Biofortification of wheat with iron through soil and foliar application of nitrogen and iron fertilizers

Increasing iron (Fe) concentration in food crops is an important global challenge due to high incidence of Fe deficiency in human populations. Evidence is available showing that nitrogen (N) fertilization increases Fe concentration in wheat grain. This positive impact of N on grain Fe was, however, not studied under varied soil and foliar applications of Fe. Greenhouse experiments were conducted to investigate a role of soil- and foliar-applied Fe fertilizers in improving shoot and grain Fe concentration in durum wheat (Triticum durum) grown under increasing N supply as Ca-nitrate. Additionally, an effect of foliar Fe fertilizers on grain Fe was tested with and without urea in the spray solution. Application of various soil or foliar Fe fertilizers had either a little positive effect or remained ineffective on shoot or grain Fe. By contrast, at a given Fe treatment, raising N supply substantially enhanced shoot and grain concentrations of Fe and Zn. Improving N status of plants from low to sufficient resulted in a 3-fold increase in shoot Fe content (e.g., total Fe accumulated), whereas this increase was only 42% for total shoot dry weight. Inclusion of urea in foliar Fe fertilizers had a positive impact on grain Fe concentration. Nitrogen fertilization represents an important agronomic practice in increasing grain Fe. Therefore, the plant N status deserves special attention in biofortification of food crops with Fe.     

Effect of foliar application of phosphorus on RNA and DNA under salt-stress in peanut plants (Arachis hypogaea L.)

Summary Peanut (Arachis hypogaea L., var. TMV-2) plants were raised in sand cultures salinized with sodium chloride at 0.4% on air dry weight basis. Phosphate was sprayed to the drip point, once daily for five days from 20th to 25th day and from 30th to 35th day. Shoot apices and mature leaves were harvested for nucleic acid analyses at 30th and 40th day, receiving one and two sets of sprays respectively. Salinity decreased RNA and DNA levels which were partially restored by foliar application of phosphate.     

Long-term effectiveness of vivianite in reducing iron chlorosis in olive trees

Iron (Fe) chlorosis is common in olive (Olea europaea L.) trees growing on highly calcareous soils in Southern Spain, where generally causes reduction in yield, size and commercial value of the olives. The objective of this research was to study the effectiveness of synthetic vivianite (Fe₃(PO₄)₂H₂O) to reduce Fe chlorosis in olive. Experiments were established in three orchards with cultivars `Hojiblanco', `Manzanillo', and `Picual'. The design was a randomised block design with two or three treatments (control with no Fe fertiliser and vivianite at one or two rates). A vivianite suspension (0.05 kg dm<sup>–3</sup> water) was injected into the soil at 10–20 points around the tree at the depth of maximum root density (25–35 cm). The rates (and times of application) were 0.5 and 1 kg tree<sup>–1</sup> for `Hojiblanco' (March 1997), 1 kg tree^–1 for `Manzanillo' (March 1998), and 2 kg tree<sup>–1</sup> for `Picual' (March 1998). The leaf chlorophyll content index (CCI) was estimated on the youngest expanded leaves by means of a Minolta apparatus (SPAD units). The colour index of the olives was estimated by visual comparison with a scale ranging from 1 (pale yellow) to 8 (normal green). For the period studied (July 1997–November 1999), the CCI of fertilised trees was, in general, significantly higher than that of control trees, and so was the case with the olive colour index. Olive yield, measured in the experimental fields with `Hojiblanco' (in 1999) and `Manzanillo' (in 1998 and 1999), was higher for the fertilised than for the control trees but differences were only significant in 1999. These results suggest that vivianite is effective to reduce Fe chlorosis for more than two seasons. Such effectiveness is probably due to the poorly crystalline Fe(III) oxides (which are good sources of Fe to plants) that result from the slow oxidation and incongruent dissolution of vivianite.     

Physiological responses and differential gene expression in Prunus rootstocks under iron deficiency conditions

Two Prunus rootstocks, the Myrobalan plum P 2175 and the interspecific peach-almond hybrid, Felinem, were studied to characterize their biochemical and molecular responses induced under iron-Deficient conditions. Plants of both genotypes were submitted to different treatments using a hydroponic system that permitted removal of Fe from the nutrient solution. Control plants were grown in 90 μM Fe (III)-EDTA, Deficient plants were grown in an iron free solution, and plants submitted to an Inductor treatment were resupplied with 180 μM Fe (III)-EDTA over 1 and 2 days after a period of 4 or 15 days of growth on an iron-free solution. Felinem increased the activity of the iron chelate reductase (FC-R) in the Inductor treatment after 4 days of iron deprivation. In contrast, P 2175 did not show any response after at least 15 days without iron. The induction of the FC-R activity in this genotype was coincident in time with the medium acidification. These results suggest two different mechanisms of iron chlorosis tolerance in both Strategy I genotypes. Felinem would use the iron reduction as the main mechanism to capture the iron from the soil, and in P 2175, the mechanism of response would be slower and start with the acidification of the medium synchronized with the gradual loss of chlorophyll in leaves. To better understand the control of these responses at the molecular level, the differential expression of PFRO2, PIRT1 and PAHA2 genes involved in the reductase activity, the iron transport in roots, and the proton release, respectively, were analyzed. The expression of these genes, estimated by quantitative real-time PCR, was different between genotypes and among treatments. The results were in agreement with the physiological responses observed.     

Characterization of phytosiderophore secretion under Fe deficiency stress in Festuca rubra

In the present study we investigated the response to iron (Fe) deficiency in two cultivars of Festuca rubra L. (Rubina and Barnica) used in correction of chlorosis of fruit trees cultivated on calcareous soils. We found that a Fe-chelating compound, identified as 2-deoxymugineic acid (DMA), was secreted from the roots in response to Fe-deficiency in both cultivars. The amount of DMA secreted into solution increased with the development of Fe-deficiency. The secretion showed a distinct diurnal rhythm characterized by a secretion peak at between 2 and 5 hours after sunrise at 20°C. However, this secretion peak was delayed by 3 hour at low temperature (<10°C) and occurred 3 h earlier at high temperature (30°C). When water used for the collection of root exudates was pre-warmed (25°C) or pre-cooled (10°C), this led to an earlier or a delayed secretion compared to control (15°C) under the same air temperature, respectively. Short-term shading treatment did not affect the secretion pattern of DMA. These results demonstrate that the secretion time of DMA from the roots is, at least partly controlled by the temperature in the root environment. Overall, these findings suggest that the ability of Festuca rubra to prevent Fe chlorosis symptoms (`re-greening effect') of associated fruit trees is partially related to the secretion of DMA which increase Fe availability in calcareous soils.     

Bulk soil pH and rhizosphere pH of peach trees in calcareous and alkaline soils as affected by the form of nitrogen fertilizers

One-year old nectarine trees [Prunus persica, Batsch var. nectarina (Ait.) Maxim.], cv Nectaross grafted on P.S.B2 peach seedlings [Prunus persica (L.) Batsch] were grown for five months in 4-litre pots filled with two alkaline soils, one of which was also calcareous. Soils were regularly subjected to fertigation with either ammonium sulphate or calcium nitrate providing a total of 550 mg N/tree. Trees were also grown in such soils receiving only deionized water, as controls. Rhizosphere pH, measured by the use of a microelectrode inserted in agar sheet containing a bromocresol purple as pH indicator and placed on selected roots, was decreased by about 2–3 units compared to the bulk soil pH in all treatments. This decrease was slightly less marked when plants were supplied with calcium nitrate rather than ammonium sulphate or control. Measurements conducted during the course of the experiment indicated that ammonium concentration was similar in the solution of soils receiving the two N fertilizers. During the experiment, soil solution nitrate-N averaged 115 mg L^–1 in soil fertilized with calcium nitrate, 68 mg L^–1 in those receiving ammonium sulphate and 1 mg L^–1 in control soils. At the end of the experiment nitrate concentrations were similar in soils receiving the two N sources and bulk soil pH was decreased by about 0.4 units by ammonium sulphate fertigation: these evidences suggest a rapid soil nitriflcation activity of added ammonium. Symptoms of interveinal chlorosis in apical leaves appeared during the course of the experiment in trees planted in the alkaline-calcareous soil when calcium nitrate was added. The slightly higher rhizosphere pH for calcium nitrate-fed plants may have contributed to this. The findings suggest that using ammonium sulphate in a liquid form (e.g. by fertigation) in high-pH soils leads to their acidification and the micronutrient availability may be improved.     

The effects of nutrient supply,predominantly addition of iron,and rhizobial inoculation on the tolerance of Lupinus pilosus genotypes to a calcareous soil

Two glasshouse experiments were conducted to examine the effects of nutrient supply and rhizobial inoculation on the performance of Lupinus pilosus genotypes differing in tolerance to calcareous soils. In experiment 1, plants were grown for 84 days in a calcareous soil (50% CaCO₃; soil water content 90% of field capacity) at four nutrient treatments (no-added nutrients, added nutrients without Fe, added nutrients with soil applied FeEDDHA, added nutrients with foliar applied FeSO₄). In experiment 2, plants were grown for 28 days with supply of NH₄NO₃ without inoculation or inoculated with Bradyrhizobium sp. (Lupinus). Chlorosis in the youngest leaves was a good indicator of the relative tolerance of the genotypes to the calcareous soil in both experiments, except the treatment with FeEDDHA at 5 mg kg^–1 soil which was toxic to all genotypes. Chlorosis scores correlated with chlorophyll meter readings and chlorophyll concentrations. The foliar application of FeSO₄ did not fully alleviate chlorotic symptoms despite concentrations of active or total Fe in the youngest leaves being increased. Adding nutrients and chemical nitrogen did not change the severity of chlorosis or improve the growth of the plant. The nutrient supply did not alter the ranking of tolerance of genotypes to the calcareous soil. The results suggest that nutrient deficiency or poor nodulation was not a major cause of poor plant growth on calcareous soils and that bicarbonate may exert a direct effect on chlorophyll synthesis. The mechanism for tolerance is likely to be related to an ability to exclude bicarbonate or prevent its transport to the leaves.     

Rational approaches to control of iron deficiency other than plant breeding and choice of resistant cultivars

Satisfactory progress has been made in recent years in preventing and correcting Fe deficiency in plants, and more can be expected in the future. Important advances include uses of acid- and Fe-fortified organic wastes and use of amended sulfur-pyrite mixes in soil. Three different approaches with organics are as an acidified matrix with Fe, as a means of chelating Fe, and as a carrier of acidifiers. Several procedures can help minimize Fe deficiency. (1) Avoid mis-management of soil physical properties and overirrigation. (2) Avoid nutrient imbalance, such as excess P or excess micronutrients. (3) Use preplant application of mildly acid-organic matter-Fe-sulfur or pyrite mixes worked into zones of soil or banded into seed rows. It is important that small bands or spots in soil be completely neutralized of CaCO₃. (4) Where foliar sprays can or need be used, especially to correct mild chlorosis, use ferrous compounds prepared to be delivered at pH 3± so that the Fe does not easily oxidize or precipitate in the solution. (5) For established trees that have become Fe deficient, inject, via slant drilling of small holes in tree trunks, dilute ferric ammonium citrate sufficient to supply not more than 100 mg kg^-1 Fe to leaves (dry weight basis). Most but not all species will respond. Procedure may be repeated in two or three weeks if necessary. (6) Iron chelates may be used in drip irrigation. If soil is sandy, soil pH not over 7.2, FeDTPA may be used; otherwise, FeEDDHA should be used. If the Fe is supplied with no other nutrients, pH may be at 4 and some FeSO₄ included to recycle the chelating agents. If Fe is used without chelating agents, the pH may be 1.0 or less and other nutrients included. (7) Urea-acid sulfate-Fe sulfate may be irrigated into soil around plants, especially if soil was polymer treated. (8) Efficiency of use of Fe chelates may be increased by making them slow release or by applying with seeds.     

Effect of foliar application of phosphorus on growth and mineral composition in peanut plants (Arachis hypogaea L.) under salt-stress

Summary Peanut (Arachis hypogaea L., Var. TMV-2) plants were raised in sand cultures under natural photoperiod. Salt treatment (0.4% NaCl on dry weight basis) was given 10 days after sowing. Nutrient solution was supplied 15 days after sowing to control and salinized plants. Phosphate (0.1% NaH₂PO₄ with 0.01% Tween-80) was sprayed to the plants to the drip point once daily from 20th to 25th day and from 30th to 35th day. The plants were harvested at 30th and 40th day for analysis. The plants grown under saline conditions showed a market reduction in growth. When phosphorus was supplied to saline plants by foliar application, there was an increase in dry weight, leaf area, stomatal frequency, and yield and the increase was more marked when the plants received phosphate for two periods. Phosphorus content decreased due to salinity which was restored by foliar spray of phosphate. With salinity, sodium accumulated while potassium and calcium were lowered. Phosphate spray decreased sodium and increased potassium and calcium in general.     

A comparison of ferric-chelate reductase and chlorophyll and growth ratios as indices of selection of quince,pear and olive genotypes under iron deficiency stress

Quince (Cydonia oblonga Mill.), pear (Pyrus communis L.) and olive (Olea europaea L.) genotypes were evaluated for their tolerance to iron deficiency stress by growing young plants in three types of aerated nutrient solutions: (1) with iron, (2) without iron or (3) low in iron and with 10 mM bicarbonate. Plants were obtained either from rooted softwood cuttings or from germination of seeds. The degree of tolerance was evaluated with several indices: (1) the chlorophyll content, (2) the root Fe³⁺ reducing capacity and (3) the whole plant relative growth. Fifteen hours before Fe³⁺ reducing capacity determination, iron was applied to the roots of plants with iron-stress, since this method resulted in increasing the reductase activity. All quince and pear genotypes increased the root Fe³⁺ reducing capacity when grown in the treatments for iron-stress, in relation to control plants of the same genotypes. In olive cultivars, the Fe³⁺ reducing capacity was lower in the iron-stress treatments than in the control one. Studying the relationship between relative growth and chlorophyll content for each genotype under iron-stress, in relation to both indices in control plants, a classification of species and genotypes was established. According to that, most olive cultivars and some pear rootstocks and cultivars appear more iron-efficient than quince rootstocks. Our study shows that in some woody species, determining root Fe³⁺ reducing capacity is not the best method to establish tolerance to iron deficiency stress.     

Fe-EDDHA对矫治秦美猕猴桃叶片失绿和营养元素组成的影响

在陕西石灰性土壤秦美猕猴桃果园施用不同剂量叶绿灵 (Fe- EDDHA) ,结果表明 ,萌芽期株施 45 g叶绿灵对矫治秦美猕猴桃叶片失绿黄化效果显著 ,可一直维持到当年落叶。同黄化对照植株相比 ,叶片叶绿素含量增加 1 82 .5 % ,活性铁含量提高 5 3 .2 % ,果实品质得到明显改善。施用叶绿灵还改善了叶片营养元素的含量     

A dioxygenase gene (Ids2) expressed under iron deficiency conditions in the roots of Hordeum vulgare

A zapII cDNA library was constructed from mRNA isolated from Fe-deficient barley roots and screened with cDNA probes made from mRNA of Fe-deficient and Fe-sufficient (control) barley roots. Seven clones were selected. Among them a clone having the putative full-length mRNA of dioxygenase as judged by northern hybridization was selected and named Ids2 (iron deficiency-specific clone 2). Using a cDNA fragment as probe, two clones from the genomic library (EMBL-III) were isolated and one was sequenced. The predicted amino acid sequence of Ids2 resembled that of 2-oxoglutarate-dependent dioxygenase. Ids2 is expressed in the Fe-deficient barley roots but is not in the leaves. The expression is repressed by the availability of Fe. Ids2 was also strongly expressed under Mn deficiency and weakly under Zn deficiency or excess NaCl (0.5%). The upstream 5-flanking region of Ids2 has a root-specific cis element of the CaMV 35S promoter and a nodule-specific element of leghemoglobin, a metal regulatory element (MRE) and several Cu regulatory elements (UAS) of yeast metallothionein (CUP1).     

Effects of foliar application of BAP on source and sink strength in four six-rowed barley (Hordeum vulgare L.) cultivars

A field experiment was conducted to investigate the effects of foliar application of a synthetic cytokinin (BAP) on source and sink strength of four different six-rowed barley (Hordeum vulgare L.) cultivars. Different spraying treatments consisting of spraying on whole plant, spraying only on leaves and spraying only on ears started at anthesis and continued for 7 days. One additional spraying was carried out on late period of grain filling. Results showed that spraying only on leaves did not affect ear weight, grain yield and 1,000-grain weight, while the two other treatments increased all above mentioned traits. Neither of treatments affected stem weight, biological yield and contribution of stem reserves in grain filling. Exogenous cytokinin did not increase photosynthetic rate and chlorophyll content in treated leaves until late period of grain filling, although there was no significant increase in final grain weight due to late application of BAP. Our results suggested that effects of foliar application of BAP were mostly due to increased sink size soon after anthesis and increased sink demand probably met by current photosynthesis of organs other than leaves, like ear green tissues. An erratum to this article is available at .     

Foliar application of zinc improves morpho-physiological and antioxidant defense mechanisms,and agronomic grain biofortification of wheat (Triticum aestivum L.) under water stress

Agronomic biofortification with zinc (Zn) may be engaged to improve the nutritious value of food crops along-with tolerance to water deficit conditions. The Zn may increase plant resistance to water stress by boosting physiological and enzymatic antioxidants defense mechanisms. Major objective of this study was to investigate the effect of foliar applied Zn on grain zin biofortification and drought tolerance in wheat. Treatments include application of Zinc at terminal growth phases (BBCH growth stage 49 and BBCH growth stage 65) with five levels: 0 (control-ck), water spray, 5, 10 and 15 mM under two levels of water regimes; well-watered (where 80% water holding capacity (WHC) was maintained in the soil) and water stress, (where 40% WHC was maintained in the soil). Results revealed that water stress significantly reduced relative water contents, gas exchange attributes, plant height, yield and yield related attributes of wheat. In contrast, hydrogen peroxide, free proline levels, activities of malondialdehyde, and concentration of soluble protein were markedly increased under water stress condition. Application of various levels of Zn significantly improved the CAT, SOD, POD and ASP activities at 40% WHC compared with control treatment. Foliarly applied 10 and 15 mM Zn predominantly reduced the damaging impact of water stress by improving the plant status in the form of plant height, RWC and gas exchange attributes. Likewise, wheat plant treated with 10 mM Zn under water stress condition increased the grain yield by improving number of grains per spike, 100 grain weight and biological yield compared with control. Moreover, increasing Zn levels also increased Zn concentration in grains and leaves. Overall, this study suggests that optimum level of Zn (10 mM) might be promising for alleviating the adverse impacts of water stress and enhance the grain biofortification in wheat.     

Management of sorghum grain mold through induced resistance mechanism using foliar spray of Pseudomonas fluorescens strains and non-conventional chemicals: a field study

Abstract

The study involved in-vitro and field studies using Pseudomonas fluorescens strains and some non-conventional chemicals for sorghum grain mold management. The culture filtrate of P. fluorescens strains PF-1 and PF-2 inhibited spore germination (p?<?0.001) and mycelial growth of grain mold fungi. Also, tested chemicals and commercial fungicides showed significant mycelial growth inhibition (p?<?0.001). PF-1, PF-2, ZnSO₄·7H₂O, salicylic acid and propiconazole were potentially effective and hence were tested in field (by foliar spraying). Two set of experiments were conducted simultaneously using naturally and artificially (Fusarium challenged) induced infected sorghum panicles. The lowest infection (score 4.2) was recorded with propiconazole. Disease severity was well controlled by PF-1 and PF-2, compared to ZnSO₄·7H₂O and salicylic acid. Furthermore, disease control was validated by determining total phenolic, flavonoid content and differentially induced phenolic acids (HPLC analysis) in harvested grains. Additionally, results of biochemical-traits, compatibility studies and antibiotic sensitivity of strains were found to be favourable.     

虹吸输铁对黄化苹果叶片光合参数及细胞器结构的影响

该研究以红富士苹果品种为材料,通过虹吸输入铁肥的方式,设置输铁液(600倍FeS04·7H20)处理并以输离子水和不输液为对照,观察了处理后苹果树的复绿情况,测定了叶绿素含量、光合参数、进行了叶绿体、线粒体超微结构的电镜扫描。结果表明:输铁后10 d,树体复绿情况明显,复绿等级由2.01变为0.53,复绿的叶片叶绿素含量有显著增加,增幅达到200%。同时,净光合速率提高了68%、蒸腾速率提高了21%、气孔导度提高了49%,胞间CO2浓度降低了100%。输铁处理后的单个细胞叶绿体个数比对照有显著增加,且个体更饱满,淀粉粒和嗜锇颗粒有明显减少;基粒片层结构规律整齐,基质清晰,被膜结构完整,线粒体内嵴清晰度高,数量增多。在输铁处理后,苹果树体复绿明显,叶绿素合成能力恢复,叶绿体、线粒体被膜结构均得到修复,表明用虹吸的方法将铁肥输入树体对苹果缺铁黄化病有很好的疗效。     

Growth promoting influence of siderophore-producing Pseudomonas strains GRP3A and PRS9 in maize (Zea mays L.) under iron limiting conditions

Maize seeds were bacterized with siderophore-producing pseudomonads with the goal to develop a system suitable for better iron uptake under iron-stressed conditions. Siderophore production was compared in fluorescent Pseudomonas spp. GRP3A, PRS9 and P. chlororaphis ATCC 9446 in standard succinate (SSM) and citrate (SCM) media. Succinate was better suited for siderophore production, however, deferration of media resulted in increased siderophore production in all the strains. Maximum siderophore level (216.23 microg/ml) was observed in strain PRS9 in deferrated SSM after 72 h of incubation. Strains GRP3A and PRS9 were used for plant growth promotion experiments. Strains GRP3A and PRS9 were also antagonistic against the phytopathogens, Colletotrichum dematium, Rhizoctonia solani and Sclerotium rolfsii. Bacterization of maize seeds with strains GRP3A and PRS9 showed significant increase in germination percentage and plant growth. Maximum shoot and root length and dry weight were observed with 10 microM Fe3+ along with bacterial inoculants suggesting application of siderophore producing plant growth promoting rhizobacterial strains in crop productivity in calcareous soil system.     

Enhanced levels of nicotianamine promote iron accumulation and tolerance to calcareous soil in soybean

Iron (Fe) is an essential nutrient in both plants and humans. Fe deficiency on calcareous soil with low Fe availability is a major agricultural problem. Nicotianamine (NA) is one of the Fe chelator in plants, which is involved in metal translocation into seeds, and serves as an antihypertensive substance in humans. In this study, soybean plants overexpressing the barley NA synthase 1 (HvNAS1) gene driven by the constitutive CaMV 35S promoter were produced using Agrobacterium-mediated transformation. The transgenic soybean showed no growth defect and grew normally. The NA content of transgenic soybean seeds was up to four-fold greater than that of non-transgenic (NT) soybean seeds. The level of HvNAS1 expression was positively correlated with the amount of NA, and a high concentration of NA was maintained in the seeds in succeeding generations. The Fe concentration was approximately two-fold greater in transgenic soybean seeds than in NT soybean seeds. Furthermore, the transgenic soybeans showed tolerance to low Fe availability in calcareous soil. Our results suggested that increasing the NA content in soybean seeds by the overexpression of HvNAS1 offers potential benefits for both human health and agricultural productivity.