بسم الله الرحمن الرحيم {وَسِعَ رَبُّنَا كُلَّ شَيْءٍ عِلْمًا ۚ عَلَى اللَّهِ تَوَكَّلْنَا ۚ رَبَّنَا افْتَحْ بَيْنَنَا وَبَيْنَ قَوْمِنَا بِالْحَقِّ وَأَنتَ خَيْرُ الْفَاتِحِينَ} الأعراف:89 صدق الله العظيم

RESEARCHER MOHAMED TAHA ABD AL-RAHMAN ZALAMA B.Sc. Agric. Sci. (Agronomy), Faculty of Agric., Mansoura University (1999) M. Sc. Agric. Sci. (Agronomy), Faculty of Agric., Mansoura University (2007) Research assistant in Seed Tech. Res. Department Field Crops Institute Agricultural Research Center, Egypt مناقشة الرسالة المقدمة من الباحث/ محمد طه زلمه للحصول علي درجة دكتوراه الفلسفة في العلوم الزراعية ( نبات زراعي)*** تحت إشراف: *أ.د/ محب طه صقر ** أ.د/ زين العابدين عبد الحميد محمد *** رئيس بحوث/ مروءة إسماعيل عطا

TITLE OF THESIS EFFECT OF SOME ANTIOXIDANTS ON SEED QUALITY AND YIELD OF FABA BEAN PLANTS UNDER SALINITY STRESS تأثير بعض مضادات الأكسدة على جودة التقاوي والمحصول في الفول البلدي تحت ظروف الإجهاد الملحي. مناقشة الرسالة المقدمة من الباحث/ محمد طه زلمه للحصول علي درجة دكتوراه الفلسفة في العلوم الزراعية ( نبات زراعي)*** تحت إشراف: *أ.د/ محب طه صقر ** أ.د/ زين العابدين عبد الحميد محمد *** رئيس بحوث/ مروءة إسماعيل عطا

Supervision committee MOHEB TAHA SAKR Prof. Dr. of Plant Physiology, Faculty of Agric., Mansoura University Zein EL-ABEDIN ABD EL- HAMID MOHAMED Prof. Dr. of Agric. Head of Botany Dept. Faculty of Agric. Mansoura University MAROUAH ISMAEL ATTA Chief Researcher Seed Tech. Res. Dept. Field Crops Institute Agric. Res. center مناقشة الرسالة المقدمة من الباحث/ محمد طه زلمه للحصول علي درجة دكتوراه الفلسفة في العلوم الزراعية ( نبات زراعي)*** تحت إشراف: *أ.د/ محب طه صقر ** أ.د/ زين العابدين عبد الحميد محمد *** رئيس بحوث/ مروءة إسماعيل عطا

Award and Discussion Committee Prof. Dr./ Moheb Taha Sakr Prof. Dr. of Plant Physiology, Faculty of Agric., Mansoura University Prof. Dr./ Hosny Mohamed Abd El-Dayem Prof. of Agric. Botany, Faculty of Agric., Moshtohor University Prof. Dr./ Mahmoud Mohamed Darwish Prof. Dr. of Plant Physiology, Faculty of Agric., Mansoura University Prof. Dr./ Zein El-Abedin A.H. Mohamed Prof. Dr. of Agric. Head of Botany Dept. Faculty of Agric. Mansoura University مناقشة الرسالة المقدمة من الباحث/ محمد طه زلمه للحصول علي درجة دكتوراه الفلسفة في العلوم الزراعية ( نبات زراعي)*** تحت إشراف: *أ.د/ محب طه صقر ** أ.د/ زين العابدين عبد الحميد محمد *** رئيس بحوث/ مروءة إسماعيل عطا

INTRODUCTION Faba bean; is a member of family Fabaceae (Leguminosae) which is the third largest family of flowering plants with 650 genera and over 18,000 species. In Egypt, area harvested of faba bean plants in 2005 was 205.661 fad. and decreased in 2011 to 136.401 fad., (FAO. 2013) Egyptian faba bean is facing several problems at the present time. Its effective on both of faba bean productivity, yield and it`s components and seed quality. one of the most important factors which causes reduction of area harvested of faba bean plants is exposure of large areas of lands for salinity as well as sensitivity of faba bean to medium levels of salinity stress. Application of antioxidants is one of the most important ways to increasing salt tolerance of plants. Treatments of [Salicylic acid (SA), Ascorbic acid (ASA), Tochopherol (TOCO), Humic acid (HA) and Yeast extract] proved effective in reducing the adverse effect of salinity on growth, yield and chemical composition of faba bean plants.

OBJECTIVES OF THE STUDY The experiment was designed to; Investigated the influence of some antioxidant materials on the harmful effects of different salinity stress levels on vegetative growth, yield and its components, biochemical constituents, nutrient element contents and protein percentage of faba bean plant (Vicia faba, L.) cv. Sakha 1 obtained from Agric. Res. Center, Ministry of Agric., Egypt.

EXPERIMENTS UNDER THIS STUDY This work includes 3 parts; 1- Pot experiment. 2- Field experiment. 3- Seed quality tests.

Pot experiments includes 3 parts:- 1- Seed presoaking. 2- Plant foliar spraying. 3- Presoaking and foliar spraying together. Pot experiments were performed twice during the growing seasons of (2010/2011 and 2011/2012)

FACTORS UNDER STUDY FIRST FACTOR:- Salinity stress levels: 1- 320 (mg/l) as (Control). 2- 2000 (mg/l). 3- 4000 (mg/l). 4- 6000 (mg/l). 5- 8000 (mg/l).

FACTORS UNDER STUDY Antioxidant materials:- SECOND FACTOR 1- Tap water (control). 2- Salicylic acid (SA) at (250 mg/l). 3- Ascorbic acid (ASA) at (250 mg/l). 4- α –Tocopherol (TOCO) at (100 mg/l). 5- Hamic acid (HA) at (1000 mg/l). 6- Yeast extracts (2000 mg/l).

STUDIED CHARACTERISTICS A- Vegetative growth observations: 1- Shoot length (cm). 2- Root length (cm). 3- Shoot fresh weight (g). 4- Shoot dry weight (g). 5- Root fresh weight (g). 6- Root dry weight (g). 7- Leaf area (cm2/plant). B- Yield and its components: 1- Number of pods / plant. 2- Pods weight / plant (g). 3- Number of seeds / plant. 4- Seeds weight / plant (g). 5- 100- Seed weight (g). C- Biochemical constituents : 1- Photosynthetic Pigments. 2- Proline concentration. 3- Total ascorbic acid. 4- Total phenols. 5- Potasium and sodium. 6- Seeds protein %.

STATISTICAL ANALYSIS All data of the experiment statistically analyzed according to the technique of analysis of variance “ANOVA” for the “Randomized Complete Block Design” by means of “MSTAT-C” Computer software package. The treatment means were compared using the Least Significant Differences “N- LSD 5%”.

Important Results مناقشة الرسالة المقدمة من الباحث/ محمد طه زلمه للحصول علي درجة دكتوراه الفلسفة في العلوم الزراعية ( نبات زراعي)*** تحت إشراف: *أ.د/ محب طه صقر ** أ.د/ زين العابدين عبد الحميد محمد *** رئيس بحوث/ مروءة إسماعيل عطا

Fig (1): Effect of salinity stress levels and applied antioxidants as (Presoaking, Foliar spraying or Presoaking and Foliar spraying together) as well as their interactions on chlorophyll (A) content in shoots of faba bean plant.

Fig (2): Effect of salinity stress levels and applied antioxidants as (Presoaking, Foliar spraying or Presoaking and Foliar spraying) as well as their interactions on chlorophyll (B) content (mg/g f.wt) in shoots of faba bean plant.

Fig (3): Effect of salinity stress levels and applied antioxidants as (Presoaking, Foliar spraying or Presoaking and Foliar spraying together) as well as their interactions on carotenoids content (mg/g. f. wt) in the shoots of faba bean plant.

Fig (4): Effect of salinity stress levels and applied antioxidants as (Presoaking, Foliar spraying or Presoaking and Foliar spraying together) as well as their interactions on proline content (mg/g. f. wt) of faba bean plant.

Fig (5): Effect of salinity stress levels and applied antioxidants as (Presoaking, Foliar spraying or Presoaking and Foliar spraying) as well as their interactions on ascorbic acid content (mg/ 100g f. wt) of faba bean plant.

Fig (6): Effect of salinity stress levels and applied antioxidants as (Presoaking, Foliar spraying or Presoaking and Foliar spraying) as well as their interactions on total phenol content (mg/ 100g f. wt) of faba bean plant.

Fig (7): Effect of salinity stress levels and applied antioxidants as (Presoaking, Foliar spraying or Presoaking and Foliar spraying together) as well as their interactions on Na+ content (mg/g. DW) in shoots of faba bean plant.

Fig (8): Effect of salinity stress levels and applied antioxidants as (Presoaking, Foliar spraying or Presoaking and Foliar spraying together) as well as their interactions on K+ content (mg/g. DW) in shoots of faba bean plant.

Fig (9): Effect of salinity stress levels and applied antioxidants as (Presoaking, Foliar spraying or Presoaking and Foliar spraying) as well as their interactions on Na+ content (mg/g. DW) in roots of faba bean plant.

Fig(10): Effect of salinity stress levels and applied antioxidants as (Presoaking, Foliar spraying or Presoaking and Foliar spraying) as well as their interactions on K+ content (mg/g. DW) in roots of faba bean plant.

Fig(12): Effect of salinity stress levels and applied antioxidants as (Presoaking, Foliar spraying or Presoaking and Foliar spraying) as well as their interactions on Na+/K+ ratio in shoots of faba bean plant.

Fig(11): Effect of salinity stress levels and applied antioxidants as (Presoaking, Foliar spraying or Presoaking and Foliar spraying) as well as their interactions on Na+/K+ ratio in roots of faba bean plant.

FIELD Experiment The experiment under study was carried out within the period of November-Marsh 2010/2011 in Tag El-Ezz Research Station, Dakahlia Governorate, Agric. Res. Center., Ministry of Agric. Egypt to investigate the role of action of some selected antioxidants on the harmful effect of soil salt stress on faba bean plant. مناقشة الرسالة المقدمة من الباحث/ محمد طه زلمه للحصول علي درجة دكتوراه الفلسفة في العلوم الزراعية ( نبات زراعي)*** تحت إشراف: *أ.د/ محب طه صقر ** أ.د/ زين العابدين عبد الحميد محمد *** رئيس بحوث/ مروءة إسماعيل عطا

FACTORS UNDER STUDY FIRST FACTOR:- SECOND FACTOR:- Salinity stress levels: 1- 1900 (mg/l). 2- 3200 (mg/l). SECOND FACTOR:- Antioxidant materials: 1- Tap water. 2- Salicylic acid (SA) at (250 mg/l). 3- Ascorbic acid (ASA) at (250 mg/l). 4- α –Tocopherol (TOCO) at (100 mg/l). 5- Hamic acid (HA) at (1000 mg/l). 6- Yeast extracts (500 mg/l).

STUDIED CHARACTERISTICS A- Vegetative growth observations: 1- Plant height (cm). 2- Plant fresh weight (g). 3- Plant dry weight (g). 4- Leaf area (cm2/plant). B- Yield and its components: 1- No. of pods/plant. 2- Weight of pods/plant (g). 3- Weight of seeds/plant (g). 4- Seed yield (Ardab/fad).

Important Results مناقشة الرسالة المقدمة من الباحث/ محمد طه زلمه للحصول علي درجة دكتوراه الفلسفة في العلوم الزراعية ( نبات زراعي)*** تحت إشراف: *أ.د/ محب طه صقر ** أ.د/ زين العابدين عبد الحميد محمد *** رئيس بحوث/ مروءة إسماعيل عطا

Fig (1): Plant height/cm of faba bean plants as influenced by salinity stress levels, applied antioxidants and their interactions during the growing seasons 2010/2011.

Fig (2): Plant fresh weight (cm) of faba bean plants as influenced by salinity stress levels, applied antioxidants and their interactions during the growing seasons 2010/2011.

Fig (3): Plant dry weight (g) of faba bean plants as influenced by salinity stress levels, applied antioxidants and their interactions during the growing seasons 2010/2011.

Fig (4): Leaf Area (cm2/plant) of faba bean plants as influenced by salinity stress levels, applied antioxidants and their interactions during the growing seasons 2010/2011.

Fig (5): No. of pods/plant as influenced by salinity stress levels, applied antioxidants and their interactions during the growing seasons 2010/2011.

Fig (6): Weight of pods/plant as influenced by salinity stress levels, applied antioxidants and their interactions during the growing season 2010/2011.

Fig (7): No. of seeds/plant as influenced by salinity stress levels, applied antioxidants and their interactions during the growing season 2010/2011.

Fig (8): Weight of seeds/plant as influenced by salinity stress levels, applied antioxidants and their interactions during the growing season 2010/2011.

CONCLUSIONS مناقشة الرسالة المقدمة من الباحث/ محمد طه زلمه للحصول علي درجة دكتوراه الفلسفة في العلوم الزراعية ( نبات زراعي)*** تحت إشراف: *أ.د/ محب طه صقر ** أ.د/ زين العابدين عبد الحميد محمد *** رئيس بحوث/ مروءة إسماعيل عطا

A: Effect of salinity stress on:- 1- Growth attributes: All pot experiments under this study recorded that, all salinity stress levels hardly decrease all growth parameters of faba bean plants when compared with unstressed treatment (control at 320 mg/l) through the two physiological development stages (45 and 90 days) during the two experimental seasons (2010/2011 and 2011/2012). In this regard, the most effective salinity stress level was (8000 mg/l) followed by (6000 mg/l,4000 mg/l and 2000 mg/l), respectively.

2-Yield and it`s components attributes: The results which were obtained showed that, there is a negative correlation between high salinity stress levels and yield of faba bean plants in all pot experiments (presoaking, foliar spraying or presoaking and foliar spraying together) during the two growing seasons. All Salinity levels recorded significant reduction in the yield and it`s components, when compared with unstressed treatment (320 mg/l). The great reduction regarding to (8000 mg/l) followed by (6000 mg/l).

3- Biochemical constituents: Photosynthetic pigments: All salinity stress levels decreased photosynthetic pigments in the shoots of faba bean plant during the two growing seasons in all pot experiments. Regarding, the great reduction was observed by salinity level (8000 mg/l). Proline, ascorbic acid and phenols: Salinity stress levels caused a slightly increase in the contents of each proline, ascorbic and phenols in the shoot of faba bean plant, while compared with unstressed treatment (control at 320 mg/l) at all pot experiments under study. The most effective salinity stress level was (8000 mg/l) followed by (6000 mg/l).

Na+, K+ content and Na+/K+ ratio:- Na+ content was increased by salinity stress levels while K+ content was decreased in the shoot and roots of faba bean plant. High salinity level (8000 mg/l) was most effective in this respect. All salinity stress levels increased Na+/K+ ratio, when compared with unstressed treatment control (320 mg/l) in all pot experiments. Regarding, the higher degree of Na+/K+ ratio caused by salinity stress level (8000 mg/l).

B- Effect of applying antioxidants on:- 1- Growth attributes: All selected antioxidants (SA, ASA, TOCO, HA and Yeast) which applied in all pot experiments (presoaking, foliar spraying or presoaking and foliar spraying together) significantly increase growth parameters of faba bean plants when compared with plants treated with control (Tap water) through the two physiological development stages (45 and 90 days) during the two growing seasons. In this respect, ASA (250 mg/l) recorded the most significant increase in growth attributes of faba bean plants at all experiments during the two growing seasons. 2- Yield and it`s components: All antioxidants which were applied in all pot experiments during the two growing seasons recorded the higher degree of yield and it`s components when compared with untreated plants (control). The most effective antioxidants in this respect were ASA (250 mg/l).

3- Biochemical constituents:- 3.1- Photosynthetic pigments: Applied antioxidants on faba bean plants as presoaking, foliar spraying or presoaking and foliar spraying together let to enhancing the content of chlorophyll A, B and carotenoids against planted treated with control (Tap water). Concerning, ASA (250 mg/l) recorded the great values. 3.2- Proline, ascorbic acid and phenols: Selected antioxidants caused synergistic effect when applied on faba bean plants when compared with control treatment (Tap water) in all pot experiments. ASA and SA was the most effective in this respect. 3.3- Na+, K+ content and Na+/K+ ratio: Applied antioxidants slightly decreased Na+ contents and increased K+ when compared with untreated plants (control treatment) in all pot experiments under study. Applied selected antioxidants clearly decreased Na+/K+ ratio when compared with untreated plants.

C: Effect of interaction between salinity stress and applying antioxidants on:- 1- Growth attributes: With regard to the interaction treatments, the data indicated that applied selected antioxidants (SA, ASA, TOCO, HA and YEAST) in all pot experiments at the two physiological stages (45 and 90 days) significantly enhanced growth attributes of faba bean plants under salinity stress levels when compared with plants treated with control (Tap water) grown under the same salinity stress levels during the two growing seasons. Concerning, treatment ASA (250 mg/l) at salinity level (320 mg/l) recorded the highest values of faba bean growth attributes. Applied antioxidants could basically mitigate the harmful effect of salinity stress on growth of faba bean plants grown under saline soil.

2- Yield and it`s attributes: With respect to the interaction treatments, the data showed that applied antioxidants increased yield and it`s attributes of faba bean plants under salinity stress levels when compared with control plants treated with tap water grown under salinity stress levels. Accordingly, applied antioxidants could be hardly alleviating the reduction caused by salinity stress on yield of faba bean plant. ASA was the most effective in this respect, then SA, Yeast, HA and TOCO, respectively.

3- Biochemical constituents: 3.1- Photosynthetic pigments: Interaction treatments of applied antioxidants with salinity stress levels hardly increased photosynthetic pigments compared with plants treated with control (Tap water) grown under the same salinity stress levels at all pot experiments, but these increases still less than control plants. In this regard applied antioxidants basically mitigate the harmful effect of salinity stress on photosynthetic pigments. ASA (250 mg/l), SA (250 mg/l) and Yeast extract (2000 mg/l) respectively, were more effective in this respect.

3.2- Proline, ascorbic acid and phenols: Regarding the interaction treatments, data cleared that applied selected antioxidants materials with the selected salinity stress levels under this study, hardly increased Proline, ascorbic acid and phenols content in faba bean shoots when compared with untreated plants (control). 3.3- Na+, K+ content and Na+/K+ ratio: Salinity stress treatment combined with applied antioxidants slightly decreased Na+ content and Na+/K+ ratio, while increased K+ content in both shoot and root of faba bean plant, when compared with applied control (Tap water) with the same salinity levels treatments. Applied antioxidants alleviate the harmful effects of salinity stress and decreased the Na+ and Na+/K+ ratio in shoots and roots of faba bean plants against control treatment.

The main obtained results of these study FIELD EXPERIMENT The main obtained results of these study A: Growth characters: All growth characters of faba bean plants significantly enhanced by applied antioxidants (Ascorbic, Salicylic, α- Tocopherol, Humic and Yeast extract) compared with treated plants with control (Tap water) in the two soils salt area (A1 and A2). The data also indicated that applied antioxidant materials were more effective in salt soil area (A1). ASA (250 mg/l) was the most effective in this respect followed by SA (250 mg/l), Yeast (2000 mg/l), HA (1000 mg/l) and TOCO (100 mg/l) respectively. مناقشة الرسالة المقدمة من الباحث/ محمد طه زلمه للحصول علي درجة دكتوراه الفلسفة في العلوم الزراعية ( نبات زراعي)*** تحت إشراف: *أ.د/ محب طه صقر ** أ.د/ زين العابدين عبد الحميد محمد *** رئيس بحوث/ مروءة إسماعيل عطا

B: Yield and it`s components: Applied antioxidants materials significantly increased yield and it`s components (No. of pods/plant, weight of pods/plant, weight of seeds/plant and seed yield (Ardab/fad) in the two salt soil areas especially (A1) compared with the control treatment (Tap water). The obtained data recorded that applied antioxidant materials could mitigate the harmful effect of high soil salt stress levels on yield and its components of faba bean plants. ASA (250 mg/l) followed by SA (250 mg/l) were more effective in this regard.

RECOMMENDATION This investigation indicated that, it can alleviate and mitigate the harmful effect of different salinity stress levels on faba bean plants by applied non-enzymatic antioxidants (Ascorbic acid, Salicylic acid, α- Tocopherol, Humic acid and Yeast extract) in experiments (presoaking, Foliar spraying or presoaking and foliar spraying together). These antioxidant materials caused an increments effect on growth and it`s attributes, yield and it`s components also chemical constituents. The Results mention previously show that the best antioxidant material is Ascorbic acid (ASA) followed by Salicylic acid (SA), Yeast extract, Humic acid and α- Tocopherol respectively. In addition, with these regard faba bean plants can be planted in wide range of saline soil in Egypt with applying natural safety antioxidants.

شكر وتقدير أتقدم بالشكر والتقدير إلى لجنة الإشراف أ.د/ محب طه صقر أ.د/ زين العابدين عبد الحميد محمد أ.د/ مروءة إسماعيل عطا كما أتقدم بالشكر للجنة المناقشة والحكم والسادة الحضور على حسن الاستماع والمتابعة. مناقشة الرسالة المقدمة من الباحث/ محمد طه زلمه للحصول علي درجة دكتوراه الفلسفة في العلوم الزراعية ( نبات زراعي)*** تحت إشراف: *أ.د/ محب طه صقر ** أ.د/ زين العابدين عبد الحميد محمد *** رئيس بحوث/ مروءة إسماعيل عطا

و السلام عليكم ورحمة الله و بركاته مناقشة الرسالة المقدمة من الباحث/ محمد طه زلمه للحصول علي درجة دكتوراه الفلسفة في العلوم الزراعية ( نبات زراعي)*** تحت إشراف: *أ.د/ محب طه صقر ** أ.د/ زين العابدين عبد الحميد محمد *** رئيس بحوث/ مروءة إسماعيل عطا