1

Yield, Yield Component and Quality of Soybean (Glicin max L.) as

Influenced by Bacterial (Bradyrhizobium japonicum) Inoculation and
Phosphorus Fertilizer Rate at Abol Woreda Gambella Region,
South West Ethiopia

A research proposal submitted to the Research Directorate of Gambella

University

Principal investigator:
Biftu Umer (Msc.in Agronomy, Gambella University)

Co-investigators:

Bezabih Addisu (Msc.in Agronomy, Gambella University)

March ,2020
Gambella
Contents
1. INTRODUCTION..................................................................................................................1
2. LITERATURE REVIEW......................................................................................................4
2.1. Soybean origin..................................................................................................................4
2.2 Agro ecology of soybean.......................................................................................................4
2.3. Soybean production in Ethiopia........................................................................................5
2.4. Uses of soybean................................................................................................................6
2.5. Phosphorous......................................................................................................................7
2.6. Role of nitrifying bacteria’s in soybean production..........................................................9
3. MATERIALS AND METHODS............................................................................................10
3.1. Description of the Study Area.............................................................................................10
3.2. Treatments and Experimental Design.................................................................................10
3.3 Experimental procedures......................................................................................................11
3.4. Soil sampling and analysis..................................................................................................12
3.5. Data to be collected.........................................................................................................12
3.6. Data analysis...................................................................................................................14
4. WORK PLAN AND LOGISTICS......................................................................................15
4.1. Work plan............................................................................................................................15
4.2. Budget and Logistics...........................................................................................................16
5. REFERENCE.......................................................................................................................20

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 List of tables

Table 1: Treatment combination....................................................................................................11

Table 2: work plan.........................................................................................................................15
Table 3Experimental materials......................................................................................................16
Table 4 Laboratory cost for Protein and Oil content of the soybean seed.....................................16
Table 5: Laboratory cost for soil analysis......................................................................................17
Table 6: Stationary cost.................................................................................................................18
Table 7: Transportation cost..........................................................................................................18
Table 8: Perdium and Daily labor costs.........................................................................................19
Table 9: Summary of Total Cost...................................................................................................19

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 1. INTRODUCTION

Soybean (Glycine max) is one of the most important food plants of the world, and seems to be
growing in importance. It can substitute for meat and to some extent for milk. It is a crop capable
of reducing protein malnutrition. In addition, soybeans are a source of high value animal feed
(Martin, 1998).

Soybean (Glycine max (L.) Merrill) is an important legume crop, which grows in the tropical,
subtropical and temperate climates (Shurtleff and Aoyagi 2007; IITA, 2009). Soybean (Glycine
max (L.) Merry), is the leading oil and protein crop of the world, which used as a source of high
quality edible oil, protein, and livestock feed (Rajcan et al., 2005). It has the highest protein
content of all food crops and is second only to groundnut in terms of oil content among food
legumes (Fekadu etal. 2009; Alghamdi, 2004).

Soybean is a source of edible oil (second most consumed oil in the world after palm oil) and is
used to produce livestock feed. Many other products with a soya basis are also directly used for
human consumption (soymilk, soy yogurt, snacks, soya sauce, protein extract and concentrates,
etc.,) (Collombet, 2013).Soybean is one of the most important oil seed crops in the world. It
contains 18 to 22% oil, highly desirable in diet and has 40 to 42 % protein.

Soybean protein provides all eight amino acids in the amount needed for human health; hence it
is the best source of protein and oil and truly claims the title of the meat/oil on plants (Ali, 2010).
In the major producing countries and particularly in Brazil, Argentina, Paraguay and the USA
soybean contributes significantly to the total value added by the agricultural sector. Among
smaller producers only India and Bolivia earn significant income from the exportation of
soybean and derived products, (Thoenes, 2014).

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Soybean has an average protein content of 40% and is more protein-rich than any of the common
food crop sources in Africa. The seeds contain about 20% oil on dry matter basis and some
appreciable amounts of vitamins and minerals (Dugje et al., 2009). Yield increased only six
percent since 2000 and contributed only 19% to the increase.

In Ethiopia soybean occupies around 95 thousand hectares of land with a corresponding gross
annual production of approximately 94, 773 metric tons (CSA, 2009). Despite the importance of
the crop in the world as a rich source of protein and oil, the yield is generally very low in
Ethiopia, i.e. below 1-ton ha-1 (CSA, 2009) as compared to the USA and Asian soybean
producing countries. Poor soil fertility status considered as one of the factors contributing to low
yield. Nitrogen (N) and phosphorus (P), in that order, are the two plants growth limiting soil
fertility factors in many soil types including those in Ethiopia. The study monitored by FAO in
38 sub-Saharan African countries, including Ethiopia showed that there are high nutrient
depletion rates in N, P and potassium in different farming systems (Stoorvogel et al., 1993).

The utilization efficiency of phosphate fertilizers by plants is only 20 to 25% largely due to its
chemical fixation in soil (Hedley et al., 1995). Unbalanced use of chemical fertilizers led to a
reduction in soil fertility and to environmental degradation (Gyaneshwar et al., 1998) and the
cost of chemical fertilizers has increased so that it is unaffordable for developing country farmers
such as Ethiopia.

Phosphorus play specific role in symbiotic N2-fixation through their effects on nodulation and
N2-fixation process (Hara et al., 2002). Symbiotic nitrogen fixation has a high P demand
because the process consumes large amounts of energy (Schulze et al., 2006) and energy
generating metabolism strongly depends upon the availability of P (Israel, 1987; Plaxton, 2004).

2
Several reports have documented that nodules are a strong P sink and nodule P concentration
normally exceeds that of roots and shoots (Sa and Israel, 1991; Drevon and Hartwig, 1997).

Among various factors that can contribute to soybean success, phosphorus and inoculation had
quite prominent effects on nodulation, growth and yield parameters (Kumaga and Ofori, 2004).
Phosphorus has important effects on photosynthesis, nitrogen fixation, root development,
flowering, seed formation, fruiting and improvement of crop quality (Brady, 2002). Symbiotic
nitrogen fixation is definitely beneficial to agriculture. It is major source of fixed nitrogen in
agricultural soils. Responses by soybean to inoculation with Bradyrhizobium japonicum
expressed in terms of both the proportion of nodules formed by the inoculum and seed yield
(Simanungkalit et al., 1995). Inoculation of soybean seeds with proper bacterial strains increased
seed production by 70-75% (Simanungkalit et al., 1996).

On nutrient deficient soils, the traditional approach is to change the soil to fit the plant, i.e.
fertilizers and other soil amendments are applied to increase soil fertility and/ or to make soil
nutrients available for plant uptake. The exploitation of plant genetic variation to increase the
efficiency with which plants absorb and use nutrients has been increasingly emerging as an
alternative approach towards low-nutrient environments (Gunawardena et al. 2006). Ethiopian
agriculture is characterized by low production per unit area due to declining of soil fertility,
rainfall variability, pest pressure, poor agronomic practices and poor accessibility to quality seed
(Katungiet al., 2010). Low soil fertility is one of the bottlenecks to sustain agricultural
production and productivity in Ethiopia.

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Among the essential nutrients, phosphorus is one of the most important nutrients for higher yield
in larger quantity (Chen et al. 1994) and controls mainly the reproductive growth of plant
(Wojnowskaet al. 1995). Generally, phosphorus is the second most crop-limiting nutrient in most
soils. Plant growth behavior is influenced by the application of phosphorus (Hajabbasi and
Schumacher, 1994; Gill et al. 1995; Kaya et al. 2001). It is needed for growth, utilization of
sugar and starch, photosynthesis, nucleus formation and cell division, fat and albumen formation.
But, until now little research was conducted on phosphorus fertilizer and bacterial inoculation on
yield of soybean. Therefore, this study will be conducted based on the following objectives:-


General Objective

To evaluate the effect of Rizobium inoculation and different levels of phosphorus on
growth and yield of soybean in abol woreda Gambella region southwest Ethiopia.
Specific objectives


To evaluate the response of soybean to different rates of P fertilizer.

To evaluate the response of soybean to bacterial inoculation.

To observe the interaction of P with rizobium inoculation on soybean.

2. LITERATURE REVIEW

2.1. Soybean origin

Soybean originated from China and the history of the crop has been reviewed in various texts
(Smith et al., 1987). The earliest documentation existing that mentions soybean as one of the five
main plant foods of China comes from the year 2700 B.C. (Hymowitz and Shurtleff, 2005).

Soybean is originated from South East Asia, from where it spread into many parts of the world
(Norman et al., 1995). The spread of soybean from its native land of origin has been mainly due
to its adaptability and predominant use as a food crop for human nutrition, source of protein for
animals, medicinal plant and lately as an industrial crop (Yusuf and Idowu, 2001).
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2.2 Agro ecology of soybean

Soybean (Glycine max, Leguminosae) a dicotyledonous annual oil crop that is photoperiod
sensitive, whose economic importance for household and industrial use is increasing.

Climate variability is an important source of risk in soybean production since it affects yield
variation and often leads to yield losses. Increased frequency of climate is expected to be
associated with future climate change (IPCC, 2007). Climate change will therefore enhance farm
risks by influencing crop growth and development, resulting in yield reduction (Adams etal.
1998).

The suitability land classification done by Verdoot (2009) indicates that even soybean is
cultivated in all agricultural zones with an altitude below 1,900 m, only large parts of the Eastern
Savanna, Eastern Plateau, Birunga and Mayaga are moderately suitable. However, correct
management can allow the crop to be cultivated in the low and middle altitude zones. In Rwanda,
soybean is generally grown in the low altitude zones (1000 -1400 masl), with rainfall of 800 –
1000 mm, and partly in the mid altitude zone (1400 – 1700 masl), with rainfall of 1000 – 1200
mm. The climate has a pronounced wet period from May to November, and a dry period from
December to April. Climate variability conditions affect the spatial and temporal distribution of
rainfall in tropical soybean-growing areas. More importantly, the interactions of precipitation
occurrence can impact the health of crops and land productivity, as unstable climate affects
important parameters on crop reproductive and vegetative growth.

2.3. Soybean production in Ethiopia

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Soybean plays a vital role in agricultural economy of Ethiopia. Yield of legumes in farmers’ field
is usually less than 0.65t/ha against the potential yield of 1.2t/ha suggesting a large yield gap
(CACC, 2002) low yield potential of legumes has made them less competitive with cereals and
other high value crops. The low productivity of the crop is due to several constraints, one among
the unbalanced important is nutrition (Sharma, etal. 1996).

Soybean grows in areas where maize and common beans are grown. It grows to a height of 60–
120 cm, maturing in 3 to 6 months depending on variety, climate, and location. Soybean is
drought tolerant. Depending on the variety, the crop can be grown from 0-2200m altitude and
under rainfall ranging from 300 to 1200mm. Altitude influences temperature that in turn affects
the initiation of flowering and maturity. At very high altitudes, flowering may not occur and the
crop remains vegetative. Soybean is therefore a crop that requires warm climates and is suitable
for low to medium altitudes (Ogema, etal, 1988). It grows best when planted in pure stands. The
presence of Rhizobium japonicum in the roots of soybean enables the crop to fix nitrogen in the
soil contributing to improved soil fertility (Kasasa, etal, 2000).

For its optimum yield realization, it is necessary to optimize the nutrient inputs. Fertilizer
application is very important practices and at present the most baffling as well. Studies carried,
out indicates that soybean shows inconsistent responses to application of nitrogen, phosphorous
and potassium. High soybean yield demand high fertilizer dosage, and applied directly to the
crop or acquired through preceding crops. Another feature of sound fertilization practices is that
all the nutrients (N, P and K) applied should be in balanced proportion to indicate the high
production efficiency. In recent years, there has been a growing interest in the use of narrow row
as well as narrow plant spacing for the production of soybean because of high labor energy and
equipment requirements for cultivation (Jordan, 2010).

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Soybean (Glycine max L. (Merr.)) is a subtropics member of the Leguminosae family, an erect
bushy annual crop with considerable morphological diversity. Soybean is one of the most
important oil seed crops in the world. It contains 18 to 22% oil, highly desirable in diet and has
40 to 42 % protein.

Soybean protein provides all eight amino acids in the amount needed for human health; hence it
is the best source of protein and oil and truly claims the title of the meat/oil on plants (Ali, 2010).
In Ethiopia soybean occupies around 95 thousand hectares of land with a corresponding gross
annual production of approximately 94, 773 metric tons (CSA, 2009). Despite the importance of
the crop in the world as a rich source of protein and oil, the yield is generally very low in
Ethiopia, i.e. below 1-ton ha-1 (CSA, 2009) as compared to the USA and Asian soybean
producing countries.

2.4. Uses of soybean

The best-known and most widely used products from soybeans are soybean oil and soybean
meal. Soybean oil is the most widely used edible oil in the world and soybean meal is the leading
protein and energy source for animal feeds.

Today, most of the world’s soybeans are processed or crushed into soybean meal and oil (Ali
2010). It is estimated that 2% of soybean production is consumed by humans directly as food
(Goldsmith 2008), which amounts to an estimated 3 MMT. Soybean seeds contain about 18% oil
and 38% protein. Of the oil fraction, 95% is consumed as edible oil with the rest used for
industrial products from cosmetics and hygiene products to paint removers and plastics (Liu
2008). Due to its high protein level, about 98% of the soybean meal is used in livestock and
aquaculture feeds. A smaller percentage is processed to make soy flour and protein for human

7
consumption. An even smaller percentage is used as a fresh vegetable known as “mao dou” in
China, “edamame” in Japan, and green vegetable soybean or edamame in the USA and other
countries (Shanmugasundaram and Yan 2010).

The vegetable soybean seeds are higher in sucrose, comparable in protein and lower in
tocopherols than grain soybeans, and are higher in protein and lower in sucrose compared to
green peas. The immature seeds (still green) are blanched, boiled or steamed and eaten in a
number of different ways including as a side dish to mix with other foods. The soybean
edamame types are selected or bred large-seeded, sweet tasting varieties that are now grown in
vegetable gardens in many parts of the world. Soybean oil is used in making numerous processed
food products like margarine and in preparation of fried foods. Consumption of soybean oil
increased dramatically with the realization of the link between animal fats and cardiovascular
disease. In order to have stable flavor and freshness for use as commercial oil, soybean oil was
hydrogenated. However, awareness in recent years of the detrimental effects of transfers initiated
the development of new soybean varieties with lower linolenic content that are more stable
without hydrogenation. Linolenic acid is an omega-3 polyunsaturated fatty acid that humans
must obtain from food. Soybean varieties with high oleic acid, up to 80% of the oil content, have
been developed recently, but commercially are not widely available.

Soybeans are unique among crop plants in that they supply protein equal in quality to that of
animal sources. For this reason, soybeans have long been consumed in Asia as a primary source
of protein in such traditional foods as tofu, soymilk, tempeh, natto, sprouts, green vegetable
soybeans, and many others. There are many functional components in soybeans, and long before
the development of the nutraceutical industry in the West, soybeans were considered to have
medicinal value (Raghuvansh and Bisht 2010). Soybeans are a rich source of nutraceutical
including but not limited to bio-flavanoids, lecithin, oligosaccharides, phytosterols, saponins, and

8
tocopherols. An important group within the bio-flavonoids is the iso-flavones, which have been
shown to benefit human health (Kumar et al. 1996).

2.5. Phosphorous

Phosphorus is an important plant nutrient involved in several energy transformation and

biochemical reactions including biological nitrogen fixation. Phosphate fertilizers have low
efficiency of utilization due to chemical fixation in soil (Gaur, 1983) and due to poor solubility of
native soil phosphorus, sometimes there is a buildup of insoluble phosphorus as a result of
chemical phosphorus application (Dubey, 1997). Phosphorus (P) is a major growth-limiting
nutrient, and unlike the case for nitrogen, there is no large atmospheric source that can be made
biologically available (Ezawaet al., 2002). Root development, stalk and stem strength, flower and
seed formation, crop maturity and production, crop quality, and resistance to plant diseases are
the attributes associated with phosphorus nutrition. Although microbial inoculants are in use for
improving soil fertility during the last century, however, a meager work has been reported on P
solublization compared to nitrogen fixation. Soil P dynamics is characterized by physicochemical
(sorption-desorption) and biological (immobilization-mineralization) processes. P is needed in
relatively large amounts by legumes for growth and nitrogen fixation and has been reported to
promote leaf area, biomass, yield, nodule number, nodule mass, etc., in a number of legumes
(Berg and Lynd, 1985; Pacovsky, et al., 1986; Kasturikrishna and Ahlawat, 1999).Phosphorus
deficiency can limit nodulation by legumes and P fertilizer application can overcome the
deficiency (Carskyet al., 2001).

Large amount of P applied as fertilizer enters in to the immobile pools through precipitation
reaction with highly reactive Aluminum (Al+) and Iron (Fe3+) in acidic, and Calcium (Ca2+) in
calcareous or normal soils (Gyaneshwaret al., 2002; Haoet al., 2002). Efficiency of P fertilizer
throughout the world is around 10 - 25 % (Isherword, 1998), and concentration of bio available

9
P in soil is very low reaching the level of 1.0 mg kg- I soil (Goldstein, 1994). Soil
microorganisms play a key role in soil P dynamics and subsequent availability of phosphate to
plants (Richardson, 2001). Microbial community inf1uences soil fertility through soil processes
viz. decomposition, mineralization, and storage/release of nutrients. Microorganisms enhance the
P availability to plants by mineralizing organic P in soil and by solubilizing precipitated
phosphates (Chen et al., 2006; Kang et al., 2002; Pradhan and Sukla, 2005).

Phosphate solubilizing bacteria (PSB) are being used as bio fertilizer since 1950s (Kudashev,
1956; Krasilinikov, 1957). Release of P by PSB from insoluble and fixed/ adsorbed forms is an
import aspect regarding P availability in soils. There are strong evidences that soil bacteria are
capable of transforming soil P to the forms available to plant. Microbial biomass assimilates
soluble P, and prevents it from adsorption or fixation (Khan and Joergesen, 2009). Nutritional
capacity of SSP was further improved by PSB inoculation due to its ability to convert sparingly
soluble inorganic organic phosphate into soluble forms by secreting organic acids (Arora and
Gaur (1979).Microorganisms through secretion of different types of organic acids like carboxylic
acid (Deubel and Merbach, 2005) and rhizosphere pH lowering mechanisms (He and Zhu, 1988)
dissociate the bound forms of phosphate like Ca 3 (PO4)2. Among various factors that can
contribute to soybean success, phosphorus and inoculation had quite prominent effects on
nodulation, growth and yield parameters (Kumaga and Ofori, 2004). It is well known that more
than two-third of phosphate fertilizers are render unavailable within a very short period of time
after its application due to fixation in soil complex as di and tri-calcium phosphate (Mandal and
Khan, 1972). Application of phosphorus along with phosphate solubilizing bacteria (PSB)
improved phosphorus uptake by plants and yields indicating that the PSB are able to solubilize
phosphates and to mobilize phosphorus in crop plants (Rogers and Wolfram, 1993). In this
respect, bio fertilization technology has taken a part to minimize production costs and at the
same time avoid the environmental hazards (Galal et al., 2001). With the above facts the study
was undertaken to investigate the best source and level of phosphorus on yield attributes, yield
and quality of seed and efficiency of phosphorus.

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 2.6. Role of nitrifying bacteria’s in soybean production

Microorganisms involved in biofertilizer production can play a major role in increasing nutrient
availability for high yield and reducing production cost by limited chemical fertilizer application.
Phosphorus biofertilizers could help increase the availability of phosphate for plant growth by
solubilization, promoting the plant growth through production of plant growth-promoting
substances, and by increasing the efficiency of biological nitrogen fixation (Kucey et al., 1989).
Although the role of phosphorus fertilizers in nodulation, nitrogen fixation, nutrient-use
efficiency and growth of soybean has been reported (Israel, 1987; Tsvetkova and Georgiev,
2003; Israel, 2006; Miao et al., 2007), information is scanty regarding the role of phosphate-
solubilizing bacteria in phosphorus bioavailability, growth promotion and also their interaction
with N2- fixing bacteria.

Co-inoculation of Bacillus strains onto soybean plants with Bradyrhizobium japonicum provided
the largest and most consistent increases in nodule number, nodule weight, shoot weight, root
weight, total biomass, total nitrogen and grain yield (Bai-Yu Ming et al., 2003).

3. MATERIALS AND METHODS

3.1. Description of the Study Area

Field experiment will be conducted during 2012 main cropping season under rain-fed conditions
at Abol woreda of Gambella region. Gambella is located 776 Km away 7 05’ N and 340 32’ west
of the capital Addis Abeba. Annual mean minimum and maximum temperatures in Gambella are
250C and 420C, respectively. Mean annual rainfall of the area varies from 800mm to 1,500 mm
with a long-term average of 1,400 mm. The site is located at an altitude of 526 m.a.s.l. Most of
the soils of the region are fluvisols (alluvial soil type) which have pH of 6.1 and it is slightly
acidic.

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3.2. Treatments and Experimental Design

Hawassa 95 soybean variety which is released from Hawassa agricultural research center is used.
This variety is well adapted to the area and gives a yield of 28 kg/ha in rain fed cropping.
(GARC).

Factorial experiment of four phosphorus level (0 kg/ha of p 2o5, 23 kg/ha of p2o5, 46 kg/ha of
p2o5 and 69 kg/ha of p2o5) with two level of inoculation L0(no inoculation) and L1(inoculated),
seed will be tested in Randomized Complete Block Design with three replications. The gross plot
sizes will be 3 × 1.5 m for the width and length of the plots, respectively. The distance between
each plot will be 1m while distance between block is 1.5 m and the total experimental area will
be 232.5 m2. A single experimental plot will have 5 rows and 100 plants with recommended
spacing of 60X5cm inter and intra row spacing.

Table 1: Treatment combination

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 No Bacterial inoculation Phosphorus level
1 I0 (non inoculated) P1 (0 kg/ha of p2o5)
2
P2(23 kg/ha of p2o5)

3 P3 (46 kg/ha of p2o5)

4 P4 (69 kg/ha of p2o5)
5 I1 (inoculated ) P1 (0 kg/ha of p2o5)
6
P2(23 kg/ha of p2o5)

7 P3 (46 kg/ha of p2o5)

8 P4 (69 kg/ha of p2o5)

3.3 Experimental procedures

The land will be ploughed once, harrowed twice and leveled to obtain the desired germination
and growth of crop. The field will be then divided into three blocks and then in to 3 m x 1.5 m
equal plots as per the treatments. Seeds of soybean used for this experiment will be obtained
from Gambella agricultural research center. The inoculant (Rhizobium japonicum) will be
brought from Adiss Ababa biotechnology institute and Inoculation will be done by adopting
standard procedures just before sowing and a control (no inoculation) will be also maintained .
The seeds will be sown in rows at the depth of 3 - 4 cm having two seeds per hill and then
thinned to one plant per hill just a week after emergence. Planting will be done on May 1/2012
E.C.

Harvesting will be done when the crop reached harvest maturity. The data will be collected from
the central rows, by leaving one border rows from each side of a plot and one plant at the two
ends of every row. The net plots will be harvested by cutting the whole plant with a sickle.
Thereafter, the pods will be picked from the main bunch and allowed to air and sundry for six
days. The dried pods will then collected on plot basis.

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3.4. Soil sampling and analysis

Pre-planting soil samples will be taken randomly in a zigzag fashion from the experimental plots
at the depth of 0-20cm. Ten soil cores will be taken by an auger from the whole experimental
field and combined to form a composited sample in a bucket. The soil will be broken in to small
crumbs and thoroughly mixed. From this mixture, a sample weighing one kg will be filled in to a
plastic bag. This will be replicated two times and prepared for the analysis.

The soil will be air dried and sieved through a 2mm sieve. Then, soil pH will be determined by
diluting the soil in a 0.01 M CaCl 2 solution in the ratio of 1 soil volume to 2.5 volume of the
CaCl2 solution. Thus, twenty-five ml of the 0.01M CaCl 2 solution will be added into soil sub
samples each weighing 10g. After equilibrating for 2-3 hours, the suspensions will be filtered
and the pH measured by a glass electrode. Texture of the soil will be determined by
sedimentation method (Hesse, 1971).

The soil samples will be analyzed for total nitrogen, exchangeable potassium, and available
phosphorous and organic carbon. Total nitrogen of the soil will be determined by the micro
Kjeldhal procedure. Organic carbon will be determined by the method of Nelsen and Sommers
(1982). Available phosphorous content of the soil will be determined by extraction with 0.5M
NaHCO3 (Olsen et al., 1954). Phosphorus in the extracts will be determined with atomic
absorption spectrophotomer calorimetrically according to the molybdenum blue color method of

Murphy and Riley (1962).

3.5. Data to be collected

Plant height (cm): the height of the plant will be measured from ground level to the tip of the
plant by using meter at physiological maturity.

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Days to 50% flowering: It will be recorded as the number of days after planting to the date
when 50% of the plants in a plot produced at least one flower.

Days to 90% maturity: It will be recorded as the number of days from planting to date when
90% the crops in a plot matured. Maturity will be determined by examining yellowing of leaves
and pods.

Number of pods per plant: It will be determined by counting number of pod per plant from five
randomly selected plants per net plot at physiological maturity

Pod length (cm): the length of the pod will be measured by using meter at physiological
maturity.

Number of nodules per plant: it will be determined by uprooting and counting the nodules
from five randomly selected plants per plot at flowering time.

Number of seeds per pod: it will be determined by selecting 20 pods from net plot and by
taking the average.

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100 seed weight (g): this will be recorded by counting hundred seeds from a bulk of shelled
seeds and weighed using a sensitive balance.

Seed yield (kg ha-1): will be determined using the following formula

seed weight(kg) of 5 plants

Seed yield (t /ha)= x 10000
Harvested plot area(m 2)

Oil content (%) and Protein content (%): this will be determined by taking sample of seeds
from all treatments which are grown at net plot and it is allowed for laboratory analysis. Crude
protein will be estimated by multiplying the estimated value of the total nitrogen by 6.25
(Sahlemedhin and Taye, 2000). While oil content will be determined by techniques and
procedures outlined by Association of Official Analytical Chemists (A.O.A.C.) (1970)

3.6. Data analysis

All the data will be examined for homogeneity of variance and normality. Then, those data
which will be found to have normal distributions will be subjected to analysis of variance using

16
SAS statistical software package 9.2 (SAS, 2008). The differences between treatment means will
be compared using least significance difference (LSD) test at 5% level of significance

4. WORK PLAN AND LOGISTICS

4.1. Work plan

Table 2: work plan

No Activities Time

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 1 Proposal write up and Defense September –October 2019
2 Site Selection March –April 2020
3 Land Preparation May 2020
4 Field lay out & Planting May 2020
5 Field Management June –August 2020
6 Data Collection June –August 2020
7 Data Compilation& Analysis July- August 2020
9 Writing up & Reporting September 2020
10 Research result defense September 2020

4.2. Budget and Logistics

Table 3Experimental materials

No Description Unit Quantit Unit cost Total

y (birr) Cost(birr)

1 Soybean seed Kg 5 100 500

2 Phosphorus fertilizer kg 30 60 1800

3 Bacteria(Bradyrhizobium japonicum) 2000

4 Sacs (50 kg) Pcs 10 10 100

18
 5 Meter (50m) Pcs 1 300 300

6 Small garment bag Pcs 24 30 720

7 Rope (50m) Pieces 2 100 200

8 Pegs (wooden) pcs 100 5 500

9 Knapsack sprayer pcs 1 1,500 1500

10 Pesticides Liters 2 200 400

(Can)

Sub Total 8,020

Table 4 Laboratory cost for Protein and Oil content of the soybean seed

No Description Cost (birr)

1 Protein content of the seed 24 samples X 80 birr = 1920

2 Total oil content of the seed 24 samples X 80 birr = 1920

Sub total 3,840

Table 5: Laboratory cost for soil analysis before sowing

No Description Cost (birr)

19
 1 Available nitrogen 5 samples X 100 birr = 500

2 Available phosphorus 5samples X 150 birr = 750

3 pH

5 samples X 100 birr = 500

4 CEC

5samples X 100 birr = 500

5 Organic carbon

5samples X 1400 birr 500

6 Soil texture 5 sample X 50 birr= 250

Sub total 3000

Laboratory cost for soil analysis after sowing

No Description Cost (birr)

1 Available nitrogen 24 samples X 100birr = 2400

2 Available phosphorus 24 samples X 150 birr = 3600

3 pH

24 samples X 100 birr = 2400

4 CEC

24 samples X 100 birr = 2400

5 Organic carbon

24 samples X 100 birr = 2400

20
 6 Soil texture 24 sample X 50 birr= 250

Sub total 14,400

Table 6: Stationary cost

No Description Quantity Unit cost Total cost (Birr)

1 Paper 3realm 120 360

2 Toner 1 number 2000 2000

3 Communication 4000

4 Clip board 1 pieces 100 100

5 Binding for submission 20 50 1000

6 Flash disk 2 400 800

Sub total 8,260

Table 7: Transportation cost

No Description Unit Total cost

. cost (Birr)
1 Air ticket (two round trips for two researchers) 2400*4 9600
First trip: for seed and bacteria purchasing

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 Second trip: soil sample delivery to laboratory before sowing
Third trip: seed and soil sample delivery to the laboratory
after harvest
Forth trip : Laboratory result collection
2 Bajaj (Gambella to Abol) 100*10 1000
Sub total 10,600

Table 8 Perdium and Daily labor costs

No Description Measurement Duratio Cost per day Total

n (days)

1 Researchers 3(in number) 40 218.40 17452

3 Labor cost 10 (in number) 5 100 5,000

4 Guards 1 (day and 120 50 2000

night)

5 Experimental land rent Area 120 250*4months 1,000

Sub Total 25,472.00

Table 9, Summary of Total Cost

No Description Total cost(Birr)

1 Experimental materials 8020
2 Laboratory cost for Protein and Oil content 3840
3 Laboratory cost for soil analysis before sowing 3000

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 4 Laboratory cost for soil analysis after sowing 14400
5 Stationary cost 8,260
6 Transportation cost 10,600
4 Perdium and Daily labor costs 25,472
7 Grand total 73,592.00

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