Roberto A. Gaxiola

Drought- and salt-tolerant plants result from overexpression of the AVP1 H+-pump

Transgenic plants overexpressing the vacuolar H+-pyrophosphatase are much more resistant to high concentrations of NaCl and to water deprivation than the isogenic wild-type strains. These transgenic plants accumulate more Na+ and K+ in their leaf tissue than the wild type. Moreover, direct measurements on isolated vacuolar membrane vesicles derived from the AVP1 transgenic plants and from wild type demonstrate that the vesicles from the transgenic plants have enhanced cation uptake. The phenotypes of the AVP1 transgenic plants suggest that increasing the vacuolar proton gradient results in increased solute accumulation and water retention. Presumably, sequestration of cations in the vacuole reduces their toxic effects. Genetically engineered drought- and salt-tolerant plants could provide an avenue to the reclamation of farmlands lost to agriculture because of salinity and a lack of rainfall.

Plant proton pumps

Chemiosmotic circuits of plant cells are driven by proton (H+) gradients that mediate secondary active transport of compounds across plasma and endosomal membranes. Furthermore, regulation of endosomal acidification is critical for endocytic and secretory pathways. For plants to react to their constantly changing environments and at the same time maintain optimal metabolic conditions, the expression, activity and interplay of the pumps generating these H+ gradients have to be tightly regulated. In this review, we will highlight results on the regulation, localization and physiological roles of these H+‐ pumps, namely the plasma membrane H+‐ATPase, the vacuolar H+‐ATPase and the vacuolar H+‐PPase.

Microbial models and salt stress tolerance in plants

Abstract Improving salt tolerance in crop plants remains an urgent issue in plant molecular biology. The adaptation of plants to NaCl involves metabolic reactions (synthesis of organic solutes) and transport phenomena (ion extrusion at the plasma membrane and vacuolar compartmentation). In addition, a plethora of salt-induced genes with a bewildering variety of suggested functions have been described. The uncertainties about the physiological roles and/or molecular bases of many of these phenomena make it difficult to select genes that could improve salt tolerance (halotolerance) in transgenic plants. We suggest that the field of salt tolerance can benefit by complementing the present phenomenological or descriptive approaches with a functional strategy directed toward isolating genes that, by overexpression of the corresponding protein, could improve salt tolerance. These halotolerance genes not only could illuminate the critical steps for salt tolerance, but also could provide the tools for improvement....

A novel and conserved salt-induced protein is an important determinant of salt tolerance in yeast.

We have isolated a novel yeast gene, HAL1, which upon overexpression improves growth under salt stress. In addition, disruption of this gene decreases salt tolerance. Therefore HAL1 constitutes a rate‐limiting determinant for halotolerance. It encodes a polar protein of 32 kDa located in the yeast cytoplasm and unrelated to sequences in data banks. The expression of this gene is increased by high concentrations of either NaCl, KCl or sorbitol. On the other hand, the growth advantage obtained by overexpression of HAL1 is specific for NaCl stress. In cells overexpressing HAL1, sodium toxicity seems to be counteracted by an increased accumulation of potassium. The HAL1 protein could interact with the transport systems which determine intracellular K+ homeostasis. The HAL1 gene and encoded protein are conserved in plants, being induced in these organisms by salt stress and abscisic acid. These results suggest that yeast serves as a convenient model system for the molecular biology of plant salt tolerance.

Expression of an Arabidopsis vacuolar H+-pyrophosphatase gene (AVP1) in cotton improves drought- and salt tolerance and increases fibre yield in the field conditions

The Arabidopsis gene AVP1 encodes a vacuolar pyrophosphatase that functions as a proton pump on the vacuolar membrane. Overexpression of AVP1 in Arabidopsis, tomato and rice enhances plant performance under salt and drought stress conditions, because up-regulation of the type I H+-PPase from Arabidopsis may result in a higher proton electrochemical gradient, which facilitates enhanced sequestering of ions and sugars into the vacuole, reducing water potential and resulting in increased drought- and salt tolerance when compared to wild-type plants. Furthermore, overexpression of AVP1 stimulates auxin transport in the root system and leads to larger root systems, which helps transgenic plants absorb water more efficiently under drought conditions. Using the same approach, AVP1-expressing cotton plants were created and tested for their performance under high-salt and reduced irrigation conditions. The AVP1-expressing cotton plants showed more vigorous growth than wild-type plants in the presence of 200 mM NaCl under hydroponic growth conditions. The soil-grown AVP1-expressing cotton plants also displayed significantly improved tolerance to both drought and salt stresses in greenhouse conditions. Furthermore, the fibre yield of AVP1-expressing cotton plants is at least 20% higher than that of wild-type plants under dry-land conditions in the field. This research indicates that AVP1 has the potential to be used for improving crops drought- and salt tolerance in areas where water and salinity are limiting factors for agricultural productivity.

Arabidopsis ALF5, a Multidrug Efflux Transporter Gene Family Member, Confers Resistance to Toxins

The Arabidopsis genome contains many gene families that are not found in the animal kingdom. One of these is the multidrug and toxic compound extrusion (MATE) family, which has homology with bacterial efflux transporters. Arabidopsis has at least 54 members of this family, which often are found in tandem repeats. Analysis of ALF5, one member of this Arabidopsis family, suggests that its function is required for protection of the roots from inhibitory compounds. Loss of ALF5 function results in the sensitivity of the root to a number of compounds, including a contaminant of commercial agar. Moreover, expression of the Arabidopsis ALF5 cDNA in yeast confers resistance to tetramethylammonium. These phenotypes are consistent with a role for ALF5 as an efflux transporter. Both transcriptional and translational fusions of ALF5 to the β-glucuronidase reporter gene show that ALF5 is expressed strongly in the root epidermis, a tissue in direct contact with the external environment. The distinct requirement for ALF5 function is remarkable because of the large number of MATE gene family members in Arabidopsis, one of which is adjacent to ALF5 and 83% identical to ALF5 at the amino acid level.

Salt tolerance and methionine biosynthesis in Saccharomyces cerevisiae involve a putative phosphatase gene.

The progressive salinization of irrigated land poses a threat to the future of agriculture in arid regions. The identification of crucial metabolic steps in salt tolerance is important for the understanding of stress physiology and may provide the tools for its genetic engineering. In the yeast Saccharomyces cerevisiae we have isolated a gene, HAL2, which upon increase in gene dosage improves growth under NaCl and LiCl stresses. The HAL2 protein is homologous to inositol phosphatases, enzymes known to be inhibited by lithium salts. Complementation analysis demonstrated that HAL2 is identical to MET22, a gene involved in methionine biosynthesis. Accordingly, methionine supplementation improves the tolerance of yeast to NaCl and LiCl. These results demonstrate an unsuspected interplay between methionine biosynthesis and salt tolerance.

Genetic Manipulation of Vacuolar Proton Pumps and Transporters

Cells expend as much as 50% of their total intracellular energy reserves to maintain gradients of ions across their membranes ([Nelson, 1994][1]). These gradients have been associated with the myriad of functions attributed to the membranes of living organisms. In the past, much of our knowledge

Genetic Manipulation of a "Vacuolar" H+ -PPase: From Salt Tolerance to Yield Enhancement under Phosphorus-Deficient Soils

Plant scientists face the difficult challenge of increasing food production without further degradation of the environment. In order to protect drinking water resources and prevent the proliferation of harmful algal blooms and “dead zones” in coastal marine ecosystems, it is imperative to reduce

A transgenic approach to enhance phosphorus use efficiency in crops as part of a comprehensive strategy for sustainable agriculture

Concerns about phosphorus (P) sustainability in agriculture arise not only from the potential of P scarcity but also from the known effects of agricultural P use beyond the field, i.e., eutrophication leading to dead zones in lakes, rivers and coastal oceans due to runoffs from fertilized fields. Plants possess a large number of adaptive responses to P(i) (orthophosphate) limitation that provide potential raw materials to enhance P(i) scavenging abilities of crop plants. Understanding and engineering these adaptive responses to increase the efficiency of crop capture of natural and fertilizer P(i) in soils is one way to optimize P(i) use efficiency (PUE) and, together with other approaches, help to meet the P sustainability challenge in agriculture. Research on the molecular and physiological basis of P(i) uptake is facilitating the generation of plants with enhanced P(i) use efficiency by genetic engineering. Here we describe work done in this direction with emphasis on the up-regulation of plant proton-translocating pyrophosphatases (H(+)-PPases).