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Journal of Applied Sciences Research, 3(3): 251-262, 2007 © 2007, INSInet Publication

Interaction Effect of some Biofertilizers and Irrigation Water Regime on Mung bean (Vigna radiata) Growth and Yield Sheteawi, S.A. and Tawfik, K.M. Botany Department, Women's College, Ain Shams University, Cairo, Egypt. Abstract: The interactive effect of biogin and nitrobin biofertilizers and compost on growth, yield and metabolic products of mung bean (Vigna radiata L.) cv. Kawmy l under different irrigation water regimes of 5d, 10d & 15d drying cycle studied. Growth and yield were suppressed in plants of water regime II (10d drying cycle), while plants under regime III (15d drying cycle severely and harmfully affected. Addition of biofertilizers mitigated the harmful effect of water stress. The greatest yield/plant was obtained in plants of water regime I (5d drying cycle) treated with biogin amounting to 282% of unfertilized plants. Plants treated with biogin under regime II (10 d drying cycle) yielded 145% of unfertilized control (regime I). Biofertilized plants exhibited higher values of leaf metabolic products than nonfertilized plants. The response of leaf metabolic products to nitrobin was more marked than for the other biofertilizers. Commonly slightly higher amounts of total carbohydrates, lipid and total crude protein contents were obtained in seeds of plants irrigated every 5 d than those of 10 d drying cycle while they were more or less comparable for the biofertilized and nonfertilized plants. Plants treated with nitrobin produced the highest level of genistein (isoflavonoid) under water regime I or II and the least level of quercetin (flavonoid). Key words: Mung bean; W ater stress; Biofertilizers; Compost effect of water stress on growth and yield of a local variety (Hama 1) in northern Syria and they concluded that biomass and yield decreased and water use efficiency was reduced under water stress. Ashraf and Ibram [9 ] recorded the effect of water stress on growth and metabolic changes in Phaseolus vulgaris and Sesbania aculeate and concluded that water deficit reduced shoot mass, while free amino acids and proline (osmoprotectant) increased in all parts under drought stress. W ater stress affects almost all aspects of mung bean growth and development [4 2 ]. Musallam et al. [4 0 ,4 1 ], Ahmed [6 ] and Podlesny [4 8 ] found that the yield of faba bean decreased under drought condition while total carbohydrates in seeds decreased and protein percentage increased. Haqqani and Pandey[2 3 ] stated that mung bean suffering water stress resulted in decreased seed yield, pod number, number of seeds/pod and 1000 seed weight. The use of natural organic manure and biofertilizers are recommended by several investigators to substitute the chemical fertilizers as they improve physical and chemical properties of soil and they are the way of clean agriculture with minimum pollution effects and reduce agriculture cost[2 0 ]. Improved management of N in low N 2 soils is critical for increased land productivity and economic sustainability [5 4 ]. Their results of rainfed rotation

INTRODUCTION Mung bean (Vigna radiata L.) is a leguminous pulse crop for its use as a vegetable protein source, animal fodder and green manure, it contains isoflavonoids having estrogen and antioxidant activities that used in prevention of much diseases such as cancer, it also exhibits antimicrobial and insecticidal activities [1 3 ,2 9 ]. Isoflavonoids are found characteristically in the Viciaceae sub-family of the Fabaceas. They were found in different parts of leguminous plants. Isoflavonoids are typically stored as 7-0 glycosides, [3 3 ]. Ye et al., [6 0 ] stated that the isoflavonoids genistein and quercetin activate different stress signaling pathways. Mung bean sprouts are valuable source of vitamins and minerals. There is great need to increase our production by expansion through reclaimed areas which represent a hope of cultivated lands. De Costa et al., [1 6 ] stated that drought is a major factor limiting yield improvement of mung bean in the dry and intermediate zones of Siri Lanka, and concluded that to maximize mung bean yield, irrigation should extend across all phenological stages, specially the pod-filling stage. Leport et al., [3 4 ] showed that early water stress affected dry matter production, biomass and seed yield in chick pea (Cicer arietinum L.). Oweis et al., [4 4 ] studied the

Corresponding Author: Sheteawi, S.A., Botany Department, Women's College, Ain Shams University, Cairo, Egypt. 251

J. Appl. Sci. Res., 3(3): 251-262, 2007 experiment, conducted in Pakistan to evaluate the effect of residue retention and N- fertilizer on N 2 fixation inputs and yield of mung bean. They found that grain yield benefits of residues were 13%, also retention of residue improved the N economy of the cropping system and enhanced crop productivity. Cook et al.,[1 4 ] stated that application of organic amendment to the soil surface is widely used in order to ameliorate top soil physical conditions, specially with respect to temperature, evaporation and water content. Hati et al., [2 4 ] found that application of farmyard manure and NPK (NPK + FYM) to soybean in central India improved the organic carbon content of soil and also increased seed yield and water use efficiency by 103% and 76%, respectively, over the control. Shen & Shen [5 5 ] reported that organic material is a good source of plant nutrients and has a positive effect on improvement of the soil physical structure. Mulching benefits yield by improving soil physical conditions including improved stability in the top soil[1 7 ] . Biofertilizers are used in biological control of soil-born diseases and stimulate to great extent, plant growth by producing growth regulators [4 3 ]. Aggarwal et al.,[5 ] maintaining that crop residues and manure on pearl millet improved soil water storage, soil nutrient availability and crop yield. The objective of the present study was to assess the role of commercial biofertilizers and compost treatments on growth, yield and metabolic products of water stressed mung bean and attempt to enhance the level of isoflavonoids which is very important products.

compost (2) or 50g biogin(3) or 50g nitrobin (4). Regime II: Irrigation once every 10 days; soil moisture content range (% of field capacity) 100%- 55%. This group comprises: control (5), pot supplied with 250g compost (6) or 50g biogin (7) or 50g nitrobin (8). Regime III: Irrigation once every 15 days; soil moisture content range (% of field capacity) 100% 40%. This group comprises: control (9), pot supplied with 250g compost (10) or 50g biogin (11) or 50g nitrobin (12). Twelve plants after seedling thinning were let to grow in each pot. Three replicate pots were kept for each treatment. Plants were harvested at 30 d (vegetative growth stage), 60d (flowering stage), 90 d (pod setting stage) and after pod maturity. Three replicates were used for determination of shoot lengths, fresh and dry masses and leaf area. Yield components (no. of pods/plant, no. of seeds/plant, yield/plant and weight of 100 seed) were determined at the end of the experiment (120-150 d of sowing). Plant Analysis: Photosynthetic Pigments: It was determined according to Metzner et al., [3 8 ]. Total Soluble Carbohydrates and Amino Acids: Total soluble carbohydrate was estimated according Dubios et al., [1 9 ] and total soluble amino acids were determined according to Sadasivum and Manickam [5 1 ].

M ATERIALS AND M ETHODS Proline Content: Proline content was determined according to Bates et al.,[1 1 ].

Pot trial was conducted on mung bean (Vigna radiata L.) during summer season 2005 in sandy loam soil at the experimental garden of Botany Department, W omen's College, Ain Shams Univ. M ung bean seeds (Kawmy 1), compost and commercial biofertilizers biogin and nitrobin were obtained from Agriculture Research Centre, Ministry of Agriculture, Giza. Mung bean seeds were sterilized with 1.5% chlorox then washed with water and the mung bean okadin was added to the seeds. Seeds were planted in big pots (50 cm diameter and 30 cm depth soil surface area = 0.2 m 2 , each pot contained 30 kg soil). Characteristics of the soil. Texture. Sandy loam; sand 82%, silt 14.5%, clay 4.5%. pH 7.9, EC 0.4 dSm -1 , organic matter 61%, field capacity 20%. Three water regimes were adopted.

Seed M etabolic Products: Total Carbohydrate Content: E xtraction and determination of total carbohydrates were carried out according to Dubois [1 9 ]. Lipid Content: lipid content was determined according to Sadasivum and Manickam [5 1 ]. Total crude protein content. The total nitrogen was estimated by the standard micro-kjeldahl method [4 6 ] and was converted into protein by using the conversion factor (6.25) A.O.A.C,[8 ]. Isoflavonoids: Extracted by Adrian et al., [4 ] and estimated by HPLC according to Stephen et al.,[5 8 ].

Regime I: Irrigation every 5 days, soil moisture content range (% of field capacity) 100%-70%. This group comprises: Control (1), pot supplied with 250g

Protein Electrophoresis: SDS-polyacrylamide gel electrophoresis was performed in 10% acrylamide slab gels according to the system of Lammli[3 2 ].

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J. Appl. Sci. Res., 3(3): 251-262, 2007 Gel Analysis: Gels were photographed scanned and analyzed using Gel Doc 2000 Bio Rad system.

III flowered later and produced fewer flowers than for regimes I & II.

Statistical Analysis: The obtained data were treated statistically using the two way analysis of variance as described by Snedecor and Cochran [5 6 ]. The means were compared by L.S.D using Spss (Statistical package for social science) Version 10.

Yield and Yield Components: In the present study regime I exhibited a significant increase in yield components as compared to regime II (Fig.2). Plants of regime III gave the least yield and yield components. Plants of regime I treated with biogin produced the highest yield and yield components (282% of nonfertilized control plants). This was followed by nitrobin treatment and then compost which produced 224% and 198% yield/ plant in respect to non-fertilized control. Non-fertilized plants of water regime II produced 49% yield/ plant as compared to control (regime I). Addition of biogin increased yield/plant to 145% of control (regime I) followed by plants treated with nitrobin then compost which produced 106% & 78% yield/ plant, respectively, as compared to control (regime I). The present study indicates the beneficial effects of biofertilizer treatment in both water regime I & II. The presented results agree with those of Haqqani and Pandey [2 3 ] who postulated that water stress decreased seed yield, pod number, no. of seed/pod and 1000 seed weight of mung and they found that seed yield was positively correlated with leaf area index. Higher yield obtained with biofertilizer depends on the concept of higher supplying rate of N and P occurring as a result of N-fixation [2 1 ]. Aggrawal et al.,[5 ] concluded that crop residue and farmyard manure incorporation increased soil water content, enhanced soil fertility status (N & P availability, organic matter and enzyme activity) and increased pearl millet grain yield. W heat straw has a beneficial effect on mung bean, due to improvement of mineral nutrition and soil physical structure in China [5 5 ].

RESULTS AND DISCUSSIONS Growth Parameters: Shoot length, fresh and dry mass and leaf area/plant decreased with water stress (Fig.1). Significantly lower values of growth criteria were obtained for plants of water regime II than plants of water regime I as noticed by 60 or 90 d age. The effect of biofertilizers mitigated growth reduction due to water stress. Plants treated with biogin of water regime I exhibited maximum growth amounting to 216% & 353% of fresh and dry mass, respectively, at 90 d age as compared to control (water regime I). Plants of water regime II exhibited suppression of growth, fresh and dry mass amounting to 54% & 60% , respectively, as compared to control (water regime I). Plants of water regime II treated with biogin produced 136% & 225% of fresh and dry mass, respectively, as compared to control (regime I). These indicate that addition of biogin or nitrobin and lastly compost mitigated the harmful effect of water stress. Plants of regime III resulted in a most severe suppression of growth. Comparable results were obtained by De and Kar [1 8 ] who stated that root, shoot length and fresh weight of mung bean seedling were declined with increasing water stress. Zayed and Zeid [6 1 ] showed that growth of mung bean seedlings were significantly reduced under stress as water stress reduces nutrient uptake by roots and transport from roots to shoots because of restricted transpiration rates and impaired active transport and membrane permeability. M ahajan and Tuteja [3 7 ] concluded that a reduction in vegetative growth, leaf expansion and leaf growth occurred under water deficit condition. Masinde et al., [3 9 ] stated that maintaining soil moisture at 60% water holding capacity would be sufficient to prevent a decline in leaf expansion, stem elongation and transpiration and sustain relatively high dry matter production in pot and field experiment on two Solanum species in Kenya. Shah et al., [5 4 ] evaluated effects of plant residue and fertilizer N on N 2 fixation in pots and growth of mung bean, and showed that residue retention increased shoot biomass yields. Flowers appeared earlier in plants of water regime I than plants of water regime II. Plants of water regime

W ater Use Efficiency (WUE). It is noticed that the water use efficiency values (W UE) of biofertilized treatments in both water regimes I or II were higher than that the values of unfertilized plants (Fig. 3). This is related to the beneficial effect of interaction of biofertilizers with both water regimes. It is also noticed a marked coordination between yield and W UE values (Fig. 2 & 3). Anyia and Herzog [7 ] stated that the water deficit treatment reduced W UE by 21%. Hubick et al.,[2 7 ] assumed that an increase in W UE is likely to increase yield. Hati et al., [2 4 ] showed that addition of farmyard manure to soybean increased W UE. This agrees with the present results concerning compost and biogin and nitrobin biofertilizers. Photosynthetic Pigments: Chlorophyll a + b increased generally in plants of water regime II than plants of water regime I notably after 30 d from sowing

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Fig. 1: Shoot growth criteria of mung bean cv. Kawmy 1 under different biofertilizers and water regimes. agreement with those obtained on Vicia faba by Abd El-Gawad [1 ] . M aamon [3 5 ] and Khalafallah [3 1 ] also demonstrated that chlorophyll content increased under water stress.

(Table 1) and at time of flowering and pod setting stages. The highest content of chlorophyll a + b was recorded in plants treated with nitrobin then biogin under water regimes I & II. The present findings are in

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J. Appl. Sci. Res., 3(3): 251-262, 2007 Table 1: Photosynthetic pigm ents of m ung bean cv. Kawm y 1 under different biofertilizers and w ater regim es. Chlorophyll a Chlorophyll b Chlorophyll a+b Carotenoids --------------------------------------------------------------------------------------------------------------------30 60 90 30 60 90 30 60 90 30 60 90 Regim e I Control 19.0 22.0 23.2 11.9 19.1 20.4 30.9 41.1 43.6 10.4 9.3 10.5 --------------------------------------------------------------------------------------------------------------------------------------------------------------Com post 20.6 29.0 30.2 15.0 28.2 29.5 35.6 57.2 59.7 10.4 12.6 14.0 --------------------------------------------------------------------------------------------------------------------------------------------------------------Biogin 33.1 36.0 38.9 23.3 30.7 33.6 56.4 66.7 72.5 16.7 16.1 19.0 --------------------------------------------------------------------------------------------------------------------------------------------------------------N itrobin 23.5 36.0 38.5 16.6 31.2 33.8 40.1 67.2 72.3 11.5 14.6 18.1 --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------Regim e II Control 29.7 31 32.1 24.3 25.3 26.4 54.0 56.3 58.5 12.6 13.4 14.3 --------------------------------------------------------------------------------------------------------------------------------------------------------------Com post 32.3 34.0 35.9 25.6 27.0 29.1 57.9 61.0 65.0 13.9 14.8 16.6 --------------------------------------------------------------------------------------------------------------------------------------------------------------Biogin 34.6 40.0 42.2 26.1 32.5 34.4 60.7 72.5 76.6 15.2 17.1 19.5 --------------------------------------------------------------------------------------------------------------------------------------------------------------N itrobin 40.0 40.0 41.7 36.7 39.0 40.4 76.7 79.0 82.1 13.1 15.0 15.0 --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------LSD 1% 1.74 1.67 2.41 2.43 1.22 1.96 3.28 4.37 3.12 1.52 1.32 1.84 --------------------------------------------------------------------------------------------------------------------------------------------------------------5% 1.17 1.05 1.98 1.54 0.84 1.47 2.35 3.12 2.20 1.04 0.95 1.02

Fig. 2: Yield and yield components of mung bean c.v Kawmy 1 under different biofertilizers and water regimes. generally increased for regime I (Fig. Biofertilized plants metabolic products

Leaves M etabolic Products: Total Soluble Carbohydrates, Total Soluble Amino Acids and Proline: It is noticed that total soluble carbohydrates, total soluble amino acids and proline 255

in plants of water regime II than 4) as response to water stress. revealed higher values of these than non fertilized plants as

J. Appl. Sci. Res., 3(3): 251-262, 2007

Fig. 3: W ater use efficiency (W UE) of mung bean c.v Kawmy 1 under different biofertilizers and water regimes.

Fig. 4: Total soluble carbohydrates, total soluble amino acids and proline of mung bean leaves c.v Kawmy 1 under different biofertilizers and water regimes.

Fig. 5: Leaves N, P and K content of mung bean c.v Kawmy 1 under different biofertilizers and water regimes.

Fig. 6: Total carbohydrates, Lipids and crude protein percentages of mung bean seeds c.v Kawmy 1 under different biofertilizers and water regimes.

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J. Appl. Sci. Res., 3(3): 251-262, 2007 response to their mitigative and stimulating effect. The response to nitrobin was more marked than biogin and compost. Bota et al.,[1 2 ] investigated that plants respond quickly to prevent the photosynthetic machinery from suffering from irreversible damages. Stomatal closure in response to water deficit stress primarily results in decline in the rate of photosynthesis. Ashraf and Foolad [1 0 ] argued that there is a p o sitiv e r e la tio n s h ip b e tw e e n p r o l in e accumulation and plant stress tolerance. Kishore et al.,[3 0 ], Hsu et al.,[2 5 ] and Ozturk and Demir [4 5 ] concluded that proline is known to occur widely in higher plants and normally accumulates in large quantities in response to environmental stress. In addition to its role as an osmolyte for osmotic adjustment, proline contributes to stabilizing subcellular structure (e.g. membranes and proteins) scavenging free radicals and buffering cellular redox potential under stress conditions. Certain metabolic processes are triggered in response to stress, which increase the net solute concentration in the cell, thereby helping the movement of water into the leaf resulting in increase in leaf turgor. Large number of compounds are synthesized, which play a key role in maintaining the osmotic equilibrium and in protection of membranes as well as macromolecules. These compounds include proline and sugars [5 0 ]. It may also function as protein-compatible hydrotrope [5 7 ]. PraveenKumar et al.,[4 9 ] postulated that incorporation of crop residue and manure in soil enriched amino acid-N and amino sugar-N in soil. Enrichment of these fractions was attributed to the conversion of N biologically fixed and partly to the conversion of unknown and non-hydrolysable N fractions to amino acid-N and amino sugar-N. Soluble carbohydrate content and proline increased in response to water stress [5 2 ,4 7 ]. Ingram and Bartels [2 8 ] stated that under water stress soluble sugars can function in two ways which are difficult to separate as osmotic agent and as osmoprotectors. As an osmotic agent, the increased sugar induced by water stress was significantly c o rrelated to osmotic ad justment and turgor maintainance. As osmoprotectors sugars stabilize protein and membranes, most likely substituting the water in the formation of hydrogen bonds with polypeptide polar residue [1 5 ] and phospholipids phosphate groups [5 9 ] . The contents of soluble saccharides, soluble proteins, amino acids and proline increased under water stress. The accumulation of such organic solutes may improve the cytoplasmic osmorgulation and thus, increase plant tolerance [6 1 ].

water regime I (Fig. 5) while the level of P decreased in stressed plants. Generally NPK contents were higher by biofertilizer treatment than non fertilized plants. Biogin treatment recorded the best stimulating response of the three biofertitizers used. Mahajan and Tuteja [3 7 ] reported that K + is required for maintaining the osmotic balance and has a role in opening and closing of stomata. Also K + is an essential co-factor for many enzymes. In stressed plants large numbers of organic or inorganic ions were accumulated which provide resistance against drought [2 6 ]. In accordance with the present results Abu-Zekry[3 ] had stated that application of different biofertilizers significantly increased P content in soybean and chickpea. Osmolytes in low concentration can protect macromolecules either by stabilizing the tertiary structure of protein or by scavenging reactive oxygen species (ROS) produce in response to drought[6 2 ]. However, higher accumulation of osmolytes can cause impaired growth in the absence of any stress, probably due to plants adaptation strategy to conserve water in acute stress [3 6 ,2 ]. Seed M etabolic Products: Total Carbohydrates, Lipid C ontent and Protein Content: Commonly highest amounts of total carbohydrate, lipid and total crude protein contents were obtained in plants treated with biogin then nitrobin and irrigated every 5 & 10 d (Fig. 6). Total crude protein increased in plants of water regime II compared to those of water regime I. A similar trend was obtained by Ahmed [6 ] who found that drought increased protein percentage. Plants under water regime I treated with biogin recorded the highest levels of NPK followed by plants treated with nitrobin then compost (Fig. 7). Plants under water regime II treated with nitrobin recorded the highest level of P and K. Isoflavonoids: Plants raised at water regime I r e c o r d e d th e h ig h e s t a m o u n t o f g e n i s t e i n (isoflavonoid) and quercetin (flavonoid) (Fig. 8). Plants treated with nitrobin produced the highest level of genistein under water regime I & II followed by plants treated with biogin, while plants treated with nitrobin produced the least level of quercetin. The isoflavonoid genistein and its' derivatives are the most abundant group in mung bean seeds [3 3 ]. Seneviratne and Harborne [5 3 ] made a survey of phytoalexine production in eight species and three subspecies of Vigna and showed that Vigna radiate contains quercetin glycosides. From the present results it is clear that addition of biofertilizers decreased the level

M ineral Contents: The level of N and K were higher in stressed plants (water regime II) than for

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Fig. 7: N, P and K of mung bean seeds c.v Kawmy 1 under different biofertilizers and water regimes.

Fig. 8: Isoflavonoids of mung bean seeds c.v Kawmy 1 under different biofertilizers and water regimes.

Fig. 9: SDS-PAGE Protein banding pattern of mung bean seeds (cv. Kawmy 1) under different biofertilizers and water regimes. of quercetin which may be desirable since quercetin causes decrease in cell proliferation and inhibit estrogen binding. The inhibition of binding is much

stronger in flavones isoflavones (genistein) [2 2 ].

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(quercetin)

than

in

the

J. Appl. Sci. Res., 3(3): 251-262, 2007 Table 2: Presence (+) and absence (-) of SD S-PA GE Protein banding pattern in seeds of m ung bean (cv. Kawm y 1) under different biofertilizers and w ater regim es. Band no. Treatm ent Regim e I Regim e II ----------------------------------------------------------------------------------------------------------------------------------------K. D a Control Com post Biogin N itrobin Control Com post Biogin N itrobin 1 136.51 + + + + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------2 116.00 + + + + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------3 104.28 + + + + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------4 95.49 + + + + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------5 83.77 + + + + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------6 77.91 + + + + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------7 66.20 + + + + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------8 63.08 + + + + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------9 60.58 + + + + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------10 56.22 + + + + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------11 52.48 + + + + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------12 45.00 + + + + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------13 43.90 + + + + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------14 42.45 + + + + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------15 40.45 + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------16 39.00 + + + + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------17 35.15 + + + + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------18 35.00 + + + + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------19 33.80 + + + + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------20 33.00 + + + + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------21 31.40 + + + + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------22 30.20 + + + + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------23 27.00 + + + + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------24 24.16 + + + + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------25 22.21 + + + + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------26 20.80 + + + + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------27 18.76 + + + + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------28 18.40 + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------29 16.27 + + + + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------30 15.27 + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------31 12.27 + + + + + + + + --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------Total 31 31 31 31 31 28 28 28 29

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J. Appl. Sci. Res., 3(3): 251-262, 2007 SDS-PAGE Protein Analysis. Fig. 9 and Table 2 demonstrated the SDS PAGE protein profile of mung bean cv. Kawmy 1 under different treatments. Low level of protein polymorphism (9.6%) was detected. Thirty one bands were scored in the seed protein p r o file , twen ty eig h t b a n d s o f th e m we re mononorphic, while the remaining three bands were considered as polymorphic ones and were recorded at molecular weights 40.45, 18.40 and 15.27 K.Da. These bands distinguished the first four treatments (water regime I) from the others.

9.

10.

11.

Conclusion: It could be concluded that biofertilizers biogin, nitrobin or compost stimulated plant growth and yield and induced drought tolerance by enhancing the accumulation of certain metabolites i.e. sugars, amino acids, proline and protein. These three biofertilizers promote the production of the medically valuable isoflavonoid genistein.

12.

13.

14. REFERENCES 1.

2.

3.

4.

5.

6.

7.

8.

Abd-El-Gawad, Z.A., 2006. Ecophysiological and molecular genetic studies and some Vicia faba L. genotype. Ph. D. Thesis, Botany Department, Fac. Girls, Ain Shims Univ. Abebe, T., A.C. Guenzi, B. Martin, J.C. C hushm an, 20 0 3 . T o lerance of mannitolaccumulating transgenic wheat to water stress and salinity, Plant Physiol., 131: 1748-1755. Abu-Zekry, S.H., 2000. Effect of Biological Fertilization on the Productivity of some Cereals and Legumes. Ph.D. Thesis, Ain Shams Univ. Adrian, A., L.J. Franke, C.M.C. Custer and K.K. Norala, 1994. Quantitation of phytoestrogens in legumes by HPLC. J.F. Agric. Food Chem., 42: 1905-1913. Aggrawal, R.K., Praveen-Kumar and J.F. Power, 1997. Use of crop residue and manure to conserve water and enhance nutrient availability and pearl millet yields in an arid tropical region. Soil & Tillage Research, 41: 43-51. Ahmed, E.A.L., 2004. Yield and seed quality of faba bean as affected by some environmental factors. Ph.D. Thesis, Department of Agronomy, Faculty of Agriculture, Cairo University. Anyia, A.O. and H. Herzog, 2004. W ater use efficiency, leaf area and leaf gas exchange of cowpeas under mid-season drought. Europ. J. Agronomy, 20: 327-339. A.O .A.C., 1975. Association of O fficial Agricultural Chemists. O fficial methods of analysis 12 th ed. Washington, D.C., USA.

15. 16.

17.

18.

19.

20.

21.

260

Ashraf, M. and A. Ibram, 2005. Drought stress induced changes in some organic substances in nodules and other plant parts of two potential legumes differing in salt tolerance. Flora, 200: 535-546. Ashraf, M. and M.R. Foolad, 2006. Roles of glycine betaine and proline in improving plant abiotic stress resistance. Environmental and Experimental B otany. Available on line at www.sciencedirect.com. Bates, L.S., R.P. W aldren and I.D. Teare, 1973. Rapid determination of free proline for waterstress studies. Plant and Soil, 39: 205-207. Bota, J., J. Flexas, H. Medrano, 2004. Is photosynthesis limited by decreased Rubisco activity and RuBP content under progressive water stress? New Phytol., 162: 671-681. Brounce fred, 2002. Soya is flavones: A new and promising ingredient for the health foods sector. Food research international, 35: 187-193. Cook, H.F., G.S. Valdes and H.C. Lee, 2006. Mulch effects on rainfall interception, soil physical characteristics and temperature under Zea mays L. Soil & Tillage Research. Available on line at www.sciencedirect.com. Crowe, J.H., F.A. Hoekstra, L.M. Crowe, 1992. Anhydrobiosis. Ann. Rev. Physiol., 54: 579-599. De Costa, W .A., K.N. Shanmigathasan and K.D. Joseph, 1999. Physiology of yield determination of mung bean (Vigna radiata L.) under various irrigation regimes in the dry and intermediate Zones of Sri Lanka. Field Crops Research, 61: 1-12. De Silva, S.H.S.A., H.F. Cook, 2003. Soil physical conditions and performance of cowpea following organic matter amelioration of sand. Commun. Soil Sci. Plant Anal., 34(7/8): 10391058. De, R. and R.K. Kar, 1995. Seed germination and seedling growth of mung bean (Vigna radiata) under water stress induced by PEG-6000. Seed Science and Technology, 23(2): 301-308. Dubois, M., K.A Gilles, J. Hamilton, R. Rebers, and F. Smith, 1956. Colorimetric method for determination of sugars and related substances. Anal. Chem. 28(3): 350-356. El-Akabawy, M.A., 2000. Effect of some biofertilizers and farmyard manure on yield and nutrient uptake of Egyptian clover Grown on loamy sand soil. Egypt. J. Agric. Res., 78(5): 1811-1819. El-Emam, M.A.A., 1999. Studies on nutrients availability from plant residues and different organic fertilizers. M. Sc. Thesis, Fac. Agric., Moshtohor, Zagazig Univ., Egypt.

J. Appl. Sci. Res., 3(3): 251-262, 2007 22. Eva Oberdörster, M.A. Clay, D.M. Cottam, F.A. W ilmot, J.A. Mclachlan and M.J. Milner, 2001. Common phytochemicals are ecdysteroid agonists and antagonists: a possible evolutionary link between vertebrate and invertebrate steroid hormones. Journal of Steroid Biochemistry & Molecular Biology, 77:229-238. 23. Haqqani, A.M . and R.K. Pandey, 1994. Response of mung bean to water stress and irrigation at various growth stages and plant densities: II. Yield and yield components. Tropical Agriculture, 71(4): 289-294. 24. Hati, K.M., K .G. Mandal, A.K. Misra, P.K. Ghosh and K.K. Bandyopadhyay, 2006. Effect of inorganic fertilizer and farmyard manure on soil physical properties, root distribution and water-use effeciecy of soybean in Vertisois of central India. Bioresource Technology, 97: 2182-2188. 25. Hsu, Z., Y.T. Hsu and C.H. Kao, 2003. The effect of po lyethelene glyco l on proline accumulation in rice leaves. Biol. Plant, 46: 7378. 26. Hoekstra, F.A., E.A. Golovina, J. Buitink, 2001. Mechanisms of plant desiccation tolerance. Trends Plant Sci., 6: 431-438. 27. Hubick, K.T., G.D. Farquhar and R. Shorter, 1986. Correlation between water use efficiency and carbon isotope discrimination in diverse peanut (Arachis) germplasm. Aust. J. Plant Physiol., 13: 803-816. 28. Ingram, J., D. Bartels, 1996. The molecular basis of dehydration tolerance in plants. Ann. Rev. Plant Physiol. Plant Mol. Biol., 47: 377-403. 29. Kaprelynts, L.V., S.V. Kisilev and E.G. Iorgachova, 2003. Soybean isoflavones and prospects of their therapeutic application. Voprosy Pitaniya, 72: 36-41. 30. Kishore, P.B.K., S. Sangam, R.N. Amrutha, P.S. Laxmi, K.B. Naidu, K.R.S.S. Bao, S. Rao, K.J. Reddy, P. Theriappan and N. Sreenivasula, 2005. Regulation of proline biosynthesis, degradation, uptake and transport in higher plants: its implications in plant growth and abiotic stress tolerance. Curr. Sci., 88: 424-438. 31. Khalafallah, A.A.M., 1997. Effect of some antitranspirents on the growth and yield of wheat plants under limited irrigation. M. Sc. Thesis. Botany department, Fac. Girls, Ain Shams University. 32. Laemmli, U.K., 1970. Cleavage of structural proteins during assembly of head bacteriophage T4. Nature, 227: 680-685.

33. Lapcik,O., M. Hill, I. Cerny, J. Lachman, N. AlMaharik, H. Adlercreutz and R. Hampl, 1999. Immuno-analysis of isoflavonoids in Pisun sativum and Vigna rediata. Plant Science, 148: 111-119. 34. Leport, L., N.C. Turner, S.L. Davies and K.H.M. Siddique, 2006. Variation in pod production and abortion among chickpea cultivars under terminal drought. Europ. J. Agronomy, 24: 236-246 35. Maamon, H.A., 1994. Physiological responses of certain forage crops to drought. M. Sc. Thesis Agron. Department, Fac. Agric., Ain Shams Univ. 36. Maggio, A., S. Miyazaki, P. Veronese, T. Fujita, J.I. Ibeas, B. Damsz, M .L. Narasimhan, P.M. Hasegawa, R.J. Joly, R.A. Bressan, 2002. Does proline accumulation play an active role in stressinduced growth reduction? Plant J., 31: 699-712. 37. Mahajan, S. and N. Tuteja, 2005. Cold, satinity and drought stresses. Archives of Biochemisty and Biophysics, 444: 139.158 38. Metzner, H., H. Rau and H. Senger, 1965. U n te rs u c hunge n Zur Sync hronis ie r-B a r k e i t einzelner pigment mangel-Mutantenvon Chlorella. Planta, 65: 186. 39. Masinde, P.W ., H . Stützel, S.G. Agong and A. Fricke, 2006. Plant growth, water relations and transpiration of two species of African nightshade (Solanum villosum Mill and S, sarrachoids) under water-limited conditions. Scientia Horticulurae, 110: 7-15. 40. Musallam, I.W ., G. Al-Karaki, K. Ereifej and A.R. Al-Tawaha, 2004a. Yield and yield components of faba bean genotypes under rainfed and irrigation conditions. Asian Journal of Plant Sciences, 3(4): 439-448. 41. Musallam, I.W ., G. Al-Karaki, K. Ereifej and A.R. Al-Tawaha, 2004b. Chemical composition of faba bean genotypes under rainfed and irrigation conditions. International Journal of Agriculture and Biology, 6(2): 369-362. 42. Nandwal, A.S., A. Hooda and D. Datta, 1998. Effect of substrate moisture and potassium on water relations and C, N and K distribution in Vigna radiata. Biologia Plantarum, 41(1): 149153. 43. O'Sullivan, D.J. and F. O'Gara, 1992. Traits of fluo rescent P seudom onas sp . Involved in suppression of plant root pathogens. Microbiol Rev., 56: 662-676. 44. Oweis, T., A. Hachum and M. Pala, 2005. Faba bean productivity under rainfed and supplemental irrigation in northern Syria. Agricultural W ater Management 73: 57-72.

261

J. Appl. Sci. Res., 3(3): 251-262, 2007 45. Ozturk, L., Y . Demir, 2002 In vivo and vitro protective role of proline. Plant growth regul., 38: 259-264. 46. Peach, K. and M.V. Tracey, 1956. Modern methods of plant analysis. Vol. Springer-Verlag, Berlin. 47. Pinheiro, C., M.M. Chaves, C.P. Ricardo, 2001. Alterations in carbon and nitrogen metabolism induced by water deficit in the stems and leaves of lupinus albus L., J. Exp. Bot., 52: 1063-1070. 48. Podlesny, J., 2001. The effect of drought on the development and yielding of two different varieties of the fodder broad bean (Vicia faba minor). Journal-of Applied-Genetics, 42(3): 283287. 49. Praveen-Kumar, K.P. Tripathi and R.K . Aggrawal, 2002. Influence of crops, crop residues and manure on amino acid and amino sugar fractions of organic nitrogen in soil. B iology & Fertility of soils, 35(3): 210-213. 50. Ramanjulu, S., D. Bartels, 2002. Drought and d e si c c a t io n in d u ced m o d u latio n of ge ne expression in plants. Plant Cell Environ., 25: 141-151. 51. Sadasivam, S. and A. Manickam, 1996. Biochemical Methods. New Agr. International (P) Limited, Publishers, London, pp 23. 52. Sánchez, F.J., M. Manzanes, E.F. De Andres, J.L. Tenorio and L. Ayerbe, 1998. Turgor maintainance, osmotic adjustment and soluble sugar and proline accumulation in 49 pea cultivars in response to water stress. Field Crops Research, 59: 225-235. 53. Seneviratne, G.I. and J.B. Harborne, 1992. Constitutive flavonoids and induced isoflavonoids as taxonomic markers in the genus Vigna. Biochemical Systematics & Ecology, 20(5): 459-467. 54. Shah, Z., S.H. Shah, M.B. Peaples, G.D. Schwenke and D.F. Herridge, 2003. Crop residue and fertilizer N effects on N fixation and yields of legume-cereal rotations and soil organic fertility. Field Crops Research, 83(1): 1-11.

55. Shen, Q.R. and Z.G. Shen, 2001. Effects of pig manure and wheat straw on growth of mung bean seedlings grown in aluminium toxicity soil. Bioresource Technology, 76: 235-240. 56. Snedecor, S.C. and W .C. Cockran, 1969. Statistical Methods 6 th Ed.lowa Univ. Press. Ames. Iowa U.S.A. 57. Srinivas, V., D. Balasubramanian, 1995. Proline is a protein-compatible hydrotrope.Langmuir, 11: 2830-2833. 58. Stephen, M. Boue, Thomas, E. W iesf, Suzanne, Nehls, Matthew E. Burow, Steven Elliott, Carol, H. Cater-W ientjes, Betty Y. Shih, John A. Mclachlan and Thomas E. Cleveland, 2003. Evaluation of the estrogenic effects of legume extracts containing phytoestrogens. J. Agric. Food Chem., 51: 2193-2199. 59. Strauss, G., H. Hauser, 1986. Stabilization of lipid bilayer vesicles by sucrose during freezing. Proc. Natl. Acad. Sci. U.S.A., 83: 2422-2426. 60. Ye, R., A.A. Goodarzi, E.U. Kurz, S. Saito, Y. Higashimoto, M.F. Lavin, E. Appella, C.W . Anderson and S.P. Lees-Miller, 2004. The isoflavonoids genistein and quercetin activate different stress signaling pathways as shown by analysis of site-specific phosphorylation of ATM, p53 and histone H2AX. DNA Repair, 3: 235244. 61. Zayed, M.A. and Zeid, I.M. (1998): Effect of water and salt stresses on growth, chlorophyll, mineral ions and organic solutes contents and enzymes activity in mung bean seedlings. Biologia Plantarum, 40(3): 351-356. 62. Zhu, J.K., 2001. Plant Salt Tolerance. Trends Plant Sci., 6: 66-71.

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