Effects of Rhizobium inoculation, organic and chemical fertilizers on ...

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The results showed that sulphur, nitrogen and chicken manure treatments significantly (p ≥ 0.05) increased yield, ... Generally, fertilization of faba bean with nitrogen, sulphur or chicken manure not only increased plant growth ... Switch Edition.
Plant Foods for Human Nutrition 51: 137–144, 1997. c 1997 Kluwer Academic Publishers. Printed in the Netherlands.

Effects of Rhizobium inoculation, organic and chemical fertilizers on yield and physical properties of faba bean seeds E.A.E. ELSHEIKH and A.A. ELZIDANY Department of Biochemistry and Soil Science, Faculty of Agriculture, Shambat, Sudan Received 23 October 1995; accepted in revised form 14 March 1997

Abstract. A field experiment was carried out to investigate the effect of Rhizobium inoculation, sulphur, nitrogen and chicken manure on yield, 100-seed weight, cookability, non-soakers, total defects and hydration coefficient of faba bean. The results showed that sulphur, nitrogen 0:05) increased yield, 100-seed weight, and chicken manure treatments significantly (p non-soakers, and hydration coefficient, in the absence of Rhizobium inoculation. The results 0:05) increased yield, 100-seed also showed that Rhizobium inoculation significantly (p weight, cookability, but decreased non-soakers. A positive correlation (r 0:90) was observed between the non-soaker percent and the total defect percent. No correlation was found between non-soakers, hydration coefficient and cookability. The results of this investigation indicate that Rhizobium inoculation is a promising fertilizer because it is cheap, easy to handle and improves plant growth and seed quality. The efficiency of inoculation could be improved with the addition of biological, chemical or organic fertilizers. Generally, fertilization of faba bean with nitrogen, sulphur or chicken manure not only increased plant growth and yield, but also improved seed quality and nutritional value.

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Key words: Cookability, Faba bean, Fertilizers, Hard-seed, Hydration coefficient Rhizobium

Introduction Faba bean (Vicia faba, L.) is one of the major leguminous crops grown in the world. It is an important source of protein for humans and animals. In Sudan, the faba bean is mainly grown for human consumption. The incidence of hard seed adversely affects cooking quality and, therefore, the market value of the products [1]. The popularity of faba bean may be due to its high protein content and availability at relatively reasonable prices. Chemical and physical properties of the seeds are indicators of its quality and nutritional value. Vigorous efforts are directed to improve yield, protein quality, cookability and to decrease tannin and hard seed percentages through breeding, fertilizers and/or genetic engineering programs [1–2]. Biofertilization receives great attention because of its minimal effect on the environment and its longerlasting effect. Biofertilizers are very important for countries like Sudan with a predominantly low-input agricultural system of production where chemical fertilizers, if available, may not be affordable [3]. Inoculation of faba bean

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138 has been found to increase the shoot and root dry weights, number of nodules per plant, as well as yield and yield components [2–4]. Undoubtedly nitrogen, sulphur, phosphorus and other fertilizers are very important factors in increasing yield of faba bean [1]. Therefore, the objective of this investigation was to study the effect of three groups of fertilizers: organic (chicken manure), chemical (nitrogen and sulphur) and biofertilizers (Rhizobium), on yield, 100-seed weight and physical properties of seeds such as non-soakers (hard seeds), hydration coefficient and cookability of faba bean cultivar Shambat 616. Materials and methods Seed and Rhizobium strain. Seeds of the faba bean cultivar Shambat 616 were supplied by Agricultural Research Corporation, Shambat, Sudan. Rhizobium leguminosarum viceae strain TAL 1400 was supplied by NifTAL Project, Paia, Hawaii, USA. The strain was maintained at 4  C on yeast extraction mannitol agar (YEMA) slopes. Field experiment. The experiment was carried out at the Demonstration Farm of the Faculty of Agriculture, Shambat, University of Khartoum (latitude 15 400 N and longitude 32 320 E) during the 1993/1994 growing season in a factorial design with four replicates. The land was prepared by deep ploughing, harrowing and leveling. Then the area was ridged and divided into 5  4 m plots. Treatments used were: 1. 2. 3. 4. 5. 6. 7. 8. 9.

Control. 25 kg S/ha (elemental sulphur). 50 kg S/ha (elemental sulphur). 100 kg S/ha (elemental sulphur). 40 kg N/ha (urea). 80 kg N/ha (urea). 3 ton chicken manure/ha (M/ha). 9 ton chicken manure/ha (M/ha). 15 ton chicken manure/ha (M/ha).

Each of these treatments was either inoculated or uninoculated with Rhizobium strain. Elemental sulphur and chicken manure treatments were applied 3 weeks before sowing to minimize the harmful effect of chicken manure and to give enough time for sulphur to react in the soil. Nitrogen was applied at sowing. Three seeds of the faba bean cultivar Shambat 616 were placed in a hole on the top of the ridge with 20 cm spacing between holes and 70 cm between ridges. Plots were immediately irrigated after sowing and then, subsequently, irrigated at 10 days intervals.

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139 Yield and 100-seed weight. At maturity, seeds were collected and the yield of each treatment was expressed on a per hectare basis. The seeds were carefully cleaned and freed from dirt, stones, chips and other extraneous grain or dirt. From each sample, 100 seeds were counted randomly in triplicate and the weight was recorded. Cookability test. Twenty grams of beans were processed in 200 ml of tap water in a Labconco apparatus at 110  C for 30 min. The sample was reweighed after processing. Cookability was calculated as follows: Cookability % =

Weight after processing , Initial weight (20 g) Initial weight (20 g)

 100

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Non-soakers and hydration coefficient. For each treatment, 100 seeds were selected at random and soaked in tap water at a ratio of 1 part to 4 parts of water for 16 hours. The percentage of non-soakers in each sample was calculated after sorting and weighing of non-soaker seeds. The non soakers percentage was calculated as follows: Non soakers % =

Weight of non-soakers Initial weight

 100

:

The hydration coefficient percentage was calculated as follows: Hydration coefficient % =

Weight of soaked beans Initial weight

 100

:

Total defects. Total defects is a term for seeds which have abnormalities such as being non-soakers, broken or physically damaged, failure to reach maturity and/or being very small in size. The total defect percentage was calculated as follows: Total defect % = other defect % + non-soaker %: Other defect % =

Weight of defect  100: Weight before soaking

Statistical analysis. Each sample was analyzed in triplicate and the figures were then averaged. Data were assessed by analysis of variance (ANOVA) [5]. The Duncan multiple range test was used to separate means. Significance was accepted at p 6 0:05. Results Effect of treatments on yield and 100-seed weight. All treatments significantly (p 6 0:001) increased the yield of the faba beans (Figure 1). The yield

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140

Figure 1. Effect of nitrogen, sulphur and chicken manure (M) on yield (Kg/ha) of faba bean in presence ( R) and absence ( R) of Rhizobium.

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of inoculated treatments were significantly (p 6 0:05) better than uninoculated treatments. Rhizobium inoculation, 40 kg N/ha and 80 kg N/ha treatments increased yield by 17.2, 17.0 and 20.4%, respectively, compared to the uninoculated control with no fertilization. This means that inoculation was as effective in increasing yield as 40 kg N/ha. The highest yield was observed when both Rhizobium and 15 ton of M/ha were used. The 100-seed weight in this study ranged from 46.2 for the uninoculated control to 57.1 g for inoculation with 15 ton M/ha (Table 1). The 100-seed weights of inoculated treatments were significantly (p 6 0:05) better than uninoculated treatments. Inoculation with 100 kg S/ha, 15 ton M/ha and 80 kg N/ha treatments increased the 100-seed weight by 13.9, 23.6 and 13.6%, respectively, compared to the uninoculated control. Non-soakers. With the exception of uninoculated nitrogen treatments, all other uninoculated treatments significantly (p 6 0:05) reduced the number of non-soakers seeds compared to the uninoculated control (Table 2). There were no significant differences between the high and low doses of S, N and manure. The range of non-soaker percentages due to all treatments was 2.9–5.4. The

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141 Table 1. Effect of nitrogen (N), sulphur (S) and chicken manure (M) on 100-seed weight (g) of faba bean cultivar Shambat 616 either uninoculated or inoculated with Rhizobium strain TAL 1400 Treatment

No Rhizobium

Rhizobium

Control 40 kg N/ha 80 kg N/ha 25 kg S/ha 50 kg S/ha 100 kg S/ha 3 ton M/ha 9 ton M/ha 15 ton M/ha

46.2a 48.6c 49.8d 47.4b 49.5cd 51.2e 52.5f 53 .7g 55.5h

48.9a 50.9b 52.5d 50.1b 51.4c 52.6d 54.5e 55.9f 57.1g



Values are means ( SD). Means not sharing a common superscript(s) in a column are sig0:05 as assessed by nificantly different at p Duncan’s multiple range test.

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Table 2. Effect of nitrogen (N), sulphur (S) and chicken manure (M) on non-soaker and total defect of seeds of faba bean cultivar Shambat 616 either uninculated or inoculated with Rhizobium strain TAL 1400 Treatment Control 40 kg N/ha 80 kg N/ha 25 kg S/ha 50 kg S/ha 100 kg S/ha 3 ton M/ha 9 ton M/ha 15 ton M/ha

Non-soaker (%) No Rhizobium 4.9 ( 4.3 ( 5.0 ( 2.9 ( 3.0 ( 2.8 ( 3.1 ( 2.5 ( 3.2 (



 2.1)a  1.3)a  1.4)a  2.2)b  0.9)b  1.3)b  4.1)b  1.4)b  2.0)b

Rhizobium 3.3 ( 4.9 ( 5.4 ( 3.2 ( 3.3 ( 2.6 ( 3.3 ( 3.3 ( 3.3 (

 0.3)a  1.6)b  1.2)c  1.8)a  0.9)a  1.8)a  1.7)a  1.5)a  1.6)a

Total defect (%) No Rhizobium 19.4 ( 18.9 ( 18.6 ( 15.3 ( 15.7 ( 14.9 ( 13.4 ( 14.6 ( 15.4 (

 1.0)a  1.9)a  1.2)a  0.8)b  1.1)b  1.9)b  1.1)b  1.2)b  1.6)b

Rhizobium 17.4 ( 21.0 ( 21.0 ( 16.3 ( 16.4 ( 16.6 ( 18.4 ( 18.5 ( 18.0 (

 1.2)a  1.1)b  1.8)b  0.8)a  1.0)a  1.4)a  1.8)a  1.5)a  1.4)a

Values are means ( SD). Means not sharing a common superscript(s) in a column are significantly different at p 0:05 as assessed by Duncan’s multiple range test.  Seeds with abnormalities such as being non-soakers, broken or physically damaged, failure to reach maturity and/or being very small in size.

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effect of sulphur and manure treatments (inoculated and uninoculated) on hard seed percentage (non-soakers) were comparable.

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142 Table 3. Effect of nitrogen (N), sulphur (S) and chicken manure (M) on hydration coefficient and cookability of seeds of faba bean cultivar Shambat 616 either uninculated or inoculated with Rhizobium strain TAL 1400 Treatment Control 40 kg N/ha 80 kg N/ha 25 kg S/ha 50 kg S/ha 100 kg S/ha 3 ton M/ha 9 ton M/ha 15 ton M/ha

Hydration coefficient (%) No Rhizobium Rhizobium

 3.1)a  2.1)a  5.9)b  1.7)b  1.3)b  3.7)b  4.0)b  2.7)b  1.6)b

193.6 ( 194.2 ( 199.4 ( 198.0 ( 198.7 ( 197.3 ( 198.6 ( 199.9 ( 200.5 (



193.6 ( 194.9 ( 199.5 ( 199.8 ( 198.0 ( 200.0 ( 199.7 ( 197.3 ( 198.8 (

 4.2)a  3.8)a  1.9)b  3.4)b  2.5)b  1.9)b  3.0)b  3.1)b  1.1)b

Cookability (%) No Rhizobium 21.5 ( 22.3 ( 23.5 ( 21.7 ( 21.8 ( 22.8 ( 27.5 ( 28.7 ( 28.2 (

 1.5)a  2.5)a  2.7)a  1.6)a  2.7)a  1.2)a  1.0)b  1.5)b  1.4)b

Rhizobium 22.5 ( 24.6 ( 24.3 ( 20.7 ( 20.8 ( 21.2 ( 28.2 ( 28.0 ( 28.9 (

 3.1)ab  1.5)bc  1.6)bc  2.3)a  1.7)a  1.1)a  1.8)d  1.7)d  1.7)d

Values are means ( SD). Means not sharing a common superscript(s) in a column are significantly different at p 0:05 as assessed by Duncan’s multiple range test.

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Total defects. The total seed defect percentages ranged between 13.4 to 21.0% due to different treatments (Table 2). The total defects followed a similar trend to that of the non-soaker percentage. In general, the total defects of this variety were low, indicating the good quality of the seeds of cultivar Shambat 616. A positive correlation (r = 0:90) was observed, for all treatments, between the non-soaker percent and the total defect percent and no correlation was found between non-soakers, hydration coefficient and cookability. Hydration coefficient %. Hydration coefficient is a very valuable attribute for both consumers and processors. It is a good indicator of seed quality because it plays a major role in defining the ability of the seeds to absorb water and, hence, become ready for the cooking process. With the exception of Rhizobium inoculation and 40 kg N/ha treatments, all other treatments significantly (p 6 0:05) increased the hydration coefficients of faba bean seeds, compared to the uninoculated control (Table 3). All sulphur and chicken manure treatments increased hydration coefficient similarly, and there was no significant differences between high and low doses. Effect of treatments on cookability. Chicken manure treatments significantly (p 6 0:001) increased the cookability of the Shambat 616 variety in the presence or absence of Rhizobium inoculation (Table 2). The highest cookability values were observed when both 15 ton/ha chicken manure and Rhizobium were applied. The percentage increment was 34.4% compared to the uninoculated control.

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143 Discussion Inoculation with Rhizobium strain TAL 1400 significantly (p 6 0:05) increased yield and 100-seed weight of the faba bean cultivar Shambat 616. Addition of 40 kg N/ha or 80 kg N/ha via urea significantly (p 6 0:05) improved the yield and 100-seed weight of faba bean plants. Elsheikh and Osman [4] found that Rhizobium inoculation significantly increased seed yield and the total nitrogen of faba bean. Also the results obtained were in agreement with those of Babiker et al. [2] who reported that application of 50 kg N/ha increased seed yield of faba bean by more than 20%. Application of sulphur significantly (p 6 0:05) improved the total plant yield and 100-seed weight, in the presence or absence of Rhizobium inoculation. The effect of high dose of sulphur (100 kg/ha) was significantly better than smaller doses 50 or 25 kg S/ha for yield and 100-seed weight. This could be due to the improved nutritional status of plants rather than the direct effect of sulphur on Rhizobium activity. Hago & Salama [6] found that application of elemental sulphur significantly increased nodulation and yield of groundnut (Arachis hypogaea). Chicken manure treatments significantly (p 6 0:001) increased yield and 100-seed weight in the presence or absence of Rhizobium inoculation. The highest dose (15 ton/ha) of chicken manure significantly (p 6 0:05) increased yield and 100-seed weight compared to the smaller dose (3 ton/ha). The beneficial effect of chicken manure could be due to its positive effect on soil structure, water and root penetration, and their complex properties which prevent penetration and fixation of many plant nutrients [7]. Unexpectedly, the non-soakers (hard seeds) were not affected by the N treatments. However, all other treatments significantly reduced non-soaker seeds compared with the control plants. This may indicate that good fertilization programs could improve seed quality. These programs should be incorporated with other breeding programs to eliminate or minimize hard seed percentage. Among the major factors that influence hard seed percentage are environmental conditions, locality, harvesting time and variety [8]. Non-soakers percentage is of great importance for both processors and consumers as it decreases the acceptability of the seed. The total defect percentage followed a similar pattern to that of the nonsoakers. Hydration coefficient is a very valuable quality factor for both the consumer and processor. Low hydration coefficient indicates that the seeds are not efficiently capable of absorbing water when soaked in water [8]. Legumes, in general, have more than double the initial weight after soaking in water. Varieties differ in their hydration coefficients when grown at different locations [8]. Hydration coefficient in this study was increased by sulphur, chicken manure and 80 kg N/ha compared with the uninoculated control.

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144 This may indicate that the hydration coefficient is improved by chemical fertilization irrespective of the type. These findings could be tested in new regions in which faba bean is being introduced in Sudan [9]. The seeds grown in these regions have been, generally, characterized by low cookability, low hydration coefficient and high percent of non-soakers. In general, hydration coefficient was found to be affected by the locality, harvesting time and genotypes [10]. Cookability percentage was only improved by chicken manure. This reflects the role of chicken manure in improving the quality of faba bean seeds. Usually the incidence of hard seed adversely affects cooking quality and, therefore, the market value of the product [1]. Cookability is known to be affected by soaking time, type of water, time of cooking, environmental factors, location and time of harvest [11].

References 1.

Salih FA, Ali AM, Elmubarak AA (1986) Effect of phosphorus application and time of harvest on the seed yield and quality of faba bean. FABIS Newsletter 15: 32–35. 2. Babiker EE, Elsheikh EAE, Osman AG, El Tinay AH (1995) Effect of nitrogen fixation, nitrogen fertilization and viral infection on yield, tannin and protein contents and in vitro protein digestibility of faba bean. Plant Foods Hum Nutr 47: 257–263. 3. Mahdi AA (1993) Biofertilizer research in the Sudan: A review. University of Khartoum, J Agric Sci 1: 137–151. 4. Elsheikh EAE, Osman AG (1995) Rhizobium leguminosarum inoculation and the decrease in damage of faba bean (Vicia faba) infected with broad bean mottle bromovirus and bean yellow mosaic potyvirus. World J Microbiol Biotech 11: 223–227. 5. Snedecor GW, Cochran WG (1987) Statistical methods, 7th edn, pp 221–222. Ames, IA, The Iowa State University Press. 6. Hago TM, Salama MA (1987) The effects of elemental sulphur on shoot dry weight nodulation and pod yield of groundnut (Arachis hypogaea) under irrigation. Exp Agric 23: 93–97. 7. El Tilib AMA, Ali AM, Abdulla MA (1993) Effect of chicken manure and salinity on growth and leaf N, P and K content of okra grown on two soil types. University of Khartoum, J Agric Sci 1: 16–33. 8. Ali AE, Ahmed GE, El Hardallou EB (1982) Faba beans and their protein diets in Sudan. In: Hawtin G and Webb I (eds), Faba beans improvement, p. 318. The Hague: Martinus Nijhoff/ICARDIA. 9. Ali AE, Salih FA, A/Galiel A (1988) Effect of time of harvest on physical and chemical composition, cookability and yield of faba beans. FABIS Newsletter 20: 33–35. 10. Salih FA, Khairi NEA (1990) Variation in testa fraction with some other seed quality attributes of faba bean grown in the new production areas in the Sudan. FABIS Newsletter 27: 30–34. 11. Elmubarak KAA, Salih FA, A/Galiel A, Ghorashi AM (1988) Effect of time of harvest on physical and chemical composition, cookability and yield of faba bean. FABIS Newsletter 20: 33–36.

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