Metagenomic analysis of rhizosphere microbiome provides insights into occurrence of iron deficiency chlorosis in field of Asian pears

Background Fe-deficiency chlorosis (FDC) of Asian pear plants is widespread, but little is known about the association between the microbial communities in the rhizosphere soil and leaf chlorosis. The leaf mineral concentration, leaf subcellular structure, soil physiochemical properties, and bacterial species community and distribution had been analysed to gain insights into the FDC in Asian pear plant. Results The total Fe in leaves with Fe-deficiency was positively correlated with total K, Mg, S, Cu, Zn, Mo and Cl contents, but no differences of available Fe (AFe) were detected between the rhizosphere soil of chlorotic and normal plants. Degraded ribosomes and degraded thylakloid stacks in chloroplast were observed in chlorotic leaves. The annotated microbiome indicated that there were 5 kingdoms, 52 phyla, 94 classes, 206 orders, 404 families, 1,161 genera, and 3,043 species in the rhizosphere soil of chlorotic plants; it was one phylum less and one order, 11 families, 59 genera, and 313 species more than in that of normal plant. Bacterial community and distribution patterns in the rhizosphere soil of chlorotic plants were distinct from those of normal plants and the relative abundance and microbiome diversity were more stable in the rhizosphere soils of normal than in chlorotic plants. Three (Nitrospira defluvii, Gemmatirosa kalamazoonesis, and Sulfuricella denitrificans) of the top five species (N. defluvii, G. kalamazoonesis, S. denitrificans, Candidatus Nitrosoarchaeum koreensis, and Candidatus Koribacter versatilis). were the identical and aerobic in both rhizosphere soils, but their relative abundance decreased by 48, 37, and 22%, respectively, and two of them (G. aurantiaca and Ca. S. usitatus) were substituted by an ammonia-oxidizing soil archaeon, Ca. N. koreensis and a nitrite and nitrate reduction related species, Ca. K. versatilis in that of chlorotic plants, which indicated the adverse soil aeration in the rhizosphere soil of chlorotic plants. A water-impermeable tables was found to reduce the soil aeration, inhibit root growth, and cause some absorption root death from infection by Fusarium solani. Conclusions It was waterlogging or/and poor drainage of the soil may inhibit Fe uptake not the amounts of AFe in the rhizosphere soil of chlorotic plants that caused FDC in this study.

Several Yeast Species Induce Iron Deficiency Responses in Cucumber Plants (Cucumis sativus L.)

Iron (Fe) deficiency is a first-order agronomic problem that causes a significant decrease in crop yield and quality. Paradoxically, Fe is very abundant in most soils, mainly in its oxidized form, but is poorly soluble and with low availability for plants. In order to alleviate this situation, plants develop different morphological and physiological Fe-deficiency responses, mainly in their roots, to facilitate Fe mobilization and acquisition. Even so, Fe fertilizers, mainly Fe chelates, are widely used in modern agriculture, causing environmental problems and increasing the costs of production, due to the high prices of these products. One of the most sustainable and promising alternatives to the use of agrochemicals is the better management of the rhizosphere and the beneficial microbial communities presented there. The main objective of this research has been to evaluate the ability of several yeast species, such as Debaryomyces hansenii, Saccharomyces cerevisiae and Hansenula polymorpha, to induce Fe-deficiency responses in cucumber plants. To date, there are no studies on the roles played by yeasts on the Fe nutrition of plants. Experiments were carried out with cucumber plants grown in a hydroponic growth system. The effects of the three yeast species on some of the most important Fe-deficiency responses developed by dicot (Strategy I) plants, such as enhanced ferric reductase activity and Fe2+ transport, acidification of the rhizosphere, and proliferation of subapical root hairs, were evaluated. The results obtained show the inductive character of the three yeast species, mainly of Debaryomyces hansenii and Hansenula polymorpha, on the Fe-deficiency responses evaluated in this study. This opens a promising line of study on the use of these microorganisms as Fe biofertilizers in a more sustainable and environmentally friendly agriculture.

Effects of Iron Deficiency Stress on Plant Growth and Quality in Flowering Chinese Cabbage and Its Adaptive Response

Iron (Fe) plays an important role in the growth and development of the human body and plants. The effects of different Fe concentrations, 1-aminocyclopropane-1-carboxylic acid (ACC), and cobalt chloride (Co2+) treatments on plant growth, quality and the adaptive response to Fe deficiency stress were investigated in flowering Chinese cabbage. The results revealed that Fe deficiency stress inhibited plant growth. The content of vitamin C (Vc), soluble protein, and soluble sugar in leaves and stalks were significantly reduced under Fe deficiency stress, while the content of cellulose and nitrate was increased. Meanwhile, Fe deficiency stress obviously reduced the net photosynthetic rate and nitrate reductase (NR) activity of leaves. The balance system of active oxygen metabolism was destroyed due to Fe deficiency, resulting in the decrease of catalase (CAT) activity, superoxide dismutase (SOD) activity of roots and leaves, and peroxidase (POD) activity of leaves, while POD activity in roots and malonaldehyde (MDA) content were significantly increased. The treatments of Fe deficiency and ACC significantly reduced pH value of the root medium, promoted release of ethylene, and increased Fe3+ reductase activity, while Co2+ treatment showed the results opposite to those of Fe deficiency and ACC treatments. Thus, Fe deficiency stress could induce nitrogen metabolism, photosynthesis, reactive oxygen metabolism, pH of root medium, and Fe3+ reductase activity that was related to physiological adaptive response and tolerance mechanisms. We also found that ethylene could involve in regulating the adaptive response to Fe deficiency stress and improve the availability of Fe in flowering Chinese cabbage.Main ConclusionFe deficiency stress could induce nitrogen metabolism, photosynthesis, reactive oxygen metabolism, pH of root medium, and Fe3+ reductase activity that was related to physiological adaptive response and tolerance mechanisms.

Short Term Elevated CO2 Interacts with Iron Deficiency, Further Repressing Growth, Photosynthesis and Mineral Accumulation in Soybean (Glycine max L.) and Common Bean (Phaseolus vulgaris L.)

Elevated CO2 (eCO2) has been reported to cause mineral losses in several important food crops such as soybean (Glycine max L.) and common bean (Phaseolus vulgaris L.). In addition, more than 30% of the world’s arable land is calcareous, leading to iron (Fe) deficiency chlorosis and lower Fe levels in plant tissues. We hypothesize that there will be combinatorial effects of eCO2 and Fe deficiency on the mineral dynamics of these crops at a morphological, biochemical and physiological level. To test this hypothesis, plants were grown hydroponically under Fe sufficiency (20 μM Fe-EDDHA) or deficiency (0 μM Fe-EDDHA) at ambient CO2 (aCO2, 400 ppm) or eCO2 (800 ppm). Plants of both species exposed to eCO2 and Fe deficiency showed the lowest biomass accumulation and the lowest root: shoot ratio. Soybean at eCO2 had significantly higher chlorophyll levels (81%, p < 0.0001) and common bean had significantly higher photosynthetic rates (60%, p < 0.05) but only under Fe sufficiency. In addition, eCO2 increased ferric chelate reductase acivity (FCR) in Fe-sufficient soybean by 4-fold (p < 0.1) and in Fe-deficient common bean plants by 10-fold (p < 0.0001). In common bean, an interactive effect of both environmental factors was observed, resulting in the lowest root Fe levels. The lowering of Fe accumulation in both crops under eCO2 may be linked to the low root citrate accumulation in these plants when grown with unrestricted Fe supply. No changes were observed for malate in soybean, but in common bean, shoot levels were significantly lower under Fe deficiency (77%, p < 0.05) and Fe sufficiency (98%, p < 0.001). These results suggest that the mechanisms involved in reduced Fe accumulation caused by eCO2 and Fe deficiency may not be independent, and an interaction of these factors may lead to further reduced Fe levels.

Analysis of the VIT1 Promoter Activity in Developing Arabidopsis thaliana Plants

Iron (Fe) deficiency in plants is one of the widespread problems limiting agricultural production. Generating crops more tolerant to Fe deficiency by genetic engineering or breeding is of great interest but challenging due to the knowledge gaps in general plant Fe homeostasis. Although several genes involved in Fe homeostasis have been identified, characterization of their roles is mainly limited to specific organs at specific developmental stages of the plant, where their mutants show the most striking phenotype. Vacuolar Iron Transporter 1 (VIT1) is a well-known gene that has been characterized for its function in the mature seed of Arabidopsis thaliana. VIT1 is an Fe transporter that determines the correct distribution of Fe storage in this organ. The study aimed to explore new physiological functions for VIT1. As a first step, Arabidopsis thaliana plants that contain PromoterVIT1: GUS constructs were used to study the temporal and spatial expression of the gene throughout the plant’s lifecycle. GUS histochemical staining revealed the VIT1 promoter is active in the mature leaves and mature reproductive organs. VIT1 promoter activity in the stamen increased developmentally and was limited to tapetum and guard cells in the pollen sac. In the female organ, VIT1 promoter activity increased as the pistil developed into a silique. Although all the silique exhibited staining, staining density was higher in the peduncle, replum, and stigma regions. Inside the developing silique, funicles were heavily stained. Furthermore, in silico analyses of VT1 transcriptome and protein levels confirmed flower and the silique are hot spots for VT1 activity. Thus, the results may suggest a possible involvement of VT1 protein in several stages of the reproductive system, specifically in the flowering and in the fruit development.

Involvement of Arabidopsis Multi-Copper Oxidase-Encoding LACCASE12 in Root-to-Shoot Iron Partitioning: A Novel Example of Copper-Iron Crosstalk

Numerous central biological processes depend on the participation of the essential elements iron (Fe) or copper (Cu), including photosynthesis, respiration, cell wall remodeling and oxidative stress protection. Yet, both Fe and Cu metal cations can become toxic when accumulated in excess. Because of the potent ligand-binding and redox chemistries of these metals, there is a need for the tight and combined homeostatic control of their uptake and distribution. Several known examples pinpoint an inter-dependence of Fe and Cu homeostasis in eukaryotes, mostly in green algae, yeast and mammals, but this is less well understood in multicellular plants to date. In Arabidopsis, Cu deficiency causes secondary Fe deficiency, and this is associated with reduced in vitro ferroxidase activity and decreased root-to-shoot Fe translocation. Here we summarize the current knowledge of the cross-talk between Cu and Fe homeostasis and present a partial characterization of LACCASE12 (LAC12) that encodes a member of the multicopper oxidase (MCO) protein family in Arabidopsis. LAC12 transcript levels increase under Fe deficiency. The phenotypic characterization of two mutants carrying T-DNA insertions suggests a role of LAC12 in root-to-shoot Fe partitioning and in maintaining growth on Fe-deficient substrates. A molecular understanding of the complex interactions between Fe and Cu will be important for combating Fe deficiency in crops and for advancing biofortification approaches.

Nitric Oxide Prevents Fe Deficiency-Induced Photosynthetic Disturbance, and Oxidative Stress in Alfalfa by Regulating Fe Acquisition and Antioxidant Defense

Iron (Fe) deficiency impairs photosynthetic efficiency, plant growth and biomass yield. This study aimed to reveal the role of nitric oxide (NO) in restoring Fe-homeostasis and oxidative status in Fe-deficient alfalfa. In alfalfa, a shortage of Fe negatively affected the efficiency of root andshoot length, leaf greenness, maximum quantum yield PSII (Fv/Fm), Fe, S, and Zn accumulation, as well as an increase in H2O2 accumulation. In contrast, in the presence of sodium nitroprusside (SNP), a NO donor, these negative effects of Fe deficiency were largely reversed. In response to the SNP, the expression of Fe transporters (IRT1, NRAMP1) and S transporter (SULTR1;2) genes increased in alfalfa. Additionally, the detection of NO generation using fluorescence microscope revealed that SNP treatment increased the level of NO signal, indicating that NO may act as regulatory signal in response to SNP in plants. Interestingly, the increase of antioxidant genes and their related enzymes (Fe-SOD, APX) in response to SNP treatment suggests that Fe-SOD and APX are key contributors to reducing ROS (H2O2) accumulation and oxidative stress in alfalfa. Furthermore, the elevation of Ascorbate-glutathione (AsA-GSH) pathway-related genes (GR and MDAR) Fe-deficiency with SNP implies that the presence of NO relates to enhanced antioxidant defense against Fe-deficiency stress.

Photoprotection during iron deficiency is mediated by the bHLH transcription factors PYE and ILR3

Iron (Fe) is an essential micronutrient whose availability is limiting in many soils. During Fe deficiency, plants alter the expression of many genes to increase Fe uptake, distribution, and utilization. In a genetic screen for suppressors of Fe sensitivity in the E3 ligase mutant bts-3, we isolated an allele of the bHLH transcription factor (TF) ILR3, ilr3-4. We identified a striking leaf bleaching phenotype in ilr3 mutants that was suppressed by limiting light intensity, indicating that ILR3 is required for phototolerance during Fe deficiency. Among its paralogs that are thought to be partially redundant, only ILR3 was required for phototolerance as well as repression of genes under Fe deficiency. A mutation in the gene-encoding PYE, a known transcriptional repressor under Fe deficiency, also caused leaf bleaching. We identified singlet oxygen as the accumulating reactive oxygen species (ROS) in ilr3-4 and pye, suggesting photosensitivity is due to a PSII defect resulting in ROS production. During Fe deficiency, ilr3-4 and pye chloroplasts retain normal ultrastructure and, unlike wild type (WT), contain stacked grana similar to Fe-sufficient plants. Additionally, we found that the D1 subunit of PSII is destabilized in WT during Fe deficiency but not in ilr3-4 and pye, suggesting that PSII repair is accelerated during Fe deficiency in an ILR3- and PYE-dependent manner. Collectively, our results indicate that ILR3 and PYE confer photoprotection during Fe deficiency to prevent the accumulation of singlet oxygen, potentially by promoting reduction of grana stacking to limit excitation and facilitate repair of the photosynthetic machinery.

IRON MAN interacts with BRUTUS to maintain iron homeostasis in Arabidopsis

IRON MAN (IMA) peptides, a family of small peptides, control iron (Fe) transport in plants, but their roles in Fe signaling remain unclear. BRUTUS (BTS) is a potential Fe sensor that negatively regulates Fe homeostasis by promoting the ubiquitin-mediated degradation of bHLH105 and bHLH115, two positive regulators of the Fe deficiency response. Here, we show that IMA peptides interact with BTS. The C-terminal parts of IMA peptides contain a conserved BTS interaction domain (BID) that is responsible for their interaction with the C terminus of BTS. Arabidopsis thaliana plants constitutively expressing IMA genes phenocopy the bts-2 mutant. Moreover, IMA peptides are ubiquitinated and degraded by BTS. bHLH105 and bHLH115 also share a BID, which accounts for their interaction with BTS. IMA peptides compete with bHLH105/bHLH115 for interaction with BTS, thereby inhibiting the degradation of these transcription factors by BTS. Genetic analyses suggest that bHLH105/bHLH115 and IMA3 have additive roles and function downstream of BTS. Moreover, the transcription of both BTS and IMA3 is activated directly by bHLH105 and bHLH115 under Fe-deficient conditions. Our findings provide a conceptual framework for understanding the regulation of Fe homeostasis: IMA peptides protect bHLH105/bHLH115 from degradation by sequestering BTS, thereby activating the Fe deficiency response.