Edward G. Barrett-Lennard

The waterlogging/salinity interaction in higher plants revisited - focusing on the hypoxia-induced disturbance to K+ homeostasis

Salinity and waterlogging (root-zone hypoxia) are abiotic stresses that often occur together on saltland. It is widely recognised that these two factors interact to increase Na+ and/or Cl- concentrations in shoots, which can have adverse effects on plant growth and survival. This review expands on this understanding, providing evidence that the adverse effects of the interaction are also associated with a disturbance to plant K+ homeostasis. This conclusion is based on a comparative analysis of changes in ion concentrations and growth reported in the literature between species (glycophytes vs halophytes) and within a single species (Hordeum marinum L.). Comparisons between species show that hypoxia under saline conditions causes simultaneous increases in Na+ and Cl- concentrations and decreases in K+ concentrations in shoots and that these changes can all be related to changes in shoot dry mass. Comparisons between accessions of a single species (Hordeum maritima L.) strengthen the argument, with increases in Na+ and decreases in K+ being related to decreases in shoot relative growth rate.

Response to non-uniform salinity in the root zone of the halophyte Atriplex nummularia: growth, photosynthesis, water relations and tissue ion concentrations

BACKGROUND AND AIMS Soil salinity is often heterogeneous, yet the physiology of halophytes has typically been studied with uniform salinity treatments. An evaluation was made of the growth, net photosynthesis, water use, water relations and tissue ions in the halophytic shrub Atriplex nummularia in response to non-uniform NaCl concentrations in a split-root system. METHODS Atriplex nummularia was grown in a split-root system for 21 d, with either the same or two different NaCl concentrations (ranging from 10 to 670 mm), in aerated nutrient solution bathing each root half. KEY RESULTS Non-uniform salinity, with high NaCl in one root half (up to 670 mm) and 10 mm in the other half, had no effect on shoot ethanol-insoluble dry mass, net photosynthesis or shoot pre-dawn water potential. In contrast, a modest effect occurred for leaf osmotic potential (up to 30 % more solutes compared with uniform 10 mm NaCl treatment). With non-uniform NaCl concentrations (10/670 mm), 90 % of water was absorbed from the low salinity side, and the reduction in water use from the high salinity side caused whole-plant water use to decrease by about 30 %; there was no compensatory water uptake from the low salinity side. Leaf Na(+) and Cl(-) concentrations were 1.9- to 2.3-fold higher in the uniform 670 mm treatment than in the 10/670 mm treatment, whereas leaf K(+) concentrations were 1.2- to 2.0-fold higher in the non-uniform treatment. CONCLUSIONS Atriplex nummularia with one root half in 10 mm NaCl maintained net photosynthesis, shoot growth and shoot water potential even when the other root half was exposed to 670 mm NaCl, a concentration that inhibits growth by 65 % when uniform in the root zone. Given the likelihood of non-uniform salinity in many field situations, this situation would presumably benefit halophyte growth and physiology in saline environments.

Aerenchymatous phellem in hypocotyl and roots enables O2 transport in Melilotus siculus

• Aerenchymatous phellem (secondary aerenchyma) has rarely been studied in roots. Its formation and role in internal aeration were evaluated for Melilotus siculus, an annual legume of wet saline land. • Plants were grown for 21 d in aerated or stagnant (deoxygenated) agar solutions. Root porosity and maximum diameters were measured after 0, 7, 14 and 21 d of treatment. Phellem anatomy was studied and oxygen (O(2)) transport properties examined using methylene blue dye and root-sleeving O(2) electrodes. • Interconnecting aerenchymatous phellem developed in hypocotyl, tap root and older laterals (but not in aerial shoots), with radial intercellular connections to steles. Porosity of main roots containing phellem was c. 25%; cross-sectional areas of this phellem were threefold greater for stagnant than for aerated treatments. Root radial O(2) loss was significantly reduced by complete hypocotyl submergence; values approached zero after disruption of hypocotyl phellem below the waterline or, after shoot excision, by covering hypocotyl phellem in nontoxic cream. • Aerenchymatous phellem enables hypocotyl-to-root O(2) transport in M. siculus. Phellem increases radially under stagnant conditions, and will contribute to waterlogging tolerance by enhancing root aeration. It seems likely that with hypocotyl submerged, O(2) will diffuse via surface gas-films and internally from the shoot system.

Plant responses to heterogeneous salinity: growth of the halophyte Atriplex nummularia is determined by the root-weighted mean salinity of the root zone

Soil salinity is generally spatially heterogeneous, but our understanding of halophyte physiology under such conditions is limited. The growth and physiology of the dicotyledonous halophyte Atriplex nummularia was evaluated in split-root experiments to test whether growth is determined by: (i) the lowest; (ii) the highest; or (iii) the mean salinity of the root zone. In two experiments, plants were grown with uniform salinities or horizontally heterogeneous salinities (10–450mM NaCl in the low-salt side and 670mM in the high-salt side, or 10mM NaCl in the low-salt side and 500–1500mM in the high-salt side). The combined data showed that growth and gas exchange parameters responded most closely to the root-weighted mean salinity rather than to the lowest, mean, or highest salinity in the root zone. In contrast, midday shoot water potentials were determined by the lowest salinity in the root zone, consistent with most water being taken from the least negative water potential source. With uniform salinity, maximum shoot growth was at 120–230mM NaCl; ~90% of maximum growth occurred at 10mM and 450mM NaCl. Exposure of part of the roots to 1500mM NaCl resulted in an enhanced (+40%) root growth on the low-salt side, which lowered root-weighted mean salinity and enabled the maintenance of shoot growth. Atriplex nummularia grew even with extreme salinity in part of the roots, as long as the root-weighted mean salinity of the root zone was within the 10–450mM range.

Improvement of salt and waterlogging tolerance in wheat: comparative physiology of Hordeum marinum-Triticum aestivum amphiploids with their H. marinum and wheat parents

Hordeum marinum Huds. is a waterlogging-tolerant halophyte that has been hybridised with bread wheat (Triticum aestivum L.) to produce an amphiploid containing both genomes. This study tested the hypothesis that traits associated with waterlogging and salinity tolerances would be expressed in H. marinum-wheat amphiploids. Four H. marinum accessions were used as parents to produce amphiploids with Chinese Spring wheat, and their responses to hypoxic and 200mM NaCl were evaluated. Relative growth rate (RGR) in the hypoxic-saline treatment was better maintained in the amphiploids (58-71% of controls) than in wheat (56% of control), but the amphiploids were more affected than H. marinum (68-97% of controls). In hypoxic-saline conditions, leaf Na+ concentrations in the amphiploids were lower than in wheat (30-41% lower) but were 39-47% higher than in the H. marinum parents. A strong barrier to radial oxygen loss formed in basal root zones under hypoxic conditions in two H. marinum accessions; this barrier was moderate in the amphiploids, absent in wheat, and was weaker for the hypoxic-saline treatment. Porosity of adventitious roots increased with the hypoxic treatments; values were 24-38% in H. marinum, 16-27% in the amphiploids and 16% in wheat. Overall, the amphiploids showed greater salt and waterlogging tolerances than wheat, demonstrating the expression of relevant traits from H. marinum in the amphiploids.

The source of nitrogen (NH4 + or NO3 - ) affects the concentration of oxalate in the shoots and the growth of Atriplex nummularia (oldman saltbush)

Atriplex nummularia Lindl. (oldman saltbush) is a halophytic shrub used widely as a forage for ruminant production in saline farming systems. However, it can contain high concentrations of oxalate in the leaves, which may cause calcium deficiency in grazing animals. We hypothesised that supplying NH4+ instead of NO3- to a clone of this species would decrease oxalate concentrations in the shoots, and also decrease plant growth. Oxalate concentrations were measured in plants in the field, and a glasshouse experiment was conducted in which plants were grown with 10mM NO3- or NH4+, with 50, 200 or 500mM NaCl. The field survey showed effects of site (P<0.001), with average oxalate concentrations in shoots varying between 2.4 and 6.4% dry mass (DM). In the glasshouse, oxalate concentrations and plant growth were both affected by N-source and salinity (P<0.001). Averaged across salinities, plants grown with NH4+ for 24 days had only 43% of the shoot DM but 25% of the oxalate concentration of plants grown with NO3-. We discuss the effects of N-source on oxalate concentrations, the implications of this for halophyte growth, and the opportunity to select halophytes with lower oxalate and higher nutritive value for livestock.

Variation in salinity tolerance, early shoot mass and shoot ion concentrations within Lotus tenuis: towards a perennial pasture legume for saline land

Perennial legumes are needed for productive pastures in saline areas. We evaluated 40 lines of Lotus tenuis for tolerance to salinity at both germination and vegetative growth stages. Salt tolerance during the early vegetative stage was assessed in a sand-tank experiment with NaCl concentrations of 0–450 mm NaCl for 5 weeks. Most L. tenuis lines were more salt tolerant and had at least 50% lower shoot Na+ plus Cl– (% dry mass (DM)) compared with some other common pasture legumes, Medicago sativa, M. polymorpha and Trifolium subterraneum. Within L. tenuis significant variation in salt tolerance was found, with C50 values (concentrations of NaCl that decreased shoot dry matter to 50% of control) ranging from ~100 to 320 mm. Shoot concentrations of Cl–, Na+ and K+ did not always correlate with salt tolerance; some tolerant lines had low shoot Na+ and Cl– (and thus better nutritive value), while others tolerated high shoot Na+ and Cl–. We also found variation within L. tenuis for salt tolerance of seeds, with lines ranging from 0 to 70% germination after recovery from a prior exposure to 800 mm NaCl for 15 days. There was no relationship between salinity tolerance of scarified seeds and subsequent growth of seedlings; therefore, testing of seeds alone would not be an appropriate screening method for salt tolerance in L. tenuis. This study of 40 L. tenuis lines has shown significant genetic variation for salt tolerance within this species, and we have identified key lines with potential to be productive in saltland pasture systems.

Application of Distributed Wireless Chloride Sensors to Environmental Monitoring: Initial Results

Over the next 30 years, it is anticipated that the world will need to source 70% more food to provide for the growing population, and it is likely that a significant amount of this will have to come from irrigated land. However, the quality of irrigation water is also important, and measuring the quality of this water will allow management decisions to be made. Soil salinity is an important parameter in crop yield, and in this paper, we describe a chloride sensor system based on a low-cost robust screen-printed chloride ion sensor, suitable for use in distributed sensor networks. Previously, this sensor has been used in controlled laboratory-based experiments, but here we provide evidence that the sensor will find application outside of the laboratory in field deployments. We report on three experiments using this sensor; one with a soil column, one using a fluvarium, and finally on an experiment in a greenhouse. All these give an insight into the movement of chloride over small distances with high temporal resolution. These initial experiments illustrate that the new sensors are viable and usable with relatively simple electronics, and although subject to ongoing development, they are currently capable of providing new scientific data at high spatial and temporal resolutions. Therefore, we conclude that such chloride sensors, coupled with a distributed wireless network, offer a new paradigm in hydrological monitoring and will enable new applications, such as irrigation using mixtures of potable and brackish water, with significant cost and resource saving.

Enhancing the germination of three fodder shrubs (Atriplex amnicola, A.nummularia, A. undulata; Chenopodiaceae): implications for the optimisation of field establishment

Saltbush (Atriplex) species are widely grown in Australia as saltland pastures. Direct seeding practices for saltbush currently result in asynchronous and unreliable seedling establishment (5% successful establishment is not uncommon from field-sown seed). In part this may stem from a limited understanding of Atriplex seed germination requirements. This paper presents findings with 3 Atriplex species, A. amnicola (Paul G. Wilson.), A. nummularia (Lindl.), and A. undulata (D. Dietr), each of which differs in germination characteristics. For A. amnicola, the presence of light (and artificial substitution of light by 1000 ppm gibberellic acid) improved germination under controlled conditions and resulted in a 4-fold increase (70% total emergence) in field emergence of seedlings. For A. undulata, removing bracteoles increased germination under controlled conditions (~15%), with a 1.5-fold improvement in field seedling emergence (55% final emergence); however, seed priming or gibberellic acid application had no significant effect. In contrast, for A. nummularia, bracteole removal and light had minor positive effects on germination under controlled conditions, but this did not translate into improved emergence in soil or in the field. Under –0.5 MPa NaCl stress, application of gibberellic acid, salicylic acid, or kinetin to the germination medium significantly increased the final germination percentage of A. amnicola seeds (58, 16, and 14%, respectively) and improved the rate at which seeds germinated. All plant signalling compounds significantly increased final germination percentage and germination rate of A. undulata, albeit with a <10% increase at –0.5 MPa NaCl. Priming seeds with plant signalling compounds had similar effects on seed germination under low water potentials compared to direct treatment of the germination media. The effects of seed priming on Atriplex seedling emergence from saline soils varied among species. Priming with water significantly increased emergence percentage of A. amnicola but had no effect on A. nummularia and A. undulata. Gibberellic acid improved A. amnicola germination parameters only, whereas salicylic acid and kinetin improved the rate of emergence in all 3 species at various levels of salinity. This study suggests that a basic understanding of seed dormancy and germination requirements has the potential to substantially improve field emergence of saltbush species.

Effects of External Concentration of (K+ + Na+) and K+/Na+ on the Growth and Ion Relations of Atriplex amnicola

Summary This paper reports on the growth and ion relations of Atriplex amnicola in response to different K+/ Na+ in nutrient solutions containing either 40 or 400 mol m-3 (K+ + Na+)C1-. Growth was substantially lower at 400 than at 40 mol m-3 external (K+ + Na+)Cl-. At 40 mol m-3 (K+ + Na+)Cl-, growth of A. amnicola did not differ at K+/Na+ of 0.025, 0.333, and 1.0 in the nutrient solution. At 400 mol m-3 (K+ + Na+)Cl , growth was substantially decreased at a K+/Na+ of 1.0 in the nutrient solution compared with lower K+/Na+. Although the [K+] and [Na+] in different plant parts were related to the K+/Na+ in the external solution, the [K+ + Na+] was generally not affected by K+/Na+ in the external medium. The single exception to this observation was at 400 mol m-3 (K+ + Na+)Cl-, when the [K+ +Na+] in the shoots was substantially higher at a K+/Na+ of 1.0 than at lower K+/Na+. In general, K+/Na+ was highest in the apical root segments (10–12 mm long), lower in the bulk roots and lowest in the leaves.