Potassium retention in leaf mesophyll as an element of salinity tissue tolerance in halophytes.

Soil salinity remains a major threat to global food security, and the progress in crop breeding for salinity stress tolerance may be achieved only by pyramiding key traits mediating plant adaptive responses to high amounts of dissolved salts in the rhizosphere. This task may be facilitated by studying natural variation in salinity tolerance among plant species and, specifically, exploring mechanisms of salinity tolerance in halophytes. The aim of this work was to establish the causal link between mesophyll ion transport activity and plant salt tolerance in a range of evolutionary contrasting halophyte and glycophyte species. Plants were grown under saline conditions in a glasshouse, followed by assessing their growth and photosynthetic performance. In a parallel set of experiments, net K+ and H+ transport across leaf mesophyll and their modulation by light were studied in control and salt-treated mesophyll segments using vibrating non-invasive ion selective microelectrode (the MIFE) technique. The reported results show that mesophyll cells in glycophyte species loses 2-6 fold more K+ compared with their halophyte counterparts. This decline was reflected in a reduced maximum photochemical efficiency of photosystem II, chlorophyll content and growth observed in the glasshouse experiments. In addition to reduced K+ efflux, the more tolerant species also exhibited reduced H+ efflux, which is interpreted as an energy-saving strategy allowing more resources to be redirected towards plant growth. It is concluded that the ability of mesophyll to retain K+ without a need to activate plasma membrane H+-ATPase is an essential component of salinity tolerance in halophytes and halophytic crop plants.

Salinity effects on chloroplast PSII performance in glycophytes and halophytes.

The effects of NaCl stress and K+ nutrition on photosynthetic parameters of isolated chloroplasts were investigated using PAM fluorescence. Intact mesophyll cells were able to maintain optimal photosynthetic performance when exposed to salinity for more than 24h whereas isolated chloroplasts showed declines in both the relative electron transport rate (rETR) and the maximal photochemical efficiency of PSII (Fv/Fm) within the first hour of treatment. The rETR was much more sensitive to salt stress compared with Fv/Fm, with 40% inhibition of rETR observed at apoplastic NaCl concentration as low as 20mM. In isolated chloroplasts, absolute K+ concentrations were more essential for the maintenance of the optimal photochemical performance (Fv/Fm values) rather than sodium concentrations per se. Chloroplasts from halophyte species of quinoa (Chenopodium quinoa Willd.) and pigface (Carpobrotus rosii (Haw.) Schwantes) showed less than 18% decline in Fv/Fm under salinity, whereas the Fv/Fm decline in chloroplasts from glycophyte pea (Pisum sativum L.) and bean (Vicia faba L.) species was much stronger (31 and 47% respectively). Vanadate (a P-type ATPase inhibitor) significantly reduced Fv/Fm in both control and salinity treated chloroplasts (by 7 and 25% respectively), whereas no significant effects of gadolinium (blocker of non-selective cation channels) were observed in salt-treated chloroplasts. Tetraethyl ammonium (TEA) (K+ channel inhibitor) and amiloride (inhibitor of the Na+/H+ antiporter) increased the Fv/Fm of salinity treated chloroplasts by 16 and 17% respectively. These results suggest that chloroplasts' ability to regulate ion transport across the envelope and thylakoid membranes play a critical role in leaf photosynthetic performance under salinity.

Ion transport in broad bean leaf mesophyll under saline conditions.

MAIN CONCLUSION: Salt stress reduces the ability of mesophyll tissue to respond to light. Potassium outward rectifying channels are responsible for 84 % of Na (+) induced potassium efflux from mesophyll cells. Modulation in ion transport of broad bean (Vicia faba L.) mesophyll to light under increased apoplastic salinity stress was investigated using vibrating ion-selective microelectrodes (the MIFE technique). Increased apoplastic Na(+) significantly affected mesophyll cells ability to respond to light by modulating ion transport across their membranes. Elevated apoplastic Na(+) also induced a significant K(+) efflux from mesophyll tissue. This efflux was mediated predominately by potassium outward rectifying channels (84 %) and the remainder of the efflux was through non-selective cation channels. NaCl treatment resulted in a reduction in photosystem II efficiency in a dose- and time-dependent manner. In particular, reductions in Fv'/Fm' were linked to K(+) homeostasis in the mesophyll tissue. Increased apoplastic Na(+) concentrations induced vanadate-sensitive net H(+) efflux, presumably mediated by the plasma membrane H(+)-ATPase. It is concluded that the observed pump's activation is essential for the maintenance of membrane potential and ion homeostasis in the cytoplasm of mesophyll under salt stress.

Xylem ionic relations and salinity tolerance in barley.

Control of ion loading into the xylem has been repeatedly named as a crucial factor determining plant salt tolerance. In this study we further investigate this issue by applying a range of biophysical [the microelectrode ion flux measurement (MIFE) technique for non-invasive ion flux measurements, the patch clamp technique, membrane potential measurements] and physiological (xylem sap and tissue nutrient analysis, photosynthetic characteristics, stomatal conductance) techniques to barley varieties contrasting in their salt tolerance. We report that restricting Na(+) loading into the xylem is not essential for conferring salinity tolerance in barley, with tolerant varieties showing xylem Na(+) concentrations at least as high as those of sensitive ones. At the same time, tolerant genotypes are capable of maintaining higher xylem K(+)/Na(+) ratios and efficiently sequester the accumulated Na(+) in leaves. The former is achieved by more efficient loading of K(+) into the xylem. We argue that the observed increases in xylem K(+) and Na(+) concentrations in tolerant genotypes are required for efficient osmotic adjustment, needed to support leaf expansion growth. We also provide evidence that K(+)-permeable voltage-sensitive channels are involved in xylem loading and operate in a feedback manner to maintain a constant K(+)/Na(+) ratio in the xylem sap.