Effects of low temperature (8 degrees C) on the hydraulic conductivity of young roots of a chilling-sensitive (cucumber, Cucumis sativus L.) and a chilling-resistant (figleaf gourd, Cucurbita ficifolia Bouche) crop have been measured at the levels of whole root systems (root hydraulic conductivity, Lp(r)) and of individual cortical cells (cell hydraulic conductivity, Lp). Exposure of roots to low temperature (LRT) for up to 6 d caused a stronger suberization of the endodermis in cucumber compared with figleaf gourd, but no development of exodermal Casparian bands in either species. Changes in anatomy after 6 d of LRT treatment corresponded with a reduction in hydrostatic root Lp(r) of cucumber roots by a factor of 24, and by a factor of 2 in figleaf gourd. In figleaf gourd, there was a reduction only in hydrostatic Lp(r) but not in osmotic Lp(r) suggesting that the activity of water channels was not much affected by LRT treatment in this species. Changes in cell Lp in response to chilling and recovery were similar to the root levels, although they were more intense at the root level. Activation energies (E(a)) and Q10 of water flow as measured at the cell level were high in cucumber (E(a)=109+/-13 kJ mol(-1); Q(10)=4.8+/-0.7; n=6-10 cells), but small in figleaf gourd (E(a)=11+/-2 kJ mol(-1); Q10=1.2+/-0.1; n=6-10 cells). Roots of figleaf gourd recovered better from LRT treatment than those of cucumber. In figleaf gourd, recovery (at both the root and cell level) often resulted in Lp and Lp(r) values which were even bigger than the original, i.e. there was an overshoot in hydraulic conductivity. These effects were larger for osmotic (representing the cell-to-cell passage of water) than for hydrostatic Lp(r). After a short-term (1 d) exposure to 8 degrees C followed by 1 d at 20 degrees C, hydrostatic Lp(r) of cucumber nearly recovered and that of figleaf gourd still remained higher due to the overshoot. By contrast, osmotic Lp(r) and cell Lp in both species remained high by a factor of 3 compared with the control, possibly due to an increased activity of water channels. After preconditioning of roots at LRT, increased hydraulic conductivity was completely inhibited by HgCl2 at both the root and cell levels. Different from figleaf gourd, recovery from chilling was not complete in cucumber after longer exposure to LRT. It is concluded that at LRT, both changes in the activity of aquaporins (AQPs) and alterations of root anatomy determine the water uptake in both species. The high temperature dependence of cell Lp in cucumber suggests conformational changes of AQPs during LRT treatment which result in channel closure and in a strong gating of AQP activity by low temperature. This mechanism is thought to be different from that in figleaf gourd where AQPs reacted in the conventional way, i.e. low temperature affected the mobility of water molecules in AQPs rather than their open/closed state, and Q(10) was low.

Water transport across root systems of young cucumber (Cucumis sativus L.) seedlings was measured following exposure to low temperature (LT, 8-13 degrees C) for varying periods of time. In addition, the amount of water transported through the stems was evaluated using a heat-balance sap-flow gauge. Following LT treatment, hydrogen peroxide was localized cytochemically in root tissue by the oxidation of cerium (III) chloride. The effects of hydrogen peroxide on the hydraulic conductivity of single cells (Lp) in root tissues, and on the H+-ATPase activity of isolated root plasma membrane, have been worked out. Cytochemical evidence suggested that exposure of roots to LT stress caused a release of hydrogen peroxide in the millimolar range in the vicinity of plasma membranes. In response to a low root temperature (8 degrees C), the hydraulic conductivity of the root (Lp(r)) decreased by a factor of 4, and the half-times of water exchange increased by a factor of 5-6. Decreasing root temperatures from 25-13 degrees C increased the half-times of water exchange in a cell by a factor of 6-9. The measurement of axial water transport with a heat-balance sap-flow gauge showed that only a small amount of water was transported when 8 degrees C was imposed on cucumber roots. Lp and the H+-ATPase activity of the isolated root plasma membrane were very sensitive to externally applied hydrogen peroxide at a concentration of 1-16 mM. These observations suggest that the accumulation of hydrogen peroxide appears to mediate decreases in water transport in cucumber roots under low temperature.

Despite the high isoform multiplicity of aquaporins in plants, with 35 homologues including 13 plasma membrane intrinsic proteins (PIPs) in Arabidosis thaliana, the individual and integrated functions of aquaporins under various physiological conditions remain unclear. To better understand aquaporin functions in plants under various stress conditions, we examined transgenic Arabidopsis and tobacco plants that constitutively overexpress Arabidopsis PIP1;4 or PIP2;5 under various abiotic stress conditions. No significant differences in growth rates and water transport were found between the transgenic and wild-type plants when grown under favorable growth conditions. The transgenic plants overexpressing PIP1;4 or PIP2;5 displayed a rapid water loss under dehydration stress, which resulted in retarded germination and seedling growth under drought stress. In contrast, the transgenic plants overexpressing PIP1;4 or PIP2;5 showed enhanced water flow and facilitated germination under cold stress. The expression of several PIPs was noticeably affected by the overexpression of PIP1;4 or PIP2;5 in Arabidopsis under dehydration stress, suggesting that the expression of one aquaporin isoform influences the expression levels of other aquaporins under stress conditions. Taken together, our results demonstrate that overexpression of an aquaporin affects the expression of endogenous aquaporin genes and thereby impacts on seed germination, seedling growth, and stress responses of the plants under various stress conditions.

When root temperature dropped below 25 degrees C, there was a sharp drop in the root pressure (P(r)) and hydraulic conductivity of excised roots (Lp(r)) of young cucumber (Cucumis sativus L.) seedlings as measured with the root pressure probe. A detailed analysis of root hydraulics provided evidence for a larger reduction in the osmotic component of Lp(r) (77%) in comparison with the hydrostatic component (34%) in response to the exposure of the root system to 13 degrees C. The activity of the plasma membrane H(+)-ATPase (EC 3.6.1.35) was reduced from 30 to 16 micro mol Pi mg(-1) protein h(-1) upon exposure to 8 degrees C for 1 day. Ultrastructural observations showed no evidence of loosening of the microstructure of endodermal cell walls in low temperature (LT)-treated roots. It is concluded that the rapid drop in the P(r) in response to LT is largely caused by a reduction in the activity of the plasma membrane H(+)-ATPase rather than by loosening of the endodermal wall which would cause substantial solute losses. On the other hand, water permeability of root cell membrane at LT was related to changes in the activity (open/closed state) of water channels.

The effects of low root temperature on growth and root cell water transport were compared between wild-type Arabidopsis (Arabidopsis thaliana) and plants overexpressing plasma membrane intrinsic protein 1;4 (PIP1;4) and PIP2;5. Descending root temperature from 25°C to 10°C quickly reduced cell hydraulic conductivity (L p ) in wild-type plants but did not affect L p in plants overexpressing PIP1;4 and PIP2;5. Similarly, when the roots of wild-type plants were exposed to 10°C for 1 d, L p was lower compared with 25°C. However, there was no effect of low root temperature on L p in PIP1;4-and PIP2;5-overexpressing plants after 1 d of treatment. When the roots were exposed to 10°C for 5 d, L p was reduced in wild-type plants and in plants overexpressing PIP1;4, whereas there was still no effect in PIP2;5-overexpressing plants. These results suggest that the gating mechanism in PIP1;4 may be more sensitive to prolonged low temperature compared with PIP2;5. The reduction of L p at 10°C in roots of wild-type plants was partly restored to the preexposure level by 5 mM Ca(NO 3 ) 2 and protein phosphatase inhibitors (75 nM okadaic acid or 1 mM Na 3 VO 4 ), suggesting that aquaporin phosphorylation/dephosphorylation processes were involved in this response. The temperature sensitivity of cell water transport in roots was reflected by a reduction in shoot and root growth rates in the wild-type and PIP1;4-overexpressing plants exposed to 10°C root temperature for 5 d. However, low root temperature had no effect on growth in plants overexpressing PIP2;5. These results provide strong evidence for a link between growth at low root temperature and aquaporin-mediated root water transport in Arabidopsis.