Kinetics of xylem loading, membrane potential maintenance, and sensitivity of K(+) -permeable channels to reactive oxygen species: physiological traits that differentiate salinity tolerance between pea and barley.

Salt sensitive (pea) and salt tolerant (barley) species were used to understand the physiological basis of differential salinity tolerance in crops. Pea plants were much more efficient in restoring otherwise depolarized membrane potential thereby effectively decreasing K(+) efflux through depolarization-activated outward rectifying potassium channels. At the same time, pea root apex was 10-fold more sensitive to physiologically relevant H2 O2 concentration and accumulated larger amounts of H2 O2 under saline conditions. This resulted in a rapid loss of cell viability in the pea root apex. Barley plants rapidly loaded Na(+) into the xylem; this increase was only transient, and xylem and leaf Na(+) concentration remained at a steady level for weeks. On the contrary, pea plants restricted xylem Na(+) loading during the first few days of treatment but failed to prevent shoot Na(+) elevation in the long term. It is concluded that superior salinity tolerance of barley plants compared with pea is conferred by at least three different mechanisms: (1) efficient control of xylem Na(+) loading; (2) efficient control of H2 O2 accumulation and reduced sensitivity of non-selective cation channels to H2 O2 in the root apex; and (3) higher energy saving efficiency, with less ATP spent to maintain membrane potential under saline conditions.

Bose J ，Shabala L ，Pottosin I ，Zeng F ，Velarde-Buendía AM ，Massart A ，Poschenrieder C ，Hariadi Y ，Shabala S ... -  《-》

Comparing Kinetics of Xylem Ion Loading and Its Regulation in Halophytes and Glycophytes.

Although control of xylem ion loading is essential to confer salinity stress tolerance, specific details behind this process remain elusive. In this work, we compared the kinetics of xylem Na+ and K+ loading between two halophytes (Atriplex lentiformis and quinoa) and two glycophyte (pea and beans) species, to understand the mechanistic basis of the above process. Halophyte plants had high initial amounts of Na+ in the leaf, even when grown in the absence of the salt stress. This was matched by 7-fold higher xylem sap Na+ concentration compared with glycophyte plants. Upon salinity exposure, the xylem sap Na+ concentration increased rapidly but transiently in halophytes, while in glycophytes this increase was much delayed. Electrophysiological experiments using the microelectrode ion flux measuring technique showed that glycophyte plants tend to re-absorb Na+ back into the stele, thus reducing xylem Na+ load at the early stages of salinity exposure. The halophyte plants, however, were capable to release Na+ even in the presence of high Na+ concentrations in the xylem. The presence of hydrogen peroxide (H2O2) [mimicking NaCl stress-induced reactive oxygen species (ROS) accumulation in the root] caused a massive Na+ and Ca2+ uptake into the root stele, while triggering a substantial K+ efflux from the cytosol into apoplast in glycophyte but not halophytes species. The peak in H2O2 production was achieved faster in halophytes (30 min vs 4 h) and was attributed to the increased transcript levels of RbohE. Pharmacological data suggested that non-selective cation channels are unlikely to play a major role in ROS-mediated xylem Na+ loading.

Zarei M ，Shabala S ，Zeng F ，Chen X ，Zhang S ，Azizi M ，Rahemi M ，Davarpanah S ，Yu M ，Shabala L ... -  《-》

Cell-Type-Specific H+-ATPase Activity in Root Tissues Enables K+ Retention and Mediates Acclimation of Barley (Hordeum vulgare) to Salinity Stress.

While the importance of cell type specificity in plant adaptive responses is widely accepted, only a limited number of studies have addressed this issue at the functional level. We have combined electrophysiological, imaging, and biochemical techniques to reveal the physiological mechanisms conferring higher sensitivity of apical root cells to salinity in barley (Hordeum vulgare). We show that salinity application to the root apex arrests root growth in a highly tissue- and treatment-specific manner. Although salinity-induced transient net Na+ uptake was about 4-fold higher in the root apex compared with the mature zone, mature root cells accumulated more cytosolic and vacuolar Na+, suggesting that the higher sensitivity of apical cells to salt is not related to either enhanced Na+ exclusion or sequestration inside the root. Rather, the above differential sensitivity between the two zones originates from a 10-fold difference in K+ efflux between the mature zone and the apical region (much poorer in the root apex) of the root. Major factors contributing to this poor K+ retention ability are (1) an intrinsically lower H+-ATPase activity in the root apex, (2) greater salt-induced membrane depolarization, and (3) a higher reactive oxygen species production under NaCl and a larger density of reactive oxygen species-activated cation currents in the apex. Salinity treatment increased (2- to 5-fold) the content of 10 (out of 25 detected) amino acids in the root apex but not in the mature zone and changed the organic acid and sugar contents. The causal link between the observed changes in the root metabolic profile and the regulation of transporter activity is discussed.

Shabala L ，Zhang J ，Pottosin I ，Bose J ，Zhu M ，Fuglsang AT ，Velarde-Buendia A ，Massart A ，Hill CB ，Roessner U ，Bacic A ，Wu H ，Azzarello E ，Pandolfi C ，Zhou M ，Poschenrieder C ，Mancuso S ，Shabala S ... -  《-》

Physiological and molecular mechanisms mediating xylem Na(+) loading in barley in the context of salinity stress tolerance.

Time-dependent kinetics of xylem Na+ loading was investigated using a large number of barley genotypes contrasting in their salinity tolerance. Salt-sensitive varieties were less efficient in controlling xylem Na+ loading and showed a gradual increase in the xylem Na+ content over the time. To understand underlying ionic and molecular mechanisms, net fluxes of Ca2+ , K+ and Na+ were measured from the xylem parenchyma tissue in response to H2 O2 and ABA; both of them associated with salinity stress signalling. Our results indicate that NADPH oxidase-mediated apoplastic H2 O2 production acts upstream of the xylem Na+ loading and is causally related to ROS-inducible Ca2+ uptake systems in the root stelar tissue. It was also found that ABA regulates (directly or indirectly) the process of Na+ retrieval from the xylem and the significant reduction of Na+ and K+ fluxes induced by bumetanide are indicative of a major role of chloride cation co-transporter (CCC) on xylem ion loading. Transcript levels of HvHKT1;5_like and HvSOS1_like genes in the root stele were observed to decrease after salt stress, while there was an increase in HvSKOR_like gene, indicating that these ion transporters are involved in primary Na+ /K+ movement into/out of xylem.

Zhu M ，Zhou M ，Shabala L ，Shabala S ... -  《-》

Xylem ionic relations and salinity tolerance in barley.

Shabala S ，Shabala S ，Cuin TA ，Pang J ，Percey W ，Chen Z ，Conn S ，Eing C ，Wegner LH ... -  《-》