Mark Tester

Mechanisms of salinity tolerance

The physiological and molecular mechanisms of tolerance to osmotic and ionic components of salinity stress are reviewed at the cellular, organ, and whole-plant level. Plant growth responds to salinity in two phases: a rapid, osmotic phase that inhibits growth of young leaves, and a slower, ionic phase that accelerates senescence of mature leaves. Plant adaptations to salinity are of three distinct types: osmotic stress tolerance, Na(+) or Cl() exclusion, and the tolerance of tissue to accumulated Na(+) or Cl(). Our understanding of the role of the HKT gene family in Na(+) exclusion from leaves is increasing, as is the understanding of the molecular bases for many other transport processes at the cellular level. However, we have a limited molecular understanding of the overall control of Na(+) accumulation and of osmotic stress tolerance at the whole-plant level. Molecular genetics and functional genomics provide a new opportunity to synthesize molecular and physiological knowledge to improve the salinity tolerance of plants relevant to food production and environmental sustainability.

Breeding technologies to increase crop production in a changing world.

To feed the several billion people living on this planet, the production of high-quality food must increase with reduced inputs, but this accomplishment will be particularly challenging in the face of global environmental change. Plant breeders need to focus on traits with the greatest potential to increase yield. Hence, new technologies must be developed to accelerate breeding through improving genotyping and phenotyping methods and by increasing the available genetic diversity in breeding germplasm. The most gain will come from delivering these technologies in developing countries, but the technologies will have to be economically accessible and readily disseminated. Crop improvement through breeding brings immense value relative to investment and offers an effective approach to improving food security.

Functional analysis of AtHKT1 in Arabidopsis shows that Na+ recirculation by the phloem is crucial for salt tolerance

Two allelic recessive mutations of Arabidopsis, sas2‐1 and sas2‐2, were identified as inducing sodium overaccumulation in shoots. The sas2 locus was found (by positional cloning) to correspond to the AtHKT1 gene. Expression in Xenopus oocytes revealed that the sas2‐1 mutation did not affect the ionic selectivity of the transporter but strongly reduced the macro scopic (whole oocyte current) transport activity. In Arabidopsis, expression of AtHKT1 was shown to be restricted to the phloem tissues in all organs. The sas2‐1 mutation strongly decreased Na+ concentration in the phloem sap. It led to Na+ overaccumulation in every aerial organ (except the stem), but to Na+ underaccumulation in roots. The sas2 plants displayed increased sensitivity to NaCl, with reduced growth and even death under moderate salinity. The whole set of data indicates that AtHKT1 is involved in Na+ recirculation from shoots to roots, probably by mediating Na+ loading into the phloem sap in shoots and unloading in roots, this recirculation removing large amounts of Na+ from the shoot and playing a crucial role in plant tolerance to salt.

Wheat grain yield on saline soils is improved by an ancestral Na+ transporter gene

The ability of wheat to maintain a low sodium concentration ([Na+]) in leaves correlates with improved growth under saline conditions. This trait, termed Na+ exclusion, contributes to the greater salt tolerance of bread wheat relative to durum wheat. To improve the salt tolerance of durum wheat, we explored natural diversity in shoot Na+ exclusion within ancestral wheat germplasm. Previously, we showed that crossing of Nax2, a gene locus in the wheat relative Triticum monococcum into a commercial durum wheat (Triticum turgidum ssp. durum var. Tamaroi) reduced its leaf [Na+] (ref. 5). Here we show that a gene in the Nax2 locus, TmHKT1;5-A, encodes a Na+-selective transporter located on the plasma membrane of root cells surrounding xylem vessels, which is therefore ideally localized to withdraw Na+ from the xylem and reduce transport of Na+ to leaves. Field trials on saline soils demonstrate that the presence of TmHKT1;5-A significantly reduces leaf [Na+] and increases durum wheat grain yield by 25% compared to near-isogenic lines without the Nax2 locus.

Shoot Na+ Exclusion and Increased Salinity Tolerance Engineered by Cell Type-Specific Alteration of Na+ Transport in Arabidopsis

Soil salinity affects large areas of cultivated land, causing significant reductions in crop yield globally. The Na+ toxicity of many crop plants is correlated with overaccumulation of Na+ in the shoot. We have previously suggested that the engineering of Na+ exclusion from the shoot could be achieved through an alteration of plasma membrane Na+ transport processes in the root, if these alterations were cell type specific. Here, it is shown that expression of the Na+ transporter HKT1;1 in the mature root stele of Arabidopsis thaliana decreases Na+ accumulation in the shoot by 37 to 64%. The expression of HKT1;1 specifically in the mature root stele is achieved using an enhancer trap expression system for specific and strong overexpression. The effect in the shoot is caused by the increased influx, mediated by HKT1;1, of Na+ into stelar root cells, which is demonstrated in planta and leads to a reduction of root-to-shoot transfer of Na+. Plants with reduced shoot Na+ also have increased salinity tolerance. By contrast, plants constitutively expressing HKT1;1 driven by the cauliflower mosaic virus 35S promoter accumulated high shoot Na+ and grew poorly. Our results demonstrate that the modification of a specific Na+ transport process in specific cell types can reduce shoot Na+ accumulation, an important component of salinity tolerance of many higher plants.

Root Plasma Membrane Transporters Controlling K+/Na+ Homeostasis in Salt-Stressed Barley

Plant salinity tolerance is a polygenic trait with contributions from genetic, developmental, and physiological interactions, in addition to interactions between the plant and its environment. In this study, we show that in salt-tolerant genotypes of barley (Hordeum vulgare), multiple mechanisms are well combined to withstand saline conditions. These mechanisms include: (1) better control of membrane voltage so retaining a more negative membrane potential; (2) intrinsically higher H+ pump activity; (3) better ability of root cells to pump Na+ from the cytosol to the external medium; and (4) higher sensitivity to supplemental Ca2+. At the same time, no significant difference was found between contrasting cultivars in their unidirectional 22Na+ influx or in the density and voltage dependence of depolarization-activated outward-rectifying K+ channels. Overall, our results are consistent with the idea of the cytosolic K+-to-Na+ ratio being a key determinant of plant salinity tolerance, and suggest multiple pathways of controlling that important feature in salt-tolerant plants.

Free oxygen radicals regulate plasma membrane Ca2+- and K+-permeable channels in plant root cells

Free oxygen radicals are an irrefutable component of life, underlying important biochemical and physiological phenomena in animals. Here it is shown that free oxygen radicals activate plasma membrane Ca2+- and K+-permeable conductances in Arabidopsis root cell protoplasts, mediating Ca2+ influx and K+ efflux, respectively. Free oxygen radicals generate increases in cytosolic Ca2+ mediated by a novel population of nonselective cation channels that differ in selectivity and pharmacology from those involved in toxic Na+ influx. Analysis of the free oxygen radical-activated K+ conductance showed its similarity to the Arabidopsis root K+ outward rectifier. Significantly larger channel activation was found in cells responsible for perceiving environmental signals and undergoing elongation. Quenching root free oxygen radicals inhibited root elongation, confirming the role of radical-activated Ca2+ influx in cell growth. Net free oxygen radical-stimulated Ca2+ influx and K+ efflux were observed in root cells of monocots, dicots, C3 and C4 plants, suggesting conserved mechanisms and functions. In conclusion, two functions for free oxygen radical cation channel activation are proposed: initialization/amplification of stress signals and control of cell elongation in root growth.

Boron-Toxicity Tolerance in Barley Arising from Efflux Transporter Amplification

Both limiting and toxic soil concentrations of the essential micronutrient boron represent major limitations to crop production worldwide. We identified Bot1, a BOR1 ortholog, as the gene responsible for the superior boron-toxicity tolerance of the Algerian barley landrace Sahara 3771 (Sahara). Bot1 was located at the tolerance locus by high-resolution mapping. Compared to intolerant genotypes, Sahara contains about four times as many Bot1 gene copies, produces substantially more Bot1 transcript, and encodes a Bot1 protein with a higher capacity to provide tolerance in yeast. Bot1 transcript levels identified in barley tissues are consistent with a role in limiting the net entry of boron into the root and in the disposal of boron from leaves via hydathode guttation.

Salt resistant crop plants

Soil salinity is a major constraint to agriculture. To improve salinity tolerance of crops, various traits can be incorporated, including ion exclusion, osmotic tolerance and tissue tolerance. We review the roles of a range of genes involved in salt tolerance traits. Different tissues and cells are adapted for specific and often diverse function, so it is important to express the genes in specific cell-types and to pyramid a range of traits. Modern biotechnology (marker-assisted selection or genetic engineering) needs to be increasingly used to introduce the correct combination of genes into elite crop cultivars. Importantly, the effects of introduced genes need to be evaluated in the field to determine their effect on salinity tolerance and yield improvement.

Control of Sodium Transport in Durum Wheat

In many species, salt sensitivity is associated with the accumulation of sodium (Na+) in photosynthetic tissues. Na+ uptake to leaves involves a series of transport steps and so far very few candidate genes have been implicated in the control of these processes. In this study, Na+ transport was compared in two varieties of durum wheat (Triticum turgidum) L. subsp. durum known to differ in salt tolerance and Na+ accumulation; the relatively salt tolerant landrace line 149 and the salt sensitive cultivar Tamaroi. Genetic studies indicated that these genotypes differed at two major loci controlling leaf blade Na+ accumulation (R. Munns, G.J. Rebetzke, S. Husain, R.A. James, R.A. Hare [2003] Aust J Agric Res 54: 627–635). The physiological traits determined by these genetic differences were investigated using measurements of unidirectional 22Na+ transport and net Na+ accumulation. The major differences in Na+ transport between the genotypes were (1) the rate of transfer from the root to the shoot (xylem loading), which was much lower in the salt tolerant genotype, and (2) the capacity of the leaf sheath to extract and sequester Na+ as it entered the leaf. The genotypes did not differ significantly in unidirectional root uptake of Na+ and there was no evidence for recirculation of Na+ from shoots to roots. It is likely that xylem loading and leaf sheath sequestration are separate genetic traits that interact to control leaf blade Na+.