Tonoplast-localized transporter ZmNRAMP2 confers root-to-shoot translocation of manganese in maize.

Almost all living organisms require manganese (Mn) as an essential trace element for survival. To maintain an irreplaceable role in the oxygen-evolving complex of photosynthesis, plants require efficient Mn uptake in roots and delivery to above-ground tissues. However, the underlying mechanisms of root-to-shoot Mn translocation remain unclear. Here, we identified an Natural Resistance Associated Macrophage Protein (NRAMP) family member in maize (Zea mays), ZmNRAMP2, which localized to the tonoplast in maize protoplasts and mediated transport of Mn in yeast (Saccharomyces cerevisiae). Under Mn deficiency, two maize mutants defective in ZmNRAMP2 exhibited remarkable reduction of root-to-shoot Mn translocation along with lower shoot Mn contents, resulting in substantial decreases in Fv/Fm and plant growth inhibition compared to their corresponding wild-type (WT) plants. ZmNRAMP2 transcripts were highly expressed in xylem parenchyma cells of the root stele. Compared to the WT, the zmnramp2-1 mutant displayed lower Mn concentration in xylem sap accompanied with retention of Mn in root stele. Furthermore, the overexpression of ZmNRAMP2 in transgenic maize showed enhanced root-to-shoot translocation of Mn and improved tolerance to Mn deficiency. Taken together, our study reveals a crucial role of ZmNRAMP2 in root-to-shoot translocation of Mn via accelerating vacuolar Mn release in xylem parenchyma cells for adaption of maize plants to low Mn stress and provides a promising transgenic approach to develop low Mn-tolerant crop cultivars.

The Intensity of Manganese Deficiency Strongly Affects Root Endodermal Suberization and Ion Homeostasis.

Manganese (Mn) deficiency affects various processes in plant shoots. However, the functions of Mn in roots and the processes involved in root adaptation to Mn deficiency are largely unresolved. Here, we show that the suberization of endodermal cells in barley (Hordeum vulgare) roots is altered in response to Mn deficiency, and that the intensity of Mn deficiency ultimately determines whether suberization increases or decreases. Mild Mn deficiency increased the length of the unsuberized zone close to the root tip, and increased the distance from the root tip at which the fully suberized zone developed. By contrast, strong Mn deficiency increased suberization closer to the root tip. Upon Mn resupply, suberization was identical to that seen on Mn-replete plants. Bioimaging and xylem sap analyses suggest that the reduced suberization in mildly Mn-deficient plants promotes radial Mn transport across the endodermis at a greater distance from the root tip. Less suberin also favors the inwards radial transport of calcium and sodium, but negatively affects the potassium concentration in the stele. During strong Mn deficiency, Mn uptake was directed toward the root tip. Enhanced suberization provides a mechanism to prevent absorbed Mn from leaking out of the stele. With more suberin, the inward radial transport of calcium and sodium decreases, whereas that of potassium increases. We conclude that changes in suberization in response to the intensity of Mn deficiency have a strong effect on root ion homeostasis and ion translocation.

Multi-Element Bioimaging of Arabidopsis thaliana Roots.

Better understanding of root function is central for the development of plants with more efficient nutrient uptake and translocation. We here present a method for multielement bioimaging at the cellular level in roots of the genetic model system Arabidopsis (Arabidopsis thaliana). Using conventional protocols for microscopy, we observed that diffusible ions such as potassium and sodium were lost during sample dehydration. Thus, we developed a protocol that preserves ions in their native, cellular environment. Briefly, fresh roots are encapsulated in paraffin, cryo-sectioned, and freeze dried. Samples are finally analyzed by laser ablation-inductively coupled plasma-mass spectrometry, utilizing a specially designed internal standard procedure. The method can be further developed to maintain the native composition of proteins, enzymes, RNA, and DNA, making it attractive in combination with other omics techniques. To demonstrate the potential of the method, we analyzed a mutant of Arabidopsis unable to synthesize the metal chelator nicotianamine. The mutant accumulated substantially more zinc and manganese than the wild type in the tissues surrounding the vascular cylinder. For iron, the images looked completely different, with iron bound mainly in the epidermis of the wild-type plants but confined to the cortical cell walls of the mutant. The method offers the power of inductively coupled plasma-mass spectrometry to be fully employed, thereby providing a basis for detailed studies of ion transport in roots. Being applicable to Arabidopsis, the molecular and genetic approaches available in this system can now be fully exploited in order to gain a better mechanistic understanding of these processes.

A chitin deacetylase PsCDA2 from Puccinia striiformis f. sp. tritici confers disease pathogenicity by suppressing chitin-triggered immunity in wheat.

Plants have the ability to recognize the essential chitin molecule present in the fungal cell wall, which stimulates the immune response. Phytopathogenic fungi have developed various strategies to inhibit the chitin-triggered immune response. Here, we identified a chitin deacetylase of Puccinia striiformis f. sp. tritici (Pst), known as PsCDA2, that was induced during the initial invasion of wheat and acted as an inhibitor of plant cell death. Knockdown of PsCDA2 in wheat enhanced its resistance against Pst, highlighting the significance of PsCDA2 in the host-pathogen interaction. Moreover, PsCDA2 can protect Pst urediniospores from being damaged by host chitinase in vitro. PsCDA2 also suppressed the basal chitin-induced plant immune response, including the accumulation of callose and the expression of defence genes. Overall, our results demonstrate that Pst secretes PsCDA2 as a chitin deacetylase involved in establishing infection and modifying the acetyl group to prevent the breakdown of chitin in the cell wall by host endogenous chitinases. Our research unveils a mechanism by which the fungus suppresses plant immunity, further contributing to the understanding of wheat stripe rust control. This information could have significant implications for the development of suitable strategies for protecting crops against the devastating effects of this disease.

Nitrate_dependent suberization regulates cadmium uptake and accumulation in maize.

In this study, effect of nitrate-dependent suberization in maize root on cadmium (Cd) uptake and accumulation was investigated. Suberization in maize roots was significantly lower in plants grown with a high nitrate supply compared with low nitrate. This decrease was seen in the total amount of suberin, in which the aliphatic suberin amount was significantly decreased, whereas no difference in aromatic suberin content between different N-treatments. RNA-sequencing showed that suberin biosynthesis genes were upregulated in low nitrate treatment, which correlated well with the increased suberin content. Bioimaging and xylem sap analysis showed that reduced exodermal and endodermal suberization in roots of plants grown under high nitrate promoted radial Cd transport along the crown root. The enhanced suberization in crown roots of plants grown in low nitrate restricted the radial transport of Cd from epidermis to cortex via decreased accessibility to Cd related transporters at the plasmalemma. Also, under low nitrate supply, the Cd transport gene ZmNramp5 was upregulated in the crown root, which may enhance Cd uptake by root tip where exodermis and endodermis were not fully suberized. These results suggest that high nitrate supply enhances Cd uptake and radial transport in maize roots by reducing exodermal and endodermal suberization.

Plant root suberin: A layer of defence against biotic and abiotic stresses.

Plant roots have important functions, such as acquiring nutrients and water from the surrounding soil and transporting them upwards to the shoots. Simultaneously, they must be able to exclude potentially harmful substances and prevent the entry of pathogens into the roots. The endodermis surrounds the vascular tissues and forms hydrophobic diffusion barriers including Casparian strips and suberin lamella. Suberin in cell walls can be induced by a range of environmental factors and contribute to against biotic and abiotic threats. Tremendous progress has been made in biosynthesis of suberin and its function, little is known about the effect of its plasticity and distribution on stress tolerance. In field conditions, biotic and abiotic stress can exist at the same time, and little is known about the change of suberization under that condition. This paper update the progress of research related to suberin biosynthesis and its function, and also discuss the change of suberization in plant roots and its role on biotic and abiotic stresses tolerance.

Hydrothermal conversion of Chinese cabbage residue for sustainable agriculture: Influence of process parameters on hydrochar and hydrolysate.

The demands on novel and sustainable techniques for vegetable waste (VW) valorization continues to increase during the past few decades due to the growing waste production under the flourishing vegetable industries. In this study, Chinese cabbage residues were hydrothermal carbonization (HTC) at 180, 200, 220 and 240 °C for 2 to 6 h to explore the impacts of process parameters on the characteristics of hydrochars and hydrolysates and their feasibility in sustainable agriculture. Results indicated that hydrothermal temperature had a greater impact on cabbage residue hydrolysis than the residence time. With the rising reaction severity, hydrochars became more alkaline with higher amount of ash and carbon (C), while the pH and dissolved organic nitrogen (DON) and NH4+-N in the hydrolysate were gradually reduced. The thermogravimetric analysis (TG-DTG) indicated that organic constitutions in the feedstock went through incomplete decomposition. Although the recalcitrance index (R50) steadily increased through HTC (0.37-0.46), hydrochars were unstable and would not applicable for carbon sequestration. Furthermore, hydrochars and hydrolysate would be optimal media for plants seedling and growth for the abundant nutrients and dissolved organic compounds but reduced phytotoxicity. In conclusion, these results showed that HTC is highly applicable for vegetable waste management for sustainable agriculture.

Bioremediation of Cd-spiked soil using earthworms (Eisenia fetida): Enhancement with biochar and Bacillus megatherium application.

In this study, we evaluated the influence of biochar and Bacillus megatherium on Cd removal from artificially contaminated soils using earthworms (Eisenia fetida). Within a 35-days remediation period, over 30% of Cd was removed by earthworms from the contaminated soil (with Cd at âˆ¼ 2.5 mg kg-1), and both additives facilitated Cd removal. Additionally, over 22% reduction in the extractable Cd contents was also achieved by earthworms. Cd accumulated in earthworms steadily increased through remediation, and the accumulated Cd decreased in the order of earthworm + biochar (T3) > earthworm + Bacillus megatherium (T4) > earthworm alone (T2). The bioaccumulation factors (BCF) were above 1, indicating the enrichment of Cd in earthworms, and there were higher BCF for both T4 (944%) and T3 (845%). The ingestion of metal-bonded biochar particle and the elevated Cd mobility would be the main reason for the enhanced Cd-remediation by earthworms under T3 and T4, respectively. Through remediation, microbiota communities in both, soil and earthworm guts, demonstrated high similarity, while a lower level of bacterial abundance was observed in earthworm guts compared with that in soils. Eventually, soils became more fertile and demonstrated higher enzyme activities after remediation. Therefore, we concluded that earthworm, alone or combined with biochar or Bacillus megatherium could be an alternative method for Cd-contaminated soil remediation.

Enhanced aqueous Cr(VI) removal using chitosan-modified magnetic biochars derived from bamboo residues.

The purpose of this study was to fabricate a chitosan modified magnetic bamboo biochar (CMBB) for Cr(VI) detoxification from aqueous solution. Results showed that chitosan modification provided more active adsorption sites on the surface of magnetic bamboo biochar (MBB), and hence enhanced Cr(VI) removal from aqueous solution. The maximum adsorption capacities of MBB and CMBB for Cr(VI) at 25 °C were 75.8 and 127 mg g-1, respectively. Increasing solution pH inbibited Cr(VI) adsorption by adsorbents. However, CMBB maintained a high Cr(VI) removal efficiencies over a broader pH range (2-10), and could attain 36% of the maximum adsorption (40 mg g-1) even at a high pH of 10.0. Rising temperature enhanced the Cr(VI) removal by two adsorbents. The escalating ionic strength and coexist substances, including Na+, Ca2+, Fe3+, Cl-, SO42-, PO43- and humate, inhibited the adsorption efficiency of Cr(VI) on adsorbents. After the fifth adsorption-desorption cycle, the adsorption efficiencies of CMBB and MBB for Cr(VI) remained above 90% and less than 50%, respectively. All these results indicated that CMBB could be a practical adsorbent that can be utilized for the detoxification of Cr(VI) ions from wastewater.

Towards single-cell ionomics: a novel micro-scaled method for multi-element analysis of nanogram-sized biological samples.

BACKGROUND: To understand processes regulating nutrient homeostasis at the single-cell level there is a need for new methods that allow multi-element profiling of biological samples ultimately only available as isolated tissues or cells, typically in nanogram-sized samples. Apart from tissue isolation, the main challenges for such analyses are to obtain a complete and homogeneous digestion of each sample, to keep sample dilution at a minimum and to produce accurate and reproducible results. In particular, determining the weight of small samples becomes increasingly challenging when the sample amount decreases. RESULTS: We developed a novel method for sampling, digestion and multi-element analysis of nanogram-sized plant tissue, along with strategies to quantify element concentrations in samples too small to be weighed. The method is based on tissue isolation by laser capture microdissection (LCM), followed by pressurized micro-digestion and ICP-MS analysis, the latter utilizing a stable µL min-1 sample aspiration system. The method allowed for isolation, digestion and analysis of micro-dissected tissues from barley roots with an estimated sample weight of only ~ 400 ng. In the collection and analysis steps, a number of contamination sources were identified. Following elimination of these sources, several elements, including magnesium (Mg), phosphorus (P), potassium (K) and manganese (Mn), could be quantified. By measuring the exact area and thickness of each of the micro-dissected tissues, their volume was calculated. Combined with an estimated sample density, the sample weights could subsequently be calculated and the fact that these samples were too small to be weighed could thereby be circumvented. The method was further documented by analysis of Arabidopsis seeds (~ 20 µg) as well as tissue fractions of such seeds (~ 10 µg). CONCLUSIONS: The presented method enables collection and multi-element analysis of small-sized biological samples, ranging down to the nanogram level. As such, the method paves the road for single cell and tissue-specific quantitative ionomics, which allow for future transcriptional, proteomic and metabolomic data to be correlated with ionomic profiles. Such analyses will deepen our understanding of how the elemental composition of plants is regulated, e.g. by transporter proteins and physical barriers (i.e. the Casparian strip and suberin lamellae in the root endodermis).