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Evaluation of yield and physicochemical properties of some lupin (Lupinus sp.) mutants induced by gamma radiation Aahmed, A. Alya., Nasr, E. Hb., Shaheen, A. Mb and Essam, M., Sallamb


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Evaluation of yield and physicochemical properties of some lupin (Lupinus sp.) mutants induced by gamma radiation

Aahmed, A. Alya ., Nasr, E.Hb., Shaheen, A. Mb. and Essam, M., Sallamb.

­­ a Home Economic Department, Faculty of Specific Education, Benha University, Egypt

bAtomic Energy Authority, Nuclear Research Center, P.No. 13759, Egypt

ABSTRACT

The objective of the present investigation was conducted to characterize physical, chemical and functional properties as well as amino acids composition of defatted lupin (Lupinus sp.) from control and mutant varieties grown in the Egyptian Atomic Energy Authority which produced by radiation mutation breeding program. Also, the functional properties of these samples were determined. The results illustrated that a highly significant differences between genotypes in plant height, stem diameter, seed yield and weight than the local two variety. In addition, radiation mutation of lupin seeds showed slight increased in protein content as the main constituent of lupin seeds, as well as total oil percentages in some mutant of these seeds in return of decreasing in total carbohydrate. Furthermore, radiation mutation had detectable effects on the total amino acids contents of lupin seeds meal which had a higher percentages on essential amino acids (E.A.A) and nonessential amino acids (N.E.A.A) and mut.3 was the highest values on E.A.A and N.E.A.A as compared to the control one. On the other hand, radiation mutation improved the protein functional properties of some mutant of lupin meal flour than the other mutant samples as compared to local commercial varieties.

Key words: lupin (Lupinus sp.), Amino acids, Protein function and radiation mutation.

INTRODUCTION

Legume seeds are second only to cereal as a source of human food and animal feed and they are 2–3 times richer in protein than cereal grains. With the increase in population expected in developing countries, It is likely that many people will not be able to afford buying animal products regularly. Legume will therefore continue to play an important dietetic part in the future. Legumes consumed by humans in many forms are excellent sources of proteins (20–40%), carbohydrates (50–60%) and are fairly good sources of thiamin, niacin, calcium and iron (Aykroyd, et al 1982).

Legumes constitute a large family of plants, many of them cultivated, such as bean (Phaseolus vulgaris), chickpea (Cicer arietinum), lentil (Lens culinaris), soya bean (Glycine max), pea (Pisum sativum), or lupin (Lupinus sp.). Nutritional value of these plants is related to the high protein, mineral and vitamin content of the seeds (Cubero & Moreno, 1983; Hebblethwaite, 1983). Among legumes, the use of lupin in human foods and livestock is limited, mainly due to the presence of toxic alkaloids in the seed. In this sense, the production of lupin protein isolates may overcome this problem because alkaloids are water soluble and removed during the preparation of protein isolates (Millan et al., 1995)

Lupin (Lupinus albus, L.) is a species of the genus Lupinus (more than 200 species) in the family Leguminosae. The species of the genus Lupinus are distributed in two centres of origin. One is the mediterranean basin and the other extends through South America (Dervas, et al., 1999; Huyghe, 1997& Swiecicki, et al., 2000). The major cultivated species of lupins are Lupinus albus, L. (white lupin), Lupinus angustifolius, L. (blue lupin), Lupinus luteus, L. (yellow lupin) and Lupinus mutabulies (pearl lupin). The first three species originate in the Mediterranean area while Lupinus mutabulies belongs to South America (Allen, 1998 & Mulayim, et al., 2002). Lupins are cultivated for three main reasons: as a ruminant feed, as a green manure contributing to improved soil structure, and for human nutrition because of their high protein and oil contents (Faluyi et al., 2000).

In addition, lupin plants are grown for use as a cut flower in the flowering period. Seeds of white lupin have a protein content ranging from 33% to 47%, according to genotype and location contrary to cereals, lupin proteins contain a high amount of lysine and a low amount of sulphur-containing amino acids (Dervas et al., 1999). Oil content varies from 6% to 13% with a high concentration of polyunsaturated fatty acid (Huyghe, 1997).

Lupin flour can be used in production of different fermented products. It can be added to pasta, crisps, bread and emulsified meat products to increase nutritional value and aroma and to modify texture. Also, protein isolate can be produced from lupin seeds. In the Middle East, lupins. In some European countries, pickle is produced from lupin seeds (Akyıldız, 1969; Papavergou, et al, 1999; Petterson, 1998; Vasilakis & Doxastakis, 1999). White lupin, which has been consumed as a food in a narrow area for a long time, was accepted for human consumption by the Australian government in 1987 and by the United Kingdom government in 1996 (Cox, 1998; Swam, 2000). 1,387,660 tons of lupin were produced in the world in 2001. Australia, which produced 89% of this amount, is the largest lupin-producing country. The other important lupin producers are Poland, France and South America (FAOSTAT, 2001).

Lupin species as L albus and L mutabilies are used as human food because they have high protein content and nutritive value (Hill, 1977). Lupin crop as also used for green maturing and medical purpose.

In recent years, useful lupine mutants induced by mutagenic agents such as high protein and free of low alkaloid forms were selected by several workers (Ripa and Orbidance, 1975 and Debelia et al., 1989)

In Egypt more effort could be done to improve yield potential, increase the protein content and or the minimize the alkaloid substances in the local lupin germ plasm. To achieve these goals, it is essential to broaden the genetic variability in the local cultivars, which mainly suffer from the narrow genetic variability. Physical and chemical mutagens have been widely used to enlarge the genetic variability of various crop plants.

It is estimated that during storage the losses of leguminous seeds are at least 10%. A modern method of food conservation is irradiation. Until now this method has been used in 54 countries for preservation of over 224 food products. The FAO and WHO published data showing that an irradiation dose up to 10 kGy was safe for human health and did not cause adverse effects on nutrition’s in food processing.

Treatment by gamma irradiation had a great value for improving agronomic characters of economic value (i.e. yield lodging resistance, disease resistance, maturity, winter hardiness, shattering resistance, ease of harvesting, seed weight, sprouting resistance, drought resistance, salinity resistance and adaptability), radiation mutation breeding program is being carried out at Plant Research Department, Radiation applications Division, Nuclear Research Center, Inshas, Atomic Energy Authority, Egypt. This program aims to improve cultivate of protein crops, among them lupin seeds.

Therefore, the objective of the present investigation was to evaluate the yield, physical, chemical and functional properties as well as amino acid composition of meal after oil extracted from control and radiation mutant of the lupin seeds (L. albus L.) grown and produced by radiation mutation breeding program at the Egyptian Atomic Energy Authority



Materials and Methods:

Materials:

Source of seeds

The commercial varieties of lupin (Lupinus sp.). Giza 1 and Giza2 seeds were obtained from Agriculture Research Center, Giza, Egypt. While the mutants lupin seeds were obtained from Plant Breeding Unit,. Plant Research Department, Radiation Applications Division, Nuclear Research Center, Inshas, Atomic Energy Authority Egypt. All seeds undertaken were carefully cleaned and tightly packed polyethylene bags and kept at room temperature until analysis.



Mutants seeds

Seeds of the lupin Giza1 and Giza2 were irradiated with gamma ray doses of 40, 80, 120 and 160 Gy and planted in an isolated field till Mut.3 generation where 40 mutant plants were selected and starting from Mut.2 generation. The mutants comprised as plant height, head diameter and seeds color and size. Those mutant characters studied in Mut.3 and the next generation.. The forty mutants lines were grouped into 12 mutant types where their characters of seeds were evaluated in the next generation . Ten mutant were selected and evaluated in this study and their characteristics are presented in table (1) in comparison with the local variety.



Methods:

Chemical analysis of seeds

Moisture, oil, protein and ash contents were determined according to the official methods described by the A.O.A.C. (1995).

Total carbohydrates, were calculated by difference according to the Egan et al., (1981) as follows: Total carbohydrates % = 100 - (% moisture + % crude protein + % total lipids + % ash).

Preparation of lupin seed meals

lupin meals which remained after oil extraction from all samples undertaken were desolventized at 60 oC under vacuum for two hr., tightly packed in polyethylene bags and kept at room temperature for further analysis.



Oil extraction

Seeds samples were ground using stainless steel mill. Then the oil was extracted by n-hexane using 2 liters capacity. Soxhelt apparatus units for 16 hr. After oil extraction, the solvent was evaporated under vacuum at 60 0C and the crude oil was dried over anhydrous sodium sulfate, filtered, packed in dark brown bottles without further purification and kept till analysis, according to A. O. C. S. (1989).



Quantitative determination of the total amino acids content except tryptophan:

Amino acids contents of control and mutant lupin meals were determined according the method described by Pellet and Young, (1980) which could be summarized as follows: A known weight of each sample containing 100 mg protein was hydrolyzed in sealed pyrex test tubes with 10 ml of 6 N HCL at 110oC for 24 hours. The hydrolyzate was quantitatively transferred to porcelain dish and the (HCL) was then evaporated to dryness at 60oC under vacuum. Five ml of distilled water was added to the hydrolyzate and then evaporated to dryness to remove the excess of HCL. The separation of amino acids was performed by (Auto Sampler Version, Analyzer, Biochrom 20 pharmacia biotech) at National Center for Radiation research and Teachnology (N.C.R.R.T), Nasr City, Cairo Egypt .



Protein function of lupin seed meals

Emulsifying and foaming properties i.e emulsion capacity, emulsion stability, foam capacity and stability were determined according to the method described by Okazie and Bello, (1988)



RESULTS AND DISCUSSION

Mean value of yield and its components for the mutants lines and the local commercial varieties are shown in table (1)

Number and weight of pods per plant:

The analysis of variance of number and weight of pods / plant (table, 1) showed a highly significant differences among the mutant lines 2,3 and 4 in comparison with the local variety G1 in the two seasons, except for mutant lines 3 and 4 for number of pods/plant in the second season the increase did not reached the level of significant in comparison with the local variety G1. mutant 2 produced the highest mean values in these characters.

AS for G2 mutant lines, the mutant 3 and 4 caused significant increase in weight op pods in comparison with the local variety G2 in the two seasons, But the increase in weight of pods/plant for mutants 1 and 2 did not reached the level of significant in comparison with G2 variety. On the other hand, it has no any significant differences for all mutant lines1, 2, 3 and 4 in comparison with the local variety G2 for number of pods/plant in the two seasons.

Weight of seeds /plant gm:

Weight of seeds /plant increased significantly for mutants 2,3 and 4 in comparison with the respective control in the two season for the two genotyes. These results are in agreement with those obtained by Atia and Shahine, (1996) and Shahine and Atia, (2002).



From these results, it could be summarized that the mutant lines 2,3 and4 generally indicated that the seed yield of those mutant exceeded that of the original parent by over 30% and well be evaluated under some agricultural.

Table (1): Mean values of the studied characters for the four lupin mutants and their commercial varsities.




No. of pods / plant

Weight of pods / plant in gm.

Weight of seeds /plant in gm.

2009

2010

2009

2010

2009

2010

Giza 1

10.77

11.13

18.40

16.51

12.72

12.30

Mut.1

11.70

11.50

18.96

16.90

12.88

12.41

2

16.10

14.77

22.60

25.5

15.97

18.70

3

14.46

11.47

23.26

19.6

16.81

13.87

4

12.46

12.56

22.61

22.67

16.10

15.27

LSD 5%

1.50

1.62

2.81

2.84

1.68

1.54

Giza 1

11.90

12.40

21.08

21.83

14.80

15.87

Mut.1

11.99

13.10

21.80

20.40

15.30

16.90

2

11.92

13.09

22.30

23.40

16.52

17.80

3

12.28

13.23

23.94

26.77

18.03

20.20

4

12.68

13.84

26.53

27.30

18.41

20.13

LSD 5%

N.S

N.S

2.4

2.4

1.8

1.72


Effect of radiation mutation on the chemical composition of lupin seeds.

The effects of mutation with gamma irradiation of control and mutant lupin seeds on the gross composition (moisture, total protein, oil content, total carbohydrates and ash contents) were determined and the obtained results are presented in table (2).

From data in Table (2), it could be noticed that the moisture, crude oil, ash content, total protein and total carbohydrates contents were 0.77, 6.41, 3.65, 45.25 and 43.69 % for Giza1 (G1) and 0.76, 6.85, 3.63, 45.68 and 43.84 % for Giza2 (G2) lupin seeds, respectively. These results are in agreement with those obtained by Jos Manuel and Arminda Martins (1997); Agnieszka Sujak et al., (2006), Elena Pasto et al., (2009), Lqari, et al., (2011) and Aleksander Siger , et al. (2012).

The same table illustrates that moisture and ash contents did not change by radiation mutation of lupin seeds. On the other hand, a noticeable increase were observed in oil and protein content of mutants Mut2 and Mut4 for G1. The same phenomena was observed in mutants Mut2, Mut.3 and Mut4 for G2 lupin seeds. On the contrary, total carbohydrate content decreased in mutants for G1 and G2 by radiation mutation of lupin seeds. These results agreed with those obtained by Abu-Heggazi,et al., (1996), Fernandez-Martinez, et al., (1997) and Ali (2003) who found that radiation mutation of lupin seeds increased oil content of these seeds.



Generally, it could be concluded that, radiation mutation of lupin seeds markedly increased oil content, as well as total protein of these seeds in return of decreasing in total carbohydrates.

Table (2) Effect of radiation mutation on the chemical composition of lupin seeds

Components (%)

Control


Mutants lupin seed (G1)

Control

Mutants lupin seed (G2)

Mut.1

Mut.2

Mut.3

Mut.4




Mut.1

Mut.2

Mut.3

Mut.4

Moisture

0.77

0.72

0.81

0.83

0.75

0.76

0.78

0.71

0.79

0.82

Total oil *

6.41

6.54

7.32

6.73

7.14

6.85

6.88

7.43

7.41

7.87

Crude protein*

45.25

45.79

46.12

45.94

46.35

45.68

45.87

46.09

46.38

46.57

Total ash *

3.65

3.67

3.72

3.62

3.70

3.63

3.75

3.67

3.72

3.65

Total carbohydrates*

44.69

44.00

42.84

43.71

42.81

43.84

43.50

42.90

42.49

41.91
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