Exogenous abscisic acid induces the formation of a suberized barrier to radial oxygen loss in adventitious roots of barley (Hordeum vulgare).

BACKGROUND AND AIMS: Internal root aeration is essential for root growth in waterlogged conditions. Aerenchyma provides a path for oxygen to diffuse to the roots. In most wetland species, including rice, a barrier to radial oxygen loss (ROL) allows more of the oxygen to diffuse to the root tip, enabling root growth into anoxic soil. Most dryland crops, including barley, do not form a root ROL barrier. We previously found that abscisic acid (ABA) signalling is involved in the induction of ROL barrier formation in rice during waterlogging. Although rice typically does not form a tight ROL barrier in roots in aerated conditions, an ROL barrier with suberized exodermis was induced by application of exogenous ABA. Therefore, we hypothesized that ABA application could also trigger root ROL barrier formation with hypodermal suberization in barley. METHODS: Formation of an ROL barrier was examined in roots in different exogenous ABA concentrations and at different time points using cylindrical electrodes and Methylene Blue staining. Additionally, we evaluated root porosity and observed suberin and lignin modification. Suberin, lignin and Casparian strips in the cell walls were observed by histochemical staining. We also evaluated the permeability of the apoplast to a tracer. KEY RESULTS: Application of ABA induced suberization and ROL barrier formation in the adventitious roots of barley. The hypodermis also formed lignin-containing Casparian strips and a barrier to the infiltration of an apoplastic tracer (periodic acid). However, ABA application did not affect root porosity. CONCLUSIONS: Our results show that in artificial conditions, barley can induce the formation of ROL and apoplastic barriers in the outer part of roots if ABA is applied exogenously. The difference in ROL barrier inducibility between barley (an upland species) and rice (a wetland species) might be attributable to differences in ABA signalling in roots in response to waterlogging conditions.

Radial Oxygen Loss Triggers Diel Fluctuation of Cadmium Dissolution in the Rhizosphere of Rice.

Cadmium (Cd) contamination poses a significant global threat to human health, primarily through dietary intake, with rice serving as a major source. While Cd predominantly resides in bound states in soil, the physiological processes by which rice facilitates Cd absorption in the rhizosphere remain largely elusive. This study delves into the mechanisms governing Cd uptake by rice plants in the rhizosphere, emphasizing the impact of daytime and nighttime fluctuations in microenvironmental conditions. Employing a microfluidic chip setup, the research reveals that radial oxygen loss from rice roots triggers dissolution of Cd in the rhizosphere. Notably, Cd mobility exhibits distinct diurnal fluctuations, peaking at 44.0 ± 4.1 nM during the daytime and dropping to 8.3 ± 1.3 nM during the nighttime. Further investigations reveal that variations in dissolved oxygen and hydroxyl radical concentrations influence Cd release, while pH changes and microbial reduction reactions play crucial roles in Cd immobilization. These findings provide insights into the intricate processes governing Cd mobilization in the rice rhizosphere, highlighting the importance of regulating these processes for effective Cd adsorption control in rice crops and safeguarding public health.

Multi-imaging platform for rhizosphere studies: Phosphorus and oxygen fluxes.

Rhizosphere is a soil volume of high spatio-temporal heterogeneity and intensive plant-soil-microbial interactions, for which visualization and process quantification is of highest scientific and applied relevance, but still very challenging. A novel methodology for quick assessment of two-dimensional distribution of available phosphorus (P) in rhizosphere was suggested, tested, and development up to the application platform. Available P was firstly trapped by an in-situ diffusive gradients in thin-films (DGT) sampler with precipitated zirconia as the binding gel, and subsequently, the loaded gel was analyzed with an optimized colorimetric imaging densitometry (CID). The imaging platform was established linking: i) DGT, ii) planar optode, and iii) soil zymography techniques to simultaneously determine available P, oxygen, and acid phosphatase in rhizosphere at sub-millimeter spatial scales. The DGT identified available P level in rice rhizosphere were spatially overlapping to the localized redox hotspots and phosphatase activity. The spatial relationship between available P and acid phosphatase activity was dependent on root development. The root radial oxygen loss (ROL) remained active during the experimental observations (2-3 days), while a flux of available P of 10 pg cm-2 s-1 was visualized within 2-3 mm of roots, confirming the correlative response of rice roots to oxygen secretion and P uptake. Summarizing, the established imaging platform is suitable to capture spatial heterogeneity and temporal dynamics of root activities, nutrient bioavailability, ROL and enzyme activities in rhizosphere.

A quantitative trait locus conferring flood tolerance to deepwater rice regulates the formation of two distinct types of aquatic adventitious roots.

A key trait conferring flood tolerance is the ability to grow adventitious roots as a response to submergence. The genetic traits of deepwater rice determining the development and characteristics of aquatic adventitious roots (AAR) had not been evaluated. We used near-isogenic lines introgressed to test the hypothesis that the impressive shoot elongation ability of deepwater rice linked to quantitative trait loci 1 and 12 also promote the development of AAR. The deepwater rice genotype NIL-12 possessed expanded regions at the stem nodes where numerous AAR developed as a response to submergence. Two types (AR1 and AR2) of roots with distinct timing of emergence and large differences in morphological and anatomical traits formed within 3 (AR1) to 7 (AR2) d of submergence. The mechanical impedance provided by the leaf sheath caused AR2 to emerge later promoting thicker roots, higher elongation capacity and higher desiccation tolerance. Upregulation of key genes suggests a joint contribution in activating the meristem in AAR enhancing the development of these in response to submergence. The morphological and anatomical traits suggested that AR2 is better adapted to long-term flooding than AR1. We therefore propose that AR2 in deepwater rice functions as an evolutionary defence strategy to tackle periodic submergence.

Responses of key root traits in the genus Oryza to soil flooding mimicked by stagnant, deoxygenated nutrient solution.

Excess water can induce flooding stress resulting in yield loss, even in wetland crops such as rice (Oryza). However, traits from species of wild Oryza have already been used to improve tolerance to abiotic stress in cultivated rice. This study aimed to establish root responses to sudden soil flooding among eight wild relatives of rice with different habitat preferences benchmarked against three genotypes of O. sativa. Plants were raised hydroponically, mimicking drained or flooded soils, to assess the plasticity of adventitious roots. Traits included were apparent permeance (PA) to O2 of the outer part of the roots, radial water loss, tissue porosity, apoplastic barriers in the exodermis, and root anatomical traits. These were analysed using a plasticity index and hierarchical clustering based on principal component analysis. For example, O. brachyantha, a wetland species, possessed very low tissue porosity compared with other wetland species, whereas dryland species O. latifolia and O. granulata exhibited significantly lower plasticity compared with wetland species and clustered in their own group. Most species clustered according to growing conditions based on PA, radial water loss, root porosity, and key anatomical traits, indicating strong anatomical and physiological responses to sudden soil flooding.

Iron plaque formation, characteristics, and its role as a barrier and/or facilitator to heavy metal uptake in hydrophyte rice (Oryza sativa L.).

The persistent bioavailability of toxic metal(oids) (TM) is undeniably the leading source of serious environmental problems. Through the transfer of these contaminants into food networks, sediments and the aquatic environmental pollution by TM serve as key routes for potential risks to soil and human health. The formation of iron oxyhydroxide plaque (IP) on the root surface of hydrophytes, particularly rice, has been linked to the impact of various abiotic and biotic factors. Radial oxygen loss has been identified as a key driver for the oxidation of rhizosphere ferrous iron (Fe2+) and its subsequent precipitation as low-to-high crystalline and/or amorphous Fe minerals on root surfaces as IP. Considering that each plant species has its unique capability of creating an oxidised rhizosphere under anaerobic conditions, the abundance of rhizosphere Fe2+, functional groups from organic matter decomposition and variations in binding capacities of Fe oxides, thus, impacting the mobility and interaction of several contaminants as well as toxic/non-toxic metals on the specific surface areas of the IP. More insight from wet extraction and advanced synchrotron-based analytical techniques has provided further evidence on how IP formation could significantly affect the fate of plant physiology and biomass production, particularly in contaminated settings. Collectively, this information sets the stage for the possible implementation of IP and related analytical protocols as a strategic framework for the management of rice and other hydrophytes, particularly in contaminated sceneries. Other confounding variables involved in IP formation, as well as operational issues related to some advanced analytical processes, should be considered.

Regulation of root adaptive anatomical and morphological traits during low soil oxygen.

Flooding causes oxygen deprivation in soils. Plants adapt to low soil oxygen availability by changes in root morphology, anatomy, and architecture to maintain root system functioning. Essential traits include aerenchyma formation, a barrier to radial oxygen loss, and outgrowth of adventitious roots into the soil or the floodwater. We highlight recent findings of mechanisms of constitutive aerenchyma formation and of changes in root architecture. Moreover, we use modelling of internal aeration to demonstrate the beneficial effect of increasing cortex-to-stele ratio on sustaining root growth in waterlogged soils. We know the genes for some of the beneficial traits, and the next step is to manipulate these genes in breeding in order to enhance the flood tolerance of our crops.

Oxygen in the air and oxygen dissolved in the floodwater both sustain growth of aquatic adventitious roots in rice.

Flooding is an environmental stress that leads to a shortage of O2 that can be detrimental for plants. When flooded, deepwater rice grow floating adventitious roots to replace the dysfunctional soil-borne root system, but the features that ensure O2 supply and hence growth of aquatic roots have not been explored. We investigate the sources of O2 in aquatic adventitious roots and relate aerenchyma and barriers for gas diffusion to local O2 gradients, as measured by microsensor technology, to link O2 distribution in distinct root zones to their anatomical features. The mature root part receives O2 exclusively from the stem. It has aerenchyma that, together with suberin and lignin depositions at the water-root and cortex-stele interfaces, provides a path for longitudinal O2 movement toward the tip. The root tip has no diffusion barriers and receives O2 from the stem and floodwater, resulting in improved aeration of the root tip over mature tissues. Local formation of aerenchyma and diffusion barriers in the mature root channel O2 towards the tip which also obtains O2 from the floodwater. These features explain aeration of floating roots and their ability to grow under water.

Genetic regulation of root traits for soil flooding tolerance in genus Zea.

Flooding stress caused by excessive precipitation and poor drainage threatens upland crop production and food sustainability, so new upland crop cultivars are needed with greater tolerance to soil flooding (waterlogging). So far, however, there have been no reports of highly flooding-tolerant upland crop cultivars, including maize, because of the lack of flooding-tolerant germplasm and the presence of a large number of traits affecting flooding tolerance. To achieve the goal of breeding flooding-tolerant maize cultivars by overcoming these difficulties, we chose highly flooding-tolerant teosinte germplasm. These flooding-tolerance-related traits were separately assessed by establishing a method for the accurate evaluation of each one, followed by performing quantitative trait locus (QTL) analyses for each trait using maize × teosinte mapping populations, developing introgression lines (ILs) or near-isogenic lines (NILs) containing QTLs and pyramiding useful traits. We have identified QTLs for flooding-tolerance-related root traits, including the capacity to form aerenchyma, formation of radial oxygen loss barriers, tolerance to flooded reducing soil conditions, flooding-induced adventitious root formation and shallow root angle. In addition, we have developed several ILs and NILs with flooding-tolerance-related QTLs and are currently developing pyramided lines. These lines should be valuable for practical maize breeding programs focused on flooding tolerance.

Constructed wetland, an eco-technology for wastewater treatment: A review on types of wastewater treated and components of the technology (macrophyte, biolfilm and substrate).

Constructed wetland (CW) represents an efficient eco-technological conglomerate interweaving water security, energy possibility and environmental protection. In the context of wastewater treatment technologies requiring substantial efficiency at reduced cost, chemical input and low environmental impact, applications of CW is being demonstrated at laboratory and field level with reasonably high contaminant removal efficiency and ecological benefits. However, along with the scope of applications, role of individual wetland component has to be re-emphasized through related research interventions. Hence, this review distinctively explores the concerns for extracting maximum benefit of macrophyte (focusing on interface of pollutant removal, root radial oxygen loss, root iron plaque, endophyte-macrophyte assisted treatment in CW, and prospects of energy harvesting from macrophyte) and role of biofilm (effect on treatment efficiency, composition and factors affecting) in a CW. Another focus of the review is on recent advances and developments in alternative low-cost substrate materials (including conventional type, industrial by-products, organic waste, mineral based and hybrid type) and their effect on target pollutants. The remainder of this review is organized to discuss the concerns of CW with respect to wastewater type (municipal, industrial, agricultural and farm wastewater). Attempt is made to analyze the practical relevance and significance of these aspects incorporating all recent developments in the areas to help making informed decisions about future directions for research and development related to CW.

Dynamics of radial oxygen loss in mangroves subjected to waterlogging.

Tidal flooding can directly result in oxygen (O2) shortage, however the functions of root aeration in flooding tolerance and O2 dynamics within mangroves are still poorly understood. Thus, in this study, the correlations among waterlogging tolerance, root porosity and O2 movement within the plants were investigated using two mangrove species (Aegiceras corniculatum and Bruguiera gymnorrhiza) and a semi-mangrove Heritiera littoralis. Based on the present data, the species A. corniculatum and B. gymnorrhiza, which possessed higher root porosity, exhibited higher waterlogging tolerance, while H. littoralis is intolerant. Increased root porosity, leaf stoma, and total ROL were observed in the roots of A. corniculatum and B. gymnorrhiza growing in stagnant solution when compared to respective aerated controls. As for ROL spatial pattern along roots, external anaerobic condition could promote ROL from apical root regions but reduce ROL from basal roots, leading to a 'tighter barrier'. In summary, the present study indicated that the plants (e.g., A. corniculatum and B. gymnorrhiza) prioritized to ensure O2 diffusion towards root tips under waterlogging by increasing aerenchyma formation and reducing O2 leakage at basal root regions.

Mixture of Pb, Zn and Cu on root permeability and radial oxygen loss in the mangrove Bruguiera gymnorrhiza.

A short term pot trail was employed to evaluate the exposure of mixed heavy metals (Cu, Pb and Zn) on growth, radial oxygen loss (ROL) and root anatomy in Bruguiera gymnorrhiza. The possible function of BgC4H, a cytochrome P450 gene, on root lignification was also discussed. The exposures of mixed Cu, Pb and Zn directly reduce O2 leakage at root surface. The reduced ROL inhibited by heavy metals was mainly ascribed by the changes in root anatomical features, such as decreased root porosity together with increased lignification within the exodermis. BgC4H was found to be up-regulated after 0.5-day metal exposure, and remained higher transcript levels within 3-day metal exposure when compared to control roots. Besides, the inhibited photosynthesis may also result in less oxygen can be transported to the underground roots. In summary, the mangrove B. gymnorrhiza appeared to react to external mixed metal contaminants by developing a lignified and impermeable exodermis, and such a root barrier induced by mixed Cu, Pb and Zn appeared to be an adaptive response to block metal ions enters into the roots.

Root adaptation in Echinodorus osiris Rataj plant under cadmium stress.

Cadmium tolerant plant, Echinodorus osiris Rataj, was selected to study its root adaptive mechanism under Cd stress. The change of root porosity, radial oxygen loss (ROL), and iron plaque formation was investigated. Results suggested that Cd treatment decreased 28.6-49.9% of ROL and reduced 13.5-23.3% of root porosity but increased 63.4-147.2% of iron plaque after 21 days, respectively. Under different Cd treatments, the uptake of Cd in root presented quick and mild models while it showed relatively consistent increase in shoot. Correlation analysis demonstrated that Cd concentrations in plant were related negatively with root porosity but had no significant correlation with ROL. There was significant positive correlation between root porosity and ROL; however, they both related negatively with root iron plaque. Moreover, the scanning electron microscopy indicates a barrier to the movement of Cd in endodermis layers.

QTLs for constitutive aerenchyma from Zea nicaraguensis improve tolerance of maize to root-zone oxygen deficiency.

Zea nicaraguensis is a wild relative of Zea mays subsp. mays (maize) that has high waterlogging tolerance. One of its traits is constitutive aerenchyma formation (CAF) in roots and this may be one of the reasons for the tolerance, but it has not yet been proven by comparing plants that differ only in CAF in the same genetic background. We therefore produced an introgression line AE24-50-44-91 (IL-AE91) possessing four quantitative trait loci for CAF from Z. nicaraguensis in the background of maize (inbred line Mi29). The degree of root CAF in IL-AE91 was intermediate between that of Mi29 and Z. nicaraguensis. Seedlings of IL-AE91 grown aerobically were more tolerant to transfer to oxygen-deficient conditions than were Mi29 seedlings. On day 2 of oxygen deficiency, the root extension rate and viability of root-tip cells in IL-AE91 were ~2.7 and ~1.3 times greater, respectively, than they were in Mi29. On day 4, the area of aerenchyma at 80 mm from the root tips was ~1.5 times greater in IL-AE91 and radial oxygen loss from the apical parts of roots was ~3.4 times higher than in Mi29. These results demonstrate that CAF reduces the stress from low external oxygen levels caused by soil waterlogging.

Root-zone hypoxia reduces growth of the tropical forage grass Urochloa humidicola in high-nutrient but not low-nutrient conditions.

BACKGROUND AND AIMS: The perennial C4 grass Urochloa humidicola is widely planted on infertile acidic and waterlogging-prone soils of tropical America. Waterlogging results in soil anoxia, and O2 deficiency can reduce nutrient uptake by roots. Interestingly, both nutrient deficiencies and soil waterlogging can enhance root cortical cell senescence, and the increased gas-filled porosity facilitates internal aeration of roots. We tested the influence of nutrient supply and root-zone O2 on root traits, leaf nutrient concentrations and growth of U. humidicola. METHODS: Plants were grown in pots in a completely randomized design under aerated or stagnant deoxygenated hydroponic conditions and six nutrient regimes, with low to high concentrations of all essential elements, for 28 d in a controlled-temperature greenhouse. The standard acid solution (SAS) used was previously designed based on infertile acidic soils of the tropical America savannas, and step increases in the concentration of SAS were used in aerated or deoxygenated 0.1 % agar solution, which mimics changes in gas composition in waterlogged soils. Measurements included shoot and root growth, root porosity, root anatomy, radial O2 loss, and leaf tissue nutrient concentrations. KEY RESULTS: Shoot dry mass was reduced for plants in stagnant compared with aerated conditions at high, but not at low, levels of mineral nutrition. In low-nutrition stagnant solution, roots were shorter, of greater porosity and had smaller radial thickness of the stele. Suberized lamellae and lignified sclerenchyma, as well as a strong barrier to radial O2 loss, were documented for roots from all treatments. Leaf nutrient concentrations of K, Mg and Ca (but not N, P and S) were higher in aerated than in stagnant conditions. CONCLUSIONS: Under low-nutrient conditions, plant growth in stagnant solution was equal to that in aerated solution, whereas under higher-nutrient regimes growth increased but dry mass in stagnant solution was less than in aerated solution. Slow growth in low-nutrient conditions limited any further response to the low O2 treatment, and greater porosity and smaller stele size in roots would enhance internal O2 movement within roots in the nutrient-limited stagnant conditions. A constitutive barrier to radial O2 loss and aerenchyma facilitates O2 movement to the tips of roots, which presumably contributes to maintaining nutrient uptake and the tolerance of U. humidicola to low O2 in the root-zone.

Dynamics of oxygen and carbon dioxide in rhizospheres of Lobelia dortmanna - a planar optode study of belowground gas exchange between plants and sediment.

Root-mediated CO2 uptake, O2 release and their effects on O2 and CO2 dynamics in the rhizosphere of Lobelia dortmanna were investigated. Novel planar optode technology, imaging CO2 and O2 distribution around single roots, provided insights into the spatiotemporal patterns of gas exchange between roots, sediment and microbial community. In light, O2 release and CO2 uptake were pronounced, resulting in a distinct oxygenated zone (radius: c. 3 mm) and a CO2 -depleted zone (radius: c. 2 mm) around roots. Simultaneously, however, microbial CO2 production was stimulated within a larger zone around the roots (radius: c. 10 mm). This gave rise to a distinct pattern with a CO2 minimum at the root surface and a CO2 maximum c. 2 mm away from the root. In darkness, CO2 uptake ceased, and the CO2 -depleted zone disappeared within 2 h. By contrast, the oxygenated root zone remained even after 8 h, but diminished markedly over time. A tight coupling between photosynthetic processes and the spatiotemporal dynamics of O2 and CO2 in the rhizosphere of Lobelia was demonstrated, and we suggest that O2 -induced stimulation of the microbial community in the sediment increases the supply of inorganic carbon for photosynthesis by building up a CO2 reservoir in the rhizosphere.

Influence of silicon treatment on antimony uptake and translocation in rice genotypes with different radial oxygen loss.

Antimony (Sb) pollution in soil may have a negative impact on the health of people consuming rice. This study investigated the effect of silicon (Si) application on rice biomass, iron plaque formation, and Sb uptake and speciation in rice plants with different radial oxygen loss (ROL) using pot experiments. The results demonstrated that Si addition increased the biomass of straw and grain, but had no obvious impact on the root biomass. Indica genotypes with higher ROL underwent greater iron plaque formation and exhibited more Sb sequestration in iron plaque. Silicon treatments increased iron levels in iron plaque from the different genotypes but decreased the total Sb concentration in root, straw, husk, and grain. In addition, Si treatment reduced the inorganic Sb concentrations but slightly increased the trimethylantimony (TMSb) concentrations in rice straw. Moreover, rice straw from hybrid genotypes accumulated higher concentrations of TMSb and inorganic Sb than that from indica genotypes. The conclusions from this study indicate that Sb contamination in rice can be efficiently reduced by applying Si treatment and selecting genotypes with high ROL.

Flood adaptive traits and processes: an overview.

Unanticipated flooding challenges plant growth and fitness in natural and agricultural ecosystems. Here we describe mechanisms of developmental plasticity and metabolic modulation that underpin adaptive traits and acclimation responses to waterlogging of root systems and submergence of aerial tissues. This includes insights into processes that enhance ventilation of submerged organs. At the intersection between metabolism and growth, submergence survival strategies have evolved involving an ethylene-driven and gibberellin-enhanced module that regulates growth of submerged organs. Opposing regulation of this pathway is facilitated by a subgroup of ethylene-response transcription factors (ERFs), which include members that require low O2 or low nitric oxide (NO) conditions for their stabilization. These transcription factors control genes encoding enzymes required for anaerobic metabolism as well as proteins that fine-tune their function in transcription and turnover. Other mechanisms that control metabolism and growth at seed, seedling and mature stages under flooding conditions are reviewed, as well as findings demonstrating that true endurance of submergence includes an ability to restore growth following the deluge. Finally, we highlight molecular insights obtained from natural variation of domesticated and wild species that occupy different hydrological niches, emphasizing the value of understanding natural flooding survival strategies in efforts to stabilize crop yields in flood-prone environments.

Arsenic dynamics in the rhizosphere and its sequestration on rice roots as affected by root oxidation.

A pot experiment was conducted to investigate the effects of root oxidation on arsenic (As) dynamics in the rhizosphere and As sequestration on rice roots. There were significant differences (P < 0.05) in pH values between rhizosphere and non-rhizosphere soils, with pH 5.68-6.16 in the rhizosphere and 6.30-6.37 in non-rhizosphere soils as well as differences in redox potentials (P < 0.05). Percentage arsenite was lower (4%-16%) in rhizosphere soil solutions from rice genotypes with higher radial oxygen loss (ROL) compared with genotypes with lower ROL (P < 0.05). Arsenic concentrations in iron plaque and rice straw were significantly negatively correlated (R = -0.60, P < 0.05). Genotypes with higher ROL (TD71 and Yinjingruanzhan) had significantly (P < 0.001) lower total As in rice grains (1.35 and 0.96 mg/kg, respectively) compared with genotypes with lower ROL (IAPAR9, 1.68 mg/kg; Nanyangzhan 2.24 mg/kg) in the As treatment, as well as lower inorganic As (P < 0.05). The present study showed that genotypes with higher ROL could oxidize more arsenite in rhizosphere soils, and induce more Fe plaque formation, which subsequently sequestered more As. This reduced As uptake in aboveground plant tissues and also reduced inorganic As accumulation in rice grains. The study has contributed to further understanding the mechanisms whereby ROL influences As uptake and accumulation in rice.

Simultaneously inhibit cadmium and arsenic uptake in rice (Oryza sativa L.) by Selenium enhanced iron plaque: Performance and mechanism.

Selenium (Se) fortification is witnessed to simultaneously inhibit absorbing Cadmium (Cd) and Arsenic (As) by rice plants, but the mechanism is unclear. Here, the effects of Se on the root morphology, iron plaque (IP) content, soil Fe2+ content, radial oxygen loss (ROL), and enzyme activities of the rice plants in the soil contaminated by Cd and As were intensively investigated through the hydroponic and soil experiments. Se effectively alleviated the toxic effects of Cd and As on the plants and the dry weight, root length, and root width were increased by 203.18%, 33.41%, and 52.81%, respectively. It also elucidated that ROL was one of the key factors to elevate IP formation by Se and the specific pathways of Se enhancing ROL were identified. ROL of the plants in the experiment group treated by Se was increased 36.76%, and correspondingly IP was magnified 50.37%, compared to the groups with Cd and As. It was owing to Se significantly increased the root porosity (62.11%), facilitating O2 transport to the roots. Additionally, Se enhanced the activities of catalase (CAT) and superoxide dismutase (SOD) to promote the catalytic degradation of ROS induced by Cd and As stress. It indirectly increased O2 release in the rhizosphere, which benefit to form more robust IP serve as stronger barrier to Cd and As. The results of our study provide a novel molecular level insight for Se promoting root IP to block Cd and As uptake by the rice plants.