001

EFFECT OF RHIZOBIUM INOCULATION AND PHOSPHORUS
FERTILIZER RATES ON YIELD COMPONENTS AND YIELD OF

FABA BEAN (Vicia faba L.) ON NITISOLS AT WOLAITA SODO,
SOUTHERN ETHIOPIA
MSc THESIS
FEKADU NIGATU
JUNE 2017
HARAMAYA UNIVERSITY, HARAMAYA
Effect of Rhizobium Inoculation and Phosphorus Fertilizer Rates on Yield
Components and Yield of Faba Bean (Vicia faba L.) on Nitisols at Wolaita
Sodo, Southern Ethiopia
A Thesis Submitted to the School of Plant Sciences

Post Graduate Program Directorate
HARAMAYA UNIVERSITY
In Partial Fulfillment of the Requirements for the Degree of

MASTER OF SCIENCE IN AGRICULTURE
(AGRONOMY)
Fekadu Nigatu
June 2017
Haramaya University, Haramaya
HARAMAYA UNIVERSITY
POSTGRADUATE PROGRAM DIRECTORATE
We hereby certify that we have read and evaluated this Thesis entitled “Effect of Rhizobium
Inoculation and Phosphorus Fertilizer Rates on Yield Components and Yield of Faba
Bean (Vicia faba L.) on Nitisols at Wolaita Sodo, Southern Ethiopia” prepared under our
guidance by Fekadu Nigatu. We recommend that it be submitted as fulfilling of the Thesis
requirement.
Jemal Abdulahi (PhD) ___________________ __________________

Major Advisor Signature Date
Tamado Tana (PhD) ___________________ __________________

Co-Advisor Signature Date
As a member of the Board of Examiners of the MSc Thesis Open Defense Examination, we
certify that we have read and evaluated the Thesis prepared by Fekadu Nigatu and examined
the candidate. We recommend that the Thesis be accepted as fulfilling the Thesis requirement
for the Degree of Master of Science in Agriculture (Agronomy).
_________________________ ___________________ __________________

Chairperson Signature Date
_________________________ ___________________ __________________

Internal Examiner Signature Date
_________________________ ___________________ __________________

External Examiner Signature Date
Final approval and acceptance of the Thesis is contingent upon the submission of its final copy
to the Council of Graduate Studies (CGS) through the candidate’s School Graduate Committee
(SGC).
ii
DEDICATION
This Thesis is dedicated to the memory of my grandfather Mr. Nigatu Melaku who was eager
to see my success but passed away, my beloved grandmother Mrs. Janame Gizaw and all my
relatives who helped and made me a man of today.
iii
STATEMENT OF THE AUTHOR
By my signature below, I declare and affirm that this Thesis is my own work. I have followed
all ethical and technical principles of scholarship in the preparation, data collection, data
analysis and compilation of this Thesis. Any scholarly matter that is included in the Thesis has
been given recognition through citation.
This Thesis is submitted in partial fulfillment of the requirements of MSc degree at Haramaya
University. The Thesis is deposited in the Haramaya University Library and is made available
to borrowers under the rule of the Library. I solemnly declare that this Thesis has not been
submitted to any other institution anywhere for the award of any academic degree, diploma or
certificate.
Brief quotations from this Thesis may be made without special permission provided that
accurate and complete acknowledgement of the source is made. Requests for permission for
extended quotations from or reproduction of this Thesis in whole or in part may be granted by
the Head of the School or Department when in his or her judgment the proposed use of the
material is in the interest of the scholarship. In all other instances, however, permission must
be obtained from the author of the Thesis.
Name: _____________________________ Signature: ____________
Date of Submission: __________________
School: ____________________________
iv
BIOGRAPHICAL SKETCH
The author, Fekadu Nigatu, was born on 13 January 1991 in Bombe town, Wolaita Zone,
southern Ethiopia. He attended his elementary and secondary education at Bombe Primary and
Secondary High Schools, respectively, from 1997 to 2008 and Preparatory program at Areka
Senior Secondary and Preparatory High School from 2008 to 2010. After passing the
Ethiopian Higher Education Entrance Certificate Examination (EHEECE) in 2010, he joined
Jimma University College of Agriculture and Veterinary Medicine (JUCAVM) and graduated
with Bachelor of Science (BSc) degree in Horticulture in June 2013.
After graduation, the author was employed by Desa Plants PLC at Mojo Ejersa in July 2013
and worked as Junior Supervisor under Production Manager of the Farm up to October 2013.
After three and half months of service at Desa Plants PLC, he was employed by Mizan-Tepi
University on 28 October 2013 to work in Horticulture Department where he served from
Graduate Assistant I to Assistant Lecturer until he joined Haramaya University, Post Graduate
Program Directorate in October 2015 to pursue study leading to Master of Science degree in
the field of Agronomy.
v
ACKNOWLEDGMENTS
Foremost, I would like to express my deepest gratitude to my major advisor Dr. Jemal
Abdulahi, for his guidance, constructive comments, close follow up by spending his time and
encouragement throughout the course of this study. I thank him also for his positive response
when I called him any time and welcome at his office and his fatherly advice. I also pay
respect and wish to express my sincere gratitude to my co-advisor, Dr. Tamado Tana for
supplying all the necessary contributions for my Thesis research, invaluable and precious
support, his intellectual guidance and advice starting from proposal development to Thesis
write up. Without his guidance and support, this work would not have been finalized
successfully. It has been a privilege to count myself as one of his honours students.
I extend my special thanks to Ministry of Education for providing finance for research work
and Mizan-Tepi University for financially supporting my post graduate study. My special
thanks go to Wolaita Sodo ATVET College for providing me land for the field experiment; all
laboratory services, inputs and equipment for field work, data collection and measurements. I
am grateful to Holleta Agricultural Research Center for providing me Rhizobium inoculants
and improved faba bean variety (Degaga) seed. I feel much pleasure to express the heartiest
gratitude to the staff and the management of Haramaya University for offering me TSP
fertilizer to undertake this research and facilitating the course work.
I would like to express my heartfelt gratitude to my grandmother Mrs. Janame Gizaw for her
nursing, support, unreserved love and keeping the belief in my ability, my mother Mrs.
Felekech Nigatu, all my family members, Mr. Afework Nigatu, Mrs. Fantaye Silalo, Mr.
Abebe Nigatu, Mr. Fikre Nigatu, Mr. Paulos Zemene, Mr. Degife Nigatu, Mr. Abenezer
Afework, Mr. Ashebir Afework and Mss. Tsigereda Tesfaye for their help, inspiration and
encouragement in my life.
Finally, I am indebted profoundly to my friends Mss. Tiblet Kasahun, Mss. Biruktait Teferi,
Mr. Endalkachew Wosenu, Mr. Simon Koroto, Mr. Endrias Chinasho, Mr. Maguje Masa, Mr.
Zewdu Paulos, Mr. Merkebu Mathewos, Mr. Dawit Yoseph, Mr. Zefine Onu, Mr. Tamene
Tadese, Mr. Terefe Hailemariam and Mr. Kalkidan Temesgen for their motivation and support
in idea, laboratory, field work and data collection during the course of study.
vi
ACRONYMS AND ABBREVIATIONS
ANOVA Analysis of Variance
ATVET Agricultural Technical, Vocational and Educational Training
BNF Biological Nitrogen Fixation
CIMMYT Centro Internacional De Mesoramiento De Maiz Y Trigo (Spanish)
International Maize and Wheat Improvement Center (English)
CSA Central Statistical Agency
EAL Ethiopian Agricultural Legumes
ETB Ethiopian Birr
HARC Holeta Agricultural Research Center
HNMA Hawassa National Meteorology Agency
IFPRI International Food Policy Research Institute
MoA Ministry of Agriculture
MRR Marginal Rate of Return
SNNPRS Southern Nations, Nationalities and Peoples’ Regional State
TSP Triple Super Phosphate
USDA United States Department of Agriculture
WSMS Wolaita Sodo Meteorology Station
vii
TABLE OF CONTENTS
STATEMENT OF THE AUTHOR iv
BIOGRAPHICAL SKETCH v
ACKNOWLEDGMENTS vi
ACRONYMS AND ABBREVIATIONS vii
LIST OF TABLES x
LIST OF TABLES IN THE APPENDIX xi
ABSTRACT xii
1. INTRODUCTION 1
2. LITERATURE REVIEW 5
2.1. Overview of Faba Bean Production 5
2.2. Bio-fertilizers and Their Importance on Faba Bean Production 6
2.3. Importance and Mechanism of Biological Nitrogen Fixation 8
2.4. Factors Limiting Biological Nitrogen Fixation 10
2.4.1. Soil temperature 10
2.4.2. Soil moisture status 11
2.4.3. Nutrient availability 12
2.4.4. Soil pH 14
2.4.5. Agronomic management 15
2.5. Importance of Phosphorus Supply on Faba Bean Production 15
2.6. Effect of Inoculation and Phosphorus Application on Yield Components and Yield
of Faba Bean 18
3. MATERIALS AND METHODS 20
3.1. Description of the Study Area 20
3.2. Experimental Materials 20
3.3. Soil Sampling and Analysis 21
3.4. Treatments and Experimental Design 21
3.5. Experimental Procedure and Plot Management 22
3.5.1. Seed treatment with Rhizobium (seed inoculation) 22
3.5.2. Management of the plots 22
3.6. Data Collection and Measurement 22
viii
TABLE OF CONTENTS (Continued)
3.6.1. Crop phenological and vegetative growth parameters 22
3.6.2. Nodulation parameters 23
3.6.3. Yield components and yield 24
3.7. Data Analysis 24
3.8. Economic Analysis 25
4. RESULTS AND DISCUSSION 26
4.1. Soil Physico-chemical Properties of the Experimental Site 26
4.2. Phenological and Vegetative Growth Parameters of Faba Bean 27
4.2.1. Days to 50% flowering 27
4.2.2. Days to 90% physiological maturity 29
4.2.3. Number of primary branches per plant 30
4.2.4. Plant height 31
4.3. Nodulation Parameters of Faba Bean 33
4.3.1. Total number of nodules per plant 33
4.3.2. Number of effective nodules per plant 35
4.3.3. Nodule volume per plant 36
4.3.4. Nodule dry weight 37
4.4. Yield Components and Yield of Faba Bean 38
4.4.1. Crop stand count at harvest 38
4.4.2. Number of pods per plant 39
4.4.3. Number of seeds per pod 41
4.4.4. Hundred seed weight 42
4.4.5. Grain yield 44
4.4.6. Above ground dry biomass yield 48
4.4.7. Harvest index 50
4.5. Economic Analysis 51
5. SUMMARY AND CONCLUSION 54
6. REFERENCES 57
7. APPENDICES 74
ix
LIST OF TABLES
Tables Page
1. Selected soil physico-chemical properties of the experimental site before planting 27
2. Days to 50% flowering of faba bean as affected by interaction of Rhizobium
inoculants and phosphorus fertilizer rate 28
3. Days to 90% physiological maturity of faba bean as influenced by the interaction of
Rhizobium inoculants and phosphorus fertilizer rate 30
4. Number of primary branches per plant of faba bean as influenced by the interaction of
Rhizobium inoculation and phosphorus fertilizer rate 31
5. Plant height (cm) of faba bean as influenced by the interaction of Rhizobium
6. Total number of nodules per plant of faba bean as influenced by the interaction of
7. Number of effective nodules per plant of faba bean as influenced by the interaction of
8. Nodule volume (ml) per plant of faba bean as affected by the interaction of Rhizobium
inoculation and phosphorus fertilizer rate 37
9. Nodule dry weight (mg) per plant of faba bean as influenced by the interaction of
10. Main effects of Rhizobium inoculation and phosphorus fertilizer rates on faba bean
stand count (plant population ha-1) at harvest 39
11. Interaction effect of Rhizobium inoculation and phosphorus fertilizer rate on number
of pods per plant of faba bean 41
12. Number of seeds per pod of faba bean as influenced by the interaction of Rhizobium
13. Hundred seed weight (g) of faba bean as influenced by the interaction of Rhizobium
14. Grain yield (kg ha-1) of faba bean as influenced by the interaction of Rhizobium
15. Above ground dry biomass yield (kg ha-1) of faba bean as influenced by the
interaction of Rhizobium inoculants and phosphorus fertilizer rate 50
16. Main effects of Rhizobium inoculation and P rates on harvest index (%) of faba bean 51
17. Summary of economic analysis of the effects of Rhizobium inoculation and
phosphorus fertilizer application on faba bean production in Wolaita Sodo 53
x
LIST OF TABLES IN THE APPENDIX
Appendix Table Page

1. Monthly total rainfall (mm) and average minimum and maximum air temperatures
(ºC) recorded in the experimental area during the growing period of test crop (July
2016 – November 2016) 74
2. Mean squares of analysis of variance showing phenological and growth parameters
of faba bean inoculation with different Rhizobium inoculants and phosphorus
fertilizer rate 74
3. Mean squares of analysis of variance for nodulation parameters of faba bean as
affected by Rhizobium inoculation and phosphorus fertilizer rates 74
4. Mean squares of ANOVA for yield components of faba bean as affected by
5. Mean squares of ANOVA for yields and harvest index of faba bean as affected by
xi
Effect of Rhizobium Inoculation and Phosphorus Fertilizer Rates on Yield
Components and Yield of Faba Bean (Vicia faba L.) on Nitisols at Wolaita
Sodo, Southern Ethiopia
ABSTRACT
Poor soil fertility, lack of indigenous soil populations of rhizobia strains and lack of adequate
information on the use of Rhizobium inoculants and phosphorus fertilizer are the major
production constraints that limit the potential yield of faba bean. Thus, field experiment was
conducted to assess the effect of Rhizobium inoculation and phosphorus fertilizer rates on
yield components and yield of faba bean; and to estimate the economically feasible
combination of inoculants and phosphorus fertilizer rate that maximizes the productivity of
faba bean in the study area during 2016 crop season at Wolaita Sodo, southern Ethiopia.
Faba bean variety Degaga was used as the test crop. Factorial combinations of three
Rhizobium inoculants (FB-1017, FB-1035 and EAL-110) and control (without Rhizobium)
and five phosphorus fertilizer levels (0, 23, 46, 69 and 92 kg P2O5 ha-1) were laid out in a
Randomized Complete Block Design (RCBD) with three replications. The result showed that
crop stand count at harvest was significantly affected by the main effect of P2O5 rates; while,
harvest index was significantly affected by the main effect of both Rhizobium inoculants and
P2O5 rates. The highest harvest index (40.11%) was obtained from Rhizobium strain FB-1017,
whereas, the highest harvest index (41.49%) was attained from 69 kg ha-1 P2O5 rate. The
interaction of Rhizobium inoculation and P2O5 rates significantly influenced the phenological
and growth parameters of faba bean. The highest number of days to flowering (55 days) was
recorded at FB-1017 and 46 kg P2O5 ha-1, whereas the highest days to physiological maturity
(111.7 days), number of primary branches per plant (3.8), total nodules (138.3) and effective
nodules (128.7) were recorded from inoculant FB-1017 at P2O5 rate of 69 kg ha-1. The tallest
plant height (137.7 cm) was obtained from inoculant FB-1035 at 69 kg P2O5 ha-1. Similarly,
the interaction of Rhizobium inoculation and P2O5 rates significantly influenced yield
components and yield of faba bean. Inoculant FB-1017 and 69 kg P2O5 ha-1 gave significantly
highest number of pods per plant (42.27), hundred seed weight (56.02 g), grain yield (4101 kg
ha-1) and above ground dry biomass (9280 kg ha-1). The economic analysis also indicated that
the highest net return (40939.80 ETB ha-1) with a marginal rate of return of 174.51% was
obtained from inoculation with Rhizobium strain FB-1017 and 69 kg P2O5 ha-1. Based on the
results of this study, it can be tentatively concluded that faba bean seed inoculation with
Rhizobium strain FB-1017 and application of P2O5 at a rate of 69 kg ha-1 found to be
appropriate for faba bean production in the study area.
Key words: Biofertilizer, Biological Nitrogen Fixation, Inoculants, Legume, Nodulation
xii
1. INTRODUCTION
Faba bean (Vicia faba L.) belongs to family Fabaceae, subfamily Faboideae, tribe Vicieae,
genus Vicia and species V. faba. It is an annual legume (Hanelt and Mettin, 1989) and known
by many common names, most of which refer to a particular subgroup rather than the whole
species. Faba bean, fava bean, broad bean, horse bean, field bean, windsor bean, tick beans,
bakela (Ethiopia), boby kurmouvje (former USSR), ful masri (Sudan), feve (French) and yeshil
bakla (Turkey) are some of the few names used in different parts of the world (Singh et al.,
2010). It is originated in the Near East and Mediterranean area and is one of the earliest
domesticated legumes after chickpea (Cicer arietinum L.) and pea (Pisum sativum L.). The
crop is grown worldwide as a protein source for food and feed (Bond et al., 1985). Ethiopia is
considered as the secondary center of diversity and also one of the nine major agro-
geographical production regions of faba bean (Asfaw et al., 1994).
Faba bean is used as human food in developing countries and as animal feed in industrialized
countries. It is widely used as a good source of protein, starch, cellulose and minerals for
human being in developing countries. It provides essential amino acids (particularly lysine)
that are not present in sufficient quantities in staple cereal crops (Giller, 2001). It is used as a
green vegetable or dried, fresh or canned in the Middle East, Mediterranean Region, China and
Ethiopia. Moreover, faba bean offers ecosystem services such as renewable inputs of nitrogen
(N) into crops and soil via biological nitrogen fixation (BNF), and a diversification of
cropping systems (Jensen et al., 2010).
Faba bean is one of the pulse crops commonly grown in Ethiopia. It is grown by subsistence
farmers in the country, during the cool main rainy season (June to September). It is the most
important pulse crop in the country, occupying about 28.43% of the total land area under
pulses (CSA, 2015). It is leading and important source of dietary protein to the majority of
population in the country and used to make various traditional dishes (Senayit and Asrat,
1994). It ranked first among cool season food legumes based on area of production and
production in quintals (CSA, 2015). The growing importance of faba bean as an export crop in
Ethiopia has led to a renewed interest by farmers to increase the area under production
(Samuel et al., 2008). Moreover, it provides large cash for producers and foreign exchange for
2
the country (Desta, 1988). The straw of the crop is also used as animal feed and soil fertility
restorer (Amanuel et al., 2000; Habtegebriel et al., 2007).
It is grown during the main rainy season on both red and black soils in Amhara, Tigray,
Oromia and Southern Nations, Nationalities and Peoples' Regional State (SNNPRS). Amhara
and Oromia regions are the major faba bean producing regions (IFPRI, 2010). According to
CSA (2015), in the country faba bean covered 538,458.21 hectares of cultivated land, from
which 991,700.28 tons production was obtained with an average productivity of 1.84 tons ha-1.
The SNNPRS is ideal for faba bean production in Ethiopia. In the region, among the total land
area of 221,974.34 hectares planted by pulses, faba bean covered 71762.58 hectares, which is
32.33% of the production area covered by all pulses grown in the region. Among all the crops
grown in the region, faba bean is the third major crop next to maize and tef (CSA, 2015). In
Wolaita Zone, faba bean is one of the major pulse crops produced for the purpose of both
home consumption and market. In the Zone out of the total land size of 33,729.25 hectares
planted by pulses, faba bean covered 1,186.40 hectares, which is 3.52% of the production area
covered by all pulses (CSA, 2015).
According to CSA (2015), in 2014/15 Ethiopian main cropping season, the productivity of
faba bean in the country was 1.84 tons ha-1, which is too low as compared to its potential yield
of 4.8 tons ha-1 (MoA, 2014). One of the principal production constraints of faba bean is poor
soil fertility, especially nitrogen deficiency. Besides, lack of improved varieties, shortage of
certified seeds, weeds, diseases like chocolate spot, rust, powdery mildew and root rot, and
insect pests such as aphids are also limiting factors (MoA, 2014). Low level of chemical and
bio-fertilizer usage and lack of effective indigenous soil populations of rhizobia strains has
limited potential yield of faba bean varieties (Asfaw and Shiferaw, 2009; Dereje et al., 2012).
In low-input cropping systems of Ethiopia, chemical fertilizers are rarely used in the
production of faba bean and other pulse crops; instead, these crops are used as a restorer of
soil fertility for the following cereal crops (Asfaw et al., 1994; Mulissa and Fassil, 2011). The
potential biological fertilizers would play key role in productivity and sustainability of soil and
also protect the environment as eco-friendly and cost effective inputs for the farmers (Tolera et
al., 2009). However, improvement of faba bean production requires selection of effective
rhizobia strains (Moawad et al., 2004).
3
Successful introduction of legumes to new farming systems has allowed by the use of
inoculants since inoculation is necessary to ensure good yield, as compatible rhizobia are
absent (Wani et al., 1995). Faba bean is a legume and is able to provide a significant level of
N2 from the soil air using a symbiotic relationship with Rhizobium bacteria. For N2 fixation to
occur, the seed or soil must be inoculated with the appropriate strain of Rhizobium. Seed
inoculation of rhizobia is one of the practices to increase the N2 fixation potential of the crops
and applications of strains improved early nodulation and increased grain yield (Carter et al.,
1998). Thus, inputs from N2 fixing bacteria may provide a sustainable alternative. However,
effective rhizobial strains may not exist in the soil, hence, application of effective rhizobial
inoculants as a bio-fertilizer is required. The success still depends on the interaction between
legume genotypes, rhizobial strain availability or application of P (Mulissa and Fassil, 2011).
In highlands of Ethiopia, phosphorus (P) is also the major yield limiting nutrients (Bereket et
al., 2011) and 70 to 75% of the agricultural soils of the highland regions of the country are P
deficient (Desta, 1982). A low level of soil P is a great obstruction to the growth and
development of leguminous crops. N fixation is very sensitive to P deficiency due to reduced
nodule mass and decreased ureide production (Sinclair and Vadez, 2002). Symbiotic N
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. It
also promotes root nodulation, nutrient-use efficiency, efficient partitioning of photosynthates
between source and sink, and biomass production (Ogola et al., 2012). Root development,
stalk and stem strength, flower and seed formation, crop maturity and production, crop quality,
and resistance to plant diseases are the traits associated with P nutrition (Ahmad et al., 2009).
The combined use of rhizobial inoculants and phosphorus supplement on legumes may play a
great practical role in cropping systems of developing countries, which can increase soil
fertility and plant productivity (Abbasi et al., 2008), thereby increasing the farm income of
farmers. Phosphorus fertilizer application along with Rhizobium inoculants influenced
nodulation and N fixation of legume crops (Bhuiyan et al., 2008). Similarly, Hussein et al.
(1997) have found that application of 37 kg P2O5 ha-1 as mono superphosphate (15% P2O5)
and inoculation with Rhizobium significantly increased faba bean seed yield by 2.23 tons ha-1,
where straw yield was also increased. Amanuel et al. (2000) have also reported that interaction
4
of P and Rhizobium inoculation (20 kg P ha-1 plus inoculation) on faba bean markedly
increased nodule number which exceeded 30 nodules plant-1. Likewise, it was reported that
application of highest concentration of P together with inoculation of effective rhizobial
isolate was found to increase growth, nodulation and yield of faba bean (Ayneabeba et al.,
2001). Hence, symbiotic N2 fixation rates and grain yield could be markedly increased by
combined application of Rhizobium and P fertilizer. However, it requires using highly
efficient, competitive and persistent strains of Rhizobium and optimizing P nutrition.
According to CSA (2015), in 2014/15 (2007) Ethiopian main cropping season, the total
production of faba bean in Wolaita Zone was 1,056.98 tons ha-1 and its productivity was 0.89 t
ha-1, which is far below the national average yield (1.84 tons ha-1). To overcome the problems
of faba bean production, the use of combination of improved varieties, inoculation with
effective Rhizobium with small amounts of mineral fertilizers is required (Palm et al., 1997).
In addition to the lack of soil rhizobial strains, P deficiency is also another significant factor
that reduces the nodulation, since both effective Rhizobium bacteria and the crop require in
larger quantity (Getachew and Rezene, 2006). Available P content of most soils in SNNPRS is
less than 5 mg kg-1, which is in the range of low P content (Kelsa et al., 1998). Particularly,
soils of many areas in Wolaita suffer from remarkable P deficiencies due to low pH, and thus,
P fixation (Eyasu and Scoones, 1999). Kassahun (2013) also reported that the available P
content in the research area of Wolaita Sodo is too low, i.e. 2.2 ppm.
Generally, adequate information on the use of Rhizobium inoculants and P fertilizer for
enhancing the yield of faba bean in the study area is lacking. Thus, conducting systematic
investigation in this line was imperative to determine their effects on production of the crop in
the study area.
In view of this, the present experiment was carried out with the following objectives:
 To assess the effect of Rhizobium inoculation and phosphorus application on yield
components and yield of faba bean; and
 To estimate the economically feasible combination of the inoculants and phosphorus
fertilizer rate that maximizes productivity of faba bean in the study area.
5
2. LITERATURE REVIEW
2.1. Overview of Faba Bean Production
Faba bean is classified as an annual grain legume, and is a vigorous pulse crop. Unlike some
other pulses, such as pea, faba bean has a thick stem that holds an increased height, up to 1.5
meters tall; stem is hollow and green, turning dark at maturity. Some other identifying
characteristics are: compound leaves, large in comparison with other pulses; flowers present at
the base of the leaf in clusters, white to pink colored, both cross and self-pollinated and
produce 1-4 pods per cluster; pods are long and green, turning dark brown-black at maturity;
three to four seeds will be formed in each pod on average and seeds vary in size and color,
depending on the variety of the faba bean (Lindsey et al., 2013).
Faba bean is one of the major leguminous crops grown in the world. It is important source of
protein for humans and animals. Faba bean has four main functions in agro-ecosystems: (1)
providing food and feed that is rich in protein; (2) supplying N to agro-ecosystems by
symbiotic N fixation with Rhizobium bacteria to increase soil fertility; (3) diversifying the crop
system to reduce constraints on growth and yield by the other crops in the rotation; and (4)
reducing fossil energy consumption for crop production (Somayeh and Hashem, 2015). The
increasing need for protein in world has stimulated scientists to research for increased yield
and quality in this crop. The faba bean is a very important source of edible protein in the diet
with a content of 18-40% for the people of the Middle East and North Africa (Haddad and
Thalji, 1988).
In Ethiopia, faba bean is the first among pulse crops cultivated and leading protein source for
the rural people and used to make various traditional dishes. It is grown as field crop
throughout the highlands and is most common in mid-lands locally called Weyna Dega,
between the altitudes 1800 and 3000 meters above sea level (m.a.s.l.). Optimum temperatures
for production range from 15 to 25°C. Rainfall of 700-1100 mm per annum evenly distributed
is ideal (MoA, 2014). The maturity period ranges from 90-220 days depending upon the
cultivars and climatic conditions (Bond et al., 1985).
In Ethiopia, over 90% of the total crop land is cultivated by small scale farmers using
traditional practices. Though, the productivity of faba bean in the country is about 1.84 tons
6
ha-1 (CSA, 2015) which is still far below its potential 4.8 tons ha-1 (MoA, 2014), due to poor
nutrient availability and lack of efficient and compatible strains of Rhizobium in the soil amid
other factors (Habtegeberial et al., 2007). However, faba bean, among the grain legumes is
reported to drive the highest percentage of nitrogen (N) from the atmosphere (Hardarson et al.,
1987), and the yield, protein content, nodulation and amount of N fixed are reported to
increase with Rhizobium inoculation and mineral nutrient applications (Shibru and Mitiku,
2000; Habtegebriel et al., 2007).
Many members of the legume plant family, such as beans, peas, alfalfa, and leucaena, have the
special ability to use biological nitrogen fixation (BNF) to meet their N needs. As a member of
this pulse family, faba bean is known for its ability to fix N. Alipour et al. (2013) reported that
faba bean acquires 79% of its required N through BNF, 20% from soil and only 1% from
fertilizers. N fixation is the result of a symbiotic relationship between the faba bean plant and
rhizobacteria. The proportions of plant N derived from BNF significantly increases with
inoculation rate (Denton et al., 2013). Hence, accumulation of enough N by the legume and
the subsequent yield effects depends on the number, efficiency and compatibility of rhizobia
and nutrient constraints that affect nodulation and N fixation (Ayneabeba et al., 2001).
2.2. Bio-fertilizers and Their Importance on Faba Bean Production
Bio-fertilizers are inputs containing microorganisms which are capable of mobilizing nutritive
elements from non-usable form to usable form through biological processes, they include
mainly the nitrogen fixing, phosphate solubilizing and plant growth-promoting
microorganisms (Goel et al., 1999). Bio-fertilizers are low cost, renewable sources of plant
nutrients which supplement chemical fertilizers. Use of bio-fertilizer is one of the important
components of integrated nutrient management, as they are cost effective and renewable
sources of plant nutrients to supplement the chemical fertilizers for sustainable agriculture.
They are one of the best modern tools for agriculture; and commonly called microbial
inoculants (Selvakumar et al., 2009).
Based on the observation that these microorganisms can have a beneficial effect on plant and
crop growth the concept of bio-fertilizer was developed (Davidson, 1988). Consequently, a
range of plant growth-promoting rhizobacteria has been identified and well characterized.
7
Direct beneficial effects can occur when the microorganisms provide the plants with useful
products. The best known cases of this are microorganisms that can directly obtain N2 from
the atmosphere and convert this into organic forms usable by plants. Such biological nitrogen
fixers include members of the genus Rhizobium, Azospirillum, and blue-green algae (Benson,
2001). Rhizobia are symbiotically associated with legumes and nitrogen fixation occurs within
root or stem nodules where the bacterium resides (Dela, 1993).
Unless fertilizer N is applied, most plants obtain their N from natural sources in the soil.
Natural reserves of soil N are normally low, so N fertilizers must be added to increase plant
growth (Silva and Uchida, 2000). The uses of bio-fertilizers effectively enrich the soil and cost
less than chemical fertilizers. For the last one-decade, bio-fertilizers are used extensively as an
eco-friendly approach to minimize the use of chemical fertilizers, improve soil fertility status
and for enhancement of crop production by their biological activity in the rhizosphere
(Selvakumar et al., 2009). Bio-microbial strains are essential since the indigenous rhizobia are
incapable of supporting adequate levels of N2 fixation. Improvement of faba bean and other
legume crops production requires selection of effective rhizobia strains (Moawad et al., 2004).
In the Rhizobium-legume symbiosis, the process of nitrogen fixation is strongly related to the
physiological state of the host plant. According to Akram et al. (2014), different varieties have
different capacity in nitrogen fixation in nodule, number of nodule formation and number of
root hairs. The beneficial effect of inoculation with rhizobia and vesicular arbuscular
mycorrhizae was reported for faba bean (Rizk and Shafeek, 2000). Inoculation had increased
total dry matter and N value of faba bean plant organs, as well as nitrogen, phosphorus and
potassium uptake (Mohamed and Gomaa, 2005).
Despite BNF being a beneficial naturally occurring process, many soils do not have sufficient
numbers of appropriate rhizobia for effective symbioses (Silva and Uchida, 2000). Thus,
inoculation of leguminous crops with appropriate and compatible rhizobia ensures maximum
BNF. The improvement in the production of these crops by inoculation with rhizobia will
therefore, contribute substantially to better human nutrition and soil health. The ability of
many legumes to form associations with bacteria that fix atmospheric nitrogen (the symbiotic
association that improve growth) is thus a big matter of ecological and economic interest
(Zahran, 2009).
8
2.3. Importance and Mechanism of Biological Nitrogen Fixation
The need of plants for N is relatively immense. For the fulfillment of this need the contribution
of biological nitrogen fixation should not be over looked. This is because N2 fixation by
micro-organisms is the major source of soil N contributing to as much as 75% of total N
content (Subba Rao, 2001). It was also stated that organic form of N is the major component
of living organisms and it is the most abundant element in the atmosphere. However, it is not
available to most organisms because of their inability to use it in the elemental form. Only a
few groups of bacteria have the ability to combine it with other elements and use it directly
and all other organisms depend on combined forms of N, mostly produced by these bacteria.
The complex process by which the rhizobia produce nitrogen for the legume is called
biological nitrogen fixation (BNF) (Benson, 2001). Only rhizobia that are specifically
compatible with a particular species of legume can stimulate the formation of root nodules, a
process called nodulation. The importance of BNF through N fixing soil organisms goes back
more than 100 years (Eaglesham, 1989). Next to photosynthesis, N fixation is the second most
important process in crop production. Photosynthesis captures sunlight and produces energy,
and N fixation uses nitrogen gas (N2) to form ammonium (NH4+). BNF is an efficient source
of N and it can provide for free up to 300-400 kg N2 ha-1 year-1 (Peoples et al., 1995).
There is a huge reservoir of N in the atmosphere; the atmosphere comprises of approximately

78% nitrogen as an inert gas, N2, which is unavailable to plants. Above every hectare of
ground there are approximately 80000 tons of this unavailable nitrogen. The gaseous N must
first be converted, or “fixed,” into forms plants can use. In order to be converted to available
form it needs to be fixed through either the industrial process (Haber Bosh Process) or through
BNF. BNF refers to the process of micro-organisms fixing atmospheric nitrogen, mostly
within subsoil plant nodules, and making it available for assimilation by plants, involves the
conversion of nitrogen to ammonia by microorganisms using a complex enzyme system
identified as nitrogenase. They may be symbiotic, associative or free-living in relation to
higher plants. Without these nitrogen-fixers, life on this planet would probably disappear
within a relatively short period of time (Benson, 2001). BNF is done by both free-living
organisms (e.g. Azotobacter, Beijerinckia, Clostridium, Bacillus, Klebsiella, Chromatium and
Rhodospirillum) and those that form symbiotic associations with other organisms (e.g.
9
Rhizobium). In agricultural settings, perhaps 80% of this biologically fixed N comes from
symbiosis involving leguminous plants (Farrand et al., 2003).
The ability of legumes to fix atmospheric N2 in symbiosis with rhizobia strains makes them
excellent colonizers of low-N environments (Graham and Vance, 2000). The rhizobacteria that
live within the root zone promote plant growth and nutrient uptake by releasing auxins and
gibberellins. Additionally, they increase indole acetic acid and gibberellic acid contents in
plants (Selvakumar et al., 2009), which are the potent growth hormones in plants. Pulse crops
have unique properties of nodulation through Rhizobium bacteria. These bacteria, through
BNF, meet about 80%–90% of the total N requirements of legumes (Verma, 1993).
Legume BNF involves a remarkable symbiosis, or mutually beneficial relationship between

the plant and N-fixing soil bacteria (Silva and Uchida, 2000). Legume nodules are special
plant organs that house N-fixing bacteria called Rhizobium. The rhizobia attracted and attach
to root hairs. The root hairs become curling and start the formation of infection thread. Root
infection is a multistep process that is initiated by pre-infection events in the rhizosphere.
Rhizobia respond by positive chemotaxis to plant root exudates and move toward localized
sites on the legume roots. For many rhizobia, primary target sites for infection are young
growing root hairs, but there are no exclusive loci for rhizobial attachment (Pieternel and Jos,
1995). Finally, rhizobia invade cortical cells, the nodule grows and fully functional nodule.
The yield of pulses in general and faba bean in particular is low, mainly limited by mineral
nutrient availability and lack of efficient and compatible strains of Rhizobium in the soil
(Habtegebrial et al., 2007). However, faba bean, among the grain legumes is reported to drive
the highest percentage of N from the atmosphere (Hardarson et al., 1987), and the yield,
protein content, nodulation and amount of N fixed are reported to increase with Rhizobium
inoculation and mineral nutrient applications (Shibru and Mitiku, 2000). Therefore,
accumulation of enough N by the legume and the subsequent yield effects depend on the
number, efficiency and compatibility of rhizobia and nutrient constraints that affect nodulation
and N-fixation (Ayneabeba et al., 2001).
Inoculation with Rhizobium can consequently lead to improved soil fertility and can reduce the
production cost of next crop through reduced input in the form of nitrogen fertilizers, which in
10
turn also minimize the health hazard effects (Noshin et al., 2008). Grain legumes have the
inherent potential to supply this important soil nutrient in cereal based cropping systems
through BNF. BNF benefits not only the legumes themselves but also any intercropped or
succeeding crop, reducing or removing the need for N fertilization. In soils with low mineral N
content, nitrogen fixing microorganisms provide ammonium into the legume biomass,
allowing faster growing than their plant competitors. In contrast, if nitrogen is abundant,
nitrogen fixing microorganisms tend to be competitively excluded by non-fixing species
because the nitrogen fixation process is bio-energetically costly (Houlton et al., 2008).
2.4. Factors Limiting Biological Nitrogen Fixation
Establishment of effective N2 fixing symbioses between legumes and their N2 fixing bacteria
is dependent upon many environmental factors, and can be greatly influenced by farm
management practices (Peoples et al., 1995). Several environmental conditions are limiting
factors to the growth and activity of the N2- fixing plants. A principle of limiting factors states
that “the level of crop production can be no higher than that allowed by the maximum limiting
factor” (Bottomely, 1991). In the Rhizobium-legume symbiosis, which is a N-fixing system,
the process of N fixation is strongly related to the physiological state of the host plant.
Therefore, a competitive and persistent rhizobial strain is not expected to express its full
capacity for nitrogen fixation if limiting factors (e.g., salinity, unfavorable soil pH, nutrient
deficiency, mineral toxicity, temperature extremes, insufficient or excessive soil moisture, and
inadequate photosynthesis) impose limitations on the vigor of the host legume (Panchali,
2011).
Despite the numerous factors that compromise nodulation and nitrogen fixation, legumes
generally assimilate 50 to 70% of their nitrogen from symbiotic nitrogen fixation (Vance,
1997). Biological processes (e.g., N fixation) capable of improving agricultural productivity
while minimizing soil loss and ameliorating adverse edaphic conditions are essential.
2.4.1. Soil temperature
Temperature plays an important role in the success of BNF due to its effect on survival and or
growth rate of microorganisms. Temperature has been reported to affect nodulation, survival
and persistence of rhizobial strains in soil (Kabahuma, 2013). High soil temperatures in
11
tropical and subtropical areas are a major problem for BNF of legume crops. High root
temperatures strongly affect bacterial infection and N fixation in several legume species,
including beans (Hungria and Franco, 1993). Temperature affects root hair infection, bacteroid
differentiation, nodule structure, and the functioning of the legume root nodule. High (not
extreme) soil temperatures will delay nodulation or restrict it to the subsurface region
(Graham, 1992). According to Drew et al. (2012), rhizobia are killed in the soil and on seed
when soil temperatures exceed 35°C.
Hernandez-Armenta et al. (1989), found that transferring nodulated bean plants from a daily
temperature of 26 to 35°C markedly inhibited N fixation. For most rhizobia, depending on
their type, exhibit an optimum growth in a temperature range of 25-30°C (Kantar et al., 2010);
and the optimum temperature range for growth in culture is 28-31°C, and many are unable to
grow at 37°C (Graham, 1992). Rhizobial survival in soil exposed to high temperature is
greater in soil aggregates than in non-aggregated soil and is favored by dry rather than moist
conditions (Graham, 1992). Ten inoculant strains of Rhizobium spp. examined by
Somasegaran et al. (1984) showed a gradual decline in population during eight weeks of
incubation at 37°C, while exposure to 46°C was lethal to all strains in less than two weeks.
2.4.2. Soil moisture status
The moisture content of the soil can have an effect on growth and survival of soil rhizobia as
well as that of the plant itself. Drought can be considered to as among the most harmful abiotic
constraints to BNF, mainly due to its effect on soil physical and biological characteristics
(Kantar et al., 2010). Drought has a pronounced effect both on the number of rhizobia and N2
fixation rates. N2 fixation is more sensitive to reductions in soil water content than other
physiological processes (Giller, 2001). The effect of drought on the soil microbial community
can be in two ways: either reducing the number of water filled pores and the thickness of water
films around soil particles or increasing the salt concentration in the soil solution. On the other
hand, also excess soil water negatively affects both the growth and survival of soil rhizobia
and plants. In conditions of water logging, the occupation of all soil pores results in limited O2
availability for both rhizobia and plant roots, thereby causing reduced respiration.
12
According to Fuhrmann et al. (1986), it is well known that some free-living rhizobia
(saprophytic) are capable of survival under drought stress or low water potential. A strain of
Prosopis (mesquite) rhizobia isolated from the desert soil survived in desert soil for one
month, whereas a commercial strain was unable to survive under these conditions (Shoushtari
and Pepper, 1985). The survival of a strain of Bradyrhizobium from Cajanus in a sandy loam
soil was very poor; this strain did not persist to the next cropping season, when the moisture
content was about 2.0 to 15.5%. Orchard and Cook (1983) described that the survival and
activity of microorganisms may depend on their distribution among microhabitats and changes
in soil moisture.
Albrech et al. (1994) reported that soil moisture deficiency has a pronounced effect on N2
fixation because nodule initiation, growth, and activity are all more sensitive to water stress
than are general root and shoot metabolism. The response of nodulation and N fixation to
water stress depends on the growth stage and species of the plants. It was found that water
stress imposed during vegetative growth was more detrimental to nodulation and nitrogen
fixation than that imposed during the reproduction stage (Albrech et al., 1994). Symbiotic N
fixation of legumes is also highly sensitive to soil water deficiency especially temperate and
tropical legumes, i.e. peas and faba bean (Guerin et al., 1990).
Numerous mechanisms have been suggested to explain the varied physiological responses of
several legumes to water stress. Morphological changes are one of the immediate responses of
rhizobia to water stress (low water potential). Mesquite Rhizobium and Rhizobium meliloti
showed irregular morphology at low water potential (Shoushtari and Pepper, 1985). The
modification of rhizobial cells by water stress will eventually lead to a reduction in infection
and nodulation of legumes. On the other hand, Ford (1984) described that legumes with a high
tolerance to water stress usually exhibit osmotic adjustment; this adjustment is partly
accounted for by changing cell turgid and by accumulation of some somatically active solutes.
2.4.3. Nutrient availability
Legume growth and the function of legume and Rhizobium symbiosis might be affected by a
multitude of nutrient disorders, including both deficiencies and toxicities (Chalk et al., 2010).
Soil salinity and acidity are usually accompanied by mineral toxicity (specific ion toxicity),
13
nutrient deficiency, and nutrient disorder. The effects of salt stress or acidity on calcium
availability and on the initial stages of nodule formation will affect the net nodulating capacity
of legumes (Graham, 1992). Aluminum and manganese toxicity and low levels of calcium
inhibit growth of rhizobia and nodulation at soil pH less than 5.0 (Drew et al., 2012).
High amount of inorganic N inhibits the Rhizobium infection process and also inhibits N2
fixation (Zahran, 1999). The former problem probably results from impairment of the
recognition mechanisms by nitrates, while the latter is probably due to diversion of
photosynthates toward assimilation of nitrates. The inhibitory effects of mineral nitrogen on
nodulation and nitrogen fixation of legumes are clear at high concentrations (>5 mM), but far
less at lower concentrations (Weisany et al., 2013). Application of large quantities of fertilizer
N inhibits N2 fixation, but low doses (<30 kg N ha-1) of fertilizer N can stimulate early growth
of legumes and increase their overall N2 fixation (Donald, 2016). The nodulation process
begins when the bacteria infect the root hairs of legumes during the early stages of the crop
after planting. At this stage, N is needed to stimulate early growth before nodulation begins in
soils with low residual N or low soil organic matter. Thereafter, legumes regulate this process
- lack of available N triggers the nodulation process and N2 fixation. The consensus is that
some level of N stress is required for symbotic relationship to fully develop. The amount of
this starter N must be defined in relation to available soil N (Graham, 1992).
Phosphorus is one of several elements which affects N fixation, and along with N, it is a
principal yield-limiting nutrient in many regions of the world (Pereira and Bills, 1989). Strains
of rhizobia differ markedly in tolerance to phosphorus deficiency. Rhizobial P deficiency
when there is a P deficiency in the soil and rhizosphere is a real possibility, especially under
acidic conditions, where dissolved phosphorus salts may be precipitated in the presence of
aluminum (Graham, 1992). Ali-Niemi et al. (1997) reported that free-living Rhizobium tropici
and bacteroids respond to P stress by increasing their P transport capacity and inducing both
acid and alkali phosphates. This P stress response occurred when the medium P concentration
decreased below 1 mM. Leguminous species differ in their phosphorus requirement for growth
from 0.8 to 3 mM (First et al., 1987).
Legume nodules are strong sinks for P and range in P content from 0.72 to 1.2%; as a
consequence, N fixation-dependent plants will require more of this element than those
14
supplied with combined nitrogen (Cassman et al., 1994). Nodulation, N fixation, and specific
nodule activity are directly related to the P supply. It has been reported that symbiotic N
fixation of the Frankia-Casuarina association requires higher P levels than those required for
plant growth, when the P concentration in soil is low (Sanginga et al., 1989).
2.4.4. Soil pH
The soil pH greatly influences rhizobia content of soils and their ability to nodulate pulse
crops (Slattery et al., 2004). Most leguminous plants require a neutral or only a slightly acid
soil pH, especially when they depend on symbiotic N2 fixation for growth with nodulation
problems to be expected once the pH falls much below 5.5 (Bordeleau and Prevost, 1994).
Soil acidity can restrict the survival and growth of rhizobia in soil and severely affect
nodulation and N2 fixation processes (Havlin et al., 2003). There exists a significant sensitivity
difference between the various rhizobial species to soil acidity. The activity of rhizobia is
affected both directly and indirectly by acidic soil conditions, the direct effect of high
hydrogen ion concentration and indirectly by the alteration of metal solubility in the soil (Tate,
2000).
Soil acidity constrains symbiotic N2 fixation in both tropical and temperate soils, limiting
Rhizobium survival and persistence in soils and reducing nodulation (Ibekwe et al., 1997).
Legumes and their rhizobia exhibit varied responses to acidity. The failure of legumes to
nodulate under acid-soil conditions is common, especially in soils of pH less than 5.0. The
inability of some rhizobia to persist under such conditions is one cause of nodulation failure,
but poor nodulation can occur even where a viable Rhizobium population can be demonstrated
(Graham, 1992).
Carter et al. (1998) found that the growth, nodulation, and yield of faba bean was improved
after inoculation with strains of Rhizobium leguminosarum bv. viciae in acid soils. It appears
that the pH sensitive stage in nodulation occurs early in the infection process and that
Rhizobium attachment to root hairs is one of the stages affected by acidic conditions in soils
(Vargas and Graham, 1988). According to Taylor et al. (1991), acidity had more severe effects
on rhizobial multiplication than did Al stress and low P conditions. The host legume appears
to be the limiting factor for establishing Rhizobium-legume symbiosis under acidic conditions.
15
Alkalinity can develop from saline soils with low calcium reserve, in lowering the water table,
soluble salts are washed down the profile and exchangeable calcium is replaced by sodium.
Soil carbon dioxide forms carbonate and bicarbonate ions and these react with sodium to raise
the pH. Very few data have been reported on the effects of high soil pH on rhizobial growth,
nodulation or legume growth. Yadav and Vyas (1971) in two surveys of 23 rhizobial isolates
from eight diverse legume species (not including Glycine max) reported that all grew well in
non-saline conditions at pH values up to 10. By contrast, none of the 17 strains of
Bradyrhizobium japonicum tested showed significant growth in liquid media at pH 8.5
(Diatloff, 1970). However, beneficial effects on root hair infection and nodulation on alfalfa
were reported on high extremes of soil pH, whereas pH above 6.0 reduced nodulation in lupins
(Tang and Robson, 1993). Eaglesham and Ayanaba (1984) are of the opinion that constraints
to rhizobial survival, nodulation and legume growth pertaining to saline-alkaline soils also
apply to saline soils where similar effects are observed.
2.4.5. Agronomic management
Establishment of effective symbiosis between rhizobia and the host plant primarily requires
optimal conditions that are necessary for growth of the host plants. In this regard, agronomic
practices have a profound influence on both the soil and the crop under consideration. For
example, the organic matter content of the soil is influenced by the agronomic management
and has several positive influences on soil fertility, moisture holding capacity and microbial
activity. Agronomic factors that influence BNF by affecting both the crop and the microbial
activity in the rhizosphere include tillage practices, selection of effective or responsive crops,
appropriate cropping systems, method of sowing, time of sowing, use of agrochemicals, use of
Rhizobium cultures and its frequency, the way of handling the inoculants and the method of
inoculation (Kantar et al., 2010; Roner and Franke, 2012).
2.5. Importance of Phosphorus Supply on Faba Bean Production
Next to nitrogen, phosphorus (P) is the most important macronutrient element for plant
growth, N fixation and adequate grain production (Brady and Weil, 2002) and making up
about 0.2-0.5% of a plant’s dry weight (Bieleski, 1973). It is a component of key molecules
such as nucleic acids, phospholipids, and adenosine triphosphate, and consequently, plants
16
cannot grow without a reliable supply of this nutrient. Phosphorus is also involved in
controlling key enzyme reactions and in the regulation of metabolic pathways (Theodorou and
Plaxton, 1993). It is vital for numerous metabolic processes both in plant and animal tissue.
Among the significant functions and qualities of plants on which P has an important effects
are: it enhances many aspects of plant physiology, including the fundamental processes of
photosynthesis, reproduction, flowering, fruiting (including seed production) and maturation;
root growth, particularly development of lateral roots and fibrous rootlets, is encouraged
several key plant structure compounds and as a catalysis in the conversion of numerous key
biochemical reactions in plants by P (Brady and Weil, 2002; Morad et al., 2013).
No soil can sustain high yields if it is deficient in P (Rao et al., 1999). Phosphorus is one of the
essential macronutrients required by plants. However, most Ethiopian soils, similar to the
agricultural soils of the other countries in the tropics, are generally low in P. This nutrient is
often limiting crop production in Ethiopia (Asgelil, 2000). Phosphorus is often a primary
factor limiting plant growth especially in tropical soils following nitrogen. Furthermore, P
deficiency is more likely to affect N dependent legumes than other species because symbiotic
nitrogen fixation is an energetically expensive process which requires more inorganic P than
mineral N assimilation. In bean, a crop widely grown in P-deficient tropical soils, production
may be highly affected by low availability of soil P, especially when the latter is accompanied
by low availability of soil N, thereby forcing plants to rely on N fixation (Israel, 1987).
Phosphorus deficiency in a tropical soil may be due to its low content or its fixation with the
formation of a complex with aluminum and iron in acidic soils, or with calcium in arid and
semi-arid soils (Tekalign and Haque, 1991). Due to this, P is also the major yield limiting
nutrients in highlands of Ethiopia (Bereket et al., 2011). A low level of soil P is a great
obstruction to the growth and development of leguminous crops. In leguminous crops, P
promotes root nodulation, N fixation, nutrient-use efficiency, efficient partitioning of
photosynthates between source and sink, and biomass production (Ogola et al., 2012). They
have an increased requirement for P over those receiving direct N fertilization, probability due
to need for nodule development and signal transduction, and to phospholipids in the large
number of bacterioids (Graham and Vance, 2000). Alternatively, faba bean requirements for P
are high due to the strong energy expenditure utilized during nodule formation and operation
17
(Kopke and Nemecek, 2010). In addition to this, it is needed to promoting the host legume
growth; it has specific roles on N fixation and nodule initiation, growth and development
(Leidi and Rodiguez-Navarro, 2000).
Although P is important to crop production and has widespread influence on both natural and
agricultural ecosystem than any other essential element, P sufficiency for crop growth does not
always exist in most soils because of losses due to erosion and high fixation (Brady and Weil,
2002). The phosphorus fertilizers also do not move readily in the soil, therefore placing the
band of phosphate near developing seedling roots of annual crops is most effective. However,
faba bean has adapted to acquire P from low P soils, and may indirectly make more P and
potassium available for subsequent crops (Kopke and Nemecek, 2010).
Yield and N fixation rate of faba bean and other legume crops can be enhanced by supplying P
fertilizer to the soil deficient in available P, since legumes require sufficient P for effective
symbiosis. The average of three different studies in Ethiopia on the need for inoculation in
faba bean showed that the response to inoculation alone was limited, but that in combination
with P fertilizer, grain yields were three times as high as in the control (Kemal et al., 2006).
Thus, an alternative source for nutritional elements is required for improvement of faba bean
production that minimizes environmental pollution. One of the options is to increase the yield
and quality of faba bean plant in combination of improving through agricultural practices such
as fertilization with phosphate and seed inoculation with different strains of biofertilizer.
The study conducted elsewhere by Zafar et al. (2011) to investigate the influence of integrated
phosphorus supply and plant growth promoting rhizobacteria on growth, nodulation, yield and
nutrient uptake in Phaseolus vulgaris indicated that two mineral phosphorus fertilizers at rate
of 60 kg P ha-1 diammonium phosphate and triple super phosphate increased plant height,
number of nodules per plant, nodule fresh weight and nodule dry weight. El-Gizawy and
Mehasen (2009) also recorded highest plant height, number of branches, 100 seed weight and
seed yield per plant of faba bean with application of 30 kg P ha-1, while Rahman et al. (2007)
observed highest number of pods per plant, pod length and pod circumference of common
bean with the treatment that received 60 kg P ha-1 as compared to 0 and 40 kg P ha-1. Serraj et
al. (1998) further specified that nodule establishment and function are important sinks for P,
and nodules usually have the highest P content in the plant. Therefore, phosphorus deficiency
18
conditions result in reduced symbiotic nitrogen fixation potential and phosphorus fertilization
would usually result in enhanced nodule number and mass, as well as greater nitrogen fixation
activity and seed yield per plant.
2.6. Effect of Inoculation and Phosphorus Application on Yield Components

and Yield of Faba Bean
Phosphorus fertilizers and their combined use with rhizobial inoculants can further increase
grain yield. Singh et al. (2008) reported that the yield of legumes is greatly influenced by
application of phosphorus and biofertilizer. Rhizobium bacteria use phosphorus as an essential
ingredient in converting atmospheric N to ammonium, a form useable by plants (Dakora and
Keya, 1997). Phosphorus fertilizer application along with Rhizobium inoculants influenced
nodulation and N fixation of legume crops (Bhuiyan et al., 2008). Hoque and Haq (1994) also
observed similar trend when they treated several legumes with Rhizobium and phosphorus and
found an increase in the number of nodules and maximum growth features.
Studies by Ndakidemi and Semoka (2006) have found that, for farmer who can afford P
fertilizers, their combined use with rhizobial inoculants can farther increase grain yield,
enhances symbiotic establishment for increased N2 fixation, and reduce the declining state of
soil fertility in Africa. This was due to the fact that seed inoculation with proper Rhizobium
strain together with minor amounts of P at early growth stage stimulated the root nodulation
and increase BNF eventually improving yield components such as number of branches per
plant, number pods per plants, number seeds per pod and seed weight in legume plants (Morad
et al., 2013). Salem and Elmasri (1986) also found that yield of faba bean increased most
when seeds were Rhizobium inoculated and plots were fertilized by phosphorus.
The combined use of inoculation and P fertilization significantly improved grain yield as 2.87
tons ha-1 and haulm yield (nutritive value of grain haulms) as 3.61 tons of dry matter ha-1 over
control and two factors independently in faba bean (Sisay et al., 2016). Amanuel et al. (2000)
also reported that P application and Rhizobium inoculation on faba bean increased the number
of nodules per plant non-significantly, but nodule number increased markedly due to the
interaction of both factors (20 kg P ha-1 plus inoculation). Correspondingly, Solomon (2016)
reported that interaction of Rhizobium inoculation and P fertilizer application (69 kg P2O5 ha-1)
19
on faba bean significantly increased total number of nodules per plant (282.42%), number of
primary branches per plant (283.33%), number of seeds per pod (53.6%), hundred seed weight
(49.86%) and grain yield (135.38%) over un-inoculated and nil phosphorus application.
Likewise, the study on other pulse crop (soybean) revealed that the interaction of biofertilizer
(Bradyrhizobium japonicum) and P levels significantly influenced growth and nodulation
parameters, such as plant height, nodule number, nodule volume, nodule dry weight and shoot
N content (Workineh et al., 2012). Similarly, the combined use of 46 kg ha-1 P2O5 and B.
japonicum significantly maximized the mean seed yield per plant of soybean by 59.2% as
compared to the control. Furthermore, the combined use of 46 kg ha-1 P2O5 and B. japonicum
increased the mean seed yield of soybean by 34.1% over the use of seed inoculation by B.
japonicum alone (Masresha and Kibebew, 2017).
Tahir et al. (2009) reported that application of Rhizobium inoculation and P fertilization
significantly increased nodule numbers from 73 in un-inoculated control (with no P
application) to 125 and 95 in Rhizobium inoculation and P treatments, respectively.
Application of P alone enhanced nodulation by 30% while its combination with Rhizobium
inoculation resulted in 47% more nodules than P alone. Correspondingly, Fatima et al. (2007)
concluded that combined application of P and Rhizobium inoculation increased nitrogenease
activity, growth, and grain yield as well as improved soil fertility for sustainable agriculture.
Likewise, the study on phosphorus and Rhizobium inoculation combination undertaken on
mung bean indicated that plant height and number of branches per plant were significantly
affected by inoculum and phosphorus application. Increased response in plant height at 35 kg
P2O5 ha-1+ Rhizobium inoculum while, the highest number of branches per plant was reported
at 65 kg P2O5 + inoculum (Muhammad et al., 2004). According to the report, the maximum
number of pods, and grains per pod were recorded at 80 kg P2O5 ha-1 + inoculum.
The results of the previous studies suggest that increasing faba bean and other pulse crops
productivity and enriching the soil with N on smallholder farms is possible and that the use of
Rhizobium inoculants with optimum phosphorus fertilizer is an appealing strategy to achieve
this. However, it requires using highly efficient, competitive and persistent strains of
Rhizobium and optimizing P nutrition.
20
3. MATERIALS AND METHODS
3.1. Description of the Study Area
The study was conducted at Wolaita Sodo ATVET College research field, Wolaita Zone of
Southern Nations, Nationalities and Peoples’ Regional State, Ethiopia. Geographically, the
area is located at 6°50'13'' N latitude and 37°45'37'' E longitude and at an altitude of 1854
meters above sea level. The area has a bimodal rainfall distribution with average annual
amount of 1572.3 mm and, the annual average minimum and maximum air temperatures are
14.9°C and 23.1°C, respectively (WSMS, 2014). The soil type of the area is Nitisols with
sandy clay loam texture and the pH is about 5.4 (Table 1).
The monthly meteorological data (amount of rainfall and maximum and minimum
temperatures) for the growing season (July to November 2016) at the experimental site is
presented in Appendix Table 1. The total rainfall in the cropping season was 480.8 mm, and
the mean maximum and minimum temperatures were 24.4°C and 15.04°C, respectively
(HNMA, 2016). The trend of rain fall of 2016 cropping season showed that it increased from
July to September, where the maximum value was recorded in September. The rain fall started
to decline as of October, which was at the normal time of physiological maturity of the crop.
3.2. Experimental Materials
The experimental materials used in this study were strains of Rhizobium (Local isolated FB-
1017, FB-1035 and EAL-110) and mineral fertilizer, Triple Superphosphate (TSP) (46% P2O5)
as source of phosphorus (P).
Improved faba bean variety namely Degaga (R-878-3), which was released by Holleta
Agricultural Research Center (HARC) in 2002 was used as the test crop. Some of the
agronomic characteristics and climatic requirements of the variety involve: altitude (1800 -
3000 meters above sea level); rainfall (800-1100 mm); temperature (18-27°C); disease
resistance (moderately resistant to chocolate spot, Botrytis fabae); days to flowering (45-62
days), days to maturity (125 days); seed size (medium); and yield at research field (2700 -
4200 kg ha-1) (MoA, 2014). It was chosen as the test variety because it is high yielder, well
adapted and widely grown in the area by smallholder farmers. Both the strains of Rhizobium
21
and the improved variety of faba bean were obtained from HARC and TSP was obtained from
Haramaya University.
3.3. Soil Sampling and Analysis
Before land preparation, soil samples were randomly taken from the experimental site in
zigzag manner from ten spots at a depth of 0-30 cm using an auger as described by Munson
and Nelson (1993), and the samples were mixed thoroughly to produce 1.0 kg of a
representative composite sample. The composite sample was put in polythene bag and taken to
Wolaita Sodo Soil Testing Laboratory for analysis of pH, organic carbon (%), total nitrogen
(%), available phosphorus (mg kg-1), cation exchange capacity (CEC) [cmol (+) kg-1] and soil
texture.
Soil organic carbon was estimated by wet digestion method (Walkley and Black, 1934). The
soil pH was measured using 1:2.5 soil-water suspensions with standard glass electrode pH
meter (Van Reeuwijk, 1992)
Total N was determined using the Kjeldhal digestion method (Jackson, 1958) with distillation
and titration as described by Cottenie (1980). Available P was determined with acidic
extracting solution using Olsen method (Olsen et al., 1954). Cation exchange capacity (CEC)
was measured using 1 M ammonium acetate, washed with ethanol and the adsorbed
ammonium was replaced by sodium (Na). Then, the CEC was determined titrimetrically by
distillation of ammonia that was displaced by Na (Sahlemedhin and Taye, 2000). Soil texture
was determined by Bouyoucos Hydrometer Method (Boyadgiev and Sayegh, 1992) using the
textural triangle of USDA system.
3.4. Treatments and Experimental Design
The experiment consisted of two factors, namely four levels of inoculation with Rhizobium
strains (Local isolated FB-1017, FB-1035 and EAL-110) and a control (without Rhizobium)
and inorganic phosphorus fertilizer at five rates (0, 23, 46, 69 and 92 kg P2O5 ha-1), making a
total of twenty treatment combinations factorially arranged in randomized complete block
design (RCBD) with three replications. The plot size was 2.4 m × 3.0 m (7.2 m2). The inter
and intra-row spacings were 40 cm and 10 cm, respectively. There were six rows per plot and
22
30 plants in each row. The outermost row in each plot and two plants at both ends of each row
were considered as border. The next one row from outermost rows was used for destructive
sampling. Thus, the net plot size was three rows of 1.2 m × 2.8 m (3.36 m2).
3.5. Experimental Procedure and Plot Management
3.5.1. Seed treatment with Rhizobium (seed inoculation)
Seeds of faba bean were inoculated using sticker (sugar solution). The seeds were divided into
three bowls for three different inoculants (FB-1017, FB-1035 and EAL-110) with each
containing about 1.0 kg of seeds. The carrier based moist peat inoculants of each strain were
applied at a rate of 10 g for 1.0 kg seed of a bowl after sprinkling sticker solution with 1:1
ratio of sugar (10% of sugar solution) and potable water. The seeds and inoculants in the
bowls were mixed carefully until seeds were coated with black film of inoculants. To maintain
the viability of inoculant cells, inoculation was done under a well shade condition and allowed
to air dry for 30 minutes.
3.5.2. Management of the plots
The experimental field was ploughed and disked by tractor and pulverized to a fine tilth by
oxen. The plots were leveled manually. The dried seeds were planted by hand at a specified
spacing by placing two seeds per hill on July 16, 2016. Plots receiving no inoculation were
planted before inoculated ones to avoid the possibility of cross contamination. The seeds were
immediately covered with soil after sowing to avoid death of cells due to the sun’s radiation.
Uniform dose of nitrogen (20 kg ha-1) was applied as a starter fertilizer (Desta, 1988) to all the
plots using urea (46% N) along with P as per the treatment. Triple super phosphate (TSP) was
applied basally at specified rate in all the respective plots at the time of planting. All the other
agronomic practices were followed as per the recommendations for the crop (MoA, 2014).
3.6. Data Collection and Measurement

3.6.1. Crop phenological and vegetative growth parameters
Days to 50% flowering: This was determined by counting the number of days from planting
to the time when first flowers appeared in 50% of the plants in a plot.
23
Days to 90% physiological maturity: It was determined as the number of days from planting
to the time when 90% of the plants started senescence of leaves (yellowing of the foliage) and
pods started to turn yellow.
Number of primary branches per plant: It was determined by counting the total number of
branches on randomly pre-tagged five plants in the net plot at physiological maturity and
averaged on per plant basis.
Plant height (cm): It was measured at physiological maturity from the base to the tip of a
plant for randomly pre-tagged five plants in harvestable rows using meter tape and averaged
on a plant basis.
3.6.2. Nodulation parameters
Total number of nodules: Nodulation was assessed at 50% flowering stage (mid flowering
stage) of the plants. Five randomly selected plants from destructive rows were carefully
uprooted for determination of nodulation by exposing roots with bulk of root mass and
nodules. Soil adhering to the roots was removed by gently washing with tap water. Nodules
from the crown and lateral roots were removed separately and spread on a sieve for some
minutes until the water was drained from the surface of the nodules. The total number of
nodules was counted and the average number of nodules per plant was calculated.
Number of effective nodules: For determination of effective number of nodules, the inside
color of nodules was observed by cutting each with the help of sharp blade and the pink
colored nodules were considered as effective nodules, while green colored were considered as
non-effective.
Nodule volume (ml): The volume of nodules collected from five sample plants was
determined by displacement of water using 20 ml graduated cylinder. The volume was divided
by the number of plants to get the mean volume of nodules per plant.
Nodule dry weight (mg): The nodules collected from five plant samples in each plot were
pooled including the dissected nodules for color determination, and their dry weight was
measured by drying in an oven at 70°C for 24 hours and averaged on a plant basis.
24
3.6.3. Yield components and yield
Crop stand count: The plants from the net plot area were counted at crop harvest to find the
plant population per hectare.
Number of pods per plant: It was recorded based on five pre-tagged plants in each net plot
area at harvest and the average was taken as number of pods per plant.
Number of seeds per pod: The total number of seeds in the pods of the five plants was
counted and divided by total number of pods to find the number of seeds per pod.
Hundred seed weight (g): The weight of 100 seeds was determined for each plot using a
sensitive balance. The weight was adjusted to a moisture content of 10%.
Grain yield (kg ha-1): Grain yield was measured by harvesting the crop from the net plot area.
The moisture was adjusted to 10%. The final yield was determined by using the formula as:
100−M
Adjusted grain yield = Recorded grain yield × ( 100−D ); where, M is the measured moisture
content and D is the designated moisture content (10%).
Above ground dry biomass yield (kg ha-1): At physiological maturity, five plants randomly
taken from the destructive rows of each plot were used to determine aboveground dry biomass
yield, which was measured after sun drying till a constant weight. The dry biomass per plant
was then multiplied by the total number of plants per net plot and was converted into kg ha-1.
This value was used to calculate the harvest index as well.
Harvest Index (HI): Harvest index was calculated by dividing grain yield per plot by the total
Seed yield
above ground dry biomass yield per plot. HI (%) = Above ground dry biomass yield × 100
3.7. Data Analysis
Data collected were subjected to analysis of variance (ANOVA) appropriate to factorial

experiment in RCBD according to the General Linear Model (GLM) of GenStat 15th edition
(GenStat, 2012) and interpretations were made following the procedure described by Gomez
and Gomez (1984). Whenever the effects of the treatments were found significant, the means
were compared using least significance difference (LSD) test at 5% level of significance.
25
3.8. Economic Analysis
An economic analysis was done using partial budget procedure described by CIMMYT
(1998). Marginal rate of return was calculated by dividing the net increase in yield of faba
bean for each phosphorus rate and Rhizobium inoculants by the cost of phosphorus and
Rhizobium inoculants applied to each treatment. Labour cost involved for inoculation of faba
bean seeds with Rhizobium and application of P2O5 fertilizer was recorded and used also for
this analysis. The current (November to December, 2016) price of the faba bean grain was
valued at an average open market price at Wolaita Sodo town which was 12 ETB kg-1. The net
returns (benefits) and other economic analysis were based on the formula developed by
CIMMYT (1998) and given as follows:
Unadjusted grain yield (UGY) (kg ha-1): is an average yield of each treatment.
Adjusted grain yield (AGY) (kg ha-1): is the average yield adjusted downward by a 10% to
reflect the difference between the experimental yield and yield of farmers.
Gross field benefit (GFB) (ETB ha-1): was computed by multiplying field/farm gate price
that farmers receive for the crop when they sell it as adjusted yield. GFB = AGY × field/farm
gate price for the crop.
Total variable cost (TVC) (ETB ha-1): was calculated by summing up the costs that vary,
including the cost of TSP fertilizer (39 ETB kg-1) and Rhizobium bio-fertilizer (160 ETB ha-1)
at the time planting (July 16, 2016), and labour cost for application of TSP (6 persons ha-1,
each 50 ETB day-1) and inoculation of seeds with bio-fertilizer (4 persons ha-1, each 50 ETB
day-1). The costs of other inputs and production practices such as labour cost for land
preparation, planting, weeding, harvesting and threshing were considered the same for all
treatments or plots.
Net benefit (NB) (ETB ha-1): was calculated by subtracting the total variable costs (TVC)
from gross field benefits (GFB) for each treatment. NB = GFB – TVC
Marginal rate of return (MRR) (%): was calculated by dividing change in net benefit
∆NB
(ΔNB) by change in total variable cost (ΔTVC). MRR = ∆TVC × 100
26
4. RESULTS AND DISCUSSION
4.1. Soil Physico-chemical Properties of the Experimental Site
Result of the soil analysis indicated that texture of the soil in the experimental site was
dominated by sand fraction, as it contained 51% sand, 28% silt and 21% clay, and classified as
sandy clay loam (Table 1). Sandy clay loam soil is grouped as medium textured soil and is
ideally suited for faba bean production (Singh et al., 2013). The soil reaction of the
experimental site was moderately acidic with a pH of 5.4 according to Tekalign (1991), who
reported that soils having pH value in the range 5.3-5.9 are moderately acid. Unkovich et al.
(1997) and Graham (2005) reported that in soils with pH lower than 5, survival of Rhizobium
becomes critical. This implies that the pH of the experimental soil was within the ranges of
optimum for nodulation.
The analysis revealed that the available P of the soil was 2.16 mg kg-1 (Table 1). Indicative
ranges of available phosphorus have been established by Olsen et al. (1954), as <5 mg kg-1
(very low), 5-15 mg kg-1 (low), 15-25 mg kg-1 (medium), and >25 mg kg-1 of soil (high). Thus,
the soils of the experimental site was considered as very low in available P content which is
unsatisfactory for optimum faba bean growth and yield. Similarly, Kassahun (2013) and
Zewde (2016) have reported that the available P content of the soil in Wolaita Sodo area is
very low, i.e. 2.2 to 2.58 mg kg-1. The low available P could be due to fixation in such acidic
soils with Aluminum (Al) and Iron (Fe) oxides. Thus, it is important to apply phosphorus
fertilizer from external sources based on the recommended rate.
Tekalign (1991) has classified soil total N content of <0.05% as very low, 0.05-0.12% as poor,
0.12-0.25% as moderate and >0.25% as high. According to this classification, the soil samples
were found to have moderate level of total N (0.133%) (Table 1), indicating that the nutrient is
a limiting factor for optimum crop growth. Thus, the level of N in the soil showed the need to
apply starter N for legumes until they start to fix atmospheric N2. In line with this, Kumar et
al. (1993) reported that incorporation of starter N together with P, regardless of the N content
in the soil, therefore, stimulates growth and enhances efficient nodulation and assimilation of
N by faba bean. Otieno et al. (2009) also indicated that N is required as starter fertilizer at the
early growth stage since there is no nodule formation where atmospheric N is fixed.
27
The soil of the study site had 1.02% of organic carbon (OC) (Table 1), which is low according
to the rating of Tekalign (1991), who rated soils having OC value in the range of 0.5-1.5% as
low, indicating low potential of the soil to supply nitrogen to plants through mineralization of
organic carbon. The low amount of SOC might be due to low addition of crop residues, less
aeration due to soil compaction of sub-soils, and slow activity of both zymogenous and
autochithnous microorganisms that play great role in organic material decomposition
(Neumann and Romheld, 1999). Thus, regular application of organic manure such as crop
residues and farm yard manure is required for the study area.
Cation exchange capacity (CEC) is an important parameter of soil, because it gives an

indication of the type of clay mineral present in the soil and its capacity to retain nutrients
against leaching. According to Landon (1991), top soils having CEC greater than 40 cmol (+)
kg-1 are rated as very high and 25-40 cmol (+) kg-1 as high, and 15-25, 5-15 and < 5 cmol (+)
kg-1 of soil are classified as medium, low and very low, respectively, in CEC. According to
this classification, the soil of the experimental site had medium/moderate CEC (20.96 cmol (+)
kg-1 soil), indicating its medium capacity to retain cations (Table 1). This medium CEC may
be attributed to the low percentage of clay (21%) and high percentage of sand (51%) and also
acidic nature of the soil.
Table 1. Selected soil physico-chemical properties of the experimental site before planting
Physical properties Chemical properties
Particle size Textural OC Total Av. P CEC
distribution (%) class pH (%) N (%) (mg kg-1) [cmol (+)
Sand Silt Clay Sandy kg-1]
51 28 21 clay loam 5.4 1.02 0.133 2.16 20.96
Moderately Low Moderate Very low Medium/
Rating acidic moderate
Where, OC= organic carbon; Total N= total nitrogen; Av. P= available phosphorus; CEC= cation exchange
capacity.
4.2. Phenological and Vegetative Growth Parameters of Faba Bean
4.2.1. Days to 50% flowering
The analysis of variance indicated that days to 50% flowering were highly significantly
(P<0.01) influenced by the main effect as well as the interaction effect of Rhizobium strains
(inoculants) and P application rate (Appendix Table 2). Inoculation of faba bean seeds with
28
Rhizobium strain FB-1017 coupled with application of 46 kg P2O5 ha-1 delayed flowering
(55.00 days) and it was statistically at par with the same inoculant at 69 kg P2O5 ha-1, while
un-inoculated plot with 92 kg P2O5 ha-1 enhanced 50% flowering in 44.67 days (Table 2).
The difference between the earliest and the latest to reach 50% flowering was 10.33 days,
which is remarkable. The possible reason for delayed flowering due to inoculation with FB-
1017 and application 46 kg P2O5 ha-1 might be enhanced nitrogen fixation and, thereby
increased N uptake by plants which might have contributed to improved vegetative growth of
the crop and availability of use more P in the soil. Such positive response of shoot
development to inoculation and P fertilization was also reported for chickpea (Verma et al.,
2013). The result is also in agreement with the finding of Abera (2015), who reported that
days to initiate 50% flowering was highly significantly (P<0.01) influenced by the interaction
of P rate and Rhizobium inoculants, where chickpea seed inoculation with inoculant Cp9 and
application of 20 kg P2O5 ha-1 delayed days to flowering (52.67 days), while plots supplied
with inoculant Cp7 and 0 kg P2O5 ha-1 reached 50% flowering in 40 days.
All inoculants gave significantly the less days to 50% flowering at 0 kg P2O5 ha-1. The reason
for this might be that nodulated legumes require high levels of P for optimal symbiotic
performance as there is close relationship between P level and symbiotic mechanism in
legumes. In line with this result, Amijee and Giller (1998) have reported that poor nodulation
and poor plant vigour have been observed in beans grown in soils low in extractable P.
Table 2. Days to 50% flowering of faba bean as affected by interaction of Rhizobium

inoculation and phosphorus fertilizer rate
P2O5 rates (kg ha-1)
Rhizobium strain 0 23 46 69 92
de ef
Control 50.00 48.67 48.33ef 46.00hi 44.67i
ghi fg
FB-1017 46.33 48.00 55.00a 54.33a 51.00cd
FB-1035 45.67i 47.67fgh 51.67bcd 53.33ab 49.00ef
i ghi
EAL-110 45.33 46.33 51.33cd 52.33bc 48.33ef
LSD (5%) 1.74
CV (%) 2.1
Where, LSD (5%) = Least significant difference at 5% level of significance; CV= Coefficient of variation; Means
followed by the same letter(s) with in columns and rows are not significantly different at 5% level of significance
29
4.2.2. Days to 90% physiological maturity
Days to 90% physiological maturity was highly significantly (P<0.01) influenced by the main
factors of Rhizobium strains (inoculants) and phosphorus application rate as well as by their
interaction (Appendix Table 2). The longest days to physiological maturity (111.7 days) were
recorded for the combination of inoculant FB-1017 and 69 kg P2O5 ha-1, whereas, the shortest
duration (101.0 days) was recorded for the control (without inoculants) along with application
of 92 kg P2O5 ha-1, which was statistically at par with no inoculation but alone 69 kg P2O5 ha-1,
and EAL-110 and FB-1035 inoculation with 0 kg P2O5 ha-1 (Table 3).
The delayed maturity due to inoculation with FB-1017 coupled with application of 69 kg P2O5
ha-1 might have been caused by the promoted vegetative growth due to enhanced supply of N
through BNF, and effective use of the applied P by the Rhizobium bacteria as an essential
ingredient in converting atmospheric N2 to ammonium (NH4+), a form useable by plants. This
result is in line with that of Abera (2015), who reported that days to 90% physiological
maturity of chickpea was highly significantly (P<0.01) affected by inoculation and P fertilizer
rate, where, the longest time to maturity (114 days) was recorded for the combination of
inoculant Cp9 and 20 kg P2O5 ha-1, while, the shortest period (102 days) was recorded for Cp7
and 0 kg P2O5 ha-1. Furthermore, Ibsa (2013) reported that days to maturity was significantly
(P<0.001) extended by the applied soil fertility treatments (Rhizobium + 46 kg P2O5 ha-1) with
the shortest maturity time observed for the control, where, inoculation in combination with P
application, resulted in the longest (104 days) growing period of chickpea than the control
(101 days). Similar effect was also reported earlier where inoculation and P application
delayed maturity time of common bean (Buttery et al., 1987) and chickpea (Gan et al., 2009).
The result of the present study also indicated that at 0 kg P2O5, inoculants EAL-110 and FB-
1035 shortest duration to maturity. Likewise, application of P alone also exerted a significant
effect on days to maturity with highest mean maturity days of 107.3 at 0 kg P2O5 ha-1, which
was significantly shorter than that of inoculation plus P treatments at 46 and 69 kg P2O5 ha-1,
but longer than that of the control (un-inoculated). The reason for this might be that nutrients
taken up by plant roots from the soil were not used for increased cell division and synthesis of
carbohydrate, which would predominantly be partitioned to the vegetative sink of the plants.
The present result agrees with the findings of Giller (2001), who has reported that deficiency
30
of P can even prevent nodulation by legumes, showing the sensitivity of the process of N
fixation to P deficiency. The result is also in conformity with that of Acharya et al. (2007) who
reported that P is important for flowering and seed formation and fastening crop maturity.
Table 3. Days to 90% physiological maturity of faba bean as influenced by the interaction of
Rhizobium inoculants and phosphorus fertilizer rate
Control 107.3de 105.3fg 104.3g 102.3ij 101.0j
FB-1017 102.7hi 104.0gh 108.7bcd 111.7a 107.7cde
FB-1035 102.0ij 104.0gh 108.0cd 110.0b 106.3ef
ij hi cd bc
EAL-110 101.7 102.7 108.3 109.0 107.3de
LSD (5%) 1.564
CV (%) 0.9
4.2.3. Number of primary branches per plant
There was a highly significant (P<0.01) effect of the main factors of seed inoculation with
Rhizobium and phosphorus fertilizer application rate as well as their interaction on number of
primary branches per plant (Appendix Table 2). The highest number of primary branches per
plant (3.80) was recorded from Rhizobium inoculant FB-1017 plus 69 kg P2O5 ha-1 and it was
statistically at par with inoculation of FB-1035 and application of 69 kg P2O5 ha-1, whereas,
plants without Rhizobium and P produced the lowest (1.933) number of branches (Table 4).
The increase in number of primary branches per plant over the control (without both
Rhizobium and P) in response to interaction of Rhizobium and phosphorus might be due to
higher vegetative growth of the plants with higher N content by BNF and due to the fact that
faba bean produces most of its primary and secondary branches during the early vegetative
growth period when there is high soil nitrogen or effective nodules. On the other hand, without
proper fertilization by phosphorus, rhizobial activities and nitrogen fixation might have been
depressed because phosphorus promotes early formation of lateral, fibrous and healthy roots.
This result is in agreement with the findings of Solomon (2016), who reported significant
(P<0.05) interaction effect of Rhizobium inoculation and P rate on number of primary
branches per plant of faba bean, where seed treatment with Rhizobium strain FB-1035 at 69 kg
P2O5 ha-1 gave the highest number of branches per plant (2.3), while, the lowest value was
31
recorded for the control (without both Rhizobium and P application). This result was also in
accordance with that of Ahmed et al. (2010), confirming positive and significant effects of
inoculation and P fertilization on number of branches of chickpea.
The possible reason for lower number of primary branches per plant at 0 kg P2O5 ha-1 might be
the fact that legumes require phosphorus for optimal symbiotic performance, as there is a close
relationship between P level and symbiotic mechanism in legumes. Similar result has also
been reported by Bildirici and Yilmaz (2005) for faba bean where small number of primary
branches was recorded at 0 kg P2O5 ha-1.
Table 4. Number of primary branches per plant of faba bean as influenced by the interaction of
Rhizobium inoculation and phosphorus fertilizer rate
Control 1.933i 2.267ghi 2.400fgh 2.533efg 2.267ghi
FB-1017 2.333gh 2.733cdef 3.000bc 3.800a 2.933cd
hi efg b a
FB-1035 2.067 2.533 3.333 3.700 2.800cde
EAL-110 2.333gh 2.400fgh 2.800cde 3.333b 2.600defg
LSD (5%) 0.3608
CV (%) 8.1
4.2.4. Plant height
Analysis of variance showed that plant height was highly significantly (P<0.01) influenced by
the main factors. The interaction of Rhizobium inoculation and P fertilizer application was also
significant (P<0.05) (Appendix Table 2), where, the maximum plant height (139.4 cm) was
observed at inoculant FB-1035 with 69 kg P2O5 ha–1, followed by inoculant FB-1017 with 69
kg P2O5 ha-1, while, the minimum value (111.9 cm) was recorded from un-inoculated and
unfertilized plots (Table 5).
The increase in plant height due to inoculation with FB-1035 and FB-1017 combined with 69
kg P2O5 ha–1 could be associated with increased availability of N from BNF by Rhizobium
inoculants and, thus, maximum vegetative growth of the plants. On the other hand, the
variation among treatments for plant height was probably due to the difference among
Rhizobium strains for effectiveness in BNF and the effect of higher rate of P on meristematic
cell activities that can play a vital role in vegetative growth of faba bean. This result is in
32
agreement with the findings of Muhammad et al. (2004), who have reported that phosphorus
and Rhizobium inoculation significantly increased plant height of mung bean. Similarly,
Ahmed et al. (2010) have observed positive and significant effects of inoculation and P
fertilization on plant height of chickpea. Furthermore, Togay et al. (2008) have reported
enhanced morphological growth due to inoculation and increased supply of nitrogen through
BNF, and direct application of phosphorus fertilizer. Pandey et al. (1998) have also reported
that inoculation with N fixers has a potential importance to improve growth and yields of the
crop, not only due to high N fixation activity, but also due to the ability of these bacteria to
produce antibiotics, growth promoting substances (phytohormones), siderophores (Fe) and the
ability to solubilize phosphate that is not available for plant use.
Moreover, the increased plant height due to Rhizobium inoculation along with P2O5
application may possibly be attributed to inoculation-induced increase in indole acetic acid
and gibberellic acid production, which subsequently increase cell division and cell elongation
(Afzal et al., 2010). Likewise, it appears that Rhizobium increases plant height by increasing
phosphorus solubility and its availability to plants. Consistent with results of the present study,
Mekki (2016) has reported that inoculation with biofertilizer and P application (34.5 kg P2O5
ha-1) significantly increased height of faba bean plant by 44.45% over the control, by 23.53%
over biofertilizer alone and by 5.16% over P alone. Masresha and Kibebew (2017) also
reported that the maximum mean plant height improvement in soybean (22.5%) was obtained
due to the interaction of Bradyrhizobium japonicum and 46 kg P2O5 ha-1.
Table 5. Plant height (cm) of faba bean as influenced by the interaction of Rhizobium
inoculants and phosphorus fertilizer rate
l ij gh cde
Control 111.9 120.6 124.1 130.0 127.1ef
FB-1017 119.5ijk 126.2fg 132.1bc 137.7a 129.3cde
jk fg cd a
FB-1035 118.6 126.1 130.3 139.4 128.8def
EAL-110 117.2k 122.1hi 130.1cde 134.5b 128.9def
LSD (5%) 2.999
CV (%) 1.4
33
4.3. Nodulation Parameters of Faba Bean
4.3.1. Total number of nodules per plant
The main effect of Rhizobium inoculation and phosphorus fertilizer application as well as their
interaction was highly significant (P<0.01) for total number of nodules per plant (Appendix
Table 3). The highest (138.3) and the lowest (80.4) number of nodules per plant were recorded
when the seeds were inoculated with Rhizobium strain FB-1017 at fertilizer rate of 69 kg P2O5
ha-1 and control (without both Rhizobium and P application), respectively (Table 6).
The increase in number of nodules due to inoculation with FB-1017 and application of 69 kg
P2O5 ha-1 could be associated with the synergistic effect of the two treatments which increased
rhizobial activities and, thus, N uptake by the plants. It might also be because of the fact that
phosphorus is known to initiate nodule formation, increasing the number of nodules, as it is
essential for the development and function of the nodules. The result of the present study is in
agreement with that of Solomon (2016), who reported that interaction between Rhizobium
inoculation and P fertilizer rate was found to significantly (P<0.05) influence the total number
of nodules per plant of faba bean, where, the highest value (126.2 plant-1) was obtained with
the combination of Rhizobium inoculation and 69 kg P2O5 ha-1, while, the lowest was recorded
at control (un-inoculated and unfertilized). This result is also in line with the findings of
Mytton and Skot (1993), who reported that the interaction between Rhizobium strain and P
fertilizer on nodule number of faba bean was found to be significant. The increase in number
of nodules per plant with P application could be due to the fact that sufficient amount of P is
required for plant growth, nodule formation and development, each process being vital for N2
fixation (Neila et al., 2012).
Such positive effects of P and inoculants on legume nodulation have also been reported for
common bean (Yoseph, 2011). Similarly, Fatima et al. (2007) have reported in soybean that
nodule formation was more sensitive to P deficiency than did plant growth and application of
phosphorus improved nodulation parameters when compared with inoculation without
phosphorous fertilizer. Likewise, Rifat et al. (2008) have reported that inoculation along with
phosphorus fertilizer had a significant effect on nodulation and % P and N in mung bean.
34
There was increasing trend in total number of nodules with increasing level of applied P from
0 to 69 kg P2O5 ha-1 that might suggest 69 P2O5 kg ha-1 is optimum for nodulation of faba bean
in the area. However, further increment in fertilizer level did not increase nodule number, as it
declined at the highest rate (92 kg P2O5 ha-1). The reason for this might be application of
excess P beyond the optimum level indirectly affected plant growth and nodulation by
reducing uptake of micronutrients, especially iron (Fe) and zinc (Zn); thus, potentially causing
deficiency of these nutrients in the plants (Ann et al., 2011). A particular high Fe requirement
exists in legumes for the heme component of hemoglobin as it is the single most abundant Fe
protein that the legume plant host makes in the nodule (Weisany et al., 2013). Thus, in
legumes Fe is required in a greater amount for total and effective nodule formation than for
host plant growth, for example in lupins and peanut (Tang and Robson, 1993).
In line with this, a reduction in specific rates of nitrogenase activity has been observed in Fe
limited peanut nodules (O’Hara et al., 1988), indicating a possible direct limitation by Fe
deficiency on nodule function and nodule mass due to excess P application. Moreover, Grant
et al. (2005) have reported that soil P supply that exceeds requirements of the crop may
preclude microbial development since, excessive P fertilizers can increase the P concentration
in plant tissue, depress symbiotic association with legumes, and affect nodulation and N
fixation. In line with the present result, Zewde (2016) has reported that the total number of
nodules in common bean increased with increased rate of application of P2O5 up to 69 kg ha-1
and declined at the highest level (92 kg P2O5 ha-1).
Table 6. Total number of nodules per plant of faba bean as influenced by the interaction of
m lm hijk gh
Control 80.4 86.9 97.3 102.9 96.9hijk
FB-1017 94.9ijk 108.0fg 130.3bc 138.3a 122.2de
jkl ghi cd ab
FB-1035 93.0 101.8 126.3 134.5 113.4f
EAL-110 91.4kl 100.1hij 128.3de 132.0abc 115.4ef
LSD (5%) 7.654
CV (%) 4.2
35
4.3.2. Number of effective nodules per plant
Results of analysis of variance revealed that the number of effective nodules per plant was
highly significantly (P<0.01) affected by the main factors of Rhizobium inoculation and P2O5
rate as well as by their interaction (Appendix Table 3). The combination of Rhizobium strain
FB-1017 with phosphorus fertilizer rate of 69 kg P2O5 ha-1 resulted in the highest number of
effective nodules per plant (128.7) which was statistically at par with inoculants FB-1035 and
EAL-110 coupled with 69 kg P2O5 ha-1. On the other hand, the control (without both
Rhizobium and P) and with 23 kg P2O5 ha-1 alone produced the lowest number of effective
nodules per plant (Table 7).
Effective nodulation is essential for a functioning legume/Rhizobium symbiosis, as plants most

susceptible to infection and capable of producing effective nodules have greater potential to
fix more atmospheric N (Kellman, 2008). The effectiveness of Rhizobium inoculation in this
study might be due to compatibility of the strain and the host plant. It is, therefore, suggested
that application of phosphorus direct to soils might be beneficial to nodule nitrogen fixation
through the prevention of the phosphorus concentration in the plants at the later growth stage.
Thus, it increased the number and size of effective nodules and the amount of nitrogen
assimilated per unit weight of nodules increasing the percent and total amount of nitrogen in
the harvested portion of the host legume and improving the density of rhizobia bacteria in the
soil surrounding the root. In line with this, Abera (2015) reported significant (P<0.01)
interaction effect of inoculation and P fertilizer application on the number of effective nodules
per plant of chickpea, where the maximum number of effective nodules per plant (184.67) was
obtained from combination of Rhizobium inoculant Cp9 with phosphorus fertilization rate of
20 kg P2O5 ha-1. Likewise, Singleton et al. (1985) have reported that, in addition to nodule
formation, deficiency of phosphorus in legumes also markedly reduced the development of
effective nodules and nodule leghaemoglobin content.
Moreover, legume seed inoculation with proper Rhizobium strain together with recommended
amount of phosphorus at early growth stage could stimulate effective root nodulation and
increase BNF, eventually giving higher yields (Morad et al., 2013). Similarly, Adejei and
Chembeiss (2002) have reported that effectiveness of a rhizobial strain is correlated to
nitrogen fixation and P fertilization. Singleton and Bohlool (1984) have also reported that, in
36
addition to the nodule formation, deficiency of phosphorus in legume also markedly affects
development of effective nodules and the nodule leghaemoglobin content, which is
responsible for the formation of dark pink or red centers inside the nodules.
Table 7. Number of effective nodules per plant of faba bean as influenced by the interaction of
Control 73.0m 79.6lm 88.1hijk 91.3hij 84.7ijkl
FB-1017 84.2ijkl 99.6fg 120.5bc 128.7a 112.5de
FB-1035 83.5jkl 93.2gh 116.1cd 125.5ab 104.9ef
kl ghi bcd abc
EAL-110 81.1 92.0 118.0 122.3 102.5f
LSD (5%) 7.971
CV (%) 4.8
4.3.3. Nodule volume per plant
Results of the analysis of variance indicated that the main effect as well as the interaction
between Rhizobium inoculation and P2O5 fertilization was highly significant (P<0.01) for
nodule volume per plant (Appendix Table 3). The combination of Rhizobium strain FB-1017
with phosphorus fertilization at a rate of 69 kg P2O5 ha-1 gave the highest nodule volume per
plant (2.767 ml), which was statistically at par with inoculants FB-1035 and EAL-110 at the
69 kg P2O5 ha-1. In contrast, the lowest nodule volume (1.207 ml plant-1) was obtained from
the control (without both Rhizobium and P) which was statistically at par with the control plus
23 kg P2O5 ha-1 and inoculation of FB-1017, EAL-110 and FB-1035 at 0 kg P2O5 ha-1 (Table
8). On the average, inoculating the seeds with FB-1017 and application of 69 kg P2O5 ha-1
increased the nodule volume per plant by 129.23% over the un-inoculated and unfertilized.
The low nodule volume for un-inoculated and 0 kg P2O5 ha-1 could be due to low number
and small size of nodules compared to those produced by Rhizobium inoculation and P
fertilization. In line with this, Mytton and Jones (1971) have reported that nodule volume in
white clover is related to size of nodules and appears to be beneficial. Similar result has
also been reported earlier, where, Rhizobium inoculation and P application significantly
affected nodule volume of faba bean, as the highest value (0.77 cm3 plant-1) was recorded
from inoculation and application of 40 kg P2O5 ha-1 (Habtegebriel et al., 2015). Besides,
37
inoculating grain legumes with efficient strains of Rhizobium and application of sufficient
amount of P fertilizer has also been widely reported to increase the number, mass and
volume of nodules (Shibiru and Mitiku, 2000; Nuruzzaman et al., 2005).
Table 8. Nodule volume (ml) per plant of faba bean as affected by the interaction of
Control 1.207j 1.307ij 1.547fg 1.660ef 1.460gh
FB-1017 1.310ij 1.507g 2.620b 2.767a 2.273c
FB-1035 1.347hij 1.427ghi 2.567b 2.673ab 2.073d
ij ghi c ab
EAL-110 1.313 1.427 2.413 2.653 1.727e
LSD (5%) 0.1453
CV (%) 4.7
4.3.4. Nodule dry weight
There was highly significant (P<0.01) effect of main factors as well as their interaction on
nodule dry weight (mg plant-1) (Appendix Table 3). The highest nodule dry weight (408.0 mg
plant-1) was recorded from faba bean inoculated with Rhizobium strain FB-1017 and fertilized
with 69 kg P2O5 ha-1, followed by Rhizobium strain FB-1035 with 69 kg P2O5 ha-1, while, the
lowest value (93.3 mg plant-1) was obtained from the control treatments (without both
Rhizobium and P2O5) (Table 9).
These results suggest that nodule dry weight has a positive relationship with nodule number.
Furthermore, it was observed that nodule dry weight significantly increased with successive
increase in phosphorus application up to 69 kg P2O5 ha-1 along with seed inoculation
treatments. Seed inoculation with all Rhizobium strains was found to be significantly better
than no inoculation at all phosphorus rates (Table 9). The lowest nodule dry weight was
observed when the seeds were not inoculated with Rhizobium strains. Thus, inoculation with
FB-1017 and FB-1035 coupled with 69 kg P2O5 ha-1 resulted in 337.7% and 319.1% increase
in nodule dry weight over un-inoculated and unfertilized, respectively, which could be
attributed to the increased total and effective number of nodules. This result is in agreement
with the findings of Workneh and Asfaw (2012), who showed that phosphorus application
levels without inoculation did not significantly influence nodule dry weight of soybean.
38
Similarly, Masresha and Kibebew (2017) have showed that soybean seed inoculated with
Bradyrhizobium japonicum and supplied with 46 kg P2O5 ha-1 increased the mean nodule dry
weight per plant (0.99 g) than the un-inoculated control. Ibsa (2013) has also reported that
application of inoculation and phosphorus fertilizer significantly enhanced nodule dry weight
per plant of chickpea.
Table 9. Nodule dry weight (mg) per plant of faba bean as influenced by the interaction of
Control 93.3l 132.0k 210.0gh 214.7g 192.0hij
FB-1017 185.3j 208.0ghi 316.7c 408.0a 270.7e
ij ghij cd a
FB-1035 190.7 200.0 300.0 391.3 247.3f
EAL-110 146.0k 185.3j 296.7d 339.3b 258.7ef
LSD (5%) 18.05
CV (%) 4.6
4.4. Yield Components and Yield of Faba Bean
4.4.1. Crop stand count at harvest
The main effect of phosphorus application was highly significant (P<0.01) for faba bean stand
count (plant population ha-1) at harvest. However, Rhizobium inoculation and its interaction
with phosphorus application was not significant (Appendix Table 4). Hence, the maximum
crop stand count at harvest (248760 plants ha-1) was recorded from application of 46 kg P2O5
ha-1, which was statistically at par with 69 kg P2O5 ha-1, while, the minimum value (243800
plants ha-1) was obtained from the unfertilized plot (Table 10).
The difference in plant population between plots received 46 kg P2O5 ha-1 and 0 kg P2O5 ha-1
was 4960 ha-1 which is notable. The reason behind this might be the positive response to
phosphorus application that created a favorable condition for nitrogen fixation and a positive
interaction with potassium, calcium and other nutrients to maintain the pH and other physical
factors in the soil. Thus, this might have created a suitable environment for the germination,
establishment, growth and development of plants to attain maximum stand count at harvest.
Similarly, Brady and Weil (2002) have reported that phosphorus enhances many aspects of
39
plant physiology, including the fundamental processes of photosynthesis, reproduction,

flowering, fruiting (including seed production) and maturation. In addition, positive responses
of phosphorus application to nitrogen fixation, growth and development of faba bean crop
have been reported by Duguma and Okali (1987).
Table 10. Main effects of Rhizobium inoculation and phosphorus fertilizer rate on faba bean
stand count (plant population ha-1) at harvest
Treatments Crop stand count at
harvest (ha-1)
Rhizobium
Control 245635
FB-1017 247024
FB-1035 246825
EAL-110 246627
LSD (5%) NS
0 243800c
23 245536bc
46 248760a
69 248016ab
92 246528abc
LSD (5%) 2760.6
CV (%) 1.4
Where, LSD (5%) = Least significant difference at 5% level of significance; CV= Coefficient of variation; NS=
non-significant; Means followed by the same letter(s) with in a columns are not significantly different at 5% level
of significance
4.4.2. Number of pods per plant
The result of analysis of variance showed that the main effect as well as interaction effect of
seed inoculation with Rhizobium and P fertilizer application was highly significant (P<0.01)
on the number of pods per plant (Appendix Table 4). The highest number of pods per plant
(42.27) was recorded with Rhizobium inoculant FB-1017 along with 69 kg P2O5 ha-1, while,
the lowest (23.47) value was recorded from plots maintained without both Rhizobium
inoculation and phosphorus fertilization (Table 11). On average, the number of pods per plant
increased by 80.10% over un-inoculated and unfertilized plots as a result of inoculating the
seeds with FB-1017 and application of 69 kg P2O5 ha-1.
The highest number of pods per plant with Rhizobium strain FB-1017 and 69 kg P2O5 ha-1 was
probably due to the supply of nitrogen through BNF and the cumulative effect of phosphorus
40
in the processes of cell division and balanced nutrition, which in turn, play an important role in
enhanced growth and assimilate accumulation, thereby improving the reproductive
performance of the plants. It might also be due to adequate availability of N and P which
might have facilitated the production of more primary and secondary branches and plant
height, which might, in turn, have contributed for the production of higher number of total
pods. The increase in number of pods per plant could also occur because of the increased leaf
area with additional N being associated with more reproductive nodes.
This result is in line with the findings of Habtegebriel et al. (2015) who reported that
Rhizobium inoculation and P application had significantly affected number of pods per
plant of faba bean as inoculation and application of 40 kg P2O5 ha-1 resulted in 112%
increase in number of pods per plant over the control. Similarly, Abdalla (2002) has showed
that treating faba bean plants with bio-fertilizer (phosphorien) and P fertilizer markedly
increased number of pods per plant and seed yield. Likewise, Ali et al. (2004) have reported
increased number of pods per plant of chickpea by seed inoculation and P fertilization. It was
explained that the increased supply of N through BNF and direct application of phosphorus
play an important role in enhancing growth and assimilate accumulation, thereby improving
the reproductive performance of the plants. Similarly, Zafar et al. (2003) have reported that
phosphorus fertilization showed significant increase in number of pods per plant of lentil due
to the cumulative effect of phosphorus in the processes of cell division and balanced nutrition.
Masresha and Kibebew (2017) have also reported that application of 46 kg P2O5 ha-1 with
Bradyrhizobium japonicum as biofertilizer resulted in 164.7% increase in mean pod number of
soybean over control.
The number of pods per plant increased with increasing levels of phosphorus up to of 69 kg
P2O5 ha-1 and, then, declined at highest rate (92 kg P2O5 ha-1). The reason behind the decreased
number of pods per plant due to high phosphorus application might be antagonistic interaction
between phosphorus and microelements. Surplus P application more than the optimum might
intensify the deficiency of Zn and Cu by reducing their uptake (Ann et al., 2011) and the
toxicity of Mn by increasing its uptake (Truong et al., 1971). In addition, there was an
indication of a Mn × P interaction in the plant which could markedly influence yield
components of some tropical legumes (James et. al., 1995). In line with this result, Meseret
41
and Amin (2014) reported significant increment in pod number per plant of common bean
with increased P application up to 20 kg P2O5 ha-1, and the pod number decreased at higher
rates of applied P (30 and 40 kg P2O5 ha-1). Similarly, Alivi et al. (2014) have reported that for
all measured yield components (number of pods per plant, number of grains per pod and 100-
grain weight), treatments with 46 kg P2O5 ha-1 had the highest values and there was no
significant difference between 46 kg P2O5 ha-1 and 69 kg P2O5 ha-1 and even, in some cases,
further increase of phosphate application decreased some yield components of faba bean.
Table 11. Interaction effect of Rhizobium inoculation and phosphorus fertilizer rate on number
of pods per plant of faba bean
k k ghij fgh
Control 23.47 24.93 28.80 31.33 29.33ghi
jk hij bcd a
FB-1017 26.07 28.40 37.67 42.27 35.33de
FB-1035 26.13jk 28.20ij 36.73cde 40.07ab 31.53fg
k ghij ef bc
EAL-110 23.80 28.73 33.93 38.40 30.47ghi
LSD (5%) 3.028
CV (%) 5.9
4.4.3. Number of seeds per pod
The number of seeds per pod is a significant constituent that directly imparts in exploiting
potential yield recovery in leguminous crops. Number of seeds per pod was highly
significantly (P<0.01) affected by the main factors of Rhizobium inoculation and phosphorus
fertilization as well as by their interaction (Appendix Table 4). The highest mean number of
grains per pod (3.717) was recorded with seed inoculation with Rhizobium inoculant FB-1017
and application of 46 kg P2O5 ha-1, while, the lowest value (2.353) was recorded with control
(without Rhizobium) at 0 kg P2O5 ha-1 (Table 12).
The result indicated that number of seeds per pod increased owing to interaction of Rhizobium
inoculation and phosphorus application, which might be due to more availability of nutrients
attributed to more root development, stem vigour, flower initiation and seed formation.
Inoculation had a significant effect on growth, N contents and uptake in shoots, as it increased
the size of the plant in order to intercept more light for photosynthesis, yield and yield
42
components of faba bean. This may probably be due to the cumulative effect of phosphorus in
the processes of cell division and balanced nutrition.
The present result is in agreement with that of Solomon (2016), who reported that the number
of seeds per pod of faba bean was significantly (P<0.01) affected due to the interaction of
Rhizobium inoculation and phosphorus rate, where the highest number of seeds per pod (3.41)
was obtained from faba bean inoculated with Rhizobium strain FB-1035 combined with 69 kg
P2O5 ha-1 application. The result is also in line with the findings of Singh et al. (2008), who
reported that the seeds and nutritional quality of legumes are greatly influenced by application
of phosphorus and bio-fertilizers. Likewise, Meena et al. (2003) reported that P fertilization
significantly increased the number of grains per pod of chickpea. Concomitantly, Malik et al.
(2006) have also found that various levels of phosphorus fertilizer and seed inoculation had
highly significant effect on the number of seeds per pod of soybean.
Furthermore, according to Zafar et al. (2003), a regulatory system through which it was
possible to direct and concentrate available nutrients to permit development of more number
of pods per plant and seeds per pod. It has also been further pointed out that phosphorus aids
in transferring photosynthates from the stalks, leaves and other growing parts to the
economically important organs like seeds making them plump and bold.
Table 12. Number of seeds per pod of faba bean as influenced by the interaction of Rhizobium
l jkl fghi bcde
Control 2.353 2.484 2.691 2.885 2.723efghi
FB-1017 2.585ijk 2.671ghij 3.717a 3.036bc 2.872cdef
jkl hijk bcdef b
FB-1035 2.482 2.633 2.882 3.069 2.831defg
EAL-110 2.460kl 2.592ijk 2.825defgh 3.002bcd 2.753efghi
LSD (5%) 0.1915
CV (%) 4.2
4.4.4. Hundred seed weight
The main effects of Rhizobium and P2O5 were highly significant (P<0.01) and significant
(P<0.05) interaction effect between Rhizobium inoculants and P2O5 fertilization rates on 100-
seed weight of faba bean (Appendix Table 4). The highest 100-seed weight (56.02 g) was
43
recorded when seeds were inoculated with Rhizobium strain FB-1017 coupled with application
of 69 kg P2O5 ha-1, while, the lowest value (45.94 g) was recorded for un-inoculated and
unfertilized, which was statistically at par with Rhizobium strain EAL-110 at 0 kg P2O5 ha-1
(Table 13). Hundred seed weight increased with increasing levels of P up to 69 kg P2O5 ha-1
and then declined at the highest rate (92 kg P2O5 ha-1).
The variation in seed weight might be due to better growth and development of the crop with
the interaction effects of inoculation and optimum P fertilizer rate that might have improved
the nutrient supply to plants, resulting in producing more assimilates which might have
partitioned more efficiently from source to sink and ultimately gained more seed weight. The
other reason might be that Rhizobium inoculation significantly increased the uptake of macro
and micro nutrients, which can favour the growth and developments of all yield related traits
of faba bean, including individual seed size. These results suggested that Rhizobium
inoculation and P fertilizer application contributed much to increase efficiency of major
nutrients and, thus, led to higher grain weight and yield.
Similarly, Makoi et al. (2013) have reported that Rhizobium inoculation significantly increased
the uptake of phosphorus (P), potassium (K), calcium (Ca) and magnesium (Mg) in different
plant organs caused by the increase in soluble phosphorus and assimilation of nutrients to the
grain, resulting in larger grains. Morad et al. (2013) have also observed that seed inoculation
with proper Rhizobium together with small amounts of P (13 kg ha-1) at early growth stage
stimulated the root nodulation and improved grain weight of common bean. Furthermore,
Shahram and Peyman (2016) reported that the interaction of inoculation with biofertilizer and
phosphorus application (69 kg P2O5 ha-1) significantly (P<0.05) influenced 100 seed weight of
faba bean, which increased by about 42% over the control (without inoculation) and nil
application of phosphorus. Similarly, Tairo and Ndakidemi (2013) found a significant increase
(91%) in hundred grain weight of soybean by inoculation and application of phosphorus over
the control treatment.
All Rhizobium inoculants produced significantly lowest hundred seed weight at 0 kg P2O5 ha-1.
This might be due to P fertilizer contributed much to increases in grains weight. Increasing P
levels up to 69 kg P2O5 ha-1 might have increased 100-seed weight probably by improving
photosynthetic activity and source sink relationship. A decrease in 100-grain weight at low
44
fertilizer rates could be resulted from the competition for nutrients and the decrease in
carbohydrate stores. Increased phosphorus content up to the optimum level increased the
amount of phytin stored in the seeds, as phyitin serves as the main source of stored P in most
grains and is an important compound for germination and seed growth with a significant
contribution to seed size and weight. This result is in line with the findings of Veeresh (2003)
who has reported increased seed weight of common bean with application of phosphorus up to
the rate 75 kg P2O5 ha-1.
Table 13. Hundred seed weight (g) of faba bean as influenced by the interaction of Rhizobium
k ijk efgh de
Control 45.94 47.16 49.11 50.45 49.52defg
hijk efgh bc a
FB-1017 47.49 48.99 52.47 56.02 49.84def
FB-1035 47.18ijk 48.52fghi 50.98cd 53.86b 49.77defg
jk ghij de bc
EAL-110 46.83 48.19 50.46 52.42 49.75defg
LSD (5%) 1.641
CV (%) 2.0
4.4.5. Grain yield
The results of analysis of variance (Appendix Table 5) revealed that grain yield of faba bean
was highly significantly (P<0.01) influenced by the main factors and significantly (P = 0.04)
affected by the interaction of Rhizobium inoculation and phosphorus fertilizer application. The
highest mean grain yield (4101 kg ha-1) was recorded for Rhizobium strain (inoculant) FB-
1017 with 69 kg P2O5 ha-1, which was statistically at par with Rhizobium strain FB-1035 with
69 kg P2O5 ha-1 (3912 kg ha-1), while, the lowest value (2700 kg ha-1) was obtained from the
control (without Rhizobium) with 0 kg P2O5 ha-1, which was also statistically at par with all
inoculants at 0 kg P2O5 ha-1 and the control (without Rhizobium) at 23 kg P2O5 ha-1 (Table 14).
On the average, inoculating the seeds with FB-1017 and application of 69 kg P2O5 ha-1
increased grain yield by 51.89% over un-inoculated and 0 kg P2O5 ha-1. All Rhizobium
inoculants with 0 kg P2O5 ha-1 produced the lowest grain yield. Similarly, un-inoculated
treatments produced lower grain yield than the inoculated ones across all the P2O5 rates, which
indicates the need for inoculation of seeds with Rhizobium and application of P for effective
nodulation and nitrogen fixation.
45
The two way interaction effect was more pronounced when inoculation and P were applied in
combination, resulting in a yield advantage over the control (Table 14). The observed yield
improvements with Rhizobium strain FB-1017 at 69 kg P2O5 ha-1 might be due to the increased
N level as a result of atmospheric nitrogen fixation by effective strain and P availability by
direct application. The yield advantage of faba bean produced by application of P and
Rhizobium inoculation could be attributed to an increase in the number of branches, which, in
turn, increased the number of productive nodes and number of pods per plant. The number of
branches and pods per plant and the number of grains per pod were found to be strongly
related with grain yield, which shows that yield components in legumes determine their grain
yields (Fageria, 2009). Similarly, Dahmardeh et al. (2010) have reported that the maximum
yield of a legume crop depends upon its yield components, such as the number of branches per
plant, pods per plant, seeds per pod and seed weight.
In line with this result, Masresha and Kibebew (2017) reported that the combined use of 46 kg
P2O5 ha-1 and Bradyrhizobium japonicum as biofertilizer on soybean significantly maximized
the mean seed yield per plant by 59.2% as compared to the control. Likewise, Habtegebriel et
al. (2015) reported that Rhizobium inoculation along with phosphorus application significantly
influenced grain yield of faba bean, where inoculation and 40 kg P2O5 ha-1 resulted in 62.5%
increase in grain yield over the control (un-inoculated) with 0 kg P2O5 ha-1. Fatima et al.
(2007) have also reported that combined application of inoculation and P increased cowpea
grain yield by 63% over the phosphorus control.
This result support the idea that inoculation could be more effective and beneficial in
enhancing grain yield in combination with phosphorus fertilizer. In agreement with this,
Amare et al. (1999) treated faba bean plants with different levels of phosphorus up to 69 kg
ha-1 and found a positive linear response of seed yield to applied phosphorus. Similarly, Jat
and Ahlawat (2004) have reported that application of phosphorus up to 26.4 kg P2O5 ha-1
significantly increased yield and total uptake of phosphorus by chickpea over the control.
Likewise, Amos et al. (2001) observed significantly higher yield due to Rhizobium inoculation
and phosphorus application in common bean, where inoculation with Rhizobium coupled with
40 kg P2O5 ha-1 gave the highest grain yield.
46
Table 14. Grain yield (kg ha-1) of faba bean as influenced by the interaction of Rhizobium
inoculants and phosphorus fertilizer rate
g g fg efg
Control 2700 2771 2989 3109 3018efg
g cde ab a
FB-1017 2716 3443 3873 4101 3578bcd
FB-1035 2702g 3090efg 3788abc 3912ab 3250def
g fg efg abc
EAL-110 2732 2870 3059 3785 3034efg
LSD (5%) 428.6
CV (%) 8.0
The observed yield increment with inoculation and P application might have resulted from the
positive effects of P on the process of N2 fixation, where, the increased supply of N through
inoculation resulted in enhanced plant growth that eventually lead to use of P fertilizer. The
present study revealed that P application with inoculant improved nodulation and increased
grain yield. In line with this, Fatima et al. (2007) have reported that mixture of Rhizobium
strains with phosphorus gave higher seed yield of soybean over inoculants without
phosphorous. Other researchers have also reported that inoculation along with phosphorus
fertilizer had a significant effect on nodulation, shoot dry matter, grain yield, % P and N of
mung bean (Rifat et al., 2008).
The increase in grain yield in response to increased rate of P application up to 69 kg P2O5 ha-1
may also be ascribed to the phenomenon that sufficient supply of phosphorus is associated
with increased root growth and leaf expansion that may lead to high dry matter accumulation
through enhanced effective exploration by the roots for the immobile nutrients such as
phosphorus (Gustfson, 2010). The result of the present study is consistent with that of Abera
(2015) who reported that grain yield of chickpea was significantly (P<0.01) affected by the
interaction of Rhizobium inoculation and phosphorus fertilization, where, the highest value
(2455 kg ha-1) was recorded for Rhizobium inoculum Cp9 and 20 kg P2O5 ha-1, and all
inoculants gave significantly lowest grain yield at 0 kg P2O5 ha-1. The present result is also in
agreement with that of Ndakidemi et al. (2006) who reported that combined application of
bacterial inoculants and P fertilizer to soybean and common bean significantly increased
biomass production and grain yield compared with the single use of nitrogen and P or
rhizobial strains alone. Malik et al. (2006) have also reported that nitrogen fixation by
47
Rhizobium hastened vegetative growth, while, phosphorus application improved yield

components of soybean, which are possible reasons for substantial increase in both biological
and grain yield.
When the rate of P2O5 increased from 69 to 92 kg ha-1, grain yield did not increase, suggesting
that 69 kg P2O5 ha-1 is the optimum rate for faba bean production in the area, but increasing the
rate of the nutrient from 0 to 69 kg P2O5 ha-1, consistently increased grain yield. The reason for
the reduction in grain yield due to application of excess P might be the nutrient interacts with
many nutrients, the most commonly observed and studied antagonistic interaction is with
micronutrients such as Zn, by decreasing their solubility and causing their deficiency due to its
toxic effect, as toxicity of one nutrient is accompanied by deficiency of another (Ann et al.,
2011). On the other hand, at higher P supply rates, excess P accumulate in vacuoles of palisade
cells of plants, which were associated with decreased photosynthetic rates and reduced yield
(Shane et al., 2004).
Similarly, Davood (2013) concluded that P fertilizer application up to certain amount can
increase yield of faba bean, but further P fertilizer not only limit yield, but also decrease it,
owing to low efficiency of P in soil due to its low solubility and absorption. Gifole et al.
(2011) have also reported that yield of common bean increased with increasing levels of P up
to 40 kg P2O5 ha-1 and declined downward for 50 and 60 kg P2O5 ha-1. Likewise, Alivi et al.
(2014) reported that grain yield of faba bean decreased when application of phosphate
fertilizer raised from 46 to 69 kg P2O5 ha-1. Similar trend was also reported by Zewde (2016)
where P application up to 69 kg P2O5 ha-1 resulted in increased grain yield, which decreased at
the highest rate of applied P (92 kg P2O5 ha-1).
The mean maximum grain yield of faba bean (4101 kg ha-1) obtained from the combination of
seed inoculation with Rhizobium and phosphorus fertilization in the current study is also
higher than the national average yield (1893 kg ha-1) and average yield of SNNPRS (1639 kg
ha-1) and Wolaita Zone (study area) (891 kg ha-1) (CSA, 2015) by about 116.64%, 150.21%
and 360.3%, respectively. The mean grain yield attained by using Degaga variety in this study,
however, is lower than the maximum yield reported for the variety at experimental station
(4200 kg ha-1) (MoA, 2014) by about 2.41%.
48
4.4.6. Above ground dry biomass yield
The result of analysis of variance indicated that main factors of Rhizobium inoculation and
phosphorus fertilization and their interaction highly significantly (P<0.01) influenced the total
above ground dry biomass yield (AGDB) of faba bean (Appendix Table 5). The highest mean
total above ground dry biomass (9280 kg ha-1) was produced with the combination of
Rhizobium inoculant FB-1017 and 69 kg P2O5 ha-1, while, the lowest value (7669 kg ha-1) was
recorded for the control (without Rhizobium) with no phosphorus (Table 15).
The substantial biomass yield increase with Rhizobium strain FB-1017 inoculation and
application of 69 kg P2O5 ha-1 could be due to sufficient nitrogen supply mainly from
biological nitrogen fixation, as nitrogen is a key factor in many biological compounds that
play a major role in photosynthetic activity and chlorophyll synthesis, which eventually result
in vigorous vegetative growth and more biomass accumulation. The total biomass of faba bean
is also of great importance in farm level nutrient cycling due to its role as either animal feed or
organic matter to be returned to the soil. The increase in above ground dry biomass yield
might also be due to inoculation of seed with appropriate Rhizobium strain and application of
sufficient amount of P, both of which enhanced availability of N through BNF for vegetative
growth of the plants. The increase in total dry biomass yield was reflected by increased
number of primary branches (Table 4) and other parameters, such as number of pods per plant
(Table 11), seeds per pod (Table 12) and 100 seed weight (Table 13) with Rhizobium
inoculation and increased rate of phosphorus application.
Consistent with the result of this study, Mekki (2016) reported that inoculation with
biofertilizer and P application (34.5 kg P2O5 ha-1) significantly increased faba bean biological
yield by 64.13% over the control, by 97.75% over faba bean received biofertilizer alone and
by 40.28% over P alone. This result is also in agreement with that of Abera (2015) who
reported that the total above ground plant dry biomass of chickpea was significantly (P<0.01)
affected by interaction of inoculation and phosphorus fertilization, where the highest mean
value (4613 kg ha-1) was produced with the combination of Rhizobium inoculant Cp9 and 20
kg P2O5 ha-1. Similarly, Abbasi et al. (2010) also pointed out that the aboveground biomass
yield of soybean was quadratically increased with Rhizobium inoculation and P application.
Ndakidemi et al. (2006) have also reported that the combined application of bacterial
49
inoculants and phosphorus fertilizer to soybean and common bean significantly increased
biomass production and grain yield compared with the single use of nitrogen and phosphorus
or rhizobial strains alone.
As indicated in Table 15, the lowest total above ground dry biomass yield was recorded from
the control and all Rhizobium inoculants with 0 kg P2O5 ha-1. The increase in dry biomass yield
in response to increased rate of phosphorus application might be ascribed to the phenomenon
that sufficient supply of phosphorus is associated with increased root growth and leaf
expansion that may lead to high dry matter accumulation by enhancing effective exploration
by the roots for the immobile nutrients such as phosphorus (Gustfson, 2010). The positive
effect of P on total biomass yield in the present study is also in agreement with the findings of
Gifole et al. (2011) for common bean, where, 40 kg P2O5 ha-1 resulted in 51.86% increase in
total biomass over the control. Likewise, Yilmaz (2008) has reported that increasing
phosphorus levels up to 75 kg P2O5 ha-1 improved aboveground biomass of narbon vetch
(Vicia narbonensis L.) at harvest by as much as 18.5%.
Application of phosphorus up to 69 kg P2O5 ha-1 showed increasing trend in above ground dry
biomass yield, as the highest value was obtained at 69 kg P2O5 ha-1, and it decreased at the
highest level of phosphorus (92 kg P2O5 ha-1), which might indicate that 69 kg P2O5 ha-1 is
optimum rate for faba bean production in the study area. The decline in above ground dry
biomass yield with respect to the highest phosphorus level might also be attributed to
imbalance of phosphorus with other nutrients, especially with nitrogen (Havlin et al., 2003). In
line with this, Meseret and Amin (2014) have reported that the dry biomass of common bean
increased by application of phosphorus fertilizer up to 20 kg P2O5 ha-1 and further rates of
phosphorus resulted in declined dry biomass yield. The same authors have reported that the
highest dry biomass yield (75 g plant-1) was obtained at 20 kg P2O5 ha-1 and decreased at 30 kg
P2O5 ha-1 (53.4 g plant-1) and 40 kg P2O5 ha-1 (50.6 g plant-1).
50
Table 15. Above ground dry biomass yield (kg ha-1) of faba bean as influenced by the
interaction of Rhizobium inoculants and P fertilizer rate
f ef de cd
Control 7669 7799 8125 8413 8358d
f bc ab a
FB-1017 7673 8825 9023 9280 8833bc
FB-1035 7670f 8131de 8904ab 9110ab 8302d
ef ef de ab
EAL-110 7743 7778 8128 9078 8267d
LSD (5%) 431.5
CV (%) 3.1
4.4.7. Harvest index
Harvest index is very useful in measuring nutrient and dry matter partitioning in crop plants,
and provides an indication of how efficiently the plant utilized acquired nutrients for grain
production. So, the higher harvest index also implies higher partitioning of dry matter into the
grain. It generally varies between 30 and 50% (Saxena, 1984).
As indicated by ANOVA, the main effects of Rhizobium inoculation and phosphorus fertilizer
rates were highly significant (P<0.01) on harvest index of faba bean. However, the interaction
of the two factors was not significant (Appendix Table 5). With respect to the Rhizobium
strains (inoculants), the highest harvest index (40.11%) was obtained from faba bean seeds
inoculated with Rhizobium strain FB-1017 and the lowest value (36.10%) was recorded from
control (without Rhizobium) (Table 16).
The increased harvest index of faba bean inoculated with Rhizobium strain FB-1017 might be
due to suitability of the strain for nodule formation (increased nodule number per plant) and
increased hundred grain weight. Similar finding was reported by Roy et al. (1995), indicating
that grain inoculation increased the nodule number per plant and gave the highest harvest
index and hundred grain weight of chickpea. Ali (1993) has also reported that inoculation
exhibited significant effect on harvest index of various legume crops. Likewise, Shahram
and Peyman (2016) have reported that inoculation of faba bean with bio-fertilizer
significantly increased the harvest index, which was increased by about 35.13% over the
control.
51
Among the phosphorus fertilizer rates, the highest harvest index (41.49%) was attained from
faba bean supplied with 69 kg P2O5 ha-1, which was statistically at par with 46 kg P2O5 ha-1,
and the lowest value (35.28%) was obtained from faba bean that received 0 kg P2O5 ha-1
(Table 16). The increment in harvest index at 69 kg P2O5 ha-1 might be ascribed to greater
photo-assimilate production and its ultimate partitioning to the grains compared to the straw.
In line with this result, Masresha and Kibebew (2017) reported that the maximum mean
harvest index per plant of soybean was obtained by application of 46 kg P2O5 ha-1, which
resulted in 19.1% increase over the control. Similarly, Zafar et al. (2003) found that
calculated values of harvest index showed an increasing trend with application of
phosphorus, while, minimum value was recorded from the control plot.
Table 16. Main effects of Rhizobium inoculation and P rates on harvest index (%) of faba bean
Treatments Harvest index (%)
Rhizobium
Control 36.10b
FB-1017 40.11a
FB-1035 39.56a
EAL-110 37.62b
LSD (5%) 1.568
-1
P2O5 rates (kg ha )
0 35.29c
23 37.22b
46 39.75a
69 41.49a
92 37.99b
LSD (5%) 1.753
CV (%) 5.5
followed by the same letter(s) with in columns are not significantly different at 5% level of significance
4.5. Economic Analysis
Production of grain-legumes is increasing significantly due to their vast use in different

situations, including food, feed and as industrial raw materials. Considering the increasing
needs for human consumption of plant products and the economic constraints of applying
fertilizer in legumes, there is a greater role for grain legumes in cropping systems, especially
in regions where affordability of fertilizer is difficult (Ndakidemi et al., 2006). The agronomic
data upon which the recommendations are based must be relevant to the farmers own agro-
52
ecological conditions and the evaluation of those information must be consistent with the
farmers goal and socio-economics circumstance (CIMMYT, 1998).
The potential response of crops to applied fertilizers and price of the fertilizer in the cropping
season ultimately determine the economic feasibility of fertilizer application (CIMMYT,
1998). To identify treatments with maximum return to the farmers’ investment, marginal rate
of return (MRR) analysis was performed on non-dominated treatments. For a treatment to be
considered as a worthwhile option to farmers, the marginal rate of return (MRR) need to be
taken as 100%. It is the relation between the costs that vary and net benefits for the non-
dominated treatments (CIMMYT, 1998).
Data presented in Table 17 indicated that the highest net benefit/return (40939.80 ETB ha-1)
was obtained from seed inoculation with Rhizobium inoculant FB-1017 and application of 69
kg P2O5 ha-1, followed by seed inoculation with Rhizobium strain FB-1017 and 46 kg P2O5 ha-1
(39374.40 ETB ha-1), while, the lowest net benefit/return (28519.20 ETB ha-1) was recorded
with EAL-110 and 92 kg P2O5 ha-1 (Table 17). In spite of the high cost of P fertilizer, the use
of inoculation and application of P up to 69 kg P2O5 ha-1 appears to be advantageous for
smallholder farmers growing faba bean in this area. Likewise, it can be inferred that
inoculation is a means to increase the economic benefits of application of P fertilizer. The
combination of inoculant FB-1017 and 69 kg P2O5 ha-1 had MRR of 174.51%, which is above
the acceptable minimum MRR of 100%, and this result suggests that for each ETB invested in
faba bean production by seed inoculation with strains FB-1017 and application of 69 kg P2O5
ha-1, the producer would get ETB 1.745 after recovering the investment cost (Table 17).
Hence, seed inoculation with Rhizobium strain FB-1017 and 69 kg P2O5 ha-1 was found to be
economically feasible.
53
Table 17. Summary of economic analysis of the effects of Rhizobium inoculation and
phosphorus fertilizer application on faba bean production in Wolaita Sodo
Treatments UGY AGY GFB TVC NB MRR
-1 -1 -1 -1 -1
Rhizobium P2O5 (kg ha ) (kg ha ) (ETB ha ) (ETB ha ) (ETB ha ) (%)
-1
Strains (kg ha )
Control 0 2700 2430 29160 0 29160
Control 23 2771 2493.9 29926.8 1197 28729.8 D
Control 46 2989 2690.1 32281.2 2094 30187.2 162.47
Control 69 3109 2798.1 33577.2 2991 30586.2 44.48
Control 92 3018 2716.2 32594.4 3888 28706.4 D
FB-1017 0 2716 2444.4 29332.8 360 28972.8 D
FB-1017 23 3443 3098.7 37184.4 1557 35627.4 555.94
FB-1017 46 3873 3485.7 41828.4 2454 39374.4 417.72
FB-1017 69 4101 3690.9 44290.8 3351 40939.8 174.51
FB-1017 92 3578 3220.2 38642.4 4248 34394.4 D
FB-1035 0 2702 2431.8 29181.6 360 28821.6 143.33
FB-1035 23 3090 2781 33372 1557 31815 250.07
FB-1035 46 3788 3409.2 40910.4 2454 38456.4 740.40
FB-1035 69 3912 3520.8 42249.6 3351 38898.6 49.30
FB-1035 92 3250 2925 35100 4248 30852 D
EAL-110 0 2732 2458.8 29505.6 360 29145.6 4.89
EAL-110 23 2870 2583 30996 1557 29439 24.51
EAL-110 46 3059 2753.1 33037.2 2454 30583.2 127.56
EAL-110 69 3785 3406.5 40878 3351 37527 774.11
EAL-110 92 3034 2730.6 32767.2 4248 28519.2 D
Where, UGY = Unadjusted grain yield; AGY = adjusted grain yield; GFB = gross field benefit; TVC = total
variable costs; NB = net benefit, MRR = marginal rate of return; ETB ha-1 = Ethiopian Birr per hectare; D =
dominated treatments. Market price of faba bean = 12 ETB kg-1; Cost of Rhizobium = 160 ETB ha-1; Cost of TSP
fertilizer = 39 ETB kg-1; Labour cost for Rhizobium inoculation = 4 persons ha-1, each 50 ETB day-1; Labour cost
for TSP fertilizer application = 6 person ha-1, each 50 ETB day-1.
54
5. SUMMARY AND CONCLUSION
Faba bean (Vicia faba L.) is a multifunctional crop, however, its productivity in Ethiopia is
still far below its potential. One of the reasons for such low yield is due to poor mineral
nutrient availability and lack of efficient and compatible strains of Rhizobium in the soil
among other constrains. By and large, there is lack of adequate information on the use of
Rhizobium inoculants and phosphorus fertilizer for enhancing the yield of faba bean in the
study area. With this background, the present study was conducted to assess the effect of
Rhizobium inoculation and P application on yield components and yield of faba bean; and to
estimate the economically feasible combination of inoculants and P rate that maximizes the
productivity of faba bean in the study area. Faba bean variety Degaga which was released by
HARC in 2002 was used as the test crop. Four levels of inoculation with Rhizobium strains
(FB-1017, FB-1035 and EAL-110) and control (without Rhizobium) and five rates of P2O5 (0,
23, 46, 69 and 92 kg ha-1) were tested in factorial arrangement in Randomized Complete Block
Design (RCBD) in three replications. The experiment was conducted at Wolaita Sodo,
southern Ethiopia, under rain fed conditions during 2016 main cropping season.
The main effect of P2O5 fertilizer rate significantly affected crop stand count at harvest, while,
harvest index was significantly affected by both Rhizobium inoculation and P fertilizer rates.
The highest harvest index (40.11%) was obtained from faba bean inoculated with Rhizobium
strain FB-1017, while, the lowest value (36.10%) was recorded with control (without
Rhizobium). The highest stand count at harvest (248760 plants ha-1) was recorded at 46 kg
P2O5 ha-1, but the highest harvest index (41.49%) was attained at 69 kg P2O5 ha-1, whereas, the
lowest value for stand count at harvest (243800 plants ha-1) and harvest index (35.28%) were
obtained from faba bean received 0 kg P2O5 ha-1.
Results of the present experiment revealed that inoculants and P fertilizer rates significantly
influenced faba bean phenology, growth, nodulation, yield components and yield parameters.
The interaction of inoculation and phosphorus fertilization significantly (P<0.05) influenced
plant height, and highly significantly (P<0.01) influenced days to 50% flowering, days to 90%
physiological maturity, number of primary branches per plant, total number of nodules,
number of effective nodules, nodule volume and nodule dry weight per plant. The maximum
number of days to flowering (55.00) was recorded from Rhizobium inoculant FB-1017 with 46
55
kg P2O5 ha-1, whereas, the maximum days to physiological maturity (111.7), number of
primary branches (3.80), total nodules (138.3), effective nodules (128.7), nodule volume
(2.767 ml plant-1) and nodule dry weight (408.0 mg plant-1) were recorded from Rhizobium
inoculant FB-1017 at 69 P2O5 ha-1. In contrary, the minimum number of days to flowering
(44.67) and physiological maturity (101.0) were recorded from the un-inoculated treatment
with 92 kg P2O5 ha-1. Similarly, the minimum number of primary branches (1.933), total
nodules (80.4), effective nodules (73.0), nodule volume (1.207 ml plant-1) and nodule dry
weight (93.3 mg plant -1) were recorded from control (un-inoculated) and nil P2O5 application.
On the other hand, the highest plant height (139.4 cm) was obtained due to Rhizobium
inoculant FB-1035 with 69 kg P2O5 ha-1, whereas, the lowest value (111.9 cm) was recorded
from without both Rhizobium inoculation and P fertilization.
There was a highly significant (P<0.01) interaction effect of Rhizobium inoculation and
phosphorus fertilizer rates on number of pods per plant, number of seeds per pod and above
ground dry biomass yield, whereas, significant (P<0.05) interaction effect of Rhizobium
inoculation and phosphorus fertilizer on hundred seed weight and grain yield of faba bean.
Accordingly, the highest number of seeds per pod (3.717) was recorded from inoculant FB-
1017 with 46 kg P2O5 ha-1, whereas, the highest number of pods per plant (42.27), hundred
seed weight (56.02 g), grain yield (4101 kg ha-1) and above ground dry biomass yield (9280 kg
ha-1) were obtained as a result of combined application of Rhizobium inoculant FB-1017 and
69 kg P2O5 ha-1. In contrast, the lowest numbers of pods per plant (23.47), seeds per pod
(2.353), 100-seed weight (45.94 g), grain yield (2700 kg ha-1) and above ground dry biomass
yield (7669 kg ha-1) were recorded from control (without both inoculant and P).
The economic analysis also indicated that the highest net benefit/return (40939.80 ETB ha-1)
was recorded from seed inoculation with FB-1017 Rhizobium strain along with application of
69 kg P2O5 ha-1. This treatment also resulted in MRR of 174.51%, which is above the
acceptable minimum MRR of 100%, while, the lowest net returns (28519.20 ETB ha-1) was
recorded with strain EAL-110 at 92 kg P2O5 ha-1.
In general, this study provided evidence that yield and economic returns of faba bean
production could be improved by inoculation with appropriate Rhizobium strains and
application of phosphorus fertilizer at optimum rates. Thus, it can be concluded from the result
56
of present study that the use of Rhizobium inoculant FB-1017 and application of 69 kg P2O5
ha-1 could be recommended to enhance the productivity of faba bean in the study area.
However, the result of the present study need to be validated and verified with different types
of Rhizobium strains and faba bean varieties in different agro-ecologies and seasons in order to
give a comprehensive recommendation for wide range of faba bean production systems.
57
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7. APPENDICES
75
Appendix Table 1. Monthly total rainfall (mm) and average minimum and maximum air
temperatures (ºC) recorded in the experimental area during the growing period of test crop
(July 2016 – November 2016)
Month Rainfall (mm) Temperature (°C)
Maximum Minimum
July 106.7 22.5 14.9
August 109.4 23.0 14.4
September 129.6 24.2 14.7
October 83.6 25.9 15.5
November 51.5 26.4 15.7
Total/Mean 480.8 24.4 15.04
Source: Hawassa National Meteorology agency (HNMA)
Appendix Table 2. Mean squares of analysis of variance showing phenological and growth
parameters of faba bean inoculation with different Rhizobium inoculants and phosphorus
fertilizer rate
Mean squares
Source of variation Df D50%F D90%PM NPBPP PH
Block 2 7.617 1.3167 0.09450 39.349
Rhizobium strain 3 30.333** 21.6611** 1.39394** 122.333**
P-rate 4 59.458** 51.8583** 2.33733** 572.767**
Rhizobium × P-rate 12 18.014** 23.5917** 0.13644** 6.734*
Error 38 1.108 0.8956 0.04766 3.292
CV (%) 2.1 0.9 8.1 1.4
Where, P-rate = Phosphorus rate; Df = degree of freedom; D50%F = days to 50% flowering; D90%PM = days to
90% physiological maturity; NPBPP = number of primary branches per plant; PH = plant height. ** and * =
highly significant and significant at 1% and 5% level of significance, respectively.
Appendix Table 3. Mean squares of analysis of variance for nodulation parameters of faba
bean as affected by Rhizobium inoculation and phosphorus fertilizer rate
Mean squares
Source of variation Df TNNPP NENPP NVPP NDWPP
Block 2 147.69 138.11 0.990002 11349.6
Rhizobium strain 3 1971.52** 1957.84** 1.310153** 36222.6**
P-rate 4 2761.89** 2587.63** 3.101090** 66616.6**
Rhizobium × P-rate 12 62.29** 82.51** 0.170361** 1715.4**
Error 38 21.44 23.25 0.007728 119.3
CV (%) 4.2 4.8 4.7 4.6
Where, P-rate = Phosphorus rate; Df = degree of freedom; TNNPP = total number of nodules per plant; NENPP =
number of effective nodules per plant; NVPP = nodule volume per plant; NDWPP = nodule dry weight per plant.
** and * = highly significant and significant at 1% and 5% level of significance, respectively.
76
Appendix Table 4. Mean squares of ANOVA for yield components of faba bean as affected by
Mean squares
Source of variation Df CSC NPPP NSPP HSW
Block 2 590513.0 1.226 0.15131 1.1284
NS
Rhizobium strain 3 5708318.0 112.334** 0.32380** 16.6914**
P-rate 4 46872056.0** 328.416** 0.71904** 70.5621**
Rhizobium × P-rate 12 910377.0NS 9.566** 0.10192** 2.2067*
Error 38 11157601.0 3.357 0.01342 0.9857
CV (%) 1.4 5.9 4.2 2.0
Where, P-rate = Phosphorus rate; Df = degree of freedom; CSC = crop stand count at harvest; NPPP = number of
pods per plant; NSPP = number of seeds per pod; HSW = hundred seed weight. **, * and NS = highly significant,
significant and non-significant at 1% and 5% level of significance, respectively.
Appendix Table 5. Mean squares of ANOVA for yields and harvest index of faba bean as
affected by Rhizobium inoculation and phosphorus fertilizer rate
Mean squares
Source of variation Df GY AGDB HI
Block 2 11104.0 26178.0 0.114
Rhizobium strain 3 1135319.0** 1235636.0** 50.630**
P-rate 4 1764269.0** 2744910.0** 67.806**
Rhizobium × P-rate 12 137892.0* 210016.0** 6.007NS
Error 38 67222.0 68148.0 4.498
CV (%) 8.0 3.1 5.5
Where, P-rate = Phosphorus rate; Df = degree of freedom; GY = grain yield; AGDB =above ground dry biomass
yield; HI = harvest index. **, * and NS = highly significant, significant and non-significant at 1% and 5% level of
significance, respectively.

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EFFECT OF RHIZOBIUM INOCULATION AND PHOSPHORUS

FERTILIZER RATES ON YIELD COMPONENTS AND YIELD OF

A Thesis Submitted to the School of Plant Sciences

In Partial Fulfillment of the Requirements for the Degree of

Jemal Abdulahi (PhD) ___________________ __________________

Tamado Tana (PhD) ___________________ __________________

_________________________ ___________________ __________________

_________________________ ___________________ __________________

_________________________ ___________________ __________________

Name: _____________________________ Signature: ____________

Date of Submission: __________________

Appendix Table Page

Key words: Biofertilizer, Biological Nitrogen Fixation, Inoculants, Legume, Nodulation

2.1. Overview of Faba Bean Production

2.2. Bio-fertilizers and Their Importance on Faba Bean Production

2.3. Importance and Mechanism of Biological Nitrogen Fixation

There is a huge reservoir of N in the atmosphere; the atmosphere comprises of approximately

Legume BNF involves a remarkable symbiosis, or mutually beneficial relationship between

2.4. Factors Limiting Biological Nitrogen Fixation

2.4.1. Soil temperature

2.4.2. Soil moisture status

2.4.3. Nutrient availability

2.4.5. Agronomic management

2.5. Importance of Phosphorus Supply on Faba Bean Production

2.6. Effect of Inoculation and Phosphorus Application on Yield Components

3. MATERIALS AND METHODS

3.1. Description of the Study Area

3.2. Experimental Materials

3.3. Soil Sampling and Analysis

3.4. Treatments and Experimental Design

3.5. Experimental Procedure and Plot Management

3.5.1. Seed treatment with Rhizobium (seed inoculation)

3.5.2. Management of the plots

3.6. Data Collection and Measurement

3.6.2. Nodulation parameters

3.6.3. Yield components and yield

content and D is the designated moisture content (10%).

3.7. Data Analysis

Data collected were subjected to analysis of variance (ANOVA) appropriate to factorial

3.8. Economic Analysis

4. RESULTS AND DISCUSSION

4.1. Soil Physico-chemical Properties of the Experimental Site

Cation exchange capacity (CEC) is an important parameter of soil, because it gives an

4.2. Phenological and Vegetative Growth Parameters of Faba Bean

4.2.1. Days to 50% flowering

Table 2. Days to 50% flowering of faba bean as affected by interaction of Rhizobium

4.2.2. Days to 90% physiological maturity

4.2.3. Number of primary branches per plant

4.2.4. Plant height

4.3. Nodulation Parameters of Faba Bean

4.3.1. Total number of nodules per plant

4.3.2. Number of effective nodules per plant

Effective nodulation is essential for a functioning legume/Rhizobium symbiosis, as plants most

4.3.3. Nodule volume per plant

4.3.4. Nodule dry weight

4.4. Yield Components and Yield of Faba Bean

4.4.1. Crop stand count at harvest

plant physiology, including the fundamental processes of photosynthesis, reproduction,

4.4.2. Number of pods per plant

4.4.3. Number of seeds per pod

4.4.4. Hundred seed weight

4.4.5. Grain yield

Rhizobium hastened vegetative growth, while, phosphorus application improved yield

4.4.6. Above ground dry biomass yield

Jemal Abdulahi (PhD) _

Tamado Tana (PhD) _

_______________ _ __________

_______________ _ __________

_______________ _ __________

Name: _________________ Signature: