الفهرس 
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Abstract
This study was carried out for the two successive seasons of 2002 and 2003 at the field of horticulture research institute, Mallawy Agricultural Research Station, Minia, Egypt to study the effect of some antioxidants on chemical constituents,antinutritional factors and yield of three vegetable legumes : phaseolus ( variety Nibrasca), pea ( variety Master B) and broad bean (variety Nubaria 1).
In the meantime, antioxidants become very important, so they were used as substitutes for synthetic auxins that causing a lot of damage to organisms. Antioxidant substances are very important in relation to disease resistance and must be known the influence of antioxidants on chemical constituents, antinutritional factors and yield of the vegetable legumes .These crops are very important as a good source of carbohydrates, mineral elements and higher level of proteins. Therefore, we should choice these vegetable legumes and treated with antioxidants to investigate the effect of them on chemical constituents, antinutritional factors and yield of the legume crops.
Plan of study:-
The first: - materials, concentrations and soaking seeds
period.
The used materials were :Salicylic acid ( SA ), citric aid ( SA ) and ascorbic acid (AA) at concentrations of 0.1,0.5,1.0,2.5and 5.0 mM.
The suitable period of soaking was half hour, 12 hour and 24 hour for phaseolus, pea and broad bean, respectively.
The second: the treatments:
1- The seeds were soaked in different concentrations of antioxidants at the suitable soaking period to each crop then the seeds were sown at the suitable time.
2- The seeds were sown then spraying the plants with antioxidants three times.
3- The seeds were soaked then spraying the plants with antioxidants times.
4- The control: the seeds and the plants were treated with water at the same condition for all treatments.
The third: Experimental design and the statically
analysis:-
A complete randomized design with three replicates was followed in this experiment. The obtained data were statically analyzed according to Duncan’s rang test. The cropping measurements and chemical constituents which obtained in this study were presented as average for two seasons as follow:-
I. the cropping measurements:-
1- Plant height ( cm ) .
2- Dry pod length ( cm ) .
3- Number of pools / plant.
4- Number of dry seeds / pod.
5- Weight of 1000 dry seeds g / plant.
6- Weight of dry pods g / plant.
7- Weight of dry seeds g / plant.
II. Chemical constituents:-
The proximate analysis were carried out for all treatments in dry seeds then they were statically analyzed and found that the best treatment was soaking + spraying treatment therefore, the other determinations were carried out at this treatment as follow :
1- Determination of protein fractionation (Albumins, goblins, prolamin and glutelins).
2- Determination of free amino acids.
3- Determination of ascorbic acid .
4- Determination of total soluble sugars .
5- Determination of total reducing sugars.
6- Determination of total non reducing sugars.
7- Determination of some antinutritional factors: -
I-Raffinose family sugars (Raffinose, stachyose and verbascose).
II-Trypsin inhibitor (T1).
III-Phytic acid.
IV-Total phenolic compounds.
From the obtained results it could be summarized as follow:-
1- By increasing the concentration of antioxidants up to 1 mM, the plant height was gradual increased then gradual decrease by increasing the concentration in comparison with seeds from untreated plants. Also it is noticed that the soaking + spraying treatment by using of citric acid gave the highest values which were 41.1, 60.1, and 140.0 cm. for phaseolus, pea and broad bean, respectively compared to the control ( 34.5, 53.0 and 122.0 cm ) , respectively.
2- The highest gradual increase in pod length of phaseolus and pea from 13.5, 9.4cm to 15.3, 10.62 cm respectively, was observed when application of soaking + spraying treatment up to 1 mM by using citric acid. While, the maximum values in broad bean were 13.4 to 15.3 ,15.1 and 15.0 cm by application citric acid at 1mM and 2.5 mM and ascorbic at 1mM with soaking + spraying treatment, respectively .
3- The highest increase in number of dry phaseolus pods / plant was recorded from 13.9 to 17.83 on citric acid. soaking + spraying treatment at 1mM while, the highest increase in corresponding values pea pods / plant and broad bean / plant were recorded on both SA and CA soaking + spraying treatment at 1.0 mM from 12.7, 12.8 to 18.7, 16.3 respectively.
4- A gradual increase in number of seeds / pod due to the treatment with ascorbic acid up to 5.0 mM. for the spraying treatment was noticed for all crops .The best results were recorded concerning the number of seeds / pod as a result of treating the plants with citric acid at 1mM ( soaking + spraying) for phaseolus and broad bean ( 5.3 and 5.2 ) compared to control ( 3.9 and 4.3 ) respectively, but it was found that salicylic acid at 1 mM. the most effective treatment for the number of pea seeds/pod (10.21) compared with control (9.0).
5- It is noticed that, there was an increase in weight of 1000 seed as a result of treating the plants with salicylic acids up to 1 mM ( soaking + spraying ) for phaseolus and broad Bean which produced the maximum weight of 1000 seed (589.7g;1610g) compared to the control (501.0g;1100g) respectively , whereas the weight of 1000 seed was increased in pea by 1.43 fold in comparison with untreated plants when using of citric acid at 1mM on soaking + spraying treatment. it seems that there was a gradual decrease in weight of 1000 seed when plants where treated whit salicylic acid and citric acid at 2.5 – 5.0 mM of all treatments for three crops while , there was a gradual increase in weight of 1000 seed for three crops by increasing ascorbic acid concentration up to 5.0 mM on spraying treatment
6- By increasing ascorbic acid concentration the weight of pods plan was increased on spraying treatment of the three legumes up to 5.0mM. Furthermore the highest values of pod Yield / plan were 1.58, 1.60 fold compared to the control for phaseolus and brood bean , respectively when treating the plants with 1mM citric acid, On soaking + spraying treatment by, salicylic acid at 1Mm. Gave the best result for the pea yield which recorded 2.05 fold comparing to control
7- Antioxidant treatments caused a considerable increase in weight of seeds / plant. The most effective. Treatment were 1mM salicylic acid with soaking + spraying treatment of pea and broad bean by 2.10 and 2.08 fold , respective compared to untreated ones soaking +spraying treatment with 1mM citric acid recorded the maximum value of seed phase us weight / plant of 2.01 compared with control .
In general it was noticed a gradual increase in seed yield of the three crops by increasing concentration of ascorbic acid with spraying treatment
The proximate analysis showed that:
1- A gradual reduction in moisture percentage and the highest reduction were 0.86, 0.91, 0.91 fold in phaseolus , pea and brood bean respectively , compared with control up to 1mM salicylic acid on soaking + spraying treatment then gradual increase in moisture percentage when treating the three plants seeds with salicylic acid at 2.5 – 5.0 mM
2- The highest gradual increase in fat percentage in phaseolus and pea due to treating with salicylic acid at 1mM (Soaking + spraying) 4.33 , 5.66 % , respectively, Also, in brood bean, the highest increases was from 2.03 to 4.22, 4.33 % on soaking + spraying treatment with salicylic acid and citric acid at 1 mM., respectively
3- Antioxidant treatments caused gradual decrease in fiber percentage for all treatment up to 1mM followed by there a gradual increase by increasing the Concentration the highest decrease was recorded by ascorbic acid soaking + spraying treatments of 0.81, 0.69 fold compared with control in phrasal us and broad bean, respectively and on treatments with salicylic acid at 2.5 mM., pea seeds recorded the highest decrease by 0.68 fold in compared with untreated ones.
4- The data showed a gradual increase in ash percentage for the three crops. Phaseolus seeds showed a highest increase on SA and AA soaking +spraying treatments at 1mm by 1.20, 1.22 folds, respectively from control. Also, the highest increase in ash was recorded for pea by treating the plants with salicylic acid soaking + spraying at 1.0mm and 2-5mm from 3.21 to 4.72 % , respectively. Broad bean seeds recorded maximum increase from 3.85 to 5.18% results from using salicylic acid at 1mm with soaking + spraying treatment.
5- Antioxidants treatment caused an increase in protein percentage, by increasing ascorbic acid concentration until 2.5 mM, the protein percentage was increased by soaking spraying treatment for the three legumes. The protein content of phaseolus and pea was marked increase from 29.0, 23.62% to 32.95, 26.45% respectively on soaking + spraying treatment with salicylic acid at 1mm. whereas, the highest increase in protein percentage of broad bean was observed from 25% to 27.85% when using that treatment with ascorbic acid at 1mM.
6- The obtained results showed that a gradual decrease in total carbohydrates percentage by increasing the concentration of all treatments until 1mM followed by a gradual increase by increasing the concentration up to 5.0mM for the three crops.
Higher increase was noticed on soaking + spraying treatment with citric acid at 5mM for phaseolus and pea, recording from 60.49 ,65.34 to 69.73, 69.67% ,respectively while, the increase in broad bean was 62.74 to 67.91, 67.90% by using citric acid at 5mm with spraying treatment and soaking one , respectively.
With respect to the effect of treatment with SA, AA and CA on some antinutritional factors, and some selected chemical constituents of tested legumes, the obtained results could be summarized as follows: -
1- Treating of pea and broad bean seeds by salicylic acid. at 1mm caused a highest gradual increase in Albumins percentage by 29.7, 17.5%, respectively compared to control and using of ascorbic acid at 1mM showed a highest increase in albumins percentage in phaseolus by 5.45%, in comparison with untreated one. Results showed gradual decrease in albumins percentage by increasing the concentration up to 1mM for all treatments in the three crops.
2- Salicylic acid at 1mM caused a highest gradual increase in globulins percentage of pea and broad bean by 6.0%, 5.2%, respectively compared to control, while the highest increase in globulins percentage of phaseolus was recorded when treating with 1m M ascorbic acid by 10.5% in comparison to untreated ones.
3- Using of antioxidants on phaseolus and brood bean caused a gradual increase in prolamines content up to 1 mM then gradual decrease by increasing the concentration .In contrast the prolamines in pea percentage were gradually decreased by increasing the concentration of antioxidant . The higher increase of prolamines content by salicylic acid at 1mM was 36.8 20.5 % in phaseolus and broad bean, respectively compared to control.
4- Obtained results indicated that the highest increase in Glutilines percentage was obtained when using salicylic acid at 1mM in pea and brood bean by 2.11, 2.53 Fold, respectively compared to control while, the highest increase in Glutilines Percentage was recorded in phaseolus seeds on treatment with citric acid at 1mM by 1.39 fold compared to control. Also the data revealed that the Glutilines content was increased as the concentration increased up to 1mMfor phaseolus and broad bean, followed by decreasing up to 5.0 m M compared to control .But pea seeds showed a decreasing after 1mM but not less than control.
5- Application of antioxidant treatment proved to be effective in decreasing in free amino acids by increasing the concentration of it up to 1Mm. The gradual increase when using 2.5 and 5.0 mM antioxidants. The maximum value of free amino acids content when treating of phaseolus, pea and broad bean with 5mM citric acid reach 1.42, 1.28 and 1.35 fold, respectively compared to control.
6- Increasing the concentration of ascorbic acid treatment increased vitamin C content of seeds. The most effective treatments were obtained from treating phaseolus, pea and broad bean with ascorbic acid at 5mM where vitamin C reached to maximum percentage of 188.8 , 109.1 and 149.0 %, respectively compared with control.
7- The method of soaking + spraying by application of antioxidant treatments proved to be effective in gradual increase in total soluble sugars percentage up to 1mM then a gradual decrease up to 5mM for all crops compared to untreated control for phaseolus pea and brood bean, respectively.
8- There is a gradual increase in total reducing sugars percentage up to 1mM for the three crops and that the highest values were recorded due to ascorbic acid applied for phaseolus and brood bean from 4.51 %, 5.22 % to 5.78 % 6.95 % respectively while, the highest increase in total reducing sugars percentage of pea from 4.12 % to 5.62 %, 5.72 % when using of salicylic acid and citric acid, respectively.
9- Ascorbic acid at 0.5 mM Gove the best result concerning in total non reducing sugars percentage for phaseolus and pea by 1.17, 1.18 fold respectively compared to control mean while, the highest increase in total non reducing sugars percentage of brood bean as a result of ascorbic acid at 0.1 mM by 1.07 fold comparing with untreated plants .
10- It is noticed that the highest increase in raffinose percentage was found when using citric acid at 1mM by 105.0, 60.0% for phaseolus and pea respectively in comparison with the check treatment. the highest increase in Raffinose percentage was recorded when treating the brood bean plants with ascorbic acid at 1mM by 57.01 over the control hen by increasing the concentration up to 1mM of it the raffinose percentage was decreased.
11- Treating phaseolus, pea and brood bean with ascorbic acid at 1mM proved caused in a gradual increase in stachyose percentage from 1.60, 2.32 and 2.20 % in control to reach 1.99, 3.08 and 2.83 %, respectively. It is clear that stachyose percentage as decreased when plants were treaded with antioxidants up to 1mM , as well as 5 mM antioxidants have a little value by increasing of antioxidants concentration up to 5.0mM.
12- Verbascose sugar percentage of phaseolus was recorded as result treating the plants with salicylic acid and ascorbic acid (1mM each) by 1.13 fold of then compared to control plants. The highest increase in verbascose sugar percentage of pea was noticed when using salicylic acid at 1mM by 2.29 fold in comparison with the check treatment. Meanwhile, in broad bean plant the highest increase in verbascose percentage when these plants were treated with all antioxidants at 0.5 and 1mM. The highest values were recorded as a result of treating the brood bean plants with citric acid at 1mM by 1.31 fold compared to control. On the other hand, the lowest values were found on antioxidant treatment at 2.5 and 5.0 mM for the three crops.
13- Trypsin inhibitor percentage decreased by increasing the antioxidants concentration up to 1mM for all crops. It is clear that citric acid at 5.0 mM gave a little value of it by 0.75, 0.71 and 0.87 fold compared with untreated plants for phaseolus, pea and brood bean, respectively.
14- All antioxidants cashed a gradual increase in phytate content mg/g defatted up to 1.0mM. The highest increase were recorded for three legumes from 4.01, 7.18 and 8.0 mg/g (control) to reach (4.46, 7.86 and 9.05), (4.28, 7.63 and 8.95) and (4.60, 7.71 and 9.39 mg/g defatted) at the same concentration of SA, CA and AA, respectively followed by a sharp decrease up to 5.0mM.
15- By increasing salicylic acid concentration, the total phenolic content of the all crops was gradual increased and the highest increase was recorded for phaseolus, pea and broad bean from 0.84, 0.52 and 1.58 % (control) to 2.25, 2.43 and 3.35%, respectively. While, citric acid and ascorbic acid treatments increased the phenolic percentage up to 1.0mM followed by a decrease by increasing the concentration for all crops.
CONCLUSION
From the results of this study, following were a raised:-
1-The necessity of using antioxidants as substitute for synthetic auxins for protecting our environment from pollution and obtained the good yield with regard to the quantity and quality for the crops used in the study (phaseolus, pea and broad bean )
2-The necessity of using three antioxidants (salicylic acid, citric acid and ascorbic acid ) for increasing weight of seed yield, plant (gm), the highest increase was at 1mM citric acid for phaseolus, at 1mM salicylic acid for pea and 1mM salicylic acid and ascorbic acid for broad bean by soaking + spraying treatment. Whereas, spraying treatment of ascorbic acid (AA) caused a gradual increase in weight of seed yield/plant (gm) for all crops by increasing the AA concentration.
3- Soaking + spraying treatment was found to be more effective than other treatments used in this study for increasing of ash, fat, protein and carbohydrate at 1mM antioxidants in the three legumes.
4- In ascending order, soaking seeds before sowing and spraying plants treatments of phaseolus, pea and brood bean crops was a marked decrease in antinutritional factors content at 2.5mM and 5.0mM, where the highest decrease was at 5.0mM.
5- The method of soaking + spraying by application of ascorbic acid treatment at 2.5mM gave somewhat a balanced results for the quantity of yield, increase in chemical constituents content of seeds (fats, ash, protein and carbohydrates) and decrease in antinutritional factors content.
6- Using of antioxidants is not coast because the antioxidants were used by a low concentrations and the yield is very high compared with its price.