countries. Providing a large pool of Zn in vegetative tissues during the grain filling (e.g., via foliar Zn spray) is an important practice to increase grain Zn and contribute to human nutrition.

Aluminium rhizotoxicity in maize grown in solutions with Al3+ or Al(OH)-4 as predominant solution Al species.

The rhizotoxicity of aluminium at low-pH with Al(3+) and at high pH with Al(OH)-(4) as the main Al species was studied. Aluminium reduced root growth to similar levels at pH 8.0 and pH 4.3, although the mononuclear Al concentration at pH 8.0 was three times lower than at pH 4.3. Al contents of root apices were much higher at pH 8 than at pH 4.3. Callose was induced only marginally at pH 8 and the formation was confined to the epidermis, whereas it proceeded through the cortex with time at pH 4.3. Well-documented genotypical differences in callose formation and Al accumulation could not be found at pH 8. The largest fraction of the root-tip Al was recovered in the cell-wall fraction independent of the solution pH. A sequential extraction of isolated cell walls suggests that most of the cell-wall Al was precipitated Al(OH)(3) at pH 8.0. This can be explained by a drastic pH reduction in the root apoplastic sap to 6.2, whereas at bulk solution pH 4.3 it rose to 5.6. Al precipitation was also confirmed by the microscopic localization of Al. At pH 8, Al could mostly be found in the epidermis, but in the apoplast of the outer cortex at pH 4.3. It is proposed here that at pH 4.3, Al(3+) inhibits root growth through binding to sensitive binding sites in the apoplast of the epidermis and the outer cortex. At pH 8, Al(OH)(3) precipitation in the epidermis causes a mechanical barrier thus impairing the root-growth control of the epidermis.

Aluminum activates a citrate-permeable anion channel in the aluminum-sensitive zone of the maize root apex. A comparison between an aluminum- sensitive and an aluminum-resistant cultivar.

In search for the cellular and molecular basis for differences in aluminum (Al) resistance between maize (Zea mays) cultivars we applied the patch-clamp technique to protoplasts isolated from the apical root cortex of two maize cultivars differing in Al resistance. Measurements were performed on protoplasts from two apical root zones: The 1- to 2-mm zone (DTZ), described as most Al-sensitive, and the main elongation zone (3-5 mm), the site of Al-induced inhibition of cell elongation. Al stimulated citrate and malate efflux from intact root apices, revealing cultivar differences. In the elongation zone, anion channels were not observed in the absence and presence of Al. Preincubation of intact roots with 90 microM Al for 1 h induced a citrate- and malate-permeable, large conductance anion channel in 80% of the DTZ protoplasts from the resistant cultivar, but only 30% from the sensitive cultivar. When Al was applied to the protoplasts in the whole-cell configuration, anion currents were elicited within 10 min in the resistant cultivar only. La3+ was not able to replace or counteract with Al3+ in the activation of this channel. In the presence of the anion-channel blockers, niflumic acid and 4, 4'-dinitrostilbene-2, 2'disulfonic acid, anion currents as well as exudation rates were strongly inhibited. Application of cycloheximide did not affect the Al response, suggesting that the channel is activated through post-translational modifications. We propose that the Al-activated large anion channel described here contributes to enhanced genotypical Al resistance by facilitating the exudation of organic acid anions from the DTZ of the maize root apex.

The role of nitrogen-efficient cultivars in sustainable agriculture.

To improve nitrogen (N) efficiency in agriculture, integrated N management strategies that take into consideration improved fertilizer, soil, and crop management practices are necessary. This paper reports results of field experiments in which maize (Zea mays L.) and oilseed rape (Brassica napus L.) cultivars were compared with respect to their agronomic N efficiency (yield at a given N supply), N uptake efficiency (N accumulation at a given N supply), and N utilization efficiency (dry matter yield per unit N taken up by the plant). Under conditions of high N supply, significant differences among maize cultivars were found in shoot N uptake, soil nitrate depletion during the growing season, and the related losses of nitrate through leaching after the growing season. Experiments under conditions of reduced N supply indicated a considerable genotypic variation in reproductive yield formation of both maize and oilseed rape. High agronomic efficiency was achieved by a combination of high uptake and utilization efficiency (maize), or exclusively by high uptake efficiency (rape). N-efficient cultivars of both crops were characterized by maintenance of a relatively high N-uptake activity during the reproductive growth phase. In rape this trait was linked with leaf area and photosynthetic activity of leaves. We conclude that growing of N-efficient cultivars may serve as an important element of integrated nutrient management strategies in both low- and high-input agriculture.

Genotypical differences in aluminum resistance of maize are expressed in the distal part of the transition zone. Is reduced basipetal auxin flow involved in inhibition of root elongation by aluminum?

Short-term Al treatment (90 microM Al at pH 4.5 for 1 h) of the distal transition zone (DTZ; 1-2 mm from the root tip), which does not contribute significantly to root elongation, inhibited root elongation in the main elongation zone (EZ; 2.5-5 mm from the root tip) to the same extent as treatment of the entire maize (Zea mays) root apex. Application of Al to the EZ had no effect on root elongation. Higher genotypical resistance to Al applied to the entire root apex, and specifically to the DTZ, was expressed by less inhibition of root elongation, Al accumulation, and Al-induced callose formation, primarily in the DTZ. A characteristic pH profile along the surface of the root apex with a maximum of pH 5.3 in the DTZ was demonstrated. Al application induced a substantial flattening of the pH profile moreso in the Al-sensitive than in the Al-resistant cultivar. Application of indole-3-acetic acid to the EZ but not to the meristematic zone significantly alleviated the inhibition of root elongation induced by the application of Al to the DTZ. Basipetal transport of exogenously applied [(3)H]indole-3-acetic acid to the meristematic zone was significantly inhibited by Al application to the DTZ in the Al-sensitive maize cv Lixis. Our results provide evidence that the primary mechanisms of genotypical differences in Al resistance are located within the DTZ, and suggest a signaling pathway in the root apex mediating the Al signal between the DTZ and the EZ through basipetal auxin transport.

Accumulation of zinc and Organic Acids in Roots of Zinc Tolerant and Non-tolerant Ecotypes of Deschampsia caespitosa.

A much higher zinc level was necessary to inhibit root elongation in the zinc tolerant ecotype as compared to the non-tolerant ecotype of Deschampsia caespitosa. In the presence of a range of high levels of zinc, zinc accumulated to a much higher concentration in the roots of the tolerant ecotype, especially in the root sap. Accumulation of citrate in the root sap was highly correlated to the accumulation of zinc. Gel filtration chromatography of the root sap showed zinc to be mainly present as zinc-citrate. This was the only zinc complex found. The malate concentration of the root sap was much lower than the concentration of citrate. However the malate content of aqueous root homogenates was comparable or even greater than the content of citrate, suggesting that malate and citrate are located in different compartments within the cell. The results are consistent with a model of zinc tolerance in which zinc is complexed with citrate in the vacuole.