Scholarly article on topic 'Biochemical evaluation of some faba bean cultivars under rainfed conditions at El-Sheikh Zuwayid'

Biochemical evaluation of some faba bean cultivars under rainfed conditions at El-Sheikh Zuwayid Academic research paper on "Biological sciences"

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{"Faba bean cultivars" / "Rainfed conditions" / "Chemical composition" / "Amino acids" / SDS–PAGE / "Total vicine" / "Growth and yield"}

Abstract of research paper on Biological sciences, author of scientific article — M.H. Hendawey, A.M.A. Younes

Abstract Two field experiments were carried out during 2009/2010 and 2010/2011 seasons at Agricultural Experimental Station of Desert Research Center (DRC) located in El-Sheikh Zuwayid region (North Sinai), Egypt, to study the biochemical evaluation of some faba bean cultivars (Giza 843, Giza 3, Sakha 4, Nubaria 1, Missr1, Sakha 2, and Sakha 3) under rainfed conditions at El-Sheikh Zuwayid. The main results could be summarized as follows: • Sakha 3 and Sakha 4 exceeded the other cultivars in plant height, fresh weight, seed yield/plant, 100-seed weight, seed yield, protein yield, and total vicine yield. On the other hand, the lowest values of such parameters were achieved by Giza 843 except protein yield under the same conditions. These results associated with increase in photosynthetic pigments under rainfed conditions at El-Sheikh Zuwayid, which related to the counteraction of faba bean cultivars to stress conditions. Concerning the yield of antinutritional factors, Sakha 4 recorded the highest value of vicine yield followed by Nubaria 1 and Sakha 3 in a descending order. • Total vicine was found in high amount in seeds of Nubaraia 1. Also, the greatest protein content was produced from Sakha 3 followed by Sakha 2, Sakha 4, and Missr1 as compared with the other cultivars. Potassium and calcium contents were found in high amounts in seeds of Sakha 4, Sakha 3, and Sakha 2. On the other hand, the lowest value of sodium content was produced from Sakha 3. In addition, Sakha 3 surpassed other faba bean cultivars in K/Na ratio and Ca/Na ratio. • Faba bean cultivars greatly varied in total vicine. The hulled seeds of all faba bean contain higher values of total vicine more than the whole seeds without any treatments (control). Approximately all treatments (processing conditions) significantly decreased total vicine in whole and milled seeds for all faba bean cultivars as compared with the control (whole seeds without any treatments). In this respect, the minimum values of such content were produced from milled seeds of all cultivars after treatment (soaking) with water, citric acid, sodium bicarbonate, and acetic acid as compared with the control. • Amino acids composition indicated the presence of 16 amino acid including essential and non-essential amino acids. In this regard, the predominant essential amino acid is arginine followed by leucine. Sakha 3 followed by Missr 1 and Sakha 2 exceeded the other cultivars in arginine content. In addition, methionine is present in low quantities comparing with other amino acids for all faba bean cultivars. In this respect, glutamic acid is the most abundant amino acid followed by aspartic and proline. Comparison between faba bean cultivars, Sakha 2 followed by Sakha 4 and Giza 3 had higher concentration of glutamic acid than other cultivars. Data also showed that non-essential amino acids are more than essential amino acids in seeds of all faba bean cultivars under investigation. • Analysis of gel (SDS–PAGE) revealed that Sakha2 and Sakha3 were resolved into 30 bands, while Giza 843, Giza 3, Nubaria 1, and Missr 1 were resolved into 31 bands. Also, Sakha 4 was resolved into 32 bands. The molecular weight of protein sub units ranged between 14 and 95kDa. The more intensive bands are presented at molecular mass 20, 41, and 43kDa. The increase in band intensities in addition to the other obtained results may be a part of response these cultivars to rainfed conditions at El-Sheikh Zuwayid. • According to the previous observed data, it could be recommended that the use of tolerance cultivars such as Sakha3 and Sakha4, which are associated with yield and biochemical constituents under rainfed conditions at El-Sheikh Zuwayid. Also, to reduce total vicine (antinutritional factors) in seeds, the study recommended soaking seeds of faba bean before human consumption in some safe chemicals such as citric acid, sodium bicarbonate, and acetic acid.

Academic research paper on topic "Biochemical evaluation of some faba bean cultivars under rainfed conditions at El-Sheikh Zuwayid"

Annals of Agricultural Science (2013) 58(2), 183-193

Faculty of Agriculture, Ain Shams University Annals of Agricultural Science

www.elsevier.com/locate/aoas

ORIGINAL ARTICLE

Biochemical evaluation of some faba bean cultivars under rainfed conditions at El-Sheikh Zuwayid

M.H. Hendawey *, A.M.A. Younes

Biochemistry Unit-Plant Genetic Resources Dept., Desert Research Center, Matarya, Cairo, Egypt

Received 20 May 2013; accepted 1 June 2013 Available online 5 September 2013

KEYWORDS

Faba bean cultivars; Rainfed conditions; Chemical composition; Amino acids; SDS-PAGE; Total vicine; Growth and yield

Abstract Two field experiments were carried out during 2009/2010 and 2010/2011 seasons at Agricultural Experimental Station of Desert Research Center (DRC) located in El-Sheikh Zuwayid region (North Sinai), Egypt, to study the biochemical evaluation of some faba bean cultivars (Giza 843, Giza 3, Sakha 4, Nubaria 1, Missr1, Sakha 2, and Sakha 3) under rainfed conditions at El-Sheikh Zuwayid. The main results could be summarized as follows:

• Sakha 3 and Sakha 4 exceeded the other cultivars in plant height, fresh weight, seed yield/plant, 100-seed weight, seed yield, protein yield, and total vicine yield. On the other hand, the lowest values of such parameters were achieved by Giza 843 except protein yield under the same conditions. These results associated with increase in photosynthetic pigments under rainfed conditions at El-Sheikh Zuwayid, which related to the counteraction of faba bean cultivars to stress conditions. Concerning the yield of antinutritional factors, Sakha 4 recorded the highest value of vicine yield followed by Nubaria 1 and Sakha 3 in a descending order.

• Total vicine was found in high amount in seeds of Nubaraia 1. Also, the greatest protein content was produced from Sakha 3 followed by Sakha 2, Sakha 4, andMissr1 as compared with the other cultivars. Potassium and calcium contents were found in high amounts in seeds of Sakha 4, Sakha 3, and Sakha 2. On the other hand, the lowest value of sodium content was produced from Sakha 3. In addition, Sakha 3 surpassed other faba bean cultivars in K/Na ratio and Ca/Na ratio.

• Faba bean cultivars greatly varied in total vicine. The hulled seeds of all faba bean contain higher values of total vicine more than the whole seeds without any treatments (control). Approximately all treatments (processing conditions) significantly decreased total vicine in whole and milled seeds for all faba bean cultivars as compared with the control (whole seeds without any treatments). In this respect, the minimum values of such content were produced from milled seeds of all cultivars after treatment (soaking) with water, citric acid, sodium bicarbonate, and acetic acid as compared with the control.

* Corresponding author. Tel.: +20 2 44441172. E-mail address: mhhendawey@yahoo.com (M.H. Hendawey). Peer review under responsibility of Faculty of Agriculture, Ain-Shams University.

0570-1783 © 2013 Production and hosting by Elsevier B.V. on behalf of Faculty of Agriculture, Ain Shams University. http://dx.doi.org/10.1016/j.aoas.2013.07.010

• Amino acids composition indicated the presence of 16 amino acid including essential and nonessential amino acids. In this regard, the predominant essential amino acid is arginine followed by leucine. Sakha 3 followed by Missr 1 and Sakha 2 exceeded the other cultivars in arginine content. In addition, methionine is present in low quantities comparing with other amino acids for all faba bean cultivars. In this respect, glutamic acid is the most abundant amino acid followed by aspartic and proline. Comparison between faba bean cultivars, Sakha 2 followed by Sakha 4 and Giza 3 had higher concentration of glutamic acid than other cultivars. Data also showed that non-essential amino acids are more than essential amino acids in seeds of all faba bean cultivars under investigation.

• Analysis of gel (SDS-PAGE) revealed that Sakha2 and Sakha3 were resolved into 30 bands, while Giza 843, Giza 3, Nubaria 1, and Missr 1 were resolved into 31 bands. Also, Sakha 4 was resolved into 32 bands. The molecular weight of protein sub units ranged between 14 and 95 kDa. The more intensive bands are presented at molecular mass 20, 41, and 43 kDa. The increase in band intensities in addition to the other obtained results may be a part of response these cultivars to rainfed conditions at El-Sheikh Zuwayid.

• According to the previous observed data, it could be recommended that the use of tolerance cul-tivars such as Sakha3 and Sakha4, which are associated with yield and biochemical constituents under rainfed conditions at El-Sheikh Zuwayid. Also, to reduce total vicine (antinutritional factors) in seeds, the study recommended soaking seeds of faba bean before human consumption in some safe chemicals such as citric acid, sodium bicarbonate, and acetic acid.

© 2013 Production and hosting by Elsevier B.V. on behalf of Faculty of Agriculture, Ain Shams

University.

Introduction

Faba bean (Vicia faba L.) is one of the oldest crops and ranks the sixth in production among the different legumes grown in the world after soybean, peanut, beans, peas, and chickpeas. Faba bean is popular legume food with high yield capacity and high nutritional value. It is widely used in the Mediterranean region as source of protein in both human and animal nutrition. And it is popular breakfast food and also used as vegetable green or fresh canned. In Egypt, faba bean is one of the most important pulse crop cultivated due to the richness of seed protein content. Also, broad bean could be eaten in several forms, i.e., stewed broad bean (Medamis), broad bean cakes (Taamia), stewed broad bean paste (Bissara), and germinated broad bean soap (Foul nabit). Therefore, there is need to increase its production by expansion through newly reclaimed areas. The most important organic components of faba bean seeds are proteins (20-41% of seed dry matter) and carbohydrates (51-68% of seed dry matter) which depend on cultivars. Most of these proteins comprise of globulins (79%), albumins (7%), and glutelins (6%) (Hossain and Mortuza, 2006). The quality of this protein appears to be limited by the low content in sulfur-containing amino acids and in tryptophan, valine, isoleucine, and threonine. The main antinutritional factors contained in the seed are tannins and two glucosides as pyrim-idine derivaties namely vicine [2,6 diamino-4,5-dihydroxy pyrimidine, 5 (B-glycopyransoide)], and convicine [2,4,5-trihy-droxy-6-amino pyrimidine, 5 (B-D glycopyransoide)]. These two glucosides are also believed to be responsible for causing favism in some genetically susceptible humans consuming faba bean that have deficiency of erythrocytic enzyme glucose-6-phosphate dehydrogenase activity (G6PD) (Corchia et al., 1995; Beutler etal., 1996). This enzyme involved in the pentose phosphate pathway (Frank, 2005). G6PD converts glucoses-phosphate into 6-phosphoglucono-S-lactone and is the rate-

limiting enzyme of this metabolic pathway that supplies reducing energy to cells by maintaining the level of the co-enzyme nicotinamide adenine dinucleotide phosphate (NADPH). The NADPH in turn maintains the supply of reduced glutathi-one in the cells that is used to form free radicals that cause oxi-dative damage. People with G6PD deficiency are therefore at risk of hemolytic anemia in states of oxidative stress (Gaskin etal., 2001). Oxidative stress can result from certain foods such as faba beans contain high levels of vicine and convicine which are oxidants. Nutritionally, it would be desirable to remove these previous substances by breeding or by technical treatments through faba bean preparation before consumption. The main goal of this work was to biochemical evaluation of seven faba bean cultivars under rainfed conditions at El-Sheikh Zuwayid.

Materials and methods

Experimental work

A field experiment was conducted at El-Sheikh Zuwayid Research Station, North Sinai, Desert Research Center for two consecutive winter seasons 2009/2010 and 2010/2011. Faba bean cultivars (Giza 843, Giza 3, Sakha 4, Nubaria 1, Missr1, Sakha 2, and Sakha 3) were obtained from the Field Crop Institute, Agricultural Research Center, Ministry of Agriculture, Egypt, and sown in second week of October in both seasons at rate of 70 kg/fad. For not late sowing date, the experiment was irrigated after planting, while different growth stages were depended on the amount of rainfall during two growing seasons. The experiments were designed in randomize design with three replicates. The plot was 6 m2, each plot was fertilized with recommended fertilization for this type of soil according to Desert Research Center leaflet. Mechanical and chemical analysis of soil and water are presented in Table 1.

Table 1 Mechanical and chemical analysis of the experimental soil and chemical analysis of irrigation water at El-Sheikh Zuwayid, North Sinai.

Characters Values

(a) Mechanical analysis of the experimental soil

Fine sand (%) 31.88

Coarse sand (%) 58.67

Clay (%) 2.28

Silt (%) 7.17

Texture class Sandy

(b) Chemical analysis of the experimental soil

pH 8.01

E.C. (mmhos/cm) 0.92

Soluble cations (meq/L)

Ca+ + 4.01

Mg+ + 0.86

Na+ 4.82

K + 0.23

Soluble anions (meq/L)

HCO^ 1.85

CP 3.15

so^ 4.92

(c) Chemical analysis of irrigation water

pH 7.90

E.C. (mmhos/cm) 4.01

Soluble cations (meq/L)

Ca+ + 6.21

Mg+ + 6.73

Na+ 28.11

K + 0.21

Soluble anions (meq/L)

HCO^ 3.31

CP 27.24

SO= 10.71

The results were determined according to Richard (1954) and Jackson (1958). Meteorological data, i.e., temperature (C0), relative humidity (%), wind speed (m/s), Sunshine hours (hour), the amount of rain (mm), and potential evapotranspiration (mm/day) for the location of El-Sheikh Zuwayid are presented in Table 2.

Two plant samples were taken. First sample was taken at 50 days after sowing, and the following growth parameters were recorded; plant height, plant fresh and dry weights and water content (%). Chemical analysis was conducted in leaves for determination of photosynthetic pigments. The second sample was taken at harvesting time (90 days after sowing) to determine yield components including, seed yield/plant (g), 100-seed weight (g), seed yield (kg/Fad.), protein yield (kg/Fad.), and vicine yield (kg/Fad.). Chemical analysis in seeds was conducted to determine chemical composition in cul-tivars, electrophoretic behavior of soluble proteins, amino acids, and total Vicine.

Chemical analysis

Photosynthetic pigments

Photosynthetic pigments in leaves were extracted and estimated according to AOAC (1990) and Wettstein (1957).

Chemical composition

Approximate chemical composition with moisture, protein, Fat, and crude fiber in seeds was determined according to the method described in AOAC (2000).

Determination of total glycosides (Total Vicine) Soaking treatments. The methods of Vidal-Valverde et al. (1998) and Hegazy and Marquardt (1983) were used to determine which of these processes might enhance the nutritional value of faba beans. Faba bean samples (whole and milled seeds) were soaking with water, 0.1% citric acid, 0.07% sodium bicarbonate, and 1% acetic acid. The proportion of seed

Table 2 Measured climatic data of El-Sheikh Zuwaid region during the period of two years (2009-2010).

Climatic data January February March April May June July August September October November December Avg.

2009 Max. temp. fC) 16.94 18.62 25.78 28.01 29.22 32.25 34.05 36.46 34.80 30.49 25.60 23.03 27.94

Min. temp. (OC) 7.60 7.76 11.62 13.31 15.28 18.83 22.31 23.00 20.68 17.07 14.18 10.05 15.14

Max. relative humidity (%) 90.24 88.24 84.36 81.36 81.39 86.31 89.18 90.27 88.22 89.62 90.17 91.38 87.56

Min. relative humidity (%) 63.32 58.18 49.24 46.83 48.78 53.66 59.83 56.98 53.77 55.27 59.50 55.66 55.09

Wind speed (m/s) 2.84 2.73 1.47 1.59 1.77 1.18 1.46 1.05 1.46 1.29 1.43 2.59 1.74

Sunshine hours (hour) 6.71 7.50 8.10 9.18 10.07 11.45 11.55 11.26 10.17 9.18 7.60 6.52 9.11

Rainfall (mm) 11.38 35.61 1.02 0.00 0.00 0.00 0.00 0.00 0.00 5.91 18.34 23.64 95.90*

ETo (mm/day) 1.64 2.14 3.14 4.27 4.91 5.40 5.68 5.61 4.84 3.43 2.24 1.99 3.78

2010 Max. temp. (OC) 23.46 23.48 28.82 30.69 33.85 35.18 37.23 37.40 37.46 35.07 30.87 21.65 31.26

Min. temp. (OC) 9.36 9.94 11.67 12.89 15.54 19.88 23.45 23.92 21.58 19.81 14.77 11.35 16.18

Max. relative humidity (%) 81.83 91.81 83.99 88.07 88.43 96.60 98.39 95.50 91.68 87.64 93.49 91.65 90.76

Min. relative humidity (%) 47.36 54.57 43.20 44.39 44.34 56.04 61.46 60.45 52.44 50.93 51.15 62.64 52.42

Wind speed (m/s) 3.03 3.58 3.00 2.62 2.16 1.01 1.34 1.46 0.75 0.82 3.15 3.95 2.24

Sunshine hours (hour) 7.52 8.40 9.07 10.28 11.28 12.83 12.94 12.61 11.39 10.28 8.51 7.30 10.20

Rainfall (mm) 26.32 32.13 12.95 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 3.47 74.87a

ETo (mm/day) 2.59 2.75 4.27 5.04 5.77 6.02 6.35 6.17 5.16 3.86 3.31 2.02 4.44

Potential evapotranspiration (ETo) was calculated by Penman-Monteith equation (Allen et al., 1998). a Total temp.: temperature, ETo: Potential evapotranspiration, Avg.: average.

to soaking solution was 1:3 (w/v). The seeds were soaked in darkness for 16 h. After this period, the soaking solution was drained off.

Extraction and determination of total vicine. Total glycosides expressed as total vicine content: total vicine content was carried out by the method of Abdallah et al. (1988). One gram of finely prepared dried seeds was homogenized in a blender with 100 ml of freshly prepared solution of 4% metaphosphoric acid for 5 min and centrifuged for 30 min at 2000 rpm, and the supernatant was filtered through a Whatman NO. 4 filter paper. Exactly 1 ml of the filtrate was diluted to 10 ml with distilled water and the absorbance measured at 273 nm using ultra violet spectrophotometer model Spectronic Genesys 5.

Protein amino acids in seeds

Protein amino acids composition in seeds was determined by amino acid analyzer apparatus model (LC 3000 Eppendorf, Central Lab. of Desert Research Center). Hydrolysis was carried out according to the method of Pellet and Young (1980).

Electrophoretic behavior of soluble proteins in seeds SDS-PAGE gel electrophoresis was performed in acrylamide slab gels following the system of Laemmli (1970) and modified by Studier (1973) to identify and separate soluble proteins in seeds.

Statistical analysis

Data were analyzed statistically according to the procedure outlined by Snedecor and Cochran (1967).Combined analysis over growing seasons for grain soaking and foliar applications was done when the homogeneity test was insignificant according to Gomez and Gomez (1984). Duncan's multiple range test was used for the comparison between means (Duncan, 1955).

Results and discussion

Growth, Yield, and its components

The combined analysis over two seasons for all traits is presented in Table 3. Data for growth parameters varied significantly in most tested cultivars. The highest values of plant height and fresh weight were produced from Sakha 3, Sakha 4, and Missr 1. On the other hand, the lowest value of such parameters was achieved by Giza 843 followed by Giza 3. It is quite clear from data that Missr1 followed by Sakha3, Sa-kha2, and Sakha4 exceeded the other cultivars in dry weight, while Giza 843 genotype recorded the minimum value under the same conditions. These cultivars differences may be due to the differences in the genetic structure. In this regard, differences among faba bean cultivars in growth parameters were noticed by El Bassiouny et al. (2005), Azooz (2009), El-Sayed (2009) and El-Saber (2010).

From the data presented in Table 3, it is clear that the differences between faba bean cultivars in yield and its components are significant in most tested cultivars. Sakha 3 and Sakha 4 recorded the highest values of seed yield/plant, 100-seed weight, seed yield, and protein yield compared with the other cultivars. In contrast, the lowest values of seed yield/

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Table 4 Concentration of photosynthetic pigments in leaves of some faba bean cultivars under rainfed conditions at El-Sheikh Zuwayid (combined analysis 2009/2010 and 2010/2011 seasons).

Faba bean cultivars Photosynthetic pigments (mg/100 g fresh wt.)

Chlorophyll (a) Chlorophyll (b) Chlorophyll (a + b) Chlorophyll (a/b) Carotenoids (a + b)/c Total pigments

Giza 843 63.99c 23.33b 87.32c 2.74bc 24.53b 3.55d 111.85c

Giza 3 69.78bc 24.25b 94.03bc 2.87bc 24.18b 3.88bc 118.21a-c

Sakha 4 73.66a 28.27a 101.93a 2.60c 23.99b 4.24a 125.92a

Nubaria 1 67.31bc 23.01b 90.32c 2.92b 26.66a 3.38de 116.98a-c

Missr 1 59.25d 28.52a 87.77c 2.07d 21.66c 4.05ab 109.43c

Sakha 2 74.32a 14.13c 88.45c 5.25a 27.25a 3.24e 115.7bc

Sakha 3 71.58ab 26.74a 98.32ab 2.67bc 25.76ab 3.81c 124.08ab

(a + b)/c = (Chlorophyll a + Chlorophyll b)/Carotenoids.

Values followed by the same letter in columns are not different at p < 0.05 by Duncan's multiple range test.

plant, 100-seed weight, and seed yield were produced from Giza 843. These findings may be due to the higher values of photosynthetic pigments in Sakha 3 and Sakha 4 than other faba bean cultivars (Table 4). In this regard, the differences of yield and its components between faba bean cultivars was noticed by Omar et al. (1998) under rainfall conditions of Maryout, El Bassiouny et al. (2005) under reclaimed sandy soil, Al Ghamdi (2007) under central region conditions of Saudi Arabia, Osman et al. (2010) in new agro-ecological regions, Ragab et al. (2010) under calcareous soil condition, El-Saber

(2010) under rainfed at Maryout conditions, Bakry et al.

(2011) under newly reclaimed sandy soil conditions, Mona et al. (2011) under calcareous soil, and Farag and Afiah (2012) under rainfed at Maryout conditions.

Concerning the yield of antinutritional factors, Sakha 4 recorded the highest value of vicine yield which reached 11.39 kg/fad., followed by Nubaria 1 and Sakha 3 (11.14 and 10.96 kg/fad, respectively) in a descending order. In another study, El Bassiouny etal. (2005) and Mona etal. (2011) noticed the differences of total vicine in seeds of faba bean cultivars.

Chemical determinations

Photosynthetic pigments

It is evident from the records in Table 4 that the greatest chlorophyll (a) was produced from cultivars Sakha 2 and Sakha 4 followed by Sakha 3 as compared with the other faba bean cul-tivars. Also, Sakha 4 followed by Sakha 3 had a higher concentration of chlorophyll (a + b) and total pigments than other cultivars. In addition, Sakha 4, Missr 1, and Sakha 3 significantly exceeded the other cultivars in Chlorophyll (b) under the same conditions. From data shown, it could be noticed that the highest values of chlorophyll (a/b) and carotenoids were recorded in leaves of Sakha 2 followed by Nubaria 1 and Sakha 3. Data for (Chlorophyll a + Chlorophyll b)/ Carotenoids varied significantly in most tested cultivars. The cultivars Sakha 4 and Missr 1 followed by Giza 3 and Sakha 3 had a higher concentration of (Chlorophyll a + Chlorophyll b)/Carotenoids than other cultivars. The increase in photosyn-thetic pigments in leaves of some faba bean plants under rain-fed conditions at El-Sheikh Zuwayid may be ascribed to the differences in the genetic structure between faba bean cultivars. In this regard, Sairam et al. (2002) and Yildirim et al. (2008) suggested that chlorophyll content is one of the important indicators of stress tolerance in crop plants. Also, Azooz (2009) found that Chlorophyll a, Chlorophyll b, and (Chloro-

phyll a + Chlorophyll b) in leaves of faba bean cultivars were (0.582-0.903 mg/g), (0.210-0.328 mg/g), and (0.792-1.231 mg/ g), respectively. Moreover, El Bassiouny et al. (2005) found that Chlorophyll a, Chlorophyll b, and (Chlorophyll a + Chlorophyll b) in leaves of faba bean cultivars were (960-1176 mg/100 g), (227-331 mg/100 g), and (239-401 mg/ 100 g), respectively, under reclaimed sandy soil.

Chemical composition in seeds

The chemical composition of the seven faba bean cultivars is shown in Table 5. Data for chemical composition in seeds varied significantly in most tested cultivars. Total vicine was found in high amounts reached 13.98 mg/g in seeds of Nuba-raia 1, while the lowest value of such content was recorded in seeds of Giza 3 under the same conditions. In this regard, El Bassiouny et al. (2005) found that vicine and convicine in seeds of faba bean were (284-472 mg/100 g) and (85-156 mg/ 100 g), respectively, under reclaimed sandy soil. Also, Mona et al. (2011) showed that green seeds of faba bean had higher amount of total vicine (32.565 mg/g) than dry seeds (13.20 mg/ g) under calcareous soil. The greatest protein content was produced from Sakha 3 followed by Sakha 2, Sakha 4, and Missr 1, while the lowest amount was produced from Giza 3. In this regard, Giza843 and Giza 3 had higher concentration of fat content than other cultivars. The range in fiber content extended from 4.61% to 6.91% for Giza 3 and Sakha 2, respectively. The differences of chemical composition between faba bean cultivars were noticed by El Bassiouny et al. (2005), Abu-sin et al., (2009) and El-Saber (2010). Also, Musalam et al. (2004) studied the chemical composition of faba bean cultivars under rainfed and irrigation conditions. They showed that ash, protein, and fiber were higher under irrigation condition, whereas the opposite was true for fat and carbohydrate.

Data listed in the same table showed that significant differences were recorded in mineral content in most tested cultivars. Potassium content was found in high amounts reached 1.22, 1.20, and 1.14 for Sakha 4, Sakha 3, and Sakha 2, respectively. On the other hand, the lowest value of sodium content was produced from Sakha 3. The maximum value of calcium content was achieved by Sakha 3, Sakha 4, and Sakha 2. It is obvious from such data in the same table that Sakha 3 surpassed other faba bean cultivars in K/Na ratio and Ca/Na ratio. In this regard, variations in these traits among different faba bean cultivars can be attributed to both genetic and environmental factors. In addition, the selectivity of high K/Na ratio in plants is important control mechanism and a selection criterion for

Table 5 Chemical composition in seeds of some faba bean cultivars under rainfed conditions at El-Sheikh Zuwayid (combined analysis 2009/2010 and 2010/2011 seasons).

Faba bean cultivars Chemical composition

Moisture Total vicine Protein Fat (g%) Fiber (g%) Minerals content

(%) (mg/g) dry wt. (g%) dry wt. dry wt. dry wt. Na (g%) K (g%) K/Na Ca (g%) Ca/Na

dry wt. dry wt. ratio dry wt. ratio

Giza 843 8.09ab 10.98b-d 24.17bc 2.21a 5.11bc 0.70ab 0.88b 1.25c 0.61de 0.87c

Giza 3 7.05b 8.55e 23.04c 2.20a 4.61c 0.61b-d 0.72c 1.18c 0.52e 0.85c

Sakha 4 7.62ab 11.85b 25.61ab 1.84bc 6.74a 0.65bc 1.22a 1.86b 0.78ab 1.20b

Nubaria 1 7.54ab 13.98a 24.11bc 1.92b 5.69b 0.58cd 0.78c 1.34c 0.65cd 1.12b

Missr 1 8.18a 10.20d 25.56ab 1.62d 5.77b 0.78a 0.94b 1.20c 0.70b-d 0.89c

Sakha 2 7.24ab 10.38cd 26.12ab 1.88bc 6.91a 0.60b-d 1.14a 1.90b 0.72a-c 1.20b

Sakha 3 7.12b 11.28bc 26.77a 1.70cd 6.88a 0.51d 1.20a 2.35a 0.81a 1.58a

Values followed by the same letter in columns are not different at p < 0.05 by Duncan's multiple range test.

Table 6 Total glycosides (total vicine) content (mg/g) and reduction% in seeds of some faba bean cultivars as affected by different processes under rainfed conditions at El-Sheikh Zuwayid (combined analysis 2009/2010 and 2010/2011 seasons).

Treatments Total vicine content and reduction% in seeds

Giza 843 Giza 3 Saka 4 Nubaria 1 Missr 1 Saka 2 Saka 3

mg/g Red% mg/g Red% mg/g Red% mg/g Red% mg/g Red% mg/g Red% mg/g Red%

WS 10.98ab - 8.55a - 11.85b - 13.98a - 10.20b - 10.38b - 11.28b -

HS 11.32a INC(3.09) 8.96a INC(4.79) 12.78a INC(7.84) 14.25a INC(1.93) 12.63a INC(23.82) 12.67a INC(22.06) 13.12a INC(16.31)

WSW 10.23bc 6.83 8.55a 00 7.76d 34.51 6.97d 50.14 9.93b 2.64 10.38b 00 7.23c 35.90

WSCA 9.45c 13.93 7.68b 10.17 7.87d 33.58 10.42c 25.46 8.77c 14.01 10.31b 0.67 6.35d 43.70

WSSB 7.12d 35.15 3.22c 62.33 10.16c 14.26 2.85e 79.61 2.88d 71.76 5.40c 47.97 5.17e 54.16

WSAA 10.98e 00 7.38b 13.68 9.56c 19.32 13.16b 5.86 9.82b 3.72 4.50d 56.64 4.35f 61.43

MSW 1.35e 87.70 1.23e 85.61 1.39e 88.27 1.05f 92.48 2.40d 76.47 1.21e 88.34 1.18gh 89.53

MSCA 1.22e 88.88 0.98e 88.53 2.11e 82.19 1.22f 91.27 1.26e 87.64 1.35e 86.99 1.42g 87.41

MSSB 1.07e 90.25 1.26e 85.26 1.86e 84.30 1.22f 91.27 1.76e 82.74 1.10e 89.40 0.76g 93.26

MSAA 1.96e 82.14 2.25d 73.68 1.39e 88.27 2.16e 84.54 1.60e 84.31 1.33e 87.18 1.34gh 88.12

Values followed by the same letter in columns are not different at p < 0.05 by Duncan's multiple range test.

Red% = Reduction%, WS = Whole seeds without any treatments (control), HS = Hulled seeds, WSW = Whole seeds + Water soaking, WSCA = Whole seeds + Citric acid soaking, WSSB = whole seeds + Sodium bicarbonate soaking, WSAA = whole seeds + acetic acid soaking, INC = increased of total vicine content. MSW = Milled seeds + Water soaking, MSCA = Milled seeds + Citric acid soaking, MSSB = Milled seeds + Sodium bicarbonate soaking, MSAA = Milled seeds + acetic acid soaking.

stress tolerance (Wenxue et al., 2003). Also, Cuin et al. (2003) concluded that high K/Na ratio is more important for many species than simply maintaining a low Na concentration.

Total glycosides (total vicine) content in seeds

From the data presented in Table 6, it is clear that the studied faba bean cultivars grown under rainfed conditions at El-Sheikh Zuwayid greatly varied in total vicine. However, it was also noticed that the hulled seeds of all faba bean cultivars contain higher values of total vicine more than the whole seeds without any treatments (control). It was reached 11.32, 8.96, 12.78, 14.25, 12.63, 12.67, and 13.12 for Giza 843, Giza 3, Sakha 4, Nubaria 1, Missr 1, Sakha 2, and Sakha 3, respectively. These results lead to a conclusion that the total vicine may be concentrated in the endosperm of the faba bean cultivars.

Approximately all treatments (processing conditions) significantly decreased total vicine in whole and milled seeds for all faba bean cultivars as compared with the control (whole

seeds without any treatments). In this respect, the minimum values of such content were produced from milled seeds of all cultivars after treatment (soaking) with water, 0.1% citric acid, 0.07% sodium bicarbonate, and 1% acetic acid as compared with the control. The decrease in total vicine in faba bean cultivars after processing conditions may be attributed to leaching out of this antinutrient into the soaking water under the influence of the concentration gradient. In addition, alkaline and acidic medium were tried as a method for improving the extractability of the antinutritional factors (especially vicine and convicine solubility) in faba bean. In this concern, Zahrah (1985) showed that acidic medium (1% acetic acid) is much better than alkaline medium (0.5% sodium bicarbonate) in removing higher quantities of total vicine in seeds of faba bean. In another study, Mona (1988) showed that the removal of vicine by using sodium bicarbonate of 0.03% is very limited in seeds of faba bean. However, El bassiouny et al. (2005) found that vicine and convicine in seeds of faba bean were (284-472 mg/100 g) and (85-156 mg/100 g), respectively, under reclaimed sandy soil.

Amino acids composition in seeds

Amino acids composition is an important feature in determining the nutritional value of faba bean seeds for human and animal diets. Table 7 and Fig. 1 indicated the 16 amino acids that detected including essential amino acids, i.e., threonine, valine, methionine, isoleucine, leucine, phenylalanine, histidine, lysine, arginine and non-essential amino acids, i.e., aspartic, serine, glutamic, proline, glycine, alanine and tyrosine.

Essential amino acids. Data cleared that the predominant essential amino acid in seeds of faba bean is arginine followed by leucine. In this regard, Sakha 3 followed by Missr 1 and Sakha 2 exceeded the other cultivars in arginine content, while Giza 843 recorded the minimum value under the same conditions. The nutritive value of proteins depends primarily on the capacity to satisfy body needs of nitrogen and essential amino acids (Pellet and Young, 1980). This implies that the evaluated cultivars would be a good source of essential amino acids and could be used for the fortification of cereal-based foods, which are particularly deficient in lysine. Arginine displays a very high increase in essential amino acids. The quality of dietary proteins can be measured in many ways. There is general acceptance that this value is a ratio of the available amino acids in the food diet compared with the daily requirements.

In addition, Sakha 2 followed by Sakha 3 and Sakha 4 were the highest cultivars where accumulated the highest values of leucine in seeds. In contrast, the lowest value of such content was produced from seeds of Giza 843. It is obvious from the data that methionine is present in low quantities comparing with other amino acids for all faba bean cultivars. In this respect, Sakha 3 surpassed other cultivars in methionine content. However, Sakha 4 recorded the second order. However, methi-onine was the limiting amino acids in the faba materials evaluated in the present study. The relatively low levels of methionine were also reported by other investigators Apata and Ologhobo (1994) in some Nigerian legume seeds, Algham-di (2009) in faba bean (Vicia faba L.) cultivars under various water regimes, El-Emery and Amin (2010) in some Egyptian cultivated soybean, and El-Saber (2010) in faba bean under rainfed at Maryout conditions.

Other essential amino acids, i.e., threonine, valine, isoleu-cine, phenylalanine, histidine, and lysine appeared to be decreased or increased depending on the faba bean cultivars under rainfed conditions at El-Sheikh Zuwayid. In this regard, lysine content of the faba bean cultivars is a very important nutritional attribute and probably more important than the total protein content, simply because it makes a significant supplementary protein to cereal based diets which are known to be deficient in lysine. In faba bean, an increased content of lysine might be advantageous, considering their use in mixed diets with cereals. As a result, there will be more products and a possible increase in demand of this crop in near future.

Non-essential amino acids. Data in the same tables showed that glutamic acid is the most abundant amino acid followed by aspartic acid and proline. Comparison between faba bean cul-tivars, Sakha 2 followed by Sakha 4 and Giza 3 had higher concentration of glutamic acid than other cultivars. The lowest value of such content was recorded in seeds of Sakha 3. However, the highest values of aspartic acid were achieved

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Fig. 1 Some amino acids composition in seeds of some faba bean cultivars under rainfed conditions at El-Sheikh Zuwayied.

Table 8 Total amino acids (TAA), essential amino acids (EAA), non-essential amino acids (NEAA), EAA/NEAA, EAA/TAA and

NEAA/TAA.

Faba bean cultivars TAA EAA NEAA EAA/NEAA EAA/TAA NEAA/TAA

Giza 843 120.29 48.55 71.74 67.67 40.36 59.63

Giza 3 139.98 57.72 82.26 70.16 41.23 58.76

Sakha 4 150.93 62.17 88.76 70.04 41.19 58.80

Nubaria 1 120.66 49.06 71.60 68.51 40.65 59.34

Missr 1 151.44 70.90 80.54 88.03 46.81 53.18

Sakha 2 164.77 77.148 87.63 88.03 46.82 53.18

Sakha 3 176.81 87.18 89.63 97.26 49.30 50.69

EAA/NEAA = = ratio of essential amino acids to non-essential amino acids.

EAA/TAA = ratio of essential amino acids to total amino acids.

NEAA/TAA = = ratio of non-essential amino acids to total amino acids.

by Sakha 4 followed by Sakha 2 and Sakha3, while Giza 843 recorded the lowest value as compared with the other cultivars. In this regard, Apata and Ologhobo (1994) in some Nigerian legume seeds, Alghamdi (2009) in faba bean cultivars, El-Emery and Amin (2010) in some Egyptian cultivated soybean and El-Saber (2010) in faba bean, showed that glutamic acid is the most abundant amino acid followed by aspartic acid. In this connection, the content of acidic amino acid (glutamic and aspartic acid) was mostly higher than other amino acids, possibly due to their being precursors for synthesis of most amino acids (Amer, 1989). Also, Zayed and Zeid (1998) found that contents of soluble proteins and amino acids were increased under drought stress. In this concern, Sakha 3 followed by Sakha 4 was the best cultivars for accumulated the highest values of proline. On the other hand, the lowest value of such content was produced from Sakha 2. Concerning tyrosine content, it was found in low quantities comparing with other non-essential amino acids and ranged between 2.14 mg/g for Giza 3 and 5.01 mg/g for Sakha 3. The present results indicate also that the amino acids, i.e., serine, glycine, and alanine were presented in moderate quantities. Also, these amino acids appeared to be decreased or increased depending on the concerned amino acid and studied cultivars under rainfed conditions at El-Sheikh Zuwayid. This is in agreement with values

Fig. 2 SDS-PAGE profiles of soluble proteins extracted from some faba bean cultivars under rainfed conditions at El-Sheikh Zuwayid.

obtained by Alghamdi (2009) in seeds of faba bean under various water regimes, El-Saber (2010) in seeds of faba bean under rainfed at Maryout condition and Mona et al. (2011) in green and dry seeds of faba bean under calcareous soil.

The amino acids composition of seven faba bean cultivars under investigation and its EAA/NEAA, EAA/TAA, and NEAA/TAA are introduced in Table 8. Sakha3 followed by Sakha 2 cultivars have a higher TAA, EAA, EAA/NEAA, and EAA/TAA than other faba bean cultivars. Also, Sakha 3 and Sakha 4 exceeded the other cultivars in NEAA. In this respect, Giza 843 and Nubaria 1 cultivars recorded the highest values of NEAA/TAA which reached 59.63 and 59.34, respectively. Meanwhile, the lowest value of TAA, EAA, NEAA, EAA/NEAA, and EAA/TAA was produced from Giza 843. Also, the lowest value of NEAA/TAA was achieved by Sakha 3 as compared with the other faba bean cultivars. Generally, the obtained data revealed that non-essential amino acids are more than essential amino acids in seeds of all faba bean cul-tivars under investigation.

Sodium Dodecyl Sulfate polyacrylamide Gel Electrophoresis (SDS-PAGE) of soluble proteins in seeds

SDS-PAGE technique showed that the differences in proteins profiles in seeds of faba bean cultivars under rainfed conditions illustrated in Fig. 2 and Table 9. Analysis of gel revealed that Sakha 2 and Sakha 3 were resolved into 30 bands, while Giza 843, Giza 3, Nubaria 1, and Missr 1 were resolved into 31 bands. Also, Sakha 4 was resolved into 32 bands. The molecular weight of protein sub units ranged between 14 and 95 kDa. The more intensive bands are presented at molecular mass 20, 41, and 43 kDa. In this regard, a band of molecular weight 95 kDa is not presented in the samples of Giza 843, Giza 3, Sakha 2, and Sakha 3. Also, the band of M.wt 52 kDa was disappeared in the sample of Nubaria 1, Missr 1, Sakha 2, and Sakha 3.

Regarding band intensity, it was increased at molecular masses 52 and 61 kDa for genotype Giza 843 as compared with the other faba bean cultivars. The same trend had been noticed at the polypeptide 57 kDa for genotype Giza 843 followed by

Table 9 SDS-PAGE patterns of soluble proteins extracted from some faba bean cultivars under rainfed conditions at El-Sheikh Zuwayid.

Band number Molecular weight (kDa) Band intensity

Faba bean cultivars

Giza 843 Giza 3 Sakha 4 Nubaria1 Missr1 Sakha 2 Sakha 3

1 95 0 0 1 1 1 0 0

2 93 2 2 2 2 2 2 2

3 83 2 2 2 2 2 2 2

4 78 1 1 1 1 1 1 1

5 71 2 2 2 2 2 2 2

6 66 2 2 2 2 2 2 2

7 64 3 3 4 4 4 4 3

8 61 4 3 3 3 3 3 3

9 59 4 3 3 1 2 2 3

10 57 4 3 3 1 1 1 1

11 54 4 3 4 4 4 4 4

12 52 4 3 3 0 0 0 0

13 50 1 1 1 1 1 1 1

14 49 3 3 3 3 3 3 2

15 47 1 1 1 1 1 1 1

16 45 1 1 1 1 1 1 1

17 43 5 5 5 5 5 5 5

18 41 5 5 5 5 5 5 5

19 38 4 4 4 4 4 4 4

20 36 2 2 2 2 2 2 2

21 35 2 2 2 2 2 2 2

22 34 1 1 1 1 1 1 1

23 32 1 1 1 1 1 1 1

24 30 2 2 2 2 2 2 2

25 27 2 2 2 2 2 2 2

26 25 2 2 2 2 2 2 2

27 24 2 2 2 2 2 2 2

28 20 5 5 5 5 5 5 5

29 18 1 1 1 1 1 1 1

30 17 1 1 1 1 1 1 1

31 16 3 3 3 3 3 3 3

32 14 3 3 3 3 3 3 3

Total 32 31 31 32 31 31 30 30

Where 5 = very deep intensity, 4 = deep intensity, 3 = moderate deep intensity, 2 = moderate intensity, 1 = pale intensity and 0 = no bands.

genotype Giza 3 and Sakha 4. Also, the increase in band intensity was at molecular weight 64 kDa for Sakha 4, Nubaria 1, Missr 1, and Sakha 2 compared to the other cultivars. Also, band of molecular weight 59 kDa for Giza 843 took the same trend followed by Giza 3, Sakha 4, and Sakha 3. In this regard, the band intensity for Giza 843, Sakha 4, Nubaria 1, Missr 1, Sakha 2, and Sakha 3 at molecular weight 54 kDa was increased as compared with the other genotype. In addition, there was an increase in band intensity for Giza 843, Giza 3, Sakha 4, Nubaria 1, Missr 1, and Sakha 2 at 49 kDa as compared with the other genotype. The increase in band intensities in addition to the other obtained results may be a part of metabolic adjustment modifications in response to rainfed conditions at El-Sheikh Zuwayid in addition to genetic background interaction. In this respect, El-Saber (2010) used sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) technique to separate proteins in seeds of faba bean (Nubaria 1) under rainfed at Maryout conditions. He found that total proteins were resolved into 28 bands and the molecular weight of protein sub units ranged between 10 and 205 kDa.

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