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Mercury (Hg) contamination in paddy soils poses a health risk to rice consumers and the environmental behavior of Hg determines its toxicity. Thus, the variations of Hg speciation are worthy of exploring. In this study, microcosm and pot experiments were conducted to elucidate Hg transformation, methylation, bioaccumulation, and risk coupled with biogeochemical cycling of key elements in a Hg-polluted alkaline paddy soil. In microcosm and pot experiments, organic- and sulfide-bound and residual Hg accounted for more than 98% of total Hg, and total contents of dissolved, exchangeable, specifically adsorbed, and fulvic acid-bound Hg were less than 2% of total Hg, indicating a low mobility and environmental risk of Hg. The decrease of pH aroused from Fe(III), SO42-, and NO3- reduction promoted Hg mobility, whereas the increase of pH caused by Fe(II), S2-, and NH4+ oxidation reduced available Hg contents. Moreover, Fe-bearing minerals reduction and organic matter consumption promoted Hg mobility, whereas the produced HgS and Fe(II) oxidation increased Hg stability. During flooding, a fraction of inorganic Hg (IHg) could be transported into methylmercury (MeHg), and during drainage, MeHg would be converted back into IHg. After planting rice in an alkaline paddy soil, available Hg was below 0.3 mg kg-1. During rice growth, a portion of available Hg transport from paddy soil to rice, promoting Hg accumulation in rice grains. After rice ripening, IHg levels in rice tissues followed the trend: root > leaf > stem > grain, and IHg content in rice grain exceed 0.02 mg kg-1, but MeHg content in rice grain meets daily intake limit (37.45 µg kg-1). These results provide a basis for assessing the environmental risks and developing remediation strategies for Hg-contaminated redox-changing paddy fields as well as guaranteeing the safe production of rice grains.

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Hexavalent chromium (Cr(VI)) is a toxic element that has negative impacts on crop growth and yield. Using plant extracts to convert toxic Cr(VI) into less toxic Cr(III) may be a more favorable option compared to chemical reducing agents. In this study, the potential effects and mechanisms of using an aqueous extract of Psidium guajava L. leaves (AEP) in reducing Cr(VI) toxicity in rice were comprehensively studied. Firstly, the reducing power of AEP for Cr(VI) was confirmed by the cyclic voltammetry combined with X-ray photoelectron spectroscopy (XPS) assays. The highest Cr(VI) reduction efficiency reached approximately 78% under 1.5 mg gallic acid equivalent (GAE)/mL of AEP and 10 mg/L Cr(VI) condition. Additionally, Cr(VI) stress had a significant inhibitory effect on rice growth. However, the exogenous application of AEP alleviated the growth inhibition and oxidative damage of rice under Cr(VI) stress by increasing the activity and level of enzymatic and non-enzymatic antioxidants. Furthermore, the addition of AEP restored the ultrastructure of root cells, promoted Cr adsorption onto root cell walls, and limited the translocation Cr to shoots. In shoots, AEP application also triggered the expression of specific genes involved in Cr defense and detoxification response, including photosynthesis pathways, antioxidant systems, flavonoids biosynthesis, and plant hormone signal transduction. These results suggest that AEP is an efficient reduction agent for Cr(VI), and exogenous application of AEP may be a promising strategy to mitigate the harm of Cr(VI) on rice, ultimately contributing to improved crop yield in Cr-contaminated environments.

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BACKGROUND: Crop recognition is the basis of intelligent agricultural machine operations. Visual perception methods have achieved high recognition accuracy. However, the reliability of such methods is difficult to guarantee because of the complex environment of paddy fields. Tactile sensing methods are not affected by background or environmental interference, and have high reliability. However, in an ideal environment, the recognition accuracy is not as high as that of the visual method. RESULTS: To balance the accuracy and reliability of rice plant recognition, a combined visual-tactile method was proposed in this study. A rice plant recognition device was developed with a poly(vinylidene fluoride) sensor embedded inside the device as a tactile perceptron and a graphic designed as a visual perceptron. The primary role of the tactile perceptron is to initially recognize rice plants and provide a time point for image capture for visual perception. The main role of the visual perceptron is to extract features from the captured images and recognize rice plants again. The results of tactile and visual recognition were eventually fused to achieve accurate recognition of rice plants. CONCLUSION: The contact speed between the recognition perceptron and rice-weed was selected for the field performance test based on the real situation of paddy field operation. The results showed that the accuracy and reliability of rice plant recognition decreased as the travelling speed of the paddy field operation machine increased. The results of this study provide a basis for intelligent farm machinery operations in rice fields. © 2024 Society of Chemical Industry.

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Selective degradation of the endoplasmic reticulum (ER) by macroautophagy/autophagy (reticulophagy) is essential for maintaining ER morphology and homeostasis under environmental stresses. Several reticulophagy receptors have been identified in mammals and yeast, but their counterparts in plants have not been extensively explored yet. Recently, we demonstrated that the HVA22-family protein OsHLP1 is a reticulophagy receptor in rice plants, and its orthologs function similarly in Arabidopsis plants. In this punctum, we discuss why the HVA22 family proteins are the reticulophagy receptors in plants and how reticulophagy is highly associated with plant immune response.

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Generation of mutant populations with high genetic diversity is key for mutant screening and crop breeding. For this purpose, the single-seed descent method, in which one mutant line is established from a single mutagenized seed, is commonly used. This method ensures the independence of the mutant lines, but the size of the mutant population is limited because it is no greater than the number of fertile M1 plants. The rice mutant population size can be increased if a single mutagenized plant produces genetically independent siblings. Here, we used whole-genome resequencing to examine the inheritance of mutations from a single ethyl methanesulfonate (EMS)-mutagenized seed (M1 ) of Oryza sativa in its progeny (M2 ). We selected five tillers from each of three M1 plants. A single M2 seed was selected from each tiller, and the distributions of mutations induced by EMS were compared. Surprisingly, in most pairwise combinations of M2 siblings from the same parent, ≥85.2-97.9% of all mutations detected were not shared between the siblings. This high percentage suggests that the M2 siblings were derived from different cells of the M1 embryo and indicates that several genetically independent lines can be obtained from a single M1 plant. This approach should allow a large reduction in the number of M0 seeds needed to obtain a mutant population of a certain size in rice. Our study also suggests that multiple tillers of a rice plant originate from different cells of the embryo.

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Melatonin (MT) has been demonstrated to provide defense against both biotic and abiotic stressors. Boron toxicity (BT) can significantly limit the growth and production of plants. However, few studies have been conducted on whether MT is effective in attenuating B toxicity in different plants. In order to evaluate the efficacy of exogenous MT treatment in reducing the negative impact of BT on rice seedlings, this study examined the influence of MT on growth, antioxidant capacity, cell wall composition, and proline metabolism in rice seedlings under hydroponics. Four treatments were established: MT (50 µM), MT + BT (50 µM MT + 800 µM B), BT (800 µM), and CK (control) in a completely randomized design. The results indicate that BT had a significant detrimental effect on the shoot length, root length, and root and shoot fresh weights of rice seedlings by 11.96%, 27.77%, 25.69%, and 18.67%, respectively as compared to the control treatment. However, exogenous MT application increased these parameters and reduced B accumulation in aboveground parts (14.05%) of the plant. Exogenous MT also increased the endogenous melatonin content and antioxidant enzyme activities (64.45%, 71.61%, 237.64%, and 55.42% increase in superoxide dismutase, ascorbate peroxidase, and peroxidase activities, respectively), while decreasing reactive oxygen species levels and oxidized forms of glutathione and ascorbic acid. Additionally, MT enhanced the biosynthesis of proline by decreasing proline dehydrogenase (ProDH) and increasing the GSH (glutathione) and ASA (ascorbic acid) contents. Exogenous MT also increased cell wall components that can increase B adsorption to the cell wall. Overall, these findings suggest that MT application can be a potential solution for strengthening the stress tolerance of rice seedlings, particularly under conditions of B toxicity. In regions where soil contains high levels of boron, the use of MT could enhance rice crop yields and quality.

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Introduction: Selenium (Se) is an essential trace element required for proper human and animal health. Methods: In this paper, we investigated the uptake and distribution characteristics of a new Se fertilizer, which comprises algal polysaccharides-selenium nanoparticles (APS-SeNPs), in rice plants in both hydroponic and pot experiments. Results: The results from the hydroponic experiments revealed that the rice root uptake of APS-SeNPs fitted the Michaelis-Menten equation, with a V max of 13.54 µg g-1 root dry weight (DW) per hour, which was 7.69 and 2.23 times those of selenite and selenate treatments, respectively. The root uptake of APS-SeNPs was inhibited by AgNO3 (64.81%-79.09%) and carbonyl cyanide 3-chlorophenylhydrazone (CCCP; 19.83%-29.03%), indicating that the uptake of APS-SeNPs by rice roots is mainly via aquaporins and is also affected by metabolic activity. Moreover, sulfur deficiency caused rice roots to absorb more APS-SeNPs, but treatment with APS-SeNPs increased the expression of the sulfate transporter OsSULTR1;2 in the roots, suggesting that OsSULTR1;2 is probably involved in the uptake of APS-SeNPs. The application of APS-SeNPs significantly increased the Se content in rice plants and the apparent Se uptake efficiency compared with selenate and selenite treatments. Most of the Se in the roots of rice plants was distributed in the cell wall, while it was primarily located in the cytosol in the shoots when treated with APS-SeNPs. The results from the pot experiments indicated that the application of Se enhanced the Se content of each rice tissue. It is worth noting that the Se content in brown rice under APS-SeNP treatment was higher than that under selenite or selenate treatment and was mainly concentrated in the embryo end, with the Se in organic form. Discussion: Our findings provide important insights into the uptake mechanism and the distribution of APS-SeNPs in rice plants.

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It is predicted that plant volatile organic compounds (VOCs) are affected by the atmospheric CO2 levels rising globally, which further affects the interaction between plants and herbivorous insects, especially the host selection behavior of herbivorous insects. In this study, the effects of elevated CO2 on the host-selection behavior of the brown planthopper (BPH) Nilaparvata lugens, and the emission of VOCs from the healthy and BPH-damaged rice plants were studied simultaneously to make clear the population occurrence of BPH under global climate change. Compared with ambient CO2, elevated CO2 significantly increased the host selection percent of BPH for the healthy (CK) and BPH-damaged rice plants, and the host selection percent of BPH for the BPH-damaged rice plants was significantly higher than that for the healthy rice plants under elevated CO2, which might be regulated by the transcription levels of OBP1, OBP2 and CSP8 in BPH due to the upregulated transcriptional levels of these three genes of BPH under elevated CO2. In addition, we analyzed and quantified the emission of VOCs in rice plants grown under ambient CO2 and elevated CO2 by GS-MS. A total of 36 VOCs from rice plants were identified into eight categories, including alkanes, alkenes, alcohols, aldehydes, ketones, esters, phenols and aromatic hydrocarbons. Elevated CO2 significantly decreased the contents of heptadecane, linalool and limonene from rice plants compared with ambient CO2. Besides, the contents of linalool, phytol, decanal, 1-methyldecalin and 2,6-diphenylphenol from BPH-damaged rice plants under ambient CO2, and undecane, hexadecane, nonanal and 2,6-diphenylphenol from BPH-damaged rice plants under elevated CO2 were all significantly higher than those from healthy rice plants. The percentage composition of phenols was positively correlated with the host selection rate of BPH. Our study indicates that elevated CO2 is beneficial to promote the host selection ability of BPH for rice plants damaged by BPHs due to the changed plant VOCs.

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The bioaccumulation of the neurotoxin methylmercury (MeHg) in rice is a significant concern due to its potential risk to humans. Thiols have been known to affect MeHg bioavailability in microorganisms, but how thiols influence MeHg accumulation in rice plants remains unknown. Here, we investigated effects of common low-molecular-weight thiols, including cysteine (Cys), glutathione (GSH), and penicillamine (PEN), on MeHg uptake and translocation by rice plants. Results show that rice roots can rapidly take up MeHg, and this process is influenced by the types and concentrations of thiols in the system. The presence of Cys facilitated MeHg uptake by roots and translocation to shoots, while GSH could only promote MeHg uptake, but not translocation, by roots. Conversely, PEN significantly inhibited MeHg uptake and translocation to shoots. Using labeled 13Cys assays, we also found that MeHg uptake was coupled with Cys accumulation in rice roots. Moreover, analyses of comparative transcriptomics revealed that key genes associated with metallothionein and SULTR transporter families may be involved in MeHg uptake. These findings provide new insights into the uptake and translocation of MeHg in rice plants and suggest potential roles of thiol attributes in affecting MeHg bioavailability and bioaccumulation in rice.

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BACKGROUND: Improvement in photosynthesis is one of the most promising approaches to increase grain yields. Transgenic rice plants overproducing Rubisco by 30% (RBCS-sense rice plants) showed up to 28% increase in grain yields under sufficient nitrogen (N) fertilization using an isolated experimental paddy field (Yoon et al. in Nat Food 1:134-139, 2020). The plant N contents above-ground sections and Rubisco contents of the flag leaves were higher in the RBCS-sense plants than in the wild-type rice plants during the ripening period, which may be reasons for the increased yields. However, some imprecise points were left in the previous research, such as contributions of photosynthesis of leaves below the flag leaves to the yield, and maintenance duration of high photosynthesis of RBCS-sense rice plants during ripening periods. RESULT: In this research, the photosynthetic capacity and canopy architecture were analyzed to explore factors for the increased yields of RBCS-sense rice plants. It was found that N had already been preferentially distributed into the flag leaves at the early ripening stage, contributing to maintaining higher Rubisco content levels in the enlarged flag leaves and extending the lifespan of the flag leaves of RBCS-sense rice plants throughout ripening periods under sufficient N fertilization. The higher amounts of Rubisco also improved the photosynthetic activity in the flag leaves throughout the ripening period. Although the enlarged flag leaves of the RBCS-sense rice plants occupied large spatial areas of the uppermost layer in the canopy, no significant prevention of light penetration to leaves below the flag leaves was observed. Additionally, since the CO2 assimilation rates of lower leaves between wild-type and RBCS-sense rice plants were the same at the early ripening stage, the lower leaves did not contribute to an increase in yields of the RBCS-sense rice plants. CONCLUSION: We concluded that improvements in the photosynthetic capacity by higher leaf N and Rubisco contents, enlarged leaf area and extended lifespan of flag leaves led to an increase in grain yields of RBCS-sense rice plants grown under sufficient N fertilization.

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Deep learning-based object counting models have recently been considered preferable choices for plant counting. However, the performance of these data-driven methods would probably deteriorate when a discrepancy exists between the training and testing data. Such a discrepancy is also known as the domain gap. One way to mitigate the performance drop is to use unlabeled data sampled from the testing environment to correct the model behavior. This problem setting is also called unsupervised domain adaptation (UDA). Despite UDA has been a long-standing topic in machine learning society, UDA methods are less studied for plant counting. In this paper, we first evaluate some frequently-used UDA methods on the plant counting task, including feature-level and image-level methods. By analyzing the failure patterns of these methods, we propose a novel background-aware domain adaptation (BADA) module to address the drawbacks. We show that BADA can easily fit into object counting models to improve the cross-domain plant counting performance, especially on background areas. Benefiting from learning where to count, background counting errors are reduced. We also show that BADA can work with adversarial training strategies to further enhance the robustness of counting models against the domain gap. We evaluated our method on 7 different domain adaptation settings, including different camera views, cultivars, locations, and image acquisition devices. Results demonstrate that our method achieved the lowest Mean Absolute Error on 6 out of the 7 settings. The usefulness of BADA is also supported by controlled ablation studies and visualizations.

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Thallium (Tl) is an extremely toxic metal, while its occurrence and fate in paddy soil environment remain understudied. Herein, the enrichment and migration mechanisms and potential health risks of Tl and metal(loid)s were evaluated in paddy soils surrounding an industrial park utilizing Tl-bearing minerals. The results showed that Tl contamination was evident (0.63-3.16 mg/kg) in the paddy soils and Tl was generally enriched in root of rice (Oryza sativa L.) with a mean content of 1.27 mg/kg. A remarkably high level of Tl(III) (30-50%) was observed in the paddy soils. Further analyses by STEM-EDS and XPS indicated that Tl(I) in the paddy soils was jointly controlled by adsorption, oxidation, and precipitation of Fe/Mn(hydr)oxide (e.g. hematite and birnessite), which might act as important stabilization mechanisms for inhibiting potential Tl uptake by rice grains. The health quotient (HQ) values indicated a potentially high Tl risk for inhabitants via consumption of the rice grains. Therefore, it is critical to establish effective measures for controlling the discharge of Tl-containing waste and wastewater from different industrial activities to ensure food safety in the rice paddy soils.

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Viruses are ubiquitous in nature and exist in a variety of habitats. The advancement in sequencing technologies has revolutionized the understanding of viral biodiversity associated with plant diseases. Deep sequencing combined with metagenomics is a powerful approach that has proven to be revolutionary in the last decade and involves the direct analysis of viral genomes present in a diseased tissue sample. This protocol describes the details of RNA extraction and purification from wild rice plant and their yield, RNA purity, and integrity assessment. As a final step, bioinformatics data analysis including demultiplexing, quality control, de novo transcriptome assembly, taxonomic allocation and read mapping following Illumina HiSeq small and total RNA sequencing are described. Furthermore, the total RNAs extraction protocol and an additional ribosomal rRNAs depletion step which are significantly important for viral genomes construction are provided.

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A new disease in rice that is characterized by leaf bleaching was recently identified in some fields in the Mekong Delta region of Vietnam. The present study was the first to isolate and identify the pathogen of this disease. We confirmed that leaf bleaching symptoms were due to infection with Methylobacterium indicum bacteria using molecular biology approaches. A full-length genome analysis of pathogenic Methylobacterium strain VL1 revealed that it comprises a single chromosome and six plasmids, with a total size of 7.05| |Mbp and GC content of 70.5%. The genomic features of VL1 were similar to those of the non-pathogenic M. indicum strain SE2.11T; however, VL1 possessed additional unique genes, including those related to homoserine lactone biosynthesis. We established a loop-mediated isothermal amplification (LAMP) assay using the unique sequences of VL1 as target sequences for the rapid and simple detection of pathogenic M. indicum strains. Our initial evaluation demonstrated that the LAMP assay successfully distinguished between pathogenic and non-pathogenic strains infecting rice plants in a rapid and sensitive manner. The present results provide insights into the pathogenesis and development of control measures for novel rice diseases.

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Thiocyanate (SCN-) present in irrigation water can have negative effects on plant growth and crop yields. Addition of plant growth regulators (PGRs) can alleviate toxic stress to plants. In the current study, we established a grey situation decision-making model (GSDM) to integrate the data of RT-qPCR analysis for screening the optimal addition of PGRs to minimise pollution stress. The effects of PGRs (i.e., jasmonic acid [JA], indole-3-acetic acid [IAA] and sodium hydrosulfide [NaHS]) on the abundance of IAA oxidation and conjugation-related genes in rice seedlings under potassium thiocyanate (KSCN) exposure was examined. The results obtained from RT-qPCR analysis can roughly present the mitigating effects of IAA, JA, and NaHS on rice seedlings under KSCN stress. Integration of RT-qPCR analysis and GSDM further quantified the regulatory effects of PGRs. Simulation results showed that the effect of NaHS on the gene expression at KSCN exposure is apparently better than that of JA and IAA. Our study provides a new simple, efficient, and cheap approach to identify the optimal plant growth regulators under the stress of environmental pollution.

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Using an in-situ experiment in Cd contaminated paddy fields in Chongqing, the absorption and distribution of Cd in rice plants was examined following the combined application of lime and organic matter, and the mechanisms driving changes in Cd fractions in soil were also studied. The results showed that pH, OM content, and CEC in the soil were significantly enhanced, and OM and CEC were significantly positively correlated, as pH showed a significant positive correlation; pH and CEC was mainly associated with a change in Cd from the acidic extractable fraction to the reducible, oxidable, and residual fractions. The percentage of acidic extractable Cd dropped by 22.92%-31.25% with the application of the amendments, thus reducing the accumulation of Cd in rice plants in the followed order:CK (control group)≫B6 (lime and maize straw)≈B4 (lime and biochar)≈B3 (lime and activated humic acidic fertilizer) > B2 (lime and humic acidic fertilizer)≈B1 (lime) > B5 (lime and oil cake). On the other hand, for both the treatments and control conditions (CK), the Cd content of brown rice was higher than Chinese standard (GB 2762-2017), while that of polished (white) rice was lower than the Chinese standard after treatment. Overall, treatment using all of the remediation agents offers some benefit for the safe utilization of agricultural contaminated soil and safe food production. The combined application of lime and oil cake proved the best measure for treating Cd contaminated acidic rice fields, yielding a reduction in acidic extractable Cd (31.25%), a reduction in the Cd content of rice plants (33.32%), and the lowest Cd content (0.13 mg·kg-1) in polished rice (a 42.17% compared to the control).

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The brown planthopper (BPH) (Nilaparvata lugens Stål) is a highly destructive pest that seriously damages rice (Oryza sativa L.) and causes severe yield losses. To better understand the physiological and metabolic mechanisms through which BPHs respond to resistant rice, we combined mass-spectrometry-based lipidomics with transcriptomic analysis and gene knockdown techniques to compare the lipidomes of BPHs feeding on either of the two resistant (NIL-Bph6 and NIL-Bph9) plants or a wild-type, BPH susceptible (9311) plant. Insects that were fed on resistant rice transformed triglyceride (TG) to phosphatidylcholine (PC) and digalactosyldiacylglycerol (DGDG), with these lipid classes showing significant alterations in fatty acid composition. Moreover, the insects that were fed on resistant rice were characterized by prominent expression changes in genes involved in lipid metabolism processes. Knockdown of the NlBmm gene, which encodes a lipase that regulates the mobilization of lipid reserves, significantly increased TG content and feeding performance of BPHs on resistant plants relative to dsGFP-injected BPHs. Our study provides the first detailed description of lipid changes in BPHs fed on resistant and susceptible rice genotypes. Results from BPHs fed on resistant rice plants reveal that these insects can accelerate TG mobilization to provide energy for cell proliferation, body maintenance, growth and oviposition.

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MAIN CONCLUSION: Higher vacuolar proton pump activity may increase plant energy and nutrient use efficiency and provide the nexus between plant inoculation with Herbaspirillum seropedicae and growth promotion. Global change and growing human population are exhausting arable land and resources, including water and fertilizers. We present inoculation with the endophytic plant-growth promoting bacterium (PGPB) Herbaspirillum seropedicae as a strategy for promoting growth, nutrient uptake and photosynthetic efficiency in rice (Oryza sativa L.). Because plant nutrient acquisition is coordinated with photosynthesis and the plant carbon status, we hypothesize that inoculation with H. seropedicae will stimulate proton (H+) pumps, increasing plant growth nutrient uptake and photosynthetic efficiency at low nutrient levels. Plants were inoculated and grown in pots with sterile soil for 90 days. Herbaspirillum seropedicae endophytic colonization was successful and, as hypothesized, inoculation (1) stimulated root vacuolar H+ pumps (vacuolar H+-ATPase and vacuolar H+-PPase), and (2) increased plant growth, nutrient contents and photosynthetic efficiency. The results showed that inoculation with the endophytic bacterium H. seropedicae can promote plant growth, nutrient uptake and photosynthetic efficiency, which will likely result in a more efficient use of resources (nutrients and water) and higher production of nutrient-rich food at reduced economic and environmental costs.

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The structural analogs, 2,4-dibromophenol (2,4-DBP) and 2,4-dibromoanisole (2,4-DBA), have both natural and artificial sources and are frequently detected in environmental matrices. Their environmental fates, especially volatilization, including both direct volatilization from cultivation solution and phytovolatilization through rice plants were evaluated using hydroponic exposure experiments. Results showed that 2,4-DBA displayed stronger volatilization tendency and more bioaccumulation in aboveground rice tissues. Total volatilized 2,4-DBA accounted for 4.74% of its initial mass and was 3.43 times greater than 2,4-DBP. Phytovolatilization of 2,4-DBA and 2,4-DBP contributed to 6.78% and 41.7% of their total volatilization, enhancing the emission of these two contaminants from hydroponic solution into atmosphere. In this study, the interconversion processes between 2,4-DBP and 2,4-DBA were first characterized in rice plants. The demethylation ratio of 2,4-DBA was 12.0%, 32.0 times higher than methylation of 2,4-DBP. Formation of corresponding metabolites through methylation and demethylation processes also contributed to the volatilization of 2,4-DBP and 2,4-DBA from hydroponic solution into the air phase. Methylation and demethylation processes increased phytovolatilization by 12.1% and 36.9% for 2,4-DBP and 2,4-DBA. Results indicate that phytovolatilization and interconversion processes in rice plants serve as important pathways for the global cycles of bromophenols and bromoanisoles.

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Due to the nonsystematic nature of previous studies on mercury (Hg) mobility with humic substances (HS) in terrestrial ecosystems and the uncertainty of Hg accumulation in plants, oxygen-rich humic acid (HA), which is the main component of HS, was used as the target in this study. Batch sorption tests and a series of pot experiments were designed to investigate the effect of HS on Hg binding and therefore Hg uptake in rice plants under extreme conditions, i.e., a high Hg/HS concentration ratio. The results showed that HA was eligible for Hg binding, though it has a tiny proportion of sulfur according to its characteristics analysis. The binding of HA and Hg was a chemisorption process in a single layer that followed the pseudo-second order and Langmuir models, and it was also verified that the pH was dependent on the ion strength associated with high Hg/HA concentration ratios. Based on the pot experiments, the performance of HA with Hg was investigated. The Hg in the toxicity characteristic leaching procedure (TCLP) leachate under high Hg/HA concentration ratios declined significantly, and accordingly, all treatments met the concentration criteria of 0.1 mg/l (GB 5085.3-2007) for wastes after 30 days of exposure. At concentration ratios of 50, 25, and 10 µg Hg/mg HA, we observed that HA application promoted rice plant growth, as reflected in the increase of fresh weight of different organs. Regarding accumulation in the soil-plant system, the degradation of HA to smaller molecules by rhizosphere microorganisms and organic acids in roots made HA available for plant uptake through the vascular bundle in roots, thus promoting Hg transformation in plants to a certain extent. However, considering the decline in available Hg in the soil, the Hg concentrations of roots, straw, and grains in the ripening stage were found to be lower than those in the standalone Hg treatments. HA clearly has a direct effect on Hg and an indirect influence on plants exposed to Hg under extreme conditions (very high Hg/HA concentration ratios); thus, the biogeochemical behavior of Hg at high Hg/HA concentration ratios should be considered and further investigated.

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