Abscisic acid in leaves and roots of willow: significance for stomatal conductance.

Excised leaves and roots of willow (Salix dasyclados Wimm.) accumulated abscisic acid (ABA) in response to desiccation. The accumulation of ABA was greater in young leaves and roots than in old leaves and roots. In mature leaves, ABA accumulation was related to the severity and duration of the desiccation treatment. Water loss equal to 12% of initial fresh weight caused the ABA content of mature leaves to increase measurably within 30 min and to double in 2.5 h. The drying treatment caused significant (P = 0.05) reductions in leaf water potential and stomatal conductance. Recovery of leaf water potential to the control value occurred within 10 min of rewatering the dehydrated leaves, but recovery of stomatal conductance took an hour or longer, depending on the interval between dehydration and rewatering. The addition of ABA to the transpiration stream of well-watered excised leaves was sufficient to cause partial stomatal closure within 1 h and, depending on ABA concentration, more or less complete stomatal closure within 3 h. When the ABA solution was replaced with water, stomatal conductance increased at a rate inversely related to the concentration of the ABA solution with which the leaves had been supplied.

Stomatal and leaf growth responses to partial drying of root tips in willow.

Root tips of intact willow (Salix dasyclados Wimm., Clone 81-090) plants were partially dried by exposure to ambient greenhouse air and then kept in water-vapor-saturated air for up to 3 days. The drying treatment increased abscisic acid (ABA) concentrations in both the root tips subjected to drying and in the xylem sap, while it reduced leaf stomatal conductance and leaf extension rate. Despite the decrease in stomatal conductance, leaf water potentials were unaffected by the root drying treatment, indicating that the treatment reduced hydraulic conductivity between roots and foliage. After roots subjected to drying were returned to a nutrient solution or excised, ABA concentrations in the remaining roots and in the xylem sap, stomatal conductance of mature leaves and extension rate of unfolding leaves all returned to values observed in control plants. The 4-fold increase in xylem sap ABA concentration following the root drying treatment was not solely the result of reduced sap flow, and thus may be considered a potential cause, not merely a consequence, of the observed reduction in stomatal conductance.

Diurnal variation in leaf extension of Salix viminalis at two nitrogen supply rates.

To investigate how nitrogen supply might affect the biophysical factors controlling diurnal variation in leaf extension, pot-grown Salix viminalis L. were supplied with nitrogen at a low relative addition rate of 0.05 g N g(-1) N day(-1) (low N) or were given free access to all nutrients (high N). Leaf extension, turgor pressure, turgor after stress relaxation and the plastic extensibility of leaf tissue were determined for growing leaves every 4 h during two days of clear skies in August. Plants in the high-N treatment had a significantly higher relative growth rate, dry weight, shoot/root ratio, leaf nitrogen concentration, total leaf area, final area of single leaves and epidermal cell size than plants in the low-N treatment. The periodicity of leaf extension was similar in both treatments with high values during the afternoon and early evening, and negligible values during the night and in the early morning. The maximum rate of leaf extension was higher in high-N than in low-N plants. Leaf water potential and leaf osmotic potential decreased in the morning and increased in the afternoon with highest values during the night. Calculated values of turgor pressure showed no consistent diurnal trend and did not correlate with the rate of leaf extension. There was no consistent difference in turgor between treatments. Turgor after stress relaxation varied diurnally. The difference between turgors before and after stress relaxation also varied diurnally and was largely in phase with the diurnal pattern of leaf extension. These data are consistent with either a causal role for growth turgor (difference between turgors before and after stress relaxation) in the regulation of cell expansion, or a diurnal variation in turgors after relaxation, attributable to different capacities for cell wall loosening at different times of day. Plastic extensibility of leaf tissue showed no diurnal pattern but consistently higher values were found in high-N than in low-N plants. We conclude that the effects of nitrogen supply on leaf water relations did not limit leaf extension, but that nitrogen supply did affect processes associated with cell wall loosening and enlargement. Nitrogen supply did not affect final values of turgor after relaxation, but it presumably affected the rate at which relaxation proceeded.

Diurnal pattern of leaf extension in Salix viminalis relates to the difference in leaf turgor before and after stress relaxation.

Diurnal variations in leaf extension and turgor relaxation were measured in young willow leaves (Salix viminalis L.). Turgor relaxation was calculated from measurements of xylem water potential and osmotic potential of whole, detached leaves that were incubated, without loss of water, under controlled conditions. There were clear diurnal variations in leaf extension and in leaf turgor relaxation and the diurnal pattern of the difference in leaf turgor before and after relaxation was in phase with that of leaf extension. The extent to which leaf turgor after relaxation may be related to threshold turgors for growth and the possible importance of leaf turgor during leaf extension are discussed.

Diurnal variation in extension growth of leaves of Salix viminalis.

Diurnal variation in leaf extension and biophysical parameters of leaf growth were measured in young leaves from a stand of Salix viminalis L. in southern Sweden over a two-day period of clear skies during late July. Leaf growth rate (irreversible extension) was greatest during the late afternoon and early evening, falling to negligible values during the night and early morning.Leaf water potential and leaf osmotic potential showed declining values in the morning with subsequent recovery in the late afternoon. Diurnal variation in osmotic potential (-1.3 to -1.7 MPa) was small compared with that of leaf water potential (-0.1 to -1.2 MPa). Calculated values of leaf turgor pressure during the night (1.2 MPa) were double the midday values. Growth rate correlated poorly with turgor, which (except on one occasion) was always above a calculated value of yield turgor at 0.53 MPa.Diurnal variation in extension growth rate was large compared with that in plastic extensibility of leaf tissue as measured by an Instron technique. Values of extensibility were low and showed little diurnal variation, which is consistent with a proposed negative feedback of expansive growth rate on extensibility. Extension growth rate correlated well with air temperature, suggesting that the rate of leaf expansion may have been limited by a temperature-dependent rate of cell wall loosening.

In vitro cell-mediated immune reactions of melanoma and colorectal carcinoma patients demonstrated by long-term 51chromium assays.

In a series of consecutive 51chromium (51Cr) assays, 21 melanoma and 14 colo-rectal carcinoma patients were tested for their in vitro cell-mediated immune reactions to melanoma, rectal adenocarcinoma, and normal fibroblast target cells. Blood lymphocytes (BL) from four individuals were included in each experiment. In 9/14 experiments the BL effects of 2 melanoma patients were compared to BL effects of 2 colorectal carcinoma patients. In two experiments, BL from 1 melanoma patient and 1 colorectal carcinoma patient were compared for cytotoxic effect with each other and with BL from 2 normal healthy donors, and in the remaining 3 experiments BL from 2 melanoma patients were compared with BL from 1 healthy donor and one patient bearing a tumor which was neither a melanoma nor a colorectal carcinoma. Eleven out of 14 experiments were performed in a criss-cross manner. Target cells in the first three tests of this series consisted of tumor cells and fibroblasts explanted from a single melanoma patient. In all of the remaining experiments, each BL population was tested for cytotoxicity against both a tumor-fibroblast target cell pair explanted from one of two melanoma patients and a tumor-fibroblast target cell pair explanted from a rectal adenocarcinoma patient. Out of 35 tumor patients, 27 (77%) demonstrated a selective cytotoxic effect on the tumor target cells compared to the corresponding fibroblasts, while 4 patients (11%) showed a selective effect on the fibroblasts, and 6/29 patients showed a selective effect on the other type of tumor cells compared to matching fibroblasts. In 8/11 experiments (including two repeat tests) tumor-specific BL effects were demonstrated in a criss-cross manner. BL separations and 51Cr tests were repeated in 11 of the 35 patients 2-4 weeks after their original tests. BL populations from these 11 patients reproduced their individual earlier effects, whether these effects showed specific, non-specific, or no cytotoxicity. In each assay, differences in sensitivity between fibroblasts and tumor target cells in matched pairs were analyzed by comparing the effects of BL from the two controls. No differences in target-cell sensitivity could be demonstrated.

A long-term 51chromium assay for in vitro cell-mediated tumor immunity. Correlation with simultaneously performed microplate assays.

In vitro assays of cell-mediated tumor immunity utilizing 51Chromium (51Cr) labelling of cultured adherent solid tumor cells were designed which allowed an effector cell/target cell incubation time of 48 h without overriding spontaneous 51Cr release. In a series of 16 consecutive experiments, blood lymphocytes from healthy human donors, from patients with tumors unrelated to the cultural tumor target cells, and from colon carcinoma and melanoma patients were tested for their cytotoxic effects on various target cell pairs, human colon carcinoma, melanoma, or skin fibroblasts. The same reagents were used in simultaneously performed microplate and 51Cr assays. Results obtained by visual counting of microplate tests and by 24-h assays of 51Cr release or 51Cr retention correlated in 20/25 effector-cell/target-cell combinations. In a series of six consecutive experiments, lymph-node cells from untreated Wistar/Furth rats, and rats bearing either chemically-induced colon carcinoma NG-W1 or polyoma virus-induced sarcoma P-W13 were tested for their cytotoxicity on syngeneic rat colon carcinoma and sarcoma target cells. Criss-cross type experiments were performed by microplate and 15Cr techniques done in parallel. Results obtained by visual counting of microplate tests and by 48 h assays of 51Cr release or 51Cr retention correlated in 15/18 effector-cell/target-cell combinations.