Guo-Qiang Wu

Puccinellia tenuiflora maintains a low Na+ level under salinity by limiting unidirectional Na+ influx resulting in a high selectivity for K+ over Na+

Puccinellia tenuiflora is a useful monocotyledonous halophyte that might be used for improving salt tolerance of cereals. This current work has shown that P. tenuiflora has stronger selectivity for K+ over Na+ allowing it to maintain significantly lower tissue Na+ and higher K+ concentration than that of wheat under short- or long-term NaCl treatments. To assess the relative contribution of Na+ efflux and influx to net Na+ accumulation, unidirectional 22Na+ fluxes in roots were carried out. It was firstly found that unidirectional 22Na+ influx into root of P. tenuiflora was significantly lower (by 31-37%) than in wheat under 100 and 150 mM NaCl. P. tenuiflora had lower unidirectional Na+ efflux than wheat; the ratio of efflux to influx was similar between the two species. Leaf secretion of P. tenuiflora was also estimated, and found the loss of Na+ content from leaves to account for only 0.0006% of the whole plant Na+ content over 33 d of NaCl treatments. Therefore, it is proposed that neither unidirectional Na+ efflux of roots nor salt secretion by leaves, but restricting unidirectional Na+ influx into roots with a strong selectivity for K+ over Na+ seems likely to contribute to the salt tolerance of P. tenuiflora.

The ZxNHX gene encoding tonoplast Na+/H+ antiporter from the xerophyte Zygophyllum xanthoxylum plays important roles in response to salt and drought

Sodium (Na(+)) has been found to play important roles in the adaptation of xerophytic species to drought conditions. The tonoplast Na(+)/H(+) antiporter (NHX) proved to be involved in the compartmentalization of Na(+) into vacuoles from the cytosol. In this study, a gene (ZxNHX) encoding tonoplast Na(+)/H(+) antiporter was isolated and characterized in Zygophyllum xanthoxylum, a succulent xerophyte growing in desert areas of northwest China. The results revealed that ZxNHX consisted of 532 amino acid residues with a conserved binding domain ((78)LFFIYLLPPI(87)) for amiloride and shared high similarity (73-81%) with the identified tonoplast Na(+)/H(+) antiporters in other plant species. Semi-quantitative RT-PCR analysis showed that the mRNA level of ZxNHX was significantly higher in the leaf than in stem or root. The transcript abundance of ZxNHX in Z. xanthoxylum subjected to salt (5-150 mM NaCl) or drought (50-15% of field water capacity (FWC)) was 1.4-8.4 times or 2.3-4.4 times that of plants grown in the absence of NaCl or 70% of FWC, respectively. Leaf Na(+) concentration in plants exposed to salt or drought was 1.7-5.2 times or 1.5-2.2 times that of corresponding control plants, respectively. It is clear that there is a positive correlation between up-regulation of ZxNHX and accumulation of Na(+) in Z. xanthoxylum exposed to salt or drought. Furthermore, Z. xanthoxylum accumulated larger amounts of Na(+) than K(+) in the leaf under drought conditions, even in low salt soil. In summary, our results suggest that ZxNHX encodes a tonoplast Na(+)/H(+) antiporter and plays important roles in Na(+) accumulation and homeostasis of Z. xanthoxylum under salt and drought conditions.

ZxNHX controls Na+ and K+ homeostasis at the whole-plant level in Zygophyllum xanthoxylum through feedback regulation of the expression of genes involved in their transport

BACKGROUND AND AIMS In order to cope with arid environments, the xerohalophyte Zygophyllum xanthoxylum efficiently compartmentalizes Na(+) into vacuoles, mediated by ZxNHX, and maintains stability of K(+) in its leaves. However, the function of ZxNHX in controlling Na(+) and K(+) homeostasis at the whole-plant level remains unclear. In this study, the role of ZxNHX in regulating the expression of genes involved in Na(+) and K(+) transport and spatial distribution was investigated. METHODS The role of ZxNHX in maintaining Na(+) and K(+) homeostasis in Z. xanthoxylum was studied using post-transcriptional gene silencing via  Agrobacterium-mediated transformation. Transformed plants were grown with or without 50 mm NaCl, and expression levels and physiological parameters were measured. KEY RESULTS It was found that 50 mm NaCl induced a 620 % increase in transcripts of ZxSOS1 but only an 80 % increase in transcripts of ZxHKT1;1 in roots of wild-type (WT) plants. Consequently, the ability of ZxSOS1 to transport Na(+) exceeded that of ZxHKT1;1, and Na(+) was loaded into the xylem by ZxSOS1 and delivered to the shoots. However, in a ZxNHX-silenced line (L7), the capacity to sequester Na(+) into vacuoles of leaves was weakened, which in turn regulated long-distance Na(+) transport from roots to shoots. In roots of L7, NaCl (50 mm) increased transcripts of ZxSOS1 by only 10 %, whereas transcripts of ZxHKT1;1 increased by 53 %. Thus, in L7, the transport ability of ZxHKT1;1 for Na(+) outweighed that of ZxSOS1. Na(+) was unloaded from the xylem stream, consequently reducing Na(+) accumulation and relative distribution in leaves, but increasing the relative distribution of Na(+) in roots and the net selective transport capacity for K(+) over Na(+) from roots to shoots compared with the WT. Silencing of ZxNHX also triggered a downregulation of  ZxAKT1 and ZxSKOR in roots, resulting in a significant decrease in K(+) accumulation in all the tissues in plants grown in 50 mm NaCl. These changes led to a significant reduction in osmotic adjustment, and thus an inhibition of growth in ZxNHX-silenced lines. CONCLUSIONS The results suggest that ZxNHX is essential for controlling Na(+), K(+) uptake, long-distance transport and their homeostasis at whole-plant level via feedback regulation of the expression of genes involved in Na(+), K(+) transport. The net result is the maintenance of the characteristic salt accumulation observed in Z. xanthoxylum and the regulation of its normal growth. A model is proposed for the role of ZxNHX in regulating the Na(+) transport system in Z. xanthoxylum under saline conditions.

The coordinated regulation of Na+ and K+ in Hordeum brevisubulatum responding to time of salt stress

Hordeum brevisubulatum, called as wild barley, is a useful monocotyledonous halophyte for soil improvement in northern China. Although previously studied, its main salt tolerance mechanism remained controversial. The current work showed that shoot Na+ concentration was increased rapidly with stress time and significantly higher than in wheat during 0-168h of 100mM NaCl treatment. Similar results were also found under 25 and 50mM NaCl treatments. Even K+ was increased from 0.01 to 50mM in the cultural solution, no significant effect was found on tissue Na+ concentrations. Interestingly, shoot growth was improved, and stronger root activity was maintained in H. brevisubulatum compared with wheat after 7days treatment of 100mM NaCl. To investigate the long-term stress impact on tissue Na+, 100mM NaCl was prolonged to 60 days. The maximum values of Na+ concentrations were observed at 7th in shoot and 14th day in roots, respectively, and then decreased gradually. Micro-electrode ion flux estimation was used and it was found that increasing Na+ efflux while maintaining K+ influx were the major strategies to reduce the Na+ concentration during long-term salt stress. Moreover, leaf Na+ secretions showed little contribution to the tissue Na+ decrease. Thereby, the physiological mechanism for H. brevisubulatum to survive from long-term salt stress was proposed that rapid Na+ accumulation occurred in the shoot to respond the initial salt shock, then Na+ efflux was triggered and K+ influx was activated to maintain a stable K+/Na+ ratio in tissues.

Co-expression of xerophyte Zygophyllum xanthoxylum ZxNHX and ZxVP1-1 confers enhanced salinity tolerance in chimeric sugar beet (Beta vulgaris L.)

Salinity is one of the major abiotic stresses that limit the growth and productivity of sugar beet (Beta vulgaris L.). To improve sugar beet’s salinity tolerance, the ZxNHX and ZxVP1-1 genes encoding tonoplast Na+/H+ antiporter and H+-PPase from xerophyte Zygophyllum xanthoxylum were co-expressed by Agrobacterium tumefaciens-mediated transformation. It is showed here that co-expression of ZxNHX and ZxVP1-1 confers enhanced salinity tolerance to the transformed sugar beet plants compared with the wild-type (WT) plants. The chimeric plants grew well in the presence of high salinity (400 mM NaCl), whereas WT plants displayed chlorosis and died within 8 days. Compared to WT plants, the chimeric plants co-expressing ZxNHX and ZxVP1-1 accumulated more proline, Na+ and K+ in their leaves and petioles when exposed to high salinity, which caused lower solute potential, retained more water and thus subjected to lesser cell membrane damage. Interestingly, the chimeric plants accumulated higher sucrose, glucose and fructose contents in their storage roots than WT plants in the absence or presence of high salinity. Our results suggested that co-expression of ZxNHX and ZxVP1-1 improved the osmoregulatory capacity in chimeric sugar beet through increased compartmentalization of ions into the vacuoles by enhancing the activity of proton pumps and thus mitigated Na+-toxicity for plants.