Excessive iron deposition in root apoplast is involved in growth arrest of roots in response to low pH.

The rhizotoxicity of protons (H+) in acidic soils is a fundamental constraint that results in serious yield losses. However, the mechanisms underlying H+-mediated inhibition of root growth are poorly understood. In this study, we revealed that H+-induced root growth inhibition in Arabidopsis depends considerably on excessive iron deposition in the root apoplast. Reducing such aberrant iron deposition by decreasing the iron supply or disrupting the ferroxidases LOW PHOSPHATE ROOT 1 (LPR) and LPR2 attenuates the inhibitory effect of H+ on primary root growth efficiently. Further analysis showed that excessive iron deposition triggers a burst of highly reactive oxygen species, consequently impairing normal root development. Our study uncovered a valuable strategy for improving the ability of plants to tolerate H+ toxicity by manipulating iron availability.

Beyond iron-storage pool: functions of plant apoplastic iron during stress.

Iron (Fe) is an essential micronutrient for plants, and its storage in the apoplast represents an important Fe pool. Plants have developed various strategies to reutilize this apoplastic Fe pool to adapt to Fe deficiency. In addition, growing evidence indicates that the dynamic changes in apoplastic Fe are critical for plant adaptation to other stresses, including ammonium stress, phosphate deficiency, and pathogen attack. In this review, we discuss and scrutinize the relevance of apoplastic Fe for plant behavior changes in response to stress cues. We mainly focus on the relevant components that modulate the actions and downstream events of apoplastic Fe in stress signaling networks.

Inhibition of shoot-expressed NRT1.1 improves reutilization of apoplastic iron under iron-deficient conditions.

Iron deficiency is a major constraint for plant growth in calcareous soils. The interplay between NO3 - and Fe nutrition affects plant performance under Fe-deficient conditions. However, how NO3 - negatively regulates Fe nutrition at the molecular level in plants remains elusive. Here, we showed that the key nitrate transporter NRT1.1 in Arabidopsis plants, especially in the shoots, was markedly downregulated at post-translational levels by Fe deficiency. However, loss of NRT1.1 function alleviated Fe deficiency chlorosis, suggesting that downregulation of NRT1.1 by Fe deficiency favors plant tolerance to Fe deficiency. Further analysis showed that although disruption of NRT1.1 did not alter Fe levels in both the shoots and roots, it improved the reutilization of apoplastic Fe in shoots but not in roots. In addition, disruption of NRT1.1 prevented Fe deficiency-induced apoplastic alkalization in shoots by inhibiting apoplastic H+ depletion via NO3 - uptake. In vitro analysis showed that reduced pH facilitates release of cell wall-bound Fe. Thus, foliar spray with an acidic buffer promoted the reutilization of Fe in the leaf apoplast to enhance plant tolerance to Fe deficiency, while the opposite was true for the foliar spray with a neutral buffer. Thus, downregulation of the shoot-part function of NRT1.1 prevents apoplastic alkalization to ensure the reutilization of apoplastic Fe under Fe-deficient conditions. Our findings may provide a basis for elucidating the link between N and Fe nutrition in plants and insight to scrutinize the relevance of shoot-expressed NRT1.1 to the plant response to stress.

Phloem iron remodels root development in response to ammonium as the major nitrogen source.

Plants use nitrate and ammonium as major nitrogen (N) sources, each affecting root development through different mechanisms. However, the exact signaling pathways involved in root development are poorly understood. Here, we show that, in Arabidopsis thaliana, either disruption of the cell wall-localized ferroxidase LPR2 or a decrease in iron supplementation efficiently alleviates the growth inhibition of primary roots in response to NH4+ as the N source. Further study revealed that, compared with nitrate, ammonium led to excess iron accumulation in the apoplast of phloem in an LPR2-dependent manner. Such an aberrant iron accumulation subsequently causes massive callose deposition in the phloem from a resulting burst of reactive oxygen species, which impairs the function of the phloem. Therefore, ammonium attenuates primary root development by insufficiently allocating sucrose to the growth zone. Our results link phloem iron to root morphology in response to environmental cues.

Knockout of FER decreases cadmium concentration in roots of Arabidopsis thaliana by inhibiting the pathway related to iron uptake.

Identifying the genes that affect cadmium (Cd) accumulation in plants is a prerequisite for minimizing dietary Cd uptake from contaminated edible parts of plants by genetic engineering. This study showed that Cd stress inhibited the expression of FERONIA (FER) gene in the roots of wild-type Arabidopsis. Knockout of FER in fer-4 mutants downregulated the Cd-induced expression of several genes related to iron (Fe) uptake, including IRT1, bHLH38, NRAMP1, NRAMP3, FRO2 andFIT. In addition, the Cd concentration in fer-4 mutant roots reduced to approximately half of that in the wild-type seedlings. As a result, the Cd tolerance of fer-4 was higher. Furthermore, increased Fe supplementation had little effect on the Cd tolerance of fer-4 mutants, but clearly improved the Cd tolerance of wild-type seedlings, showing that the alleviation of Cd toxicity by Fe depends on the action of FER. Taken together, the findings demonstrate that the knockout of FER might provide a strategy to reduce Cd contamination and improve the Cd tolerance in plants by regulating the pathways related to Fe uptake.

[Effects of dicyandiamide on cadmium accumulation in pakchoi under instant soluble nitrogen fertilizers]. / é€Ÿæ•ˆæ°®è‚¥é æ–½åŒæ°°èƒºå¯¹å‡æŽ§å°ç™½èœé•‰ç§¯ç´¯çš„ä½œç”¨.

We investigated the effects of dicyandiamide (DCD) on the growth and Cd concentrations in pakchoi cultivated under different instant soluble N fertilizers [ammonium sulfate, ammonium sulfate and sodium nitrate (1:1, ammonium/nitrate), and urea] in Cd-contaminated soils. The results showed that the fresh weight of the edible parts of Cd-stressed pakchoi were increased by 583.3%, 41.5%, and 206.8% under ammonium, ammonium/nitrate, and urea treatments in the presence of DCD, respectively compared with control, and the tolerance index and photosynthetic rate significantly increased, whereas no significant changes were observed under nitrate supply. Under all N treatments with DCD, the MDA and H2O2 contents and the superoxide radical production rate in the leaves of pakchoi were decreased, with the highest reduction occurred in ammonium and urea treatments. Cd concentrations in the leaves of pakchoi fertilized with ammonium, ammonium/nitrate, and urea were lowered by 58.3%, 34.0%, and 44.5% and those in the petioles were lowered by 61.8%, 29.4%, and 55.6%, respectively. Cd concentration in the leaves and petioles of pakchoi in the nitrate treatment did not differ significantly from control. These changes could be attributable to the reduction in the acidification of rhizosphere soil in response to the combined application of N fertilizer and DCD. Accordingly, in Cd-contaminated soils with a low buffering capacity, the application of DCD combined with ammonium, ammonium/nitrate, or urea N fertili-zers could alleviate Cd-induced growth stress and inhibit photosynthesis in pakchoi plants and effectively minimize the Cd accumulation.

Ammonium aggravates salt stress in plants by entrapping them in a chloride over-accumulation state in an NRT1.1-dependent manner.

Global climate change has exacerbated flooding in coastal areas affected by soil salinization. Ammonium (NH4+) is the predominant form of nitrogen in flooded soils, but the role played by NH4+ in the plant response to salt stress has not been fully clarified. We investigated the responses of Arabidopsis thaliana, Oryza sativa, and Nicotiana benthamiana plants fed with NH4+. All species were hypersensitive to NaCl stress and accumulated more Cl- and less Na+ than those fed with NO3-. Further investigation of A. thaliana indicated that salt hypersensitivity induced by the presence of NH4+ was abolished by removing the Cl- but was not affected by the removal of Na+, suggesting that excess accumulation of Cl- rather than Na+ is involved in NH4+-conferred salt hypersensitivity. The expression of nitrate transporter NRT1.1 protein was also up-regulated by NH4+ treatment, which increased root Cl- uptake due to the Cl- uptake activity of NRT1.1 and the absence of uptake competition from NO3-. Knockout of NRT1.1 in plants decreased their root Cl- uptake and retracted the NH4+-conferred salt hypersensitivity. Our findings revealed that NH4+-aggravated salt stress in plants is associated with Cl- over-accumulation through the up-regulation of NRT1.1-mediated Cl- uptake. These findings suggest the significant impact of Cl- toxicity in flooded coastal areas, an issue of ecological significance.

Knockdown of BTS may provide a new strategy to improve cadmium-phytoremediation efficiency by improving iron status in plants.

The identification of the key genes related to cadmium (Cd) tolerance and accumulation is a major element in genetically engineering improved plants for Cd phytoremediation. Owing to the similarity between the ionic hydrated radius of Cd2+ and Fe2+, this study investigated how the Cd tolerance and accumulation of Arabidopsis plants was affected by the knockdown of BTS, a gene that negatively regulates Fe nutrition. After exposure to 40 µM Cd, the BTS-knockdown mutant, bts-1, exhibited greater Fe nutrition and better growth than wild-type plants. In addition, the Cd concentration in both roots and shoots was approximately 50% higher in the bts-1 mutant than in wild-type plants. Consequently, the bts-1 mutant accumulated approximately 100% and 150% more Cd in the roots and shoots, respectively, than wild-type plants. Further study showed that Fe removal from the growth medium and inhibition of the Fe transporter gene, IRT1, removed the differences observed in the growth and Cd concentration of the bts-1 and wild-type plants, respectively. These results demonstrated that BTS knockdown improved Cd tolerance and accumulation in plants by improving Fe nutrition; thus, the knockdown of BTS via biotechnological pathways may represent a valuable strategy for the improvement in the efficiency of Cd phytoremediation.

Inhibition of DNA demethylation enhances plant tolerance to cadmium toxicity by improving iron nutrition.

Although the alteration of DNA methylation due to abiotic stresses, such as exposure to the toxic metal cadmium (Cd), has been often observed in plants, little is known about whether such epigenetic changes are linked to the ability of plants to adapt to stress. Herein, we report a close linkage between DNA methylation and the adaptational responses in Arabidopsis plants under Cd stress. Exposure to Cd significantly inhibited the expression of three DNA demethylase genes ROS1/DML2/DML3 (RDD) and elevated DNA methylation at the genome-wide level in Col-0 roots. Furthermore, the profile of DNA methylation in Cd-exposed Col-0 roots was similar to that in the roots of rdd triple mutants, which lack RDD, indicating that Cd-induced DNA methylation is associated with the inhibition of RDD. Interestingly, the elevation in DNA methylation in rdd conferred a higher tolerance against Cd stress and improved cellular Fe nutrition in the root tissues. In addition, lowering the Fe supply abolished improved Cd tolerance due to the lack of RDD in rdd. Together, these data suggest that the inhibition of RDD-mediated DNA demethylation in the roots by Cd would in turn enhance plant tolerance to Cd stress by improving Fe nutrition through a feedback mechanism.

Sulfide alleviates cadmium toxicity in Arabidopsis plants by altering the chemical form and the subcellular distribution of cadmium.

Several sulfur compounds are thought to play important roles in the plant tolerance to cadmium (Cd), but the role of inorganic sulfide in Cd tolerance remains largely unknown. In this study, we found that Cd exposure increased the accumulation of soluble sulfide in Arabidopsis plants. When exogenous sulfide, in the form of NaHS, was foliarly applied, Cd-induced growth inhibition and oxidative stress were alleviated. In addition, although the foliar application of sulfide did not affect the total Cd levels, it significantly decreased the soluble Cd fractions in plants. Furthermore, foliar applications of sulfide decreased Cd distribution in the cytoplasm and organelles, but increased Cd retention in the cell wall, which is a less sensitive compartment. These results suggest that the Cd-induced accumulation of soluble sulfide alleviates Cd toxicity in plants by inactivating Cd and sequestering it into the cell wall.

Ethylene and nitric oxide interact to regulate the magnesium deficiency-induced root hair development in Arabidopsis.

Nitric oxide (NO) and ethylene respond to biotic and abiotic stresses through either similar or independent processes. This study examines the mechanism underlying the effects of NO and ethylene on promoting root hair development in Arabidopsis under magnesium (Mg) deficiency. The interaction between NO and ethylene in the regulation of Mg deficiency-induced root hair development was investigated using NO- and ethylene-related mutants and pharmacological methods. Mg deficiency triggered a burst of NO and ethylene, accompanied by a stimulated development of root hairs. Interestingly, ethylene facilitated NO generation by activation of both nitrate reductase and nitric oxide synthase-like (NOS-L) in the roots of Mg-deficient plants. In turn, NO enhanced ethylene synthesis through stimulating the activities of 1-aminocyclopropane-1-carboxylate (ACC) oxidase and ACC synthase (ACS). These two processes constituted an NO-ethylene feedback loop. Blocking either of these two processes inhibited the stimulation of root hair development under Mg deficiency. In conclusion, we suggest that Mg deficiency increases the production of NO and ethylene in roots, each influencing the accumulation and role of the other, and thus these two signals interactively regulate Mg deficiency-induced root hair morphogenesis.

Roles of chemical signals in regulation of the adaptive responses to iron deficiency.

Iron is an essential micronutrient for plants but is not readily accessible in most calcareous soils. Although the adaptive responses of plants to iron deficiency have been well documented, the signals involved in the regulatory cascade leading to their activation are not well understood to date. Recent studies revealed that chemical compounds, including sucrose, auxin, ethylene and nitric oxide, positively regulated the Fe-deficiency-induced Fe uptake processes in a cooperative manner. Nevertheless, cytokinins, jasmonate and abscisic acid were shown to act as negative signals in transmitting the iron deficiency information. The present mini review is to briefly address the roles of chemical signals in regulation of the adaptive responses to iron deficiency based on the literatures published in recent years.

Increased Sucrose Accumulation Regulates Iron-Deficiency Responses by Promoting Auxin Signaling in Arabidopsis Plants.

Previous studies have identified that auxins acts upstream of nitric oxide in regulating iron deficiency responses in roots, but the upstream signaling molecule of auxins remains unknown. In this study, we showed that Fe deficiency increased sucrose (Suc) level in roots of Arabidopsis (Arabidopsis thaliana). Exogenous application of Suc further stimulated Fe deficiency-induced ferric-chelate-reductase (FCR) activity and expression of Fe acquisition-related genes FRO2, IRT1, and FIT in roots. The opposite patterns were observed in the dark treatment. In addition, FCR activity and expression of Fe acquisition-related genes were higher in the Suc high-accumulating transgenic plant 35S::SUC2 but were lower in the Suc low-accumulating mutant suc2-5 compared with wild-type plants under Fe-deficient conditions. Consequently, Fe deficiency tolerance was enhanced in 35S::SUC2 but was compromised in suc2-5. Exogenous Suc also increased root ß-glucuronidase (GUS) activity in auxin-inducible reporter DR5-GUS transgenic plants under Fe deficiency. However, exogenous Suc failed to increase FCR activity and expression of Fe acquisition-related genes in the auxin transport-impaired mutants aux1-7 and pin1-1 as well as in the wild-type plants treated with an auxin transport inhibitor under Fe deficiency. In summary, we found that increased Suc accumulation is required for regulating Fe deficiency responses in plants, with auxins acting downstream in transmitting the Fe deficiency signal.

Modification of nitrate uptake pathway in plants affects the cadmium uptake by roots.

NRT1.1 is a dual-affinity nitrate (NO3(-)) transporter involved in both high- and low-affinity NO3(-) uptake in Arabidopsis plants. In a recent study, we showed that, under cadmium (Cd) exposure, blocking the NRT1.1-mediated NO3(-) uptake reduces Cd entry into roots, thus lowing Cd levels in plants and improving plant growth. In addition, we also found that the Cd levels in edible parts of 11 Chinese cabbage (Brassica rapa L. ssp. pekinensis) cultivars correlated well with the NO3(-) uptake rates of their roots. These results suggested that the NO3(-) uptake of roots negatively regulate Cd uptake. Modification of NO3(-) uptake in crops by modulating NO3(-) uptake pathway might provide a biological engineering approach to reducing Cd accumulation in edible organs, thus improving food safety.

Oxalate synthesis in leaves is associated with root uptake of nitrate and its assimilation in spinach (Spinacia oleracea L.) plants.

BACKGROUND: Excessive accumulation of oxalate in numerous vegetables adversely affects their quality as food. While it is known that nitrate could effectively stimulate oxalate accumulation in many vegetables, little information is available about the mechanism of nitrate-induced oxalate accumulation. In this study, we examined the association of oxalate synthesis with nitrate uptake and assimilation in two genotypes of spinach (Spinacia oleracea L.), Heizhenzhu and Weilv. RESULTS: Increasing nitrate levels resulted in enhanced synthesis of oxalate, as well as increased root uptake of nitrate and leaf activities of nitrate reductase (NR) and glutamine synthetase (GS) for both genotypes. Correlation analysis revealed that oxalate accumulation in spinach leaves was positively related with rate of nitrate uptake by roots, as well as leaf activities of NR and GS. Addition of plasmalemma H(+)-ATPase inhibitor sodium vanadate (Na3VO4) significantly decreased leaf oxalate accumulation in both genotypes. Presence of NR or GS inhibitors led to reduction of leaf oxalate contents, GS/NR activities and decreased nitrate uptake rate. Significantly higher levels of nitrate root uptake, leaf NR and GS activities were observed in the high-oxalate genotype Heizhenzhu than in Weilv. CONCLUSION: Oxalate synthesis in leaves of spinach is not only positively associated with root uptake of nitrate, but also with its assimilation within the plants.

Inhibition of nitrate transporter 1.1-controlled nitrate uptake reduces cadmium uptake in Arabidopsis.

Identification of mechanisms that decrease cadmium accumulation in plants is a prerequisite for minimizing dietary uptake of cadmium from contaminated crops. Here, we show that cadmium inhibits nitrate transporter 1.1 (NRT1.1)-mediated nitrate (NO3 (-)) uptake in Arabidopsis (Arabidopsis thaliana) and impairs NO3 (-) homeostasis in roots. In NO3 (-)-containing medium, loss of NRT1.1 function in nrt1.1 mutants leads to decreased levels of cadmium and several other metals in both roots and shoots and results in better biomass production in the presence of cadmium, whereas in NO3 (-)-free medium, no difference is seen between nrt1.1 mutants and wild-type plants. These results suggest that inhibition of NRT1.1 activity reduces cadmium uptake, thus enhancing cadmium tolerance in an NO3 (-) uptake-dependent manner. Furthermore, using a treatment rotation system allowing synchronous uptake of NO3 (-) and nutrient cations and asynchronous uptake of cadmium, the nrt1.1 mutants had similar cadmium levels to wild-type plants but lower levels of nutrient metals, whereas the opposite effect was seen using treatment rotation allowing synchronous uptake of NO3 (-) and cadmium and asynchronous uptake of nutrient cations. We conclude that, although inhibition of NRT1.1-mediated NO3 (-) uptake by cadmium might have negative effects on nitrogen nutrition in plants, it has a positive effect on cadmium detoxification by reducing cadmium entry into roots. NRT1.1 may regulate the uptake of cadmium and other cations by a common mechanism.

An underground tale: contribution of microbial activity to plant iron acquisition via ecological processes.

BACKGROUND: Iron (Fe) deficiency in crops is a worldwide agricultural problem. Plants have evolved several strategies to enhance Fe acquisition, but increasing evidence has shown that the intrinsic plant-based strategies alone are insufficient to avoid Fe deficiency in Fe-limited soils. Soil micro-organisms also play a critical role in plant Fe acquisition; however, the mechanisms behind their promotion of Fe acquisition remain largely unknown. SCOPE: This review focuses on the possible mechanisms underlying the promotion of plant Fe acquisition by soil micro-organisms. CONCLUSIONS: Fe-deficiency-induced root exudates alter the microbial community in the rhizosphere by modifying the physicochemical properties of soil, and/or by their antimicrobial and/or growth-promoting effects. The altered microbial community may in turn benefit plant Fe acquisition via production of siderophores and protons, both of which improve Fe bioavailability in soil, and via hormone generation that triggers the enhancement of Fe uptake capacity in plants. In addition, symbiotic interactions between micro-organisms and host plants could also enhance plant Fe acquisition, possibly including: rhizobium nodulation enhancing plant Fe uptake capacity and mycorrhizal fungal infection enhancing root length and the nutrient acquisition area of the root system, as well as increasing the production of Fe(3+) chelators and protons.

Exogenous abscisic acid application decreases cadmium accumulation in Arabidopsis plants, which is associated with the inhibition of IRT1-mediated cadmium uptake.

Cadmium (Cd) contamination of agricultural soils is an increasingly serious problem. Measures need to be developed to minimize Cd entering the human food chain from contaminated soils. We report here that, under Cd exposure condition, application with low doses of (0.1-0.5 µM) abscisic acid (ABA) clearly inhibited Cd uptake by roots and decreased Cd level in Arabidopsis wild-type plants (Col-0). Expression of IRT1 in roots was also strongly inhibited by ABA treatment. Decrease in Cd uptake and the inhibition of IRT1 expression were clearly lesser pronounced in an ABA-insensitive double mutant snrk2.2/2.3 than in the Col-0 in response to ABA application. The ABA-decreased Cd uptake was found to correlate with the ABA-inhibited IRT1 expression in the roots of Col-0 plants fed two different levels of iron. Furthermore, the Cd uptake of irt1 mutants was barely affected by ABA application. These results indicated that inhibition of IRT1 expression is involved in the decrease of Cd uptake in response to exogenous ABA application. Interestingly, ABA application increased the iron level in both Col-0 plants and irt1 mutants, suggesting that ABA-increased Fe acquisition does not depend on the IRT1 function, but on the contrary, the ABA-mediated inhibition of IRT1 expression may be due to the elevation of iron level in plants. From our results, we concluded that ABA application might increase iron acquisition, followed by the decrease in Cd uptake by inhibition of IRT1 activity. Thus, for crop production in Cd contaminated soils, developing techniques based on ABA application potentially is a promising approach for reducing Cd accumulation in edible organs in plants.

[A review on current situations of steroid estrogen in the water system].

Steroid estrogens are a group of biologically active endocrine disruptors. An extremely low level of steroid estrogens is sufficient to result in reproduction and behavior disorders, and larval mutation of both human being and animals. The ecological and environmental effects of steroid estrogens should be more direct on organisms in water system as compared with those in other environment systems. Therefore, in this paper we reviewed the physical and chemical properties of some frequently-used steroid estrogens, and their contamination status, origins and transfers in water system, so as to provide a theoretical basis for developing estrogen removal strategies.

The 14-3-3 protein GENERAL REGULATORY FACTOR11 (GRF11) acts downstream of nitric oxide to regulate iron acquisition in Arabidopsis thaliana.

Here we report the function of a general regulatory factor, GENERAL REGULATORY FACTOR11 (GRF11), in terms of the iron (Fe) deficiency response. Physiological and molecular responses of the loss-of-function Arabidopsis thaliana grf11 mutant to Fe supply were investigated. Genes involved in posttranscriptional regulation of FER-LIKE FE DEFICIENCY-INDUCED TRANSCRIPTION FACTOR (FIT) were also analyzed. In addition, the molecular link between the signaling molecule nitric oxide (NO) and Fe deficiency responses was further dissected. Our results suggest that GRF11 is necessary for induction of Fe-deficiency-tolerance mechanisms. The FIT protein can bind to the promoter of GRF11, which contains an E-box motif. GRF11 also positively affects FIT transcription but has no influence on the genes involved in posttranscriptional regulation of FIT. Furthermore, NO positively regulates GRF11 induction upon the onset of Fe deficiency. We propose that, upon the onset of Fe deficiency, induction of FIT expression is dependent on GRF11, which acts downstream of NO to mediate Fe deficiency responses.