In flooded soils, an efficient internal aeration system is essential for root growth and plant survival. Roots of many wetland species form barriers to restrict radial O₂ loss (ROL) to the rhizosphere. The formation of such barriers greatly enhances longitudinal O₂ diffusion from basal parts towards the root tip, and the barrier also impedes the entry of phytotoxic compounds produced in flooded soils into the root. Nevertheless, ROL from roots is an important source of O₂ for rhizosphere oxygenation and the oxidation of toxic compounds. In this paper, we review the methodological aspects for the most widely used techniques for the qualitative visualization and quantitative determination of ROL from roots. Detailed methodological approaches, practical set-ups and examples of ROL from roots with or without barriers to ROL are included. This paper provides practical knowledge relevant to several disciplines, including plant–soil interactions, biogeochemistry and eco-physiological aspects of roots and soil biota.

Floods and droughts are becoming more frequent as a result of climate change and it is imperative to find ways to enhance the resilience of staple crops to abiotic stresses. This is crucial to sustain food production during unfavourable conditions. Here, we analyse the current knowledge about suberised and lignified outer apoplastic barriers, focusing on the functional roles of the barrier to radial O2 loss formed as a response to soil flooding and we discuss whether this trait also provides resilience to multiple abiotic stresses. The barrier is composed of suberin and lignin depositions in the exodermal and/or sclerenchyma cell walls. In addition to the important role during soil flooding, the barrier can also restrict radial water loss, prevent phytotoxin intrusion, salt intrusion and the main components of the barrier can impede invasion of pathogens in the root. However, more research is needed to fully unravel the induction pathway of the outer apoplastic barriers and to address potential trade-offs such as reduced nutrient or water uptake. Nevertheless, we suggest that the outer apoplastic barriers might act as a jack of all trades providing tolerance to multiple abiotic and/or biotic stressors.

Rice production worldwide represents a major anthropogenic source of greenhouse gas emissions. Nitrogen fertilization and irrigation practices have been fundamental to achieve optimal rice yields, but these agricultural practices together with by-products from plants and microorganisms, facilitate the production, accumulation and venting of vast amounts of CO₂, CH₄ and N₂O. We propose that the development of elite rice varieties should target root traits enabling an effective internal O₂ diffusion, via enlarged aerenchyma channels. Moreover, gas tight barriers impeding radial O₂ loss in basal parts of the roots will increase O₂ diffusion to the root apex where molecular O₂ diffuses into the rhizosphere. These developments result in plants with roots penetrating deeper into the flooded anoxic soils, producing higher volumes of oxic conditions in the interface between roots and rhizosphere. Molecular O₂ in these zones promotes CH₄ oxidation into CO₂ by methanotrophs and nitrification (conversion of NH₄⁺ into NO₃^-), reducing greenhouse gas production and at the same time improving plant nutrition. Moreover, roots with tight barriers to radial O₂ loss will have restricted diffusional entry of CH₄ produced in the anoxic parts of the rhizosphere and therefore plant-mediated diffusion will be reduced. In this review, we describe how the exploitation of these key root traits in rice can potentially reduce greenhouse gas emissions from paddy fields.

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. 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. 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. 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.

Lack of O₂ and high concentrations of Fe and Mn commonly occur in waterlogged soils. The development of a barrier to impede radial O₂ loss (ROL) is a key trait improving internal O₂ transport and waterlogging tolerance in plants. We evaluated the ability of the barrier to ROL to impede the entry of excess Fe into the roots of the waterlogging tolerant grass Urochloa humidicola. Plants were grown in aerated or stagnant deoxygenated nutrient solution with 5 or 900 µM Fe. Quantitative X-ray-microanalysis was used to determine cell-specific Fe concentrations at two positions behind the root apex in relation to ROL and the formation of apoplastic barriers. At a mature zone of the root, Fe was ‘excluded’ at the exodermis where a suberized lamella was evident, a feature also associated with a strong barrier to ROL. By contrast, the K concentration was similar in all root cells, indicating that K uptake was not affected by apoplastic barriers. The hypothesis that the formation of a tight barrier to ROL impedes the apoplastic entry of toxic concentrations of Fe into the mature zones of roots was supported by the significantly higher accumulation of Fe on the outer-side of the exodermis.

In recent years, research on flooding stress and hypoxic responses in plants has gathered increasing attention due to climate change and the important role of O2 in metabolism and signalling. This Collection of Functional Plant Biology on ‘Flooding stress and responses to hypoxia in plants’ presents key contributions aimed at progressing our current understanding on how plants respond to low-O2 conditions, flooding stress and a combination of stresses commonly found in flooded areas. The Collection emphasises the characterisation of diverse plant responses across different developmental stages, from seed germination to fully developed plants, and under different water stress conditions ranging from waterlogging to complete submergence, or simply low-O2 conditions resulting from limited O2 diffusivity in bulky tissues. Additionally, this Collection highlights diverse approaches, including eco-physiological characterisation of plant responses, detailed descriptions of root anatomical characteristics and their surrounding microenvironments, evaluation of the seed microbiota under flooding stress, the modification of gene expression, and evaluations of diverse germplasm collections.