Barley grain cultivars were obtained from the breeding program of seeds station, Beni-Suef, Egypt. Barley grains were surface sterilized by immersion in a mixture of ethanol 96% and H₂O₂ (1:1) for 3 minutes, followed by several washings with sterile distilled water, seeds were grown in 1 kg pots in Botany and Microbiology Department garden. Barley plant was considered as one of the most important crop plants in Egypt because of its contribution as nutrient foods for people. Barley is a major cereal grain commonly found in bread, beverages and various cuisines of every culture. It remains one of the most widely consumed grains, globally, source of dietary. Fiber, vitamins and minerals are not found in refined and enriched grains. Five seeds were sown in each pot and soil was brought to field capacity. The seedlings were left to grow under the desired soil moisture content levels (90%, 70%, 50% and 30%). Soil moisture content was measured by calculate the soil field capacity, this consider as 100% moisture content and so could be determine the other lower soil moisture content. The clay soil comprise four components minerals and soil organic matter make up the solid fraction, whereas air and water comprise the pore space fraction. A typical agricultural soil is usually around 50% solid particles and 50% pores (Adapted from Brady and Weil, 2002) [16]. Soil particle of clay is <0.002 invisible to naked eye. Considerations of working in controlled environments were followed by Tibbitts & Langhans [17].

The previous treatments were repeated for treatment with spraying vegetative parts with 0.5 mM SA as second groups. Three replicate was made for each treatment and plants were grown in natural conditions for crop yield production at 120-days.

At the end of the experimental period (120 days) plant height, dry matter yield of the different organs (shoot, root and spike) were determined. Plant height was determined by direct measurement from soil surface to the tip of the flag leaf. Determination of the dry matter involved harvesting and careful separation of fresh organs. Fresh organs were then dried in an oven at 80˚C. Successive weighing was carried out until a constant dry weight was recorded. The plants were uprooted, roots carefully separated from the soil, washed and the length of roots were measured. From determining the shoot and root weight, was calculated. Leaf area was determined by measuring leaf length and maximum width and applying the formula; Leaf area = k (leaf length * leaf maximum width) Cm² plant⁻¹. This formula provided a simple way for determination of leaf area particularly in the field where large leaves had to be measured. The coefficient k was calculated and assigned different values for different grasses [18] [19] , and recently reviewed and given a value of 0.75 for maize [20]. Dry matter was determined after drying plants in an aerated oven at 70˚C to constant mass. Pigment content was measured by Metzner [21]. Saccharides were determined by the anthrone-sulfuric acids method [22]. Soluble protein was measured according to Lowry et al. (1951) [23]. Amino acids and proline were measured by Moore and Stein (1948) and Bates et al. (1973) [24] [25]. Sodium and potassium were determined by flam photometric method [26] , and calcium and magnesium by the versene titration method [27].

The experimental data were subjected to the one way analysis of variances (ANOVA test) using the SPSS version 11.0 to quantify and evaluate the source of variation and the means were separated by the least significant differences, L. S. D. at P level of 0.05% [28]. Experimental data were subjected to one way analysis of variance and the means were separated by the least significant differences, L. S. D. [28]. Correlation coefficients were calculated using statgraphics 5.0 software.

In cv. Giza 123 decreasing moisture content induced three position the firstly was more or less have the same value of control 100% in shoot fresh matter, secondly a marked increase in shoot dry matter was detected up to 50% M. C., thirdly in root there is a slight decreasing effect in fresh and dry matter up to 70% (Table 1(a)). After that level, a significant reduction in fresh and dry matter was induced reach a low level at 30% M. C., the percent of reduction was 84% and 72.4%. Length of both organs showed a decreasing effect reach a 20% at 30% M. C. compared with control plants (Table 1(a)). Fresh and dry matter of shoot and root of Giza 2000 generally become more or less unchanged up to 70%, after that a marked reduction was recorded (Table 1(b)). The percent of reduction at 30% M. C. was 69.3%, 81.7%, 70.5%, 67.2% in fresh and dry matter of shoot and root respectively. Length in both organs was decreased with decreasing moisture content. The percent of reduction was 58.8% and 75% in shoot and root compared with control plants (Table 1(b)). A lowering effect was recorded with decreasing moisture content in fresh and dry matter of shoot, root and length of cv. Giza 124, this effect was more obvious in root than in shoot (Table 1(c)). The percent of reduction in fresh, dry matter and length at 30% M. C. was 63.9%, 63.4%, 43.7%, 66.6%, 65.4% and 72%. Also decreasing soil moisture content induced a significant reduction in fresh, dry matter and length of Giza 129 from 70% to 30% M. C. The percent of reduction was 53.2%, 51.4%, 39%, 34.7%, 51.9% and 48.4% in fresh, dry and length of shoot and root at 30% M. C. (Table 1(d)). Leaf area smoothly decreased at all M. C. levels in cv. Giza 123 moreover a marked reduction was recorded at 50% and 30% in both Giza 2000 and Giza 124, however a significant reduction was recorded in cv. Giza 129. The percent of reduction at 30% M. C. was 81.8%, 60.7%, 54.9%, 53.3% as compared with control plants (Figure 1). Chlorophyll a, b and carotenoids were significantly decreased as decreasing drought stress in four barley genotypes compared with reference control plants (Figure 2(a) & Figure 2(b)). The percent of reduction in Chl. a, b and c at 30% M. C. level was 80.3%, 98.9%, 67.4%, in Giza 132 plant,

87.6%, 58.3%, 98.1 in Giza 2000 plant, 54.3%, 50%, 88.6%, in Giza 124 plant, 54.1%, 38.9%, 74.1% in Giza 129 plant. Water content was reduced in shoot and root as decreasing M. C. levels in four tested genotypes, and the low values were recorded at 30% M. C. level (Table 2). The highest value in water content was recorded in Giza 123 while the lower one was observed in Giza 129. The percent of reduction at 30% M. C. level was 97.9%, 85.3%, 66.7%, 71.4%, 85.8%, 75%, 39.2% and 39.3% in Giza 124, Giza 2000, Giza 124 and Giza 129 respectively (Table 2).

Soluble sugar and soluble protein significantly accumulated as decreasing moisture content in both shoot and root of Giza 123 (Table 3(a)). This more obvious in case of soluble sugar in root than in shoot and in case of soluble protein in shoot than in root. The high value was recorded at 30% M. C. in both contents, the percent of increase at that level was 114%, 160%, 150%, and 200% compared with control pants. However amino acids become around the value of control in

shoot at all moisture content levels, in root this effect was observed at 70% M. C. level, afterthat a smooth reduction was recorded in Giza 123 (Table 3(a)). There is a significant reduction in proline content in shoot, whereas a huge accumulation

was induced in root Giza 123 plant reach 3-folds at 30% M. C. levels (Table 3(a)). Decreasing moisture content induced no significant change in soluble sugar in Giza 2000 till 50% M. C. level and then a slight increase was recorded in both shoot and root (Table 3(b)). However a significant accumulation in soluble protein was detected in shoot and root. The maximum percent value was recorded at 30% M. C. level reach a 150% and 206% compared with control plants. While decreasing moisture content induced an increasing effect in amino acids content in shoot while a slight decreasing effect was induced in root (Table 3(b)). There is a two situation in case of proline content, a decreasing effect in shoot was induced and in root an increasing effect was exhibited (Table 3(b)). While a marked decreasing effect was occurred in soluble sugar in shoot and root of cv. Giza 124, an increasing effect was induced in soluble protein of both organs reach a high percent value at 30% M. C. level 186% and 265% (Table 3(c)). Amino acids give the value of control up to 50% M. C. level, then a smooth increasing effect in shoot and a reduction in root was recorded (Table 3(c)). A surprise position was detected in proline content decreasing effect was observed in shoot and root of Giza 124 plant (Table 3(c)). The data in Table 3(d) revealed that soluble sugar in shoot and root of Giza 129 was significantly decreased as decreasing moisture content reach a low value at 30% M. C. level. This reduction was more pronounced in root than shoot organ. Soluble protein and amino acids were markedly elevated with decreasing moisture content. The percent of increase at 30% M. C. level was 162%, 139%, 159% and 125% of both content in shoot and root compared with control plants control. Proline content was markedly accumulated as decreasing soil moisture content in shoot and root of Giza 129, reach a high value at 30% M. C. level 2-folds compared with control plants (Table 3(d)). K⁺, Ca⁺⁺, and Mg⁺⁺ were generally remain unchanged in shoot and root of both barley Giza 123 and Giza 2000 (Figure 3(a)). Except of this trend root K⁺⁺ and Ca⁺⁺ in root of Giza 2000 and Ca⁺⁺ in both organs of Giza 123 tended to increase as decreasing M. C. level. K⁺ content tended to decrease in both shoot and root of Giza 124 (Figure 3(b)). While Ca⁺⁺ content decreased in shoot, increase in root of Giza 124, moreover Mg⁺⁺ do not change with increasing drought stress in shoot while tended to increase in root (Figure 3(b)). In Giza 129 K⁺ content was lowered as decreasing M. C. level while Ca⁺⁺ was increased in shoot, in root this content remain unchanged (Figure 3(b)). Mg⁺⁺ content tended to decreased in shoot while tended to increase in root.

Soluble sugar, soluble protein, amino acids and proline tended to smoothly increase especially at 50% and 70% M. C. in spikes of Giza 123 (Table 4(a)). In spikes of Giza 2000 drought stress increased soluble sugar and soluble protein while amino acids tended to exhibit an irregular pattern and a reduction in proline content was recorded (Table 4(a)). A slight accumulation in soluble sugar, soluble protein and amino acids in spikes of Giza 124 (Table 4(b)) was induced. Moreover a hug increases in soluble sugar and proline content was recorded in spikes of Giza 129 (Table 4(b)). The activation was reached 2.5-folds in soluble sugar and 8-folds in proline content in spikes of Giza 129 (Table 4(b)). On the other side a small increase in soluble protein and amino acids in spikes of Giza 124 was induced. In spikes of Giza 123 both K⁺ and Mg⁺⁺ were markedly increased as increasing drought stress, reached 2-folds at 50% M. C. level in case of K⁺ and at 30% M. C. level in case of Mg⁺⁺ (Table 5(a)). While Ca⁺⁺ increased at 70% M. C., decreased at 50% and 30% M. C. levels compared with control plants (Table 5(a)). Ca⁺⁺ and Mg⁺⁺ were significantly increased in Giza 2000 especially at 70% and 50% M. C. and gave the same values, however K⁺ was smoothly decreased at 70% and 50% M. C. levels and dramatically reduced at 30% M. C. level (Table 4(a)). Drought stress induced reduction in K⁺, Ca⁺⁺ in both Giza 124 and Giza 129 and Mg⁺⁺ in Giza 124 especially at lower moisture content (50% and 30% M. C.) (Table 5(b)). Except of this trend Mg⁺⁺ in Giza 129 significantly increased compared with control plant (Table 5(b)). Osmotic pressure was increased in shoot, root and spike of four barley tested genotypes with varied degree, moreover root organ was the lowest values than shoot and spike organs (Figures 4(a)-(d)). The percent of increase at 30% M. C. was 132.6%, 117.7% and 115.5% in shoot, root and spike of Giza 123, in Giza 2000 it was 136.4%, 126.3%, 127.8%, in Giza 124 it was 134.5%, 105.5%, 106.3% and finally in Giza 129 this percent was 135.2%, 113.0% and 139.9% compared with control plants.

SA application was markedly enhanced the production of fresh, dry mater, length of shoot and root with varied degree according to each tested barley genotype (Tables 1(a)-(d)). The medium percent of different moisture content levels in fresh, dry matter and length of shoot and root after spraying with 0.5 mM

SA was 109.5%, 132.3%, 129.9%, 120.3%, 96.3%, 97.3%, in Giza 123. In Giza 2000 this percent of increase was 142.5, 136.7, 112.9%, 103.9%, 90.3%, 85.6%. However in Giza 124 it was 101.6%, 100.7%, 102.4%, 99.9%, 94.4%, 105.5%, and in Giza 129 this increasing percent was 99.2%, 87.9%, 102.2%, 80.5%, 121.1%,

114.9% compared with unsprayed barley plants. SA increased water content and leaf area at all tested barley cultivars, this effect was more pronounced in Giza 129, Giza 2000, and Giza 124 than Giza 123 especially in leaf area parameter (Table 2 and Figure 1). SA application enhanced the synthesis of photosynthetic pigments in four barley genotypes (Figure 2(a) & Figure 2(b)). This activation effect was more obvious in case of Chl. a in Giza 123 and Giza 2000 (Figure 2(a)) and in case of production Chl. a and Chl.b in both Giza 124 and Giza 129 (Figure 2(b)). Spraying vegetative parts with SA was markedly increased the soluble sugar, soluble protein and amino acids of both shoot and root of four barley cultivars. The medium percent of increase of the previous parameters in plants spraying with SA in Giza 123 was 146.5%, 162.3%, 146.5%, 157%, 125%, 115.3%, was in shoot and root (Table 3(a)). This value in Giza 2000 was 136.8%, 114%, 129.5%, 142.8%, 125.5%, 112.8%, (Table 3(b)). In Giza124 it was 90%, 124.5%, 213%, 176.8%, 113.8%, 114.5% (Table 3(c)). In Giza 129 it was 115.3%, 72.5%, 191%, 151.8%, 143.3%, 127.3% compared with control plants (Table 3(d)). On the other side SA application lowered the accumulation of proline in shoot and root of four barley genotypes (Tables 3(a)-(d)). SA application induced unchanged in soluble sugar, a slight accumulation in soluble protein and amino acids, while a reduction in proline in spikes of Giza 123 was recorded (Table 4(a)). Activation in the accumulation of soluble sugar and amino acids especially at 30% M. C. and a reduction in proline was recorded in spikes of Giza 2000 (Table 4(a)). SA treatment increased soluble sugar, unchanged in amino acids content while a reduction was induced in proline in spikes of Giza 124 (Table 4(b)). SA activated the accumulation of following contents, soluble sugar reached 3-folds, protein and proline this with unchanged effet in amino acids as compared with 90% M. C. (Table 4(b)). While spraying vegetative parts with SA induced no significant change in K⁺, Ca⁺⁺, and Mg⁺⁺ in shoot and root of Giza 123. While significantly increased K⁺, Ca⁺⁺ and Mg⁺⁺ in shoot and root of Giza 2000 was reorded (Table 5(a) & Table 5(b)). SA application enhanced accumulation of K⁺, Ca⁺⁺ in both shoot and root of Giza 124 and K⁺, Ca⁺⁺ and Mg⁺⁺ in Giza 129. SA increased K⁺ reached 3-folds at 50% M. C. level. Ca⁺⁺ reached 2-folds at 30% M. C. and Mg⁺⁺ in spikes of Giza 123 (Table 5(a)). SA application resulted unchanged in K⁺, Ca⁺⁺ and Mg⁺⁺ in both Giza 124 and Giza 129 (Table 5(b)). Except of this position Mg⁺⁺ increased as decreasing M. C. in spikes of Giza 129, the high value was recorded at 30% M. C. (Table 5(b)). SA treatment was activated the value of OP in four tested barley genotypes, this effect was more pronounced in shoot and spike of Giza 129 than other organs of other genotypes (Figures 4(a)-(d)). The percent of activation at lowest value of moisture content (30% M. C.) was 144.2%, 137.9%, 119.4% in shoot, root and spike of Giza 123. In Giza 2000, it was 144.7%, 170.6%, 146.3%, in Giza 124 it was 151.4%, 131.2%, 113.4 and finally in Giza 129 this percent was 177.9%, 125.3% and 156.9% as compared with control plants.