Boosting underwater germination in Echinochloa colona seeds: the impact of high amplitude alternating temperatures and potassium nitrate osmopriming.

Underwater germination could risk seedling survival, suggesting the need for control through seed perception of environmental cues. These cues include diurnally alternating temperatures tied to drained soils or shallow water tables. We examined high-amplitude alternating temperatures impact on underwater germination. Besides, the conditions experimented by seeds in the soil (e.g. hydration/dehydration phases) change their germinability so we tested if osmopriming could affect underwater germination. We worked with Echinochloa colona seedlots from extensive crop fields, exposing seeds to sequential submergence and drained treatments in combination with cues that promote germination. While a 10°C difference between maximum and minimum daily temperatures maximised germination in drained conditions, higher amplitudes (>15°C) alternating temperatures promoted E. colona underwater germination under hypoxic water (pO2 <4.1kPa). KNO3 osmopriming in drained conditions promoted later underwater germination even under hypoxic water; however, PEG 6000 osmopriming induced seeds to enter secondary dormancy inhibiting underwater germination. KNO3 improved E. colona underwater germination under air-equilibrated floodwater (pO2 : 16.5-17.4kPa) yet not under hypoxic conditions. This suggests that germination can proceed in flooded nitrate-fertile soils as long as it remains aerobic. Hypoxic submergence did not inhibit the induction of hypersensitivity to light in E. colona seeds. This research expands our understanding of wetland seed germination ecophysiology, shedding light on the inducible nature of underwater germination in hydrophyte weeds.

Shaking off the blow: plant adjustments during submergence and post-stress growth in Lotus forage species.

Flooding significantly hampers global forage production. In flood-prone regions, Lotus tenuis and Lotus corniculatus are common forage legumes, yet little is known about their responses to partial or complete submergence. To address this, we evaluated 10 Lotus accessions subjected to 11days of either partial or complete submergence, analysing growth traits related to tolerance and recovery after de-submergence. Principal component analyses revealed that submergence associated growth parameters were linked to L. corniculatus accessions, whereas recovery was associated with L. tenuis accessions. Notably, in L. tenuis , recovery from complete submergence positively correlated with leaf mass fraction but negatively with root mass fraction, showing an opposite pattern than in L. corniculatus . Encouragingly, no trade-off was found between inherent growth capacity and submergence tolerance (both partial and complete) or recovery ability, suggesting genetic selection for increased tolerance would not compromise growth potential. L. tenuis exhibited accessions with both partial and complete submergence tolerance, making them versatile for flood-prone environments, whereas L. corniculatus accessions were better suited for partial submergence. These findings offer valuable insights to enhance forage production in flood-prone areas and guide the selection of appropriate Lotus accessions for specific flood conditions.

Understanding plant responses to saline waterlogging: insights from halophytes and implications for crop tolerance.

MAIN CONCLUSION: Saline and wet environments stress most plants, reducing growth and yield. Halophytes adapt with ion regulation, energy maintenance, and antioxidants. Understanding these mechanisms aids in breeding resilient crops for climate change. Waterlogging and salinity are two abiotic stresses that have a major negative impact on crop growth and yield. These conditions cause osmotic, ionic, and oxidative stress, as well as energy deprivation, thus impairing plant growth and development. Although few crop species can tolerate the combination of salinity and waterlogging, halophytes are plant species that exhibit high tolerance to these conditions due to their morphological, anatomical, and metabolic adaptations. In this review, we discuss the main mechanisms employed by plants exposed to saline waterlogging, intending to understand the mechanistic basis of their ion homeostasis. We summarize the knowledge of transporters and channels involved in ion accumulation and exclusion, and how they are modulated to prevent cytosolic toxicity. In addition, we discuss how reactive oxygen species production and cell signaling enhance ion transport and aerenchyma formation, and how plants exposed to saline waterlogging can control oxidative stress. We also address the morphological and anatomical modifications that plants undergo in response to combined stress, including aerenchyma formation, root porosity, and other traits that help to mitigate stress. Furthermore, we discuss the peculiarities of halophyte plants and their features that can be leveraged to improve crop yields in areas prone to saline waterlogging. This review provides valuable insights into the mechanisms of plant adaptation to saline waterlogging thus paving the path for future research on crop breeding and management strategies.

High Recovery from Either Waterlogging or Drought Overrides Any Beneficial Acclimation of Chloris gayana Facing a Subsequent Round of Stress.

Climate models predict that plants will face extreme fluctuations in water availability in future global change scenarios. Then, forage production will be more frequently subjected to the destabilizing pressure of sequentially occurring waterlogging and drought events. While the isolated effects of drought (D) and waterlogging (WL) are well characterized, little is known about the consequences when both stresses occur sequentially. We hypothesized that plants sequentially subjected to opposite water scenarios (D followed by WL or vice versa) are less stress tolerant than plants experiencing repetitions of the same type of water stress (i.e., D + D or WL + WL) due to contrasting acclimation and allocation to either shoots (WL) or roots (D). Chloris gayana (a tropical forage grass capable of tolerating either D and WL) plants were randomly assigned to nine treatments (a sequence of two stress rounds-WL or D-each followed by a recovery phase at field capacity). Relative growth rates and allometric responses were measured after each stress round and recovery period. In the first round of stress, both WL and D reduced plant RGR similarly, despite their allocation being opposite-prioritizing shoots or roots under WL and D, respectively. The high recovery displayed after either WL or D overrode any possible acclimation of the plants facing a second round of water stress. We conclude that the tolerance of C. gayana to sequential water stress (either for WL or D) is likely to depend more heavily on its recovery ability than on its previous adjustment to any stress scenario that may evoke memory responses. Knowledge like this could help improve forage grass breeding and the selection of cultivars for poorly drained soils subject to sequential stress events.

Plant Responses to Hypoxia: Signaling and Adaptation.

Molecular oxygen deficiency leads to altered cellular metabolism and can dramatically reduce crop productivity. Nearly all crops are negatively affected by lack of oxygen (hypoxia) due to adverse environmental conditions such as excessive rain and soil waterlogging. Extensive efforts to fully understand how plants sense oxygen deficiency and their ability to respond using different strategies are crucial to increase hypoxia tolerance. It was estimated that 57% of crop losses are due to floods [1]. Progress in our understanding has been significant in the last years. This topic deserved more attention from the academic community; therefore, we have compiled a Special Issue including four reviews and thirteen research articles reflecting the advancements made thus far.[...].

Submergence tolerance and recovery in Lotus: Variation among fifteen accessions in response to partial and complete submergence.

Several Lotus species are perennial forage legumes which tolerate waterlogging, but knowledge of responses to partial or complete shoot submergence is scant. We evaluated the responses of 15 Lotus accessions to partial and complete shoot submergence and variations in traits associated with tolerance and recovery after de-submergence. Accessions of Lotus tenuis, L. corniculatus, L. pedunculatus and L. japonicus were raised for 43 d and then subjected to aerated root zone (control), deoxygenated stagnant root zone with shoots in air (stagnant), stagnant root zone with partial (75 %) and complete submergence of shoots, for 7 d. The recovery ability from complete submergence was also assessed. We found inter- and intra-specific variations in the stem extension responses (i.e. promoted or restricted compared to controls) depending on water depth. Eight of 15 accessions promoted the stem extension when in partial submergence, while three of those eight (all L. tenuis accessions) had a restricted stem extension when under complete submergence. Two accessions (belonging to L. corniculatus and L. penduculatus species) also promoted the stem extension under complete submergence. The accessions that attained better recovery in terms of leaves produced after de-submergence, were those that had high leaf and root sugar concentration at de-submergence, and high thickness and persistence of gas films on leaves during submergence (all L. tenuis accessions). We conclude that all Lotus accessions were able to tolerate 7 d of partial and complete shoot submergence, despite adopting different stem extension responses.

Anatomical, morphological and growth responses of Thinopyrum ponticum plants subjected to partial and complete submergence during early stages of development.

Seedling recruitment and growth of forage grasses in flood-prone grasslands is often impaired by submergence. We evaluate the responses of Thinopyrum ponticum (Podp.) Barkw. & Dewey to partial and complete submergence at two early stages of development. Two greenhouse experiments were carried out with plants at three expanded leaves (Experiment 1) or five expanded leaves stage (Experiment 2). In each case, three treatments were applied for 14 days: control (C), partial submergence (PS; water level to half plant height), and complete submergence (CS; water level to 1.5 times plant height). Submergence was followed by a recovery period of 14 days at well drained conditions. Assessments included plant survival, height, leaf blade and pseudostem length, soluble carbohydrates in pseudostem, and shoot and root dry mass accumulation at the beginning and end of the submergence, and at the end of the recovery period. Root aerenchyma formation was determined on day 14 in both experiments. Under PS all plants survived, and the impact of the stress was related to the plants' developmental stage. However, plants with five expanded leaves increased total plant biomass with respect to control by 48%, plants with three expanded leaves reduced it by the same percentage. This response could be related to a higher ability to form root aerenchyma (17 vs 10%), and an enhanced leaf de-submergence capacity due to promoted leaf blade and pseudostem lengthening. Complete submergence treatment compromised the survival of 70% of the individuals with three expanded leaves but did not affect the survival at the five expanded leaves stage. In any developmental stage (three or five expanded leaves) plants fail to promote enough elongation of leaf blades or pseudostems to emerge from the water, so that always remained below the water surface. Root aerenchyma was not increased by CS at either of these two plant developmental stages. The high amount and concentration of pseudostem total soluble carbohydrates of the larger (five expanded leaves) plants facilitated their recovery growth after submergence. Our results predict the successful introduction of this species in areas where water excesses can cause soil waterlogging or shallow-partial plant submergence, but suggest avoidance of areas prone to suffer high-intensity flooding that lead to full plant submergence as this would highly constrain plant recruitment.

Eco-Physiological Traits Related to Recovery from Complete Submergence in the Model Legume Lotus japonicus.

Submergence is a severe form of stress for most plants. Lotus japonicus is a model legume with potential use in assisting breeding programs of closely related forage Lotus species. Twelve L. japonicus genotypes (10 recombinant inbred lines (RILs) and 2 parental accessions) with different constitutive shoot to root dry mass ratios (S:R) were subjected to 7 days of submergence in clear water and allowed to recover for two weeks post-submergence; a set of non-submerged plants served as controls. Relative growth rate (RGR) was used to indicate the recovery ability of the plants. Leaf relative water content (RWC), stomatal conductance (gs), greenness of basal and apical leaves, and chlorophyll fluorescence (Fv/Fm, as a measure of photoinhibition) were monitored during recovery, and relationships among these variables and RGR were explored across genotypes. The main results showed (i) variation in recovery ability (RGR) from short-term complete submergence among genotypes, (ii) a trade-off between growth during vs. after the stress indicated by a negative correlation between RGR during submergence and RGR post-submergence, (iii) an inverse relationship between RGR during recovery and S:R upon de-submergence, (iv) positive relationships between RGR at early recovery and RWC and gs, which were negatively related to S:R, suggesting this parameter as a good estimator of plant water balance post-submergence, (v) chlorophyll retention allowed fast recovery as revealed by the positive relationship between greenness of basal and apical leaves and RGR during the first recovery week, and (vi) full repair of the submergence-damaged photosynthetic apparatus occurred more slowly (second recovery week) than full recovery of plant water relations. The inclusion of these traits contributing to submergence recovery in L. japonicus should be considered to speed up the breeding process of the closely related forage Lotus spp. used in current agriculture.

Tolerance to partial and complete submergence in the forage legume Melilotus siculus: an evaluation of 15 accessions for petiole hyponastic response and gas-filled spaces, leaf hydrophobicity and gas films, and root phellem.

Background and Aims: Submergence is a severe stress for most plants. Melilotus siculus is a waterlogging- (i.e. root zone hypoxia) tolerant annual forage legume, but data were lacking for the effects of partial and full submergence of the shoots. The aim was to compare the tolerance to partial and full submergence of 15 M. siculus accessions and to assess variation in traits possibly contributing to tolerance. Recovery ability post-submergence was also evaluated. Methods: A factorial experiment imposed treatments of water level [aerated root zone with shoots in air as controls, stagnant root zone with shoots in air, stagnant root zone with partial (75 %) or full shoot submergence] on 15 accessions, for 7 d on 4-week-old plants in a 20/15 °C day/night phytotron. Measurements included: shoot and root growth, hyponastic petiole responses, petiole gas-filled spaces, leaflet sugars, leaflet surface hydrophobicity, leaflet gas film thickness and phellem area near the base of the main root. Recovery following full submergence was also assessed. Key Results: Accessions differed in shoot and root growth during partial and full shoot submergence. Traits differing among accessions and associated with tolerance were leaflet gas film thickness upon submergence, gas-filled spaces in petioles and phellem tissue area near the base of the main root. All accessions were able to re-orientate petioles towards the vertical under both partial and full submergence. Petiole extension rates were maintained during partial submergence, but decreased during full submergence. Leaflet sugars accumulated during partial submergence, but were depleted during full submergence. Growth resumption after full submergence differed among accessions and was positively correlated with the number of green leaves retained at desubmergence. Conclusions: Melilotus siculus is able to tolerate partial and full submergence of at least 7 d. Leaflet surface hydrophobicity and associated gas film retention, petiole gas-filled porosity and root phellem abundance are important traits contributing to tolerance. Post-submergence recovery growth differs among accessions. The ability to retain green leaves is essential to succeed during recovery.

Flooding tolerance of forage legumes.

We review waterlogging and submergence tolerances of forage (pasture) legumes. Growth reductions from waterlogging in perennial species ranged from >50% for Medicago sativa and Trifolium pratense to <25% for Lotus corniculatus, L. tenuis, and T. fragiferum. For annual species, waterlogging reduced Medicago truncatula by ~50%, whereas Melilotus siculus and T. michelianum were not reduced. Tolerant species have higher root porosity (gas-filled volume in tissues) owing to aerenchyma formation. Plant dry mass (waterlogged relative to control) had a positive (hyperbolic) relationship to root porosity across eight species. Metabolism in hypoxic roots was influenced by internal aeration. Sugars accumulate in M. sativa due to growth inhibition from limited respiration and low energy in roots of low porosity (i.e. 4.5%). In contrast, L. corniculatus, with higher root porosity (i.e. 17.2%) and O2 supply allowing respiration, maintained growth better and sugars did not accumulate. Tolerant legumes form nodules, and internal O2 diffusion along roots can sustain metabolism, including N2 fixation, in submerged nodules. Shoot physiology depends on species tolerance. In M. sativa, photosynthesis soon declines and in the longer term (>10 d) leaves suffer chlorophyll degradation, damage, and N, P, and K deficiencies. In tolerant L. corniculatus and L. tenuis, photosynthesis is maintained longer, shoot N is less affected, and shoot P can even increase during waterlogging. Species also differ in tolerance of partial and complete shoot submergence. Gaps in knowledge include anoxia tolerance of roots, N2 fixation during field waterlogging, and identification of traits conferring the ability to recover after water subsides.

No escape? Costs and benefits of leaf de-submergence in the pasture grass Chloris gayana under different flooding regimes.

Elongation-induced leaf emergence is one way for plants to deal with complete submergence by 'escaping' from water. This growth strategy is hypothesised to be more beneficial under single long-term submergence than under repeated short-term submergence events (i.e. fluctuating environment), as costs of repeated plant 'adjustment' would exceed the initial benefits of shoot elongation. To test this idea, 2-week-old plants of Chloris gayana Kunth. cv. Fine Cut (a submergence-tolerant cultivar first selected by a screening experiment) were grown for 4 weeks under (i) control conditions, (ii) two 1-week submergence cycles, or (iii) one 2-week submergence cycle. Additionally, a set of plants were placed below nettings to assess the cost of remaining forcedly submerged. Impeding leaves emergence through nettings did not compromise survival when submergence was 1-week long, but determined the death of all plants when extended to 2 weeks. Growth as affected by flooding regime revealed that under one 2-week submergence event, plants accumulated a 2.9-fold higher dry mass than when they experienced the same submergence duration in separate events along 1week. The 'escape' strategy in the grass C. gayana, by which leaf contact with air is re-established, is essential for its survival, and it is more beneficial for plant growth under long-term submergence than under repeated short-term submergence cycles.

Mechanisms of waterlogging tolerance in wheat--a review of root and shoot physiology.

We review the detrimental effects of waterlogging on physiology, growth and yield of wheat. We highlight traits contributing to waterlogging tolerance and genetic diversity in wheat. Death of seminal roots and restriction of adventitious root length due to O2 deficiency result in low root:shoot ratio. Genotypes differ in seminal root anoxia tolerance, but mechanisms remain to be established; ethanol production rates do not explain anoxia tolerance. Root tip survival is short-term, and thereafter, seminal root re-growth upon re-aeration is limited. Genotypes differ in adventitious root numbers and in aerenchyma formation within these roots, resulting in varying waterlogging tolerances. Root extension is restricted by capacity for internal O2 movement to the apex. Sub-optimal O2 restricts root N uptake and translocation to the shoots, with N deficiency causing reduced shoot growth and grain yield. Although photosynthesis declines, sugars typically accumulate in shoots of waterlogged plants. Mn or Fe toxicity might occur in shoots of wheat on strongly acidic soils, but probably not more widely. Future breeding for waterlogging tolerance should focus on root internal aeration and better N-use efficiency; exploiting the genetic diversity in wheat for these and other traits should enable improvement of waterlogging tolerance.

Oxygen deficiency and salinity affect cell-specific ion concentrations in adventitious roots of barley (Hordeum vulgare).

Oxygen deficiency associated with soil waterlogging adversely impacts root respiration and nutrient acquisition. We investigated the effects of O2 deficiency and salinity (100 mM NaCl) on radial O2 concentrations and cell-specific ion distributions in adventitious roots of barley (Hordeum vulgare). Microelectrode profiling measured O2 concentrations across roots in aerated, aerated saline, stagnant or stagnant saline media. X-ray microanalysis at two positions behind the apex determined the cell-specific elemental concentrations of potassium (K), sodium (Na) and chloride (Cl) across roots. Severe O2 deficiency occurred in the stele and apical regions of roots in stagnant solutions. O2 deficiency in the stele reduced the concentrations of K, Na and Cl in the pericycle and xylem parenchyma cells at the subapical region. Near the root apex, Na declined across the cortex in roots from the aerated saline solution but was relatively high in all cell types in roots from the stagnant saline solution. Oxygen deficiency has a substantial impact on cellular ion concentrations in roots. Both pericycle and xylem parenchyma cells are involved in energy-dependent K loading into the xylem and in controlling radial Na and Cl transport. At root tips, accumulation of Na in the outer cell layers likely contributed to reduction of Na in inner cells of the tips.

Differential growth of Spartina densiflora populations under saline flooding is related to adventitious root formation and innate root ion regulation.

Global change anticipates scenarios of sea level rise that would provoke long lasting floods, especially in lowland areas of salt marshes. Our aim was to evaluate the morpho-physiological adjustment ability to deal with continuous saline flooding of Spartina densiflora Brogn. plants from lowlands and uplands along a subtle topographical gradient (0.2m differential altitude). Plants from both origins were subjected to continuous saline flooding (300mM NaCl) for 35 days. Responses associated to adventitious rooting, aerenchyma formation, concentration of Na+, K+ and Cl- in roots and shoots tissues, tillering and growth were assessed. Root responses differentiated populations given that lowland plants showed higher ability for adventitious root formation and innate superior root ion regulation than upland plants. High constitutive K+ concentration plus high Na+ exclusion in root tissues led to significant low values of Na+:K+ ratios in lowland plants. Better root functioning was, in turn, related with more consistent shoot performance as lowland plants maintained plant tiller number and shoot relative growth rate unaltered while upland plants decreased both parameters by 35 and 18%, respectively, when in saline flooding. The superior performance of lowland plants indicates that locally adapted populations can be promoted in salt marsh habitats with subtle differences at topographic level.

Phenomic networks reveal largely independent root and shoot adjustment in waterlogged plants of Lotus japonicus.

Waterlogging imposes severe stress to the plant, and the interplay between root and aerial organs in the adjustment to this stress is poorly understood. A set of recombinant inbred lines (RILs) of Lotus japonicus (Gifu B-129 × Miyakojima MG-20) was subjected to control and waterlogging conditions for 21 d, and 12 traits related to leaf physiological functioning, root aerenchyma formation, shoot and root morphology, and dry mass accumulation were assessed to generate phenomic networks. The phenomic network became more complex under waterlogging as a result of the incorporation of root aerenchyma and dark-adapted Fv/Fm. Significant waterlogging-induced variation was found for stomatal conductance, dark-adapted Fv/Fm and aerenchyma. The RILs with stronger induction of aerenchyma in response to waterlogging tended to show reduced negative impact of this stress on root growth but suffered average impact on shoot growth. The RILs that retained higher stomatal conductance under waterlogging tended to retain higher dark-adapted Fv/Fm and shoot growth under waterlogging conditions but showed average impact on root traits. We propose a model where, although the stress experienced by the roots during waterlogging is transmitted to the shoot, shoots and roots deal with waterlogging in a less interdependent manner than often assumed.

Radial oxygen loss and physical barriers in relation to root tissue age in species with different types of aerenchyma.

Plant root aeration relies on aerenchyma and barrier formation in outer cortex influencing the radial oxygen loss (ROL) from roots towards the rhizosphere. Plant species display large variation in strategies for both responses. We investigated the impacts of root-zone hypoxia on aerenchyma formation and development of ROL apoplastic barriers in the outer cortex as a function of root tissue age using three lowland grassland species, each with alternative aerenchyma structure. All species increased root aerenchyma and continued with root elongation after imposing hypoxia. However, ROL barrier development differed: (i) Rumex crispus L. displayed only 'partial' barrier to ROL evidenced at older tissue ages, (ii) Cyperus eragrostis Lam. initiated a 'tighter' barrier to ROL following exposure to hypoxia in tissues older than 3 days, and (iii) Paspalidium geminatum (Forssk.) Stapf demonstrated highly effective inhibition of ROL under aerated and hypoxic conditions at all tissue ages related to constitutive 'tight' apoplastic barriers in outer cortex. Thus, hypoxic conditions affected root elongation and 'tightness' of apoplastic barriers depending on species. The physiological implications of the different ROL responses among species in relation to the differential formation of barriers are discussed.