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The concentration of salt ions accumulated in the leaves was determined from analyses of leaves sampled from the previous flush of growth. These leaves were identified as originating from lignifying stem segments occurring directly behind the youngest, light green leaves on solid green stems. Consistency of tissue maturity has been shown to be an important characteristic for obtaining comparable results (Mills and Jones 1996). Leaf tissue-sampling events occurred on 17 October 2005, 9 January 2006, 18 May 2006, 22 September 2006 and 15 January 2007. The experiment was terminated shortly after the fifth sampling. Both proximal (P) and distal (D) leaf blade sections were collected on each date. The distal portions of leaves were removed GW3965 manufacturer first; the halfway cut point was determined visually. Then the Target Selective Inhibitor Library basal sections of the cut leaves were removed by cutting them as closely to the stem as possible. A minimum of 1.5 g dry weight (3.8 g fresh weight, 39 % dry: fresh weight ratio) was collected for each sample. The dried samples were analysed for % Ca+, % Cl? and % Na+ by using the ��Nitric/Perchloric Wet Ashing Open Vessel�� (P �C 3.10) technique, and Cl was analysed using the ��2 % Acetic Acid Extraction�� (P �C 4.20) technique by Dellavalle? Laboratory, Inc. (Fresno, CA, USA). Ion accumulation rates were evaluated for the different concentrations within each salinity treatment type (e.g. 1.0 dS m?1 NaCl vs. 6.0 dS m?1 NaCl), as well as within treatment concentration level between the various salinity treatment types (e.g. 1.0 dS m?1 NaCl vs. 1.0 dS m?1 CaCl2). Statistical analysis The experimental design represented a randomized complete block (RCB) with every treatment appearing in either block. Blocking of experimental groups in the greenhouse was used to control for variances that may have originated from the environmental conditions. The treatment and block effects were treated as random variables. Analysis of variance (ANOVA) tests were used to evaluate if differences in stem (height) growth and stem diameter growth could be confidently attributed (P Adenine type, a linear regression was used to determine the influence increasing salinity concentrations (EC) had on height and diameter growth responses (Fig.?2). Tukey's HSD were used as a post-hoc pairwise comparison of means within treatments when statistical differences were attributable to the treatment variables (salt or concentration). Relative comparisons of the mean ion concentrations, collected in proximal and distal tissues, were used to understand the differences attributable to salinity type and salinity concentration (Figs?3?3�C5). Figure?2. (A) Irrespective of salt ion the relative diameter growth was inversely proportional to the increase in salinity (R2 = 0.
