Michael B. Jackson

Ethylene-promoted adventitious rooting and development of cortical air spaces (aerenchyma) in roots may be adaptive responses to flooding in Zea mays L

The roots and stem base of intact, 10 day old maize (Zea mays L. cv. LG11) plants, grown in nutrient solution, were continuously aerated either with ethylene (5 μl l-1) in air or with air alone. Ethylene treatment hastened the emergence of adventitious (nodal) roots from the base of the shoot, but slowed their subsequent extension. Ethylene also promoted the collapse of cells in the cortex of these roots, with lysigenous development of prominent air spaces (aerenchyma). Non-aeration of the nutrient solution caused endogenously produced ethylene to accumulate in the roots, and stimulated both the emergence of adventitious roots and the formation of cortical air spaces in them. With non-aeration the concentration of oxygen did not fall below 1% in the equilibrium gas phase (air=20.8%). Complete deoxygenation of the nutrient solution, produced by passing oxygen-free nitrogen gas, prevented both air space formation and the evolution of ethylene by root segments.These results suggest that adventitious rooting and cortical air space formation in nodal roots in Zea mays may be stimulated by enhanced concentrations of endogenous ethylene arising either from entrapment of the gas by unstirred water layers around the roots and/or by increased biosynthesis. These responses are considered conducive to survival in waterlogged soil.

Characterisation of programmed cell death during aerenchyma formation induced by ethylene or hypoxia in roots of maize ( Zea mays L.)

Abstract. Aerenchyma is a tissue type characterised by prominent intercellular spaces which enhance flooding tolerance in some plant species by facilitating gas diffusion between roots and the aerial environment. Aerenchyma in maize roots forms by collapse and death of some of the cortical cells in a process that can be promoted by imposing oxygen shortage or by ethylene treatment. Maize roots grown hydroponically in 3% oxygen, 1 μl l−1 ethylene or 21% oxygen (control) were analysed by a combination of light and electron microscopy. Use of in-situ terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL) suggested internucleosomal cleavage of DNA. However, chromatin condensation detectable by electron microscopy was preceded by cytoplasmic changes including plasma membrane invagination and the formation of vesicles, in contrast to mammalian apoptosis in which chromatin condensation is the first detectable event. Later, cellular condensation, condensation of chromatin and the presence of intact organelles surrounded by membrane resembling apoptotic bodies were observed. All these events were complete before cell wall degradation was apparent. Therefore, aerenchyma formation initiated by hypoxia or ethylene appears to be a form of programmed cell death that shows characteristics in part resembling both apoptosis and cytoplasmic cell death in animal cells.

Hormones from roots as signals for the shoots of stressed plants

Intracellular signalling processes that translate hormone perception into genetic, metabolic and developmental change are the focus of much current plant hormone research. By characterizing these processes it is hoped to establish an understanding of the regulated and integrated behaviour of whole plants at the molecular level. However, for a complete picture, the extracellular signalling processes that coordinate the behaviour and development of the various plant organs must also be uncovered. Roots may use hormones, or their precursors, to provide shoots with early warning of deteriorating soil conditions in ways that increase resilience to stress. Recent progress springs from integrated studies involving sensitive and specific methods of hormone analysis, plant water relations and the use of genetically transformed plants.

Inhibition by silver ions of gas space (aerenchyma) formation in adventitious roots of Zea mays L. subjected to exogenous ethylene or to oxygen deficiency.

We have studied the role of ethylene in accelerating the lytic formation of gas spaces (aerenchyma) in the cortex of adventitious roots of maize (Zea mays L.) growing in poorly aerated conditions. Such roots had previously been shown to contain increased concentrations of ethylene. Ten day-old maize plants bearing seminal roots and one whorl of emerging adventitious roots were grown in nutrient solution bubbled with air, ethylene in air (0.1 to 5.0 μl l−1), or allowed to become oxygen-deficient in nonaerated (but not completely anaerobic) solution. Additions of 0.1 μl l−1 ethylene or more promoted the formation of aerenchyma, with lysis of up to 47% of the cortical cells. The effects of non-aeration were similar to those of exogenous ethylene. When silver ions, an ethylene antagonist, were present at low, non-toxic concentrations (circa 0.6 μM), aerenchyma formation was prevented in ethylene treated roots and in those exposed to oxygen deficiency. Silver ions also blocked the inhibiting effect of exogenous ethylene on root extension. By contrast, the suppression of aerenchyma formation by silver ions under oxygendeficient conditions was associated with a retardation of root extension, indicating the importance of aerenchyma for root growth in poorly aerated media. Rates of production of ethylene by excised roots were stimulated by a previous non-aeration treatment. The effectiveness of Ag+ in inhibiting equally the action on cortical cells of exogenous ethylene and of non-aeration, supports the view that gas space (aerenchyma) formation in adventitious roots ‘adpted’ to oxygendeficient environments is mediated by increased concentrations of endogenous ethylene. The possibility that extra ethylene could arise from increased biosynthesis of a precursor in root tissues with a restricted oxygen supply is discussed.

Stomatal Closure in Flooded Tomato Plants Involves Abscisic Acid and a Chemically Unidentified Anti-Transpirant in Xylem Sap

We address the question of how soil flooding closes stomata of tomato (Lycopersicon esculentum Mill. cv Ailsa Craig) plants within a few hours in the absence of leaf water deficits. Three hypotheses to explain this were tested, namely that (a) flooding increases abscisic acid (ABA) export in xylem sap from roots, (b) flooding increases ABA synthesis and export from older to younger leaves, and (c) flooding promotes accumulation of ABA within foliage because of reduced export. Hypothesis a was rejected because delivery of ABA from flooded roots in xylem sap decreased. Hypothesis b was rejected because older leaves neither supplied younger leaves with ABA nor influenced their stomata. Limited support was obtained for hypothesis c. Heat girdling of petioles inhibited phloem export and mimicked flooding by decreasing export of [14C]sucrose, increasing bulk ABA, and closing stomata without leaf water deficits. However, in flooded plants bulk leaf ABA did not increase until after stomata began to close. Later, ABA declined, even though stomata remained closed. Commelina communis L. epidermal strip bioassays showed that xylem sap from roots of flooded tomato plants contained an unknown factor that promoted stomatal closure, but it was not ABA. This may be a root-sourced positive message that closes stomata in flooded tomato plants.

Export of Abscisic Acid, 1-Aminocyclopropane-1-Carboxylic Acid, Phosphate, and Nitrate from Roots to Shoots of Flooded Tomato Plants (Accounting for Effects of Xylem Sap Flow Rate on Concentration and Delivery)

We determined whether root stress alters the output of physiologically active messages passing from roots to shoots in the transpiration stream. Concentrations were not good measures of output. This was because changes in volume flow of xylem sap caused either by sampling procedures or by effects of root stress on rates of whole-plant transpiration modified concentrations simply by dilution. Thus, delivery rate (concentration x sap flow rate) was preferred to concentration as a measure of solute output from roots. To demonstrate these points, 1-aminocyclopropane-1-carboxylic acid (ACC), abscisic acid, phosphate, nitrate, and pH were measured in xylem sap of flooded and well-drained tomato (Lycopersicon esculentum Mill., cv Ailsa Craig) plants expressed at various rates from pressurized detopped roots. Concentrations decreased as sap flow rates were increased. However, dilution of solutes was often less than proportional to flow, especially in flooded plants. Thus, sap flowing through detopped roots at whole-plant transpiration rates was used to estimate solute delivery rates in intact plants. On this basis, delivery of ACC from roots to shoots was 3.1-fold greater in plants flooded for 24 h than in well-drained plants, and delivery of phosphate was 2.3-fold greater. Delivery rates of abscisic acid and nitrate in flooded plants were only 11 and 7%, respectively, of those in well-drained plants.

Submergence tolerance in rainfed lowland rice: physiological basis and prospects for cultivar improvement through marker-aided breeding

Two important factors influencing rice plant survival during submergence are limitations to gas diffusion under water, and reduced irradiance that impair photosynthesis and efficient utilization of carbohydrates. Thus, survival during submergence may largely depend on accumulation of high carbohydrate concentrations prior to submergence and a capacity for maintaining energy production through rapid alcoholic fermentation under oxygen shortage. During flash flooding, a third factor thought to affect survival is the aerobic shock during the post-submergence period when floodwaters recede. Changes in the level of antioxidants and enzymes such as superoxide dismutase (SOD) suggest that tolerant rice cultivars develop protective systems to air after exposure to hypoxic or anoxic environments. These responses are similar to other wetland plants. The capacity to survive submergence depends not only on specific environmental factors, but also on the strategy that plants have evolved for adoption to particular flood-prone environments. In rice the two main strategies are to elongate and escape, or not to elongate and conserve resources. For rainfed lowland rice exposed to flash flooding, elongation growth during complete submergence has major adverse effects on survival, presumably since this competes with maintenance processes which require carbohydrates and energy. Selection for minimal elongation during submergence is currently being exploited as a trait for submergence tolerance by rainfed lowland rice breeders in south and southeast Asia. Gene mapping for submergence tolerance has been useful in identifying one prominent locus for submergence tolerance. Fine scale gene mapping and sequencing may facilitate further progress in the physiology and genetics of submergence tolerance. Recently published data demonstrate that improving submergence tolerance may be possible through up-regulation of genes for particular traits such as pyruvate decarboxylase (PDC) for alcoholic fermentation. Validation of appropriate mechanisms in other cultivars for target environments, and development and utilization of molecular markers to follow these traits in breeding programs, will therefore be high priorities for future work on submergence tolerance of rice.

Increased 1-Aminocyclopropane-1-Carboxylic Acid Oxidase Activity in Shoots of Flooded Tomato Plants Raises Ethylene Production to Physiologically Active Levels

Soil flooding increased 1-aminocyclopropane-1-carboxylic (ACC) acid oxidase activity in petioles of wild-type tomato (Lycopersicon esculentum L.) plants within 6 to 12 h in association with faster rates of ethylene production. Petioles of flooded plants transformed with an antisense construct to one isoform of an ACC oxidase gene (ACO1) produced less ethylene and had lower ACC oxidase activity than those of the wild type. Flooding promoted epinastic curvature but did so less strongly in plants transformed with the antisense construct than in the wild type. Exogenous ethylene, supplied to well-drained plants, also promoted epinastic curvature, but transformed and wild-type plants responded similarly. Flooding increased the specific delivery (flux) of ACC to the shoots (picomoles per second per square meter of leaf) in xylem sap flowing from the roots. The amounts were similar in both transformed and wild-type plants. These observations demonstrate that changes in ACC oxidase activity in shoot tissue resulting from either soil flooding or introducing ACC oxidase antisense constructs can influence rates of ethylene production to a physiologically significant extent. They also implicate systemic root to shoot signals in regulating the activity of ACC oxidase in the shoot.

Roots of willow (Salix viminalis L.) show marked tolerance to oxygen shortage in flooded soils and in solution culture

Responses to soil flooding and oxygen shortage were studied in field, glasshouse and controlled environment conditions. Established stools ofSalix viminalis L., were compared at five field sites in close proximity but with contrasting water table levels and flooding intensities during the preceding winter. There was no marked effect of site on shoot extension rate, time to half maximum length or final length attained. When rooted cuttings were waterlogged for 4 weeks in a glasshouse, soil redox potentials quickly decreased to below zero. Shoot extension was slowed after a delay of 20 d, while, in the upper 100 mm of soil, formation and outgrowth of unbranched adventitious roots with enhanced aerenchyma development was promoted after 7 d. At depths of 100–200 mm and 200–300 mm, extension by existing root axes was halted by soil flooding, while adventitious roots from above failed to penetrate these deeper zones. After 4 weeks waterlogging, all arrested root tips recommenced elongation when the soil was drained; their extension rates exceeding those of roots that were well-drained throughout. Growth in fresh mass was also stimulated. The additional aerenchyma found in adventitious roots in the upper 100 mm of soil may have been ethylene regulated since gas space development was inhibited by silver nitrate, an ethylene action inhibitor. The effectiveness of aerenchyma was tested by blocking the entry of atmospheric oxygen into plants with lanolin applied to lenticels of woody shoots of plants grown in solution culture. Root extension was halved, while shoot growth remained unaffected. H Lambers Section editor

Hormones and developmental change in plants subjected to submergence or soil waterlogging

Abstract The morphology of many plants spcies is modified markedly by waterlogging of the soil or by submerging part, or all, of the shoot system. Effects can begin within a few minutes of inundation and some may increase tolerance to more prolonged inundation. Responses take place in tissues that experience poor aeration (Class I responses) and also in parts located some distances from the site of impended aeration (Class II responses). Evidence is reviewed for harmonal mediation in both classes of response, especially by ethylene (ethene).