Agronomic and symbiotic characteristics of chickpea, Cicer arietinum (L.

), as
influenced by Rhizobium inoculation and phosphorus fertilization under farming
systems of Wolaita area, Ethiopia

Ibsa Aliyi Abdula

MSc Thesis Plant Production Systems
``June 2013

Agronomic and symbiotic characteristics of chickpea, Cicer arietinum (L.), as

influenced by Rhizobium inoculation and phosphorus fertilization under farming
systems of Wolaita area, Ethiopia

Ibsa Aliyi Abdula (Reg. No = 841118003050)

MSc thesis Plant Production Systems
PPS 80436
June 2013

Supervisors: Dr. Katrien Descheemaeker

Plant Production Systems
Wageningen University
Prof.Dr. Ken Giller
Head, Plant Production Systems Chair Group
Co-supervisor: Dr. Endalkachew W/Meskel
Hawassa University, Ethiopia

Examiners:

Prof.Dr. Ken Giller

Dr. Katrien Descheemaeker

Acknowledgements
I am highly indebted to my supervisors Dr. Katrien Descheemaeker, Dr. Endalkachew
Wolde-meskel and Prof. Dr. Ken Giller for their instruction, guidance and scientific support
starting from proposal writing phase until this final report. I would also like to thank Esther
Ronner for her support and facilitation of the required inputs throughout the implementation
of the experiment. My deepest gratitude is also extended to Endalkachew Wolde-meskel of
Hawassa University, Ethiopia and his staff members without whom execution of this work
could have not been a success. His staff members have been providing me with useful
technical support throughout this study. Ones again, my special thanks are for Prof. Dr. Ken
Giller for initiating this study and funding the entire experiment through N2Africa project.
Dr. Katrien Descheemaeker was always keen to answer my questions, arrange facilities, read
my document and allocate her time for discussion whenever I needed. She was so friendly
during the course of my stay at the chair group and she played a crucial role for the successful
accomplishment of this work, thank you Katrien.
The innocent and invaluable support and cooperation of farmers in Taba kebele of Wolaita
zone is highly appreciated. I deep heartedly thank my friends; Dereje Wube, Micheal
Mohamed, Bulti Tesso, Bacha Daba and many others who were always there to share my joy
and sorrow. All members of my family are also gratefully acknowledged.
I would also like to thank my wife Etifwork and my son Naol for their understanding and
patience even when I was totally devoted to this work.

ii

Acronyms
BNF

biological nitrogen fixation

CSA

Central Statistics Authority, Ethiopia

NDWPP

nodule dry weight per plant

NNPP

number of nodules per plant

NNPPM

number of nodules per plant at maturity

DAS

days after sowing

NSPP

nodule score per plant

NSPPM

nodule score per plant at maturity

NPPP

number of pods per plant

NoSPP

number of seeds per pod

NBPP

number of branches per plant

SDWPP

shoot dry weight per plant

SFWPP

shoot fresh weight per plant

TN

total nitrogen

AP

available phosphorus

FAOSTAT

Food and Agricultural Organization statistics

1000sw

thousand seeds weight

GY

grain yield

SNNPRS

Southern Nations, Nationalities and Peoples Regional State

MPN

most probable number

STN

straw total nitrogen

STP

straw total phosphorus

iii

Table of Contents
Acknowledgements.................................................................................................................................ii
Acronyms...............................................................................................................................................iii
TableofContents...................................................................................................................................iv
Foreword..............................................................................................................................................viii
Summary.................................................................................................................................................x
1.Introduction........................................................................................................................................7
1.1.Chickpea.....................................................................................................................................10
1.1.1.Origin,distributionandglobalproductiontrend................................................................10
1.1.2.ProductionandrolesofchickpeainEthiopia.....................................................................10
1.2.Phosphorusanditsimportanceincropproduction..................................................................13
1.3.Nitrogenanditsroleincropproduction...................................................................................14
1.4.BiologicalNitrogenFixation.......................................................................................................15
1.5.FactorsaffectingBiologicalNitrogenFixation...........................................................................16
1.5.1.SoilMineralNitrogen..........................................................................................................16
1.5.2.SoilPhosphorus...................................................................................................................17
1.5.3.Competitionbetweenrhizobia...........................................................................................17
1.5.4.Soiltemperature,pHandsalinity.......................................................................................17
1.5.5.Soilmoisturestatus.............................................................................................................18
1.5.6.Agronomicmanagement....................................................................................................18
2.MaterialsandMethod......................................................................................................................19
2.1.Descriptionofthestudyareaandtheprevailingfarmingsystem.............................................19
2.2.FarmSelection...........................................................................................................................22
2.3. Estimation of indigenous rhizobial population of the study soils..............................................23
2.4.Experimentaldetailsoffieldexperiment...................................................................................24
2.4.1.Cropcultivar........................................................................................................................24
2.4.2.TreatmentsandLayout.......................................................................................................24
2.4.3.Rhizobiumpreparationandinoculation.............................................................................24
2.5.RainfallData...............................................................................................................................25
2.6.Soilsamplingandanalysis..........................................................................................................25
2.7.Planttissueanalysis...................................................................................................................26
2.8.Managementpractices..............................................................................................................26
2.9.ObservationsandDatacollection..............................................................................................27
2.9.1.Cropphenology...................................................................................................................27
2.9.2.Nodulation..........................................................................................................................27
iv


2.9.3.Shootweight.......................................................................................................................27
2.9.4.Grainyield,yieldrelatedtraitsandtotalbiomassproduction...........................................28
2.9.5.Dataonhouseholdcharacteristics......................................................................................28
2.10.Statisticalanalysis....................................................................................................................28
3.Results...............................................................................................................................................29
3.1.Soilcharacteristicsofthestudyfarms.......................................................................................29
3.1.1.Physicochemicalproperties...............................................................................................29
3.1.2. Nativerhizobiapopulations................................................................................................30
3.2.Rainfalldata...............................................................................................................................31
3.3.Effectsofsoiltypesonnodulation,growth,podproductionanddaystomaturityofchickpea
(CicerarietinumL.)............................................................................................................................32
3.4.EffectsofRhizobiuminoculationand/orPhosphorusfertilizationonnodulationofchickpea
(CicerarietinumL.)............................................................................................................................33
3.4.1.Nodulenumberplant1.......................................................................................................33
3.4.2.Nodulescoreplant1............................................................................................................34
3.4.3.Noduledryweightplant1at45DAS,(NDWPPat45DAS).................................................34
3.5.EffectsofRhizobiuminoculationand/orphosphorusfertilizationonphenologicaland
agronomictraitsofchickpea(CicerarietinumL.).............................................................................36
3.5.1.Shootfreshweightplant1..................................................................................................36
3.5.2.Daystomaturity..................................................................................................................36
3.5.3.Numberofbranchesplant1...............................................................................................36
3.5.4.Plantheight(cm).................................................................................................................36
3.6.EffectsofRhizobiuminoculationand/orPhosphorusfertilizationonyieldandyieldrelated
traitsofchickpea(CicerarietinumL.)...............................................................................................38
3.6.1.Numberofpodsplant1(NPPP)andnumberofseedspod1(NSPP)...................................38
3.6.2.Totalbiomassproduction(tonha1)...................................................................................38
3.6.3.Grainyield(tonha1)...........................................................................................................38
3.6.4.ThousandSeedsweight(gram)...........................................................................................38
3.7.EffectsofRhizobiuminoculationand/orphosphorusfertilizationonstrawtotalnitrogen(STN,
%)andphosphoruscontent(STP,mgkg1)andtotalNuptakeofchickpea.....................................40
3.8.Interactioneffectsbetweensoilfertilitytreatmentsandsoiltypesongrowthand
performanceofchickpea..................................................................................................................41
3.8.1.Nodulenumberandscoreplant1at45daysaftersowing(45DAS)..................................41
3.8.2.Daystomaturity..................................................................................................................41
3.9.Correlationsamongthesymbioticandagronomicvariablesofchickpea.................................43
3.10.Acrossfarmvariabilityinresponsetothesoilfertilitytreatments.........................................44
v


3.10.1.Yieldandtotalbiomassproduction..................................................................................44
3.10.2.Nodulenumberplant1andshootfreshweightplant1at45DAS....................................45
3.11.Acrossfarmvariabilityinnodulation,biomass,yieldandtheirresponsetothesoilfertility
treatmentsunderthetwosoiltypes................................................................................................46
3.11.1.Nodulenumberplant1......................................................................................................46
3.11.2.Yieldandbiomass.............................................................................................................47
3.11.3.Yieldresponsetothesoilfertilitytreatmentsofthetwosoiltypes................................48
3.12.Theeffectofinitialsoilfertilityonyieldandmagnitudeofyieldresponsetothesoilfertility
treatments........................................................................................................................................49
3.13.Householdcharacteristics........................................................................................................51
4.Discussion..........................................................................................................................................53
4.1.Nodulation.................................................................................................................................53
4.2.Morphologicalandphenologicaltraitsofchickpea...................................................................54
4.3.Yieldandyieldrelatedtraitsofchickpea...................................................................................55
4.4.StrawqualityandtotalNuptake...............................................................................................57
4.5.Acrossfarmvariability...............................................................................................................58
4.6.Costbenefitanalysis..................................................................................................................59
5.Conclusions.......................................................................................................................................61
6.Implicationsoftheresultsforpolicyandresearch...........................................................................63
7.References........................................................................................................................................64
8.Appendices........................................................................................................................................69

vi

vii

Foreword
This experiment was conducted at Taba kebele of Damot Galle woreda located in Wolaita
zone, Ethiopia from August 2012 till January 2013. The study was initiated and fully funded
by the N2Africa project and executed as part of MSc thesis research for fulfilment of Masters
of sciences in Agronomy/ plant sciences. N2AFRICA is a large scale research project focused
on putting nitrogen fixation to work for smallholder farmers growing legume crops in Africa.
The project came into action in 2010 and it is funded by the Bill & Melinda Gates Foundation
and the Howard G. Buffet Foundation through a grant to Plant Production Systems chair
group of Wageningen University in the Netherlands. It is led by Wageningen University,
CIAT-TSBF and IITA along with many partners in 13 African countries, including five
countries with which a new partnership was established recently. Ethiopia is one of these five
countries. The aims of the project are to raise average grain legume yields by 954 kg ha-1 in
four crops i.e. groundnut, cowpea, soybean and common bean, increase average biological
nitrogen fixation by 46 kg/ha, and increase household income by $465, directly benefitting
225,000 households in eight sub-Saharan African countries. The first phase of the project
ends in October this year, and based on the results obtained so far, the project seems to be in
a good position to meet the set target.
In Ethiopia, partnerships have been established with various research and development
organizations, among which Hawassa University. N2Africa considers the newly joining
Ethiopia as a potential improvement area as far as productivity of grain legumes of
smallholder farmers is concerned and it was within this setting that this experiment was
conducted on farmers field. The aim of the experiment was to assess the response of
chickpea to the packages of N2Africa, being phosphorus fertilizers and Rhizobium inoculants,
under farming conditions of Wolaita area, one of the high potential areas for Ethiopian
chickpea production. Accordingly, the experiment was conducted with full financial support
of the N2Africa project and technical assistance of the soil microbiology laboratory of the
School of Plants Sciences, Hawassa University of Ethiopia. The results obtained are used in
this MSc thesis report to be submitted to the Plant Production Systems group, Wageningen
University.

viii

ix

Summary
Chickpea is the third most widely cultivated food grain legume in Ethiopia and it is known as
a multifunctional crop that fits well in rotation with cereals like maize and tef. Ethiopia is the
largest chickpea grower in Africa with a share of 39% of the total production in 2011.
Despite its potential yield of more than 3 ton ha-1, current chickpea productivity is only 1.6
ton ha-1 in Ethiopia. In this regard, there is a need to improve the current productivity through
improved technologies and agronomic practices. N2Africa, a large research project focused
on putting biological nitrogen fixation to work for smallholder farmers growing legume crops
in Africa, provides Rhizobium inoculants and P fertilizer technologies to farmers in several
African countries and achieved satisfactory results so far. This experiment was conducted
with the aim of evaluating the response of chickpea to inoculation, P fertilizer, and their
combined application under farming conditions in the Wolaita area of Ethiopia, which is
among the major food insecure and densely populated regions in the country.
For this purpose, Mesorhizobium ciceri strain CP 41 was obtained from the soil microbiology
laboratory of the School of Plant and Horticultural Sciences of Hawassa University, Ethiopia
and used along with a chickpea variety Natoli. Four treatments, including control, inoculants,
P-fertilizer (TSP) and the combination of inoculants + P-fertilizer, were tested on twenty
farms spread over two soil types. The experiment was conducted from September 2012 to
January 2013 at Taba Kebele of Damot Gale woreda in Wolaita zone, where the majority of
farmers grow chickpea at the end of the main rainy season. To measure the response of
chickpea to the treatments, data were collected on various symbiotic, phenological, yield and
yield related traits of chickpea and subjected to analysis of variance using statistical software
GENSTAT 15.
Compared with the control, inoculation and P fertilizer significantly improved nodulation,
growth and yield of chickpea, with a more prominent improvement with combined
application of inoculation and P. Inoculation, P and their combined application increased
grain yield of chickpea by 26%, 19% and 33% over the control respectively. Similarly, the
total nitrogen content of the straw increased by 56% and 82% compared to control due to
inoculation and P + inoculation treatments respectively. Positive effects the soil fertility
treatments were also observed for the total N uptake of chickpea as eventually 28, 53, 42, and
65 kg N ha-1 was recorded for the control, inoculation, P, and P + inoculation treatments
respectively.
x

For total straw P content, the use of P and P + inoculation resulted in 41% and 28% more
total P uptake.
On the other hand, the result of the most probable number test showed that the soils of this
area were inhabited by a very small population of resident rhizobia (< 10 gram-1 of soil).
For almost all of the variables studied a large variation across farms was observed. On 85%
of the farms, the soil fertility treatments gave higher grain yield compared to the control
treatment and the yield range was 0.5-2.5 ton ha-1, 0.5-2.8 ton ha-1, 0.7-3.0 ton ha-1 and 0.72.9 ton ha-1 for the control, inoculation, P, and inoculation + P treatments respectively with
their respective overall average yield of 1.6, 2, 1.9 and 2.1 ton ha-1. The magnitude of yield
response to the soil fertility treatments showed a negative but non-significant correlation with
initial soil fertility of the farms. Although yields for all treatments were larger on black than
on red soils, the magnitude of the yield response to the treatments was greater on the red than
on the black soils. Generally, the observed improvement in chickpea performance can be
attributed to the increased supply of nitrogen, through enhanced biological nitrogen fixation,
and phosphorus, which were both present in low supply in the soils.
These positive effects of the soil fertility treatments will, if adopted by the local farmers,
contribute to increasing chickpea productivity and household income. Given the small
landholdings (on average 0.6 ha) of the households, the observed improvement may play a
modest role in improving the livelihood of the smallholders in this area. The use of
inoculation, P fertilization, and their combination on 25% of the land would increase the
household income by 63, 33, and 63 USD respectively. However, the unavailability of P
fertilizers and inoculants on the market in the country might greatly constrain the promotion
of these technologies. Therefore, solutions need to be sought by working closely with private
and governmental organizations and stimulate their interest to get involved in the business.

xi

1. Introduction
The concern of soil fertility decline in Africa has been raised since many years, influencing
top policy priorities to establish a range of initiatives with the aim of alleviating the problem
(Elias and Scoones 1999). Mineral mining due to continuous cultivation, lack of inorganic
fertilizer inputs, and limited application of organic inputs are some of the reasons for the
depletion of soil fertility in developing countries. Large number of African farmers generally
consider soil fertility decline as the major limitation to farming (Smaling et al. 1997). Poor
soil fertility has resulted in low crop productivity and is one of the causes of food insecurity
in the region. In Ethiopia, where the agricultural sector is dominated by smallholder farmers,
the soil fertility situation is said to be desperate (Pound et al. 2005). The food insecurity
problem is aggravated by human population growth, with decreasing farm sizes as a
consequence. Therefore, there is a need to improve soil fertility and increase productivity
thereby producing sufficient food for the expanding population. To achieve this, fertilizer use
is proposed as a solution (Foth and Ellis 1997).
In African agriculture, the nitrogen requirement is by far the greatest of all major nutrients
(Woldeyohannes et al. 2007) and projected to rise significantly (FAO 2012). However, in
developing countries small-scale farmers often cannot afford costly inorganic fertilizer. The
situation is further aggravated by poor infrastructure and lack of credit services. Therefore,
there is a need to reduce the dependence on nitrogen fertilizer thereby minimizing
environmental degradation while improving crop productivity. As stated by Shamseldin and
Werner (2004), biological nitrogen fixation (BNF) can reduce the need for fertilizer. Each
season, grain legumes fix about 15 - 210 kg N ha-1 in Africa (Dakora and Keya 1997). More
specifically, chickpea can fix up to 140 kg N ha-1 in a growing season (Kumar and Abbo
2001) and its inclusion in crop rotations of cereal dominated farming systems can also reduce
disease severity of non-legume crops through disrupting the disease cycle.
In soils where native rhizobial population densities are low, BNF can be enhanced by
inoculation of legume seeds with an effective and persistent Rhizobium strain. In this regard,
Rhizobium inoculation of chickpea seed may substitute costly N fertilizers in chickpea
production (Kosgey 1994). Several studies demonstrated improved yields with inoculation:
Hailemariam and Tsige (2003, cited by Yoseph 2011) reported a yield advantage of 10 to
50% for faba bean, field pea and lentil inoculated with Rhizobium leguminosarum.

Similarly, the same study showed that the inoculation of chickpea seed with an appropriate
strain increased yields up to 38% over the control. The development of rhizobial inoculants is
seen as the greatest success achieved in scientific research conducted on BNF (Giller and
Cadisch 1995). Depending on the native Rhizobium population in the soil, inoculation is
proven worthy as it is economically cheap, environmentally sound, and ecologically
beneficial.
However, the process of nitrogen fixation is sensitive to P deficiency, which results in
reduced nodule mass and low ureide production. P plays an important role in N cycling as
adenosine triphosphate is required in large quantities for legumes to undergo N2 fixation
(Vance 2001; Sinclair and Vadez 2002). Unfortunately, low soil P availability is limiting crop
production both at country level and in the Southern Nations, Nationalities and Peoples
Region (SNNPR) of Ethiopia (Mamo and Haque 1987; Schulze et al. 2006). Particularly soils
of many areas in Wolaita suffer from remarkable P deficiencies due to P fixation (Elias and
Scoones 1999).
Ethiopia is the largest chickpea producer in Africa, with a share of about 39% of total
chickpea production of the continent in 2011 (FAOSTAT 2012). In SNNPR more than 6000
ha of land were planted with chickpea in the year 2008. In the same year, Wolaita Zone
stood third in the region as far as chickpea production area is concerned (Kassie et al. 2009).
In Wolaita chickpea is one of the most important grain legumes produced by small scale
farmers and it serves as a source of food and cash. Given its ability to grow on residual
moisture, chickpea plays an important role in the farming system by fitting in the crop
rotation. As a result, growing chickpea allows the farmers to produce extra crop each year.
However, in spite of its potential to produce about 3 ton ha-1, the average productivity of
chickpea in Ethiopia for 2011 was only 1.6 ton ha-1 (FAOSTAT 2012). In Wolaita, like in
other parts of the country, productivity of chickpea and other crops is constrained by poor soil
fertility. With the aim of improving crop productivity, the Ethiopian government has been
providing and promoting various technologies and improved practices to the farmers
including improved cultivars and fertilizer.
Also, given the potential of BNF, several effective strains of Rhizobium have been identified
for chickpea. However, these strains were identified through on-station research and hardly
tested on farmers fields. Moreover, only limited efforts have been made to distribute the
identified strains to farmers so that they could benefit from this technology.
8

The present study, therefore, was conducted with the following objectives:
To determine the status of resident indigenous rhizobia in the soils of the study area
To investigate the effect of Rhizobium inoculation, P fertilization and their
combination on nodule development, plant growth, grain yield and residue production
and quality of chickpea sown on farmers fields in Wolaita area
To investigate the effect of soil type on the potential benefit of Rhizobium inoculation
and P fertilization on chickpea yield
To understand the variation in response to the soil fertility treatments across farms

1.1. Chickpea
1.1.1. Origin, distribution and global production trend

Chickpea, Cicer arietinum (L.), one of the most important cool season crops, is believed to
have originated in present-day south eastern Turkey and adjoining Syria where several of its
natural species are found (Saxena and Singh 1987). The crop later spread to India, Europe
and subsequently reached Africa, Latin and central American countries. Chickpea is a selfpollinated annual crop that can complete its life cycle in 90 to 180 days depending on the
prevailing meteorological conditions. Chickpea can be grown under a wide range of agroclimatic conditions around the world (Singh and Diwakar 1995). Chickpea growing areas can
be classified into the following four major geographical regions: Indian subcontinent; West
Asia, North Africa, and southern Europe; Ethiopia and East Africa; and the Americas and
Australia (Smithson 1985; Singh and Diwakar 1995).
Currently, chickpea is produced worldwide and it is the worlds third most important food
legume next to haricot bean and soybean (Namvar and Sharifi 2011). In 2011, chickpea was
grown on about 13.2 million hectares of land across the world with an average productivity of
0.9 ton ha-1. Globally, during 1994 2005, the annual chickpea production increased by
1.87% (Kassie et al. 2009). Chickpea is among the widely cultivated pulse crops by small
scale farmers of the semi-arid tropics (Anbessa and Bejiga 2002). Generally, Desi and Kabuli
types are the two major types of chickpea grown in the world with major differences in seed
size, seed colour, surface and thickness of the seed coat. The Desi type is characterized by
small seeds with angular appearance, sharp edges and varying colours but usually light
brown. On the other hand, the Kabuli type produces large round seeds of white or pale cream
or yellow colour.
1.1.2. Production and roles of chickpea in Ethiopia

In Ethiopia, chickpea is mainly grown in the central, northern and eastern highland areas of
the country at an altitude of 1400-2300 m.a.s.l., where annual rainfall ranges between 700
and 2000 mm (Bejiga 1994; Anbessa and Bejiga 2002). During the 2010/11 cropping year,
Ethiopia produced 322,839 ton of chickpea on 208,389 ha of land and stood sixth on the
global ranking of chickpea producing countries (FAOSTAT 2012). The average productivity
of Ethiopian chickpea in 2011 was 1.6 ton ha-1 which is nearly half of the 3 ton ha-1 that can
be expected under prevailing conditions. In this year, chickpea contributed about 17% of

10

Ethiopias total food legume production and stood third after faba bean and haricot bean
(Fig.1B) (FAOSTAT 2012).
In the same year, Ethiopias share in African chickpea production was 39% (Fig.1A)
(FAOSTAT 2012). Over the past years there has been an increasing trend in total area of
production, the quantity of chickpea produced and productivity of chickpea in Ethiopia
(Abate et al. 2011). During 1995-2005, Ethiopian chickpea planted area and chickpea
production showed annual growth rate of 2.1% and 7.6% respectively (Kassie et al. 2009).
A

Others
11%

Tanzania
14%

Lentils
4%
Ethiopia
39%

Soya
beans
1%
G.peas
11%

Faba
beans
36%

Chickpea
17%

Morroco
15%

Haricot
beans
18%

Malawi
21%

Field
peas
13%

Figure1:MajorAfricanchickpeaproducingcountrieswiththeirshareoftotalproductioninAfricaduring2011(A)and
majorpulsecropsinEthiopiawiththeirshareoftotalgrainlegumeproductionin2011(B)(adaptedfromCSAand
FAOSTAT,2012)

The crop can be grown on different soil types as long as good drainage is ensured. However,
to achieve optimum growth well drained black soils (usually Vertisols) are identified as the
most suitable soil type (Kassie et al. 2009). In Ethiopia, chickpea is mostly grown on vertisols
which have good water holding capacity. Ethiopian farmers essentially grow chickpea on
residual moisture after the end of the main rainy season, which is usually in September October. Of the two types of chickpea, traditionally the Desi types are more widely cultivated
in Ethiopia (Kassie et al. 2009).
Chickpea, a multi-functional crop, has an important role in the diet of the Ethiopian smallscale farmers households and also serves as protein source for the rural poor who cannot
afford to buy animal products. The crop also serves as a source of cash income and plays a
major role in Ethiopias foreign exchange earnings through export to Asia and Europe. Its
straw is also used for animal feed and due to its capacity of biological nitrogen fixation,
11

chickpea can improve the soil fertility status (Pundir and Mengesha 1995). In spite of all
these virtues and benefits, the productivity of this crop remains very low in Ethiopian
agriculture.
Over the past 38 years, Ethiopian chickpea research has focused mainly on breeding and
selection of improved cultivars with better yield and disease resistance. Through this
approach, Ethiopian Agricultural Research Organization (EARO) released 11 improved
chickpea varieties (6 Kabuli types and 5 Desi types) from 1974 to 2005 (Kassie et al. 2009).
However, this did not result in the desired level of productivity as the average yield was still
below 1.6 ton ha-1 in 2011. Variety development can be seen as a component of a package
through which crop yield can be improved and it has to be supported by appropriate
agronomic management including optimum fertilizer rate, proper weeding, planting at a right
time, inoculation, and disease and pest control measures. Therefore, one way of improving
yield of leguminous crops is inoculation of their seeds with Rhizobium bacteria that has
already shown remarkable result in other African countries (Woomer 2012).

12

1.2. Phosphorus and its importance in crop production

Next to nitrogen, phosphorus is the most important element for adequate grain production.
The evolution of science, particularly in the past century, has clearly demonstrated the
significance of phosphorus for all animal and plant life on the earth (Ryan et al. 2012). Rock
phosphate is the global source of raw material for P fertilizer and it is wise to efficiently use
this finite resource. Substantial amount of P is found in different parts of plants. Large
quantities of P are found in seed and fruit, and it is considered essential for seed formation
(Gidago et al. 2012). Especially in the early stages of plant development, adequate supply of
P is required for development of the reproductive parts and P has a positive effect on root
growth, early maturity, and reduced disease incidence. Despite the presence of a large P pool,
Africa is suffering from shortage of available P, which remains a yield limiting factor. Also
in the highlands of Ethiopia, available P is often a limiting element in crop production (Mamo
and Haque 1987), with 70 to 75% of the agricultural soils deficient in P. Similarly, available
P content of most soils in SNNPR of Ethiopia is low (Gidago et al. 2012).
Application of small amounts of P fertilizer (26 kg P ha-1) dramatically increased nodulation,
N accumulation, and seed yields of haricot bean grown on farmers fields in northern
Tanzania (Giller et al. 1998). From data obtained from multi location experiments, consistent
yield response of faba bean to P fertilization was observed (Ghizaw et al. 1999). Likewise,
application of 10 kg P ha-1 significantly improved grain yield and biological yield of haricot
bean planted at Areka research station, SNNPR-Ethiopia (Gidago et al. 2012). The same
study revealed that application of P enhanced physiological maturity and yield components,
though not in a statistically significant way. Similarly, application of P fertilizer enhanced
mid-season dry matter accumulation and tissue P accumulation of both Desi and Kabuli
chickpea, but grain yield was increased only modestly for Desi chickpea, while yield of
Kabuli chickpea was not affected by application of P (Walley et al. 2005)

13

1.3. Nitrogen and its role in crop production

Nitrogen is known to be an essential nutrient for plant growth and development (Werner and
Newton 2005) due to its role in biochemical, physiological and morphological processes of
plant production (Novoa and Loomis 1981).
Although this critically important element is abundant in the atmosphere, nitrogen is the most
limiting element for crop growth worldwide. In sub-Saharan Africa both yield and quality of
crops are highly constrained by low nitrogen availability. Application of mineral fertilizer,
addition of organic material and enhancing biological N2 fixation are the main ways of
improving the nitrogen availability to the plants.
Application of nitrogen fertilizer is reported to have positive effects on morphological traits
and yield of crops in general and chickpea in particular. The usage of 100 kg urea ha-1, for
example, resulted in the highest biomass production and grain yield of chickpea as compared
to the control and lower rates of fertilizer on a silty loam soil in Iran. Compared to the
control, addition of 100 kg urea ha-1 improved grain yield of chickpea by 36% (Namvar et al.
2011). Similarly, application of 30 or 45 kg N ha-1 significantly increased grain yield of Desi
type chickpea by 221 kg ha-1 relative to the 1.6 ton ha-1 of control treatment when averaged
across locations on clay loam soils of Saskatchewan, Canada (Walley et al. 2005). The same
study revealed that nitrogen application increased shoot P accumulation of Desi type
chickpea by 1.1 kg ha-1.

14

1.4. Biological Nitrogen Fixation

Biological nitrogen fixation (BNF) is a process by which N2 in the atmosphere is reduced into
a biologically useful, combined form of N-ammonia by living organisms (Hardy and Burns
1968; Giller 2001). The greatest proportion of N found on the earth is located in the
atmosphere, as N2. Nevertheless, the majority of organisms cannot utilize this free and
abundant, but highly stable source of N because they can only use N which is combined with
other atoms into plant usable forms, such as ammonium, nitrates and ammonia (Giller 2001;
Giller and Cadisch 1995). The process of making N2 available constitutes a specialized and
intricately evolved interaction of soil microbes (bacteria) and higher plants via the formation
of nodules (Sessitsch et al. 2002). Nodules are formed on roots or, in some cases, stems
(Tamimi and Timko 2003).
Each year, about 175 million ton of N is contributed by BNF globally, of which nearly 79%
is accounted for by terrestrial fixation. Therefore, symbiotic nitrogen fixation is of great
importance not only in the production of leguminous crops but also in the global nitrogen
cycle (Ben Romdhane et al. 2008). The most important N2 fixing agents in agricultural
systems are the symbiotic associations between legumes and the microsymbiont rhizobia
bacteria (Giller 2001).
Depending on the availability and effectiveness of the native rhizobia, one way of improving
N2 fixation in grain legumes is inoculation of the crop seeds with effective strains of rhizobia.
Despite being mentioned by some as a promiscuous host (Rivas et al. 2007), there is
consensus that both nodulation and growth of chickpea can be improved by inoculation
(Giller 2001).The growth and yield of chickpea can be improved by inoculating seeds with
competitive strains of rhizobia and this can be an economically feasible way of increasing
productivity of chickpea (Ben Romdhane et al. 2008). Across 16 on-station trials inoculation
of chickpea with Rhizobium increased grain yield by an average 342 kg ha-1 (Wani et al.
1995). Likewise, depending on cultivar, effectiveness of bacterial strain and environmental
factors the association of chickpea and Mesorhizobium Cicer produced up to 176 kg N ha-1
annually in India (Rupela et al. 1987a).

15

1.5. Factors affecting Biological Nitrogen Fixation

The ability of symbiotic nitrogen fixing agents to fix N2 is strongly influenced by the
prevailing environmental conditions that can mainly be categorized as physical factors,
chemical factors, and nutrient deficiencies (Giller 2001). In addition, Rhizobium species
producing nodules in chickpea are specific only to this crop and inoculation with effective
strains is advised in soils with no or weak bacterial presence (Rupela et al. 1987a;
Somasegaran et al. 1988).
The survival of Rhizobium sp. in the soil is influenced by a combination of factors such as
nitrogen and phosphorus availability, acidity, salinity, alkalinity, concentration of
micronutrients, soil temperature, moisture, soil structure, and fertility status of the soil (Giller
2001; Slattery et al. 2001). These factors may influence the growth of microorganisms in the
free-living state, the process of plant infection or nodule development, and the fixation of N2
after the symbiosis has been established (Giller 2001). Depending on their type, rhizobia
exhibit an optimum growth in a temperature range of 25 30C (Kantar et al. 2010).
However, some chickpea rhizobia have shown their maximum growth at 20C (Rodrigues et
al. 2006). Similarly, saline conditions may limit the symbiosis through affecting survival and
proliferation of Rhizobium spp. in the soil and rhizosphere, inhibiting the infection process,
affecting root nodule function directly or indirectly by reducing plant growth and its other
physiological processes (Singleton et al. 1982).
1.5.1. Soil Mineral Nitrogen

Depending on the accessible quantity present, the mineral nitrogen content of the soil can
have both positive and negative effects on yield and growth response of chickpea to
inoculation. Usually a higher mineral nitrogen content in the rhizosphere leads to poor N2
fixation through inhibition of nodulation of chickpea (Namvar et al. 2011). On the other
hand, small amounts of soil or fertilizer N often have a stimulatory effect on nodulation and
N2 fixation which is principally due to the positive effect of N on growth and plant
establishment during the period between root emergence and the onset of active N2 fixation
(Giller and Cadisch 1995).

16


1.5.2. Soil Phosphorus

Usually, suboptimal availability of P leads to limited root growth, reduction of photosynthesis

and translocation of sugars, thus affecting N fixation. Moreover, being a building block of a
plant energy source, P is important in N cycling because adenosine triphosphate is required in
large quantities by legumes to develop nodules and undergo the fixation process (Sessitsch et
al. 2002). As N2 fixation is an energy demanding process (Giller 2001), larger P quantities are
needed by N2 fixing plants than by mineral N supplied plants. In this regard, poor nodulation
and poor plant vigour have been observed in beans grown in soils low in extractable P
(Amijee and Giller 1998) while acute deficiency of phosphorus can even prevent nodulation
by legumes (Giller 2001), showing the sensitivity of the process of N2 fixation to P
deficiency.
1.5.3. Competition between rhizobia

Indigenous rhizobia in soils can vary in population density and infectivity from place to place
ranging from < 10 to 107 cells g-1 of soil. Competition in case of rhizobia is mostly used to
refer to the competition for nodule occupancy (Giller 2001), which is a complex and
controversial area in the study of the legume-rhizobium symbiosis (Thies et al. 1992).
However, competition between inoculated and indigenous rhizobia is most strongly
influenced by the size of indigenous rhizobial populations whereas environmental factors did
not play a major role other than affecting the size of the native rhizobial population.
Decreasing nodule occupancy by inoculant strains was observed with increasing number of
indigenous rhizobia for lima bean and cowpea (Thies et al. 1992).
1.5.4. Soil temperature, pH and salinity

Temperature plays an important role in the success of BNF due to its effect on survival and or
growth rate of microorganisms. Both extremely high and low temperatures have either
depressive or killing effect. Maximum soil temperatures in tropics regularly exceed 40 C at
5cm and 50C at 1cm depth (Hungria and Vargas 2000). Similarly, the soil pH greatly
influences rhizobia content of soils and their ability to nodulate pulse crops (Slattery et al.
2004). Soil acidity reduces nitrogen fixation in legumes, particularly affecting Rhizobium
survival in soil and reducing nodulation. Nodulation of soya bean and haricot bean was
drastically reduced at pH of 4.5, whereas a pH of 5.2 resulted in good nodulation as well as
satisfactory N2 fixation (Hungria and Vargas 2000).
17

Moreover, production of grain legumes is severely reduced in salt affected soils mainly due
to the impairing effect of both salinity and sodicity on the plant ability to form and maintain
nitrogen fixing nodules (Rao et al. 2002). For chickpea, very small nodule dry mass was
recorded for all the genotypes tested under highly saline soil (Rao et al. 2002). With
increasing salinity, a sharp decrease in both nodule number and nodule biomass was observed
for all chickpea genotypes tested.
1.5.5. 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 be 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.
1.5.6. 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).

18

2. Materials and Method

2.1. Description of the study area and the prevailing farming system
The experiment was conducted on twenty farmers fields at Taba kebele1 close to Boditi town
of Wolaita zone, Southern Ethiopia, from September 2012 to January 2013. The study area is
located in Damot Galle, one of the 12 districts of Wolaita zone, which is found at about 350
km south-west of Addis Ababa, and 135 km from Hawassa, capital of Southern Nations,
Nationalities, and Peoples Regional State (SNNPRS) of Ethiopia. The area is situated at 07
00N and 37 54 E at an altitude of 1900 meters above sea level (masl).The soils are
dominantly Vertisols and Nitosols with pH values of 5-6.

Figure2:Mapofthestudyarea(Adaptedfrom(Jufare2008))

For the majority of the household in Wolaita agriculture is the main source of income with a
significant supplement from off-farm activities. Agriculture is dominated by subsistence
farming where limited usage of improved technologies and agricultural inputs significantly

1
NB: Kebele is Amharic term for Peasant association and it is similar to a village.

19

limits productivity and per capita income. A very high population density of up to 746
persons per square kilometre has reduced the average land holding of the area to about 0.25-1
ha per household (Jufare 2008). The farming system is characterized by small-scale mixed
production of crops and livestock (Eyasu 1998).
The area receives rainfall during a bimodal rainy season, which extends from March to
September with April, July, August and September receiving peak amounts of rain. Average
annual rainfall is 1200-1300 mm and the mean monthly temperature varies from 11 to 26C.
Meher is the main cropping season receiving rainfall from June to October whereas
Belg, the second cropping season, lasts from February to June. Almost all cereal crop
production of the area takes place during the Meher season. Cereals, root crops, pulses, and
spices are the main crop categories produced in the area. Due to the bimodal rainfall pattern,
resulting in a long growing season, multiple cropping practices such as intercropping, relay
cropping and double cropping are common.
In spite of its relatively green coverage, the region experiences frequent food shortage and is
known as the land of green famine (Elias and Scoones 1999). Currently, the Wolaita zone
represents one of the major food deficit and famine prone areas in Ethiopia (Jufare 2008).
Farmers consider soil fertility as the second most limiting factor for farming only next to
shortage of draught oxen (Elias 1998). The soils in many areas of Wolaita have been depleted
and degraded through continuous cultivation and the effects of leaching and erosion (Elias
and Scoones 1999), so that soil fertility has been declining over time. In addition to lack of
inorganic fertilizer, severe organic matter depletion, driven by competing uses for crop
residues and manure, plays an important role in the decline of soil fertility. Compared to
other regions of Ethiopia, the average livestock holding in this area of 1-2 heads per
household is small. This has changed over time as two decades ago an average household
used to keep 7 - 8 heads of cattle (Amede et al. 2001). Shortage of feed, conversion of
grazing areas to cropland, and diseases are some of the reasons for the decline of livestock.
Farmers of Wolaita area normally divide their land into several parts and use each part for
different purposes such as grazing, growing perennials like enset (false banana) and coffee,
and growing maize, sweet potato and other tuber crops. The homestead plots typically
contain crops such as enset, coffee, vegetables and also serves as seed bed to raise seedlings
of trees and pepper. Usually farmers prefer to apply manure and other organic wastes to the
fields in or closer to their homestead.
20

As a result, soils in the outer fields are less fertile and require mineral fertilizers to produce a
reasonable yield. Maize, tef, potatoes, haricot bean, sorghum and pepper are crops mostly
planted on the main/outer fields. Some plots are used for cut and carry fodder production for
livestock feeding.
Damot Galle in general and Taba kebele in particular are the densest areas in the region.
Unlike in previous times, the majority of farmers of this area are not food self-sufficient and
they attribute this to the declining rainfall and delay in onset of rainfall, sharp increase of
population size, and decreasing soil fertility. In this area, rain used to begin in December and
its amount was enough to produce crops twice a year. Nowadays rainfall starts as late as early
March and is received in a more erratic distribution. Consequently, the majority of the
household heads are forced to flee to adjacent towns to find job opportunities and send some
food to the family. Especially at the time of wheat harvest, which is around November, many
farmers travel to the Arsi area, where they are employed in harvesting activities.
Taba is one of the leading kebeles in SNNRPS with regard to total area allocated to chickpea
production and served as the centre for chickpea research in the region. Maize, sorghum, and
tef are the main cereal crops whereas chickpea and haricot bean are the most widely
cultivated pulse crops (Table1).
Table1.Listofannualcropsandtheircultivatedareaintheyear2011/12atTaba,excluding
perennialslikeensetandcoffee(Source:Tababureauofagricultureanddevelopment)
Crop
Areaofcultivation(ha)
Maize
175.5
Potato
100
Haricotbean
81
Tef
80
Chickpea
37
Sweetpotato
32
Pepper
25
Sorghum
10
Cassava
2
Abeshacabbage(localcabbage)
2
Boyya(rootcrop)
1.5
Total
546ha

Many of the farmers in this site grow chickpea at least on part of their farm. In Taba kebele,
chickpea is the fifth and second widely cultivated crop and food legume, respectively (Table
1). Chickpeas drought tolerance enables the farmers to prioritize it in rotation with cereals,
thereby practicing efficient utilization of the already scarce land.
21

Chickpea is used in many ways such as the direct consumption of its green pod and dry
beans, which are an important source of protein in the diet. Farmers also sell chickpea on the
local markets where it can fetch a good price.
However, due to the need for cash to buy food and other items, farmers are often forced to
sell most of the produced chickpea immediately after harvest, when prices are low. Also the
straw of chickpea is used as high quality livestock feed, rich in crude protein. Another crucial
reason why strengthening chickpea production in the area could be beneficial is related to its
ability to grow on residual moisture and produce something in seasons with insufficient
rainfall for more exigent crops, such as potato. Due to its lower input requirements than
cereals, its multi-functionality and flexibility, chickpea is considered an important crop for
smallholder, food-insecure farmers.
However, farmers often plant chickpea only on a small part of their land and leave the rest
unplanted, mainly because of the lack of chickpea seed and fertilizer at the time of planting.
Therefore, the provision of inputs at the time of planting would help in increasing
productivity and enabling efficient use of the scarce land.

2.2. Farm Selection

Damot Gale as a district and Taba as a kebele are among the leading chickpea growing areas
in SNNPRS, and were therefore chosen for this experiment. 20 farms located on two different
soil types were selected. To make the selection, we visited the farms in the area well ahead of
planting time to get insights on the farming system and soil differences. Farmers in the area
classify their soil based on colour and there were two types of soil in the kebele, which was
also confirmed by the Taba Office of agriculture and development. Farms found on the
uplands were characterized by black soil and towards the valley area the soil was red. Ten
farms, which frequently grow chickpea, were selected in each landscape position in
consultation with the local development agent, Mr. Mulatu Chafo. Care was taken to include
farmers from different wealth categories (poor, medium and rich). The Taba Office of
agriculture and development classifies farmers on the bases of assets like herd size and land
size. However, we were informed that rich farmers are hardly present in this site and poor
farmers are much more numerous.
To represent this distribution, three farmers were picked from the medium wealth group and
seven from the resource poor category for each landscape position and therefore soil type.
22

A clear discussion with the selected individuals on duties and responsibilities of the two
parties (researchers and farmers) was conducted and they agreed to provide 100 m2 area of
land for the experiment.

2.3. Estimation of indigenous rhizobial population of the study soils

For the purpose of determining the presence of native and viable rhizobia cells in the soils of
the study area, a test was conducted by implementing most probable number (MPN) method
(Somasegaran and Hoben 1985). For this purpose, soils were sampled two days ahead of
planting from the top 15 cm at five locations and bulked to form one composite sample per
farm. The samples were brought to the laboratory and stored in a fridge at 4C until the date
of inoculation (see next paragraph).
Chickpea, cv Natoli, was used as an indicator plant. Seeds of uniform size and high viability
were selected. The seeds were surface sterilized with ethanol [CH3CH2OH] for 5-8 minutes
and then transferred to Sodium hypochlorite for 10 seconds. Petri dishes were prepared and
their bottom covered with tissue paper, after which they were autoclaved and then used to
pre-germinate the seeds. The number of seeds was intentionally larger than required just in
case some failed to germinate. Accordingly, the required number of growth pouches (two for
each plant where the bottom one served for nutrient and water supply) were prepared. The
upper pouch was holed and cotton thread was inserted so that water and nutrients could be
taken up from the bottom pouch. The pouches were filled with sand, covered tightly with
aluminium foil and autoclaved at 121C for three hours. The pre-germinated seeds were
aseptically transferred into the growth pouches.
Meanwhile, any root or dirt material was removed from the soil samples being kept in the
fridge. A tenfold serial dilution was prepared by adding 10 gram of soil into 90 ml of distilled
water and sequentially diluting 1 in 10 to give a dilution series to 10-10. When all plants had
germinated, the pouches were inoculated with each of the ten serial dilutions from the soil by
using 1ml aliquots. The plants were frequently inspected and water and nutrients provided
periodically. When the plants were about four weeks old, they were carefully uprooted and
nodulated units were recorded. This was done for all the four replications and the noninoculated control.

23

2.4. Experimental details of field experiment

2.4.1. Crop cultivar

The experiment was carried out with chickpea, cv Natoli. While selecting the variety,
consideration was given to its market demand, farmers interest, and availability of seeds.
Accordingly, the improved chickpea variety Natoli was selected because it is among the five
top yielding chickpea varieties used by farmers of the study area and has a market potential
too.
2.4.2. Treatments and Layout

The experiment included four treatments that were applied on all farms. The four treatments
were control (un-inoculated plot), Rhizobium inoculated plot (I), non-inoculated but
phosphorus fertilized plot (P), and Rhizobium inoculated and phosphorus fertilized plot (I+P).
For the P and I+P treatments, Triple Super Phosphate (TSP), with its composition of 0-46-0
for NPK respectively, was used at a rate of 100 kg TSP ha-1 or 46 kg P ha-1.
On each of the 20 selected farms, a 100 m2 plot was divided in to four equal subplots with
gross and net size of 20.25 m2 and 17.55 m2 respectively. A distance of 1 metre was left
between each subplot to enable easy management and data collection. During planting, rows
were kept 30 cm apart and 10 cm intra row spacing was used. Each plot consisted of 15 rows
out of which the outer two rows were considered as border rows and each row consisted of 45
plants. Treatments were assigned randomly to the subplots. The experiment was not
replicated on each farm, but farms were considered as replicates.
2.4.3. Rhizobium preparation and inoculation

Mesorhizobium ciceri strain CP 41 was obtained from the soil microbiology laboratory of the
School of Plant and Horticultural Sciences, Hawassa University. This strain has been proven
to enhance the nodulation capacity of chickpea seeds under wide ecological conditions and is
considered the best strain of the laboratory as far as agronomic and yield performance of
chickpea is concerned.
Strain multiplication was undertaken prior to planting in a sufficient amount at this laboratory
and sterile peat was obtained from Legume Technology Ltd. in the UK. During both
inoculum preparation and inoculation, all the necessary aseptic measures were given due
attention. Accordingly, peat based inoculation was done at the recommended rate of 10g kg
seed-1.
24

Before planting, inoculation of the seeds was done on-farm using the peat as a carrier and
sugar as adhesive material to stick the inoculum on the seeds. In order to maintain viability of
the rhizobia, inoculation was done in the shade to avoid direct sun light. The inoculated seeds
were kept in the shade for a few minutes to let them air dry before planting. In all farms, noninoculated seeds were sown first and followed by the inoculated seeds to avoid cross
contamination. Moreover, measure was taken to divide the labour into two so that one group
planted the non-inoculated plots and the other group planted the inoculated plots.

2.5. Rainfall Data

With the aim of assessing the amount and variability of rainfall across the two landscape
positions (upland and valley), two ordinary rain gauges were manufactured at the local metal
workshop and used to record daily rainfall data.
The diameter of the rain collecting cylinder was 18 cm whereas the above ground height of
the rain gauge was 1.5 m. The rain gauge was fixed to be 100% level. On each soil type, one
household with an individual capable of writing and reading figures was selected. The rain
gauges were fixed at a convenient position near to the houses, but care was taken to keep
them away from tall objects. Accordingly, the daily rainfall was recorded for each soil type
throughout the whole growing period.

2.6. Soil sampling and analysis

Before planting, soil samples were taken from each subplot to a depth of 0-30 cm using an
auger. Samples from the four subplots of each farm were bulked together and a composite
sample was prepared for physico-chemical analysis. Soils were air dried to a constant weight,
grinded and mixed thoroughly and passed through a 2 mm sieve.
In the laboratory, the soils were analysed for particle size, total N, available P, organic
carbon, pH, CEC, Ca, Mg, K, and Na. Standard procedures were followed for each
parameter. CEC was determined using the ammonium distillation method whereas pH was
measured with the potentiometric water extraction method. Available P was measured by the
Olsen et al. (1954) method and for the determination of the total N, the Kjeldahl (1883)
method was employed. Textural class and organic carbon content of the soil were determined
using hydrometer and the wet combustion method of Walkley and Black (1934), respectively.
Determination of exchangeable Na and K was carried out using ammonium acetate extraction
and flame photometry after wet combustion.
25

For the determination of exchangeable Ca and Mg, ammonium acetate extraction was
followed by flame atomic absorption spectrometry (AAS). The conductometric method was
used for measuring electrical conductivity.

2.7. Plant tissue analysis

Doing the plant analysis for all the treatments on 20 farms was costly and a solution was
found by randomly selecting three farms for each soil type and five plants per treatment from
the middle rows. The plants were sampled at the time of harvest and separated into grain and
straw.
The straw part was oven dried at 70C for 48 hours, milled with mechanical miller and
passed through a 1mm sieve. Subsequently, the plant samples were analysed for total N, P
and ash content.
The straw total N uptake was first calculated as a product of the straw yield and its N content
obtained from the lab analysis. Grain N content was assumed to be about twice of that of the
straw (Kassa 2009). Hence, this grain N content was multiplied by the grain yield to give
grain total N uptake. Consequently, total N uptake was calculated as a summation of straw
and grain total N uptake.

2.8. Management practices

In the preceding season, five types of crops had been grown on the experimental fields of the
twenty farms. Haricot bean, potato, maize, tef and sweet potato were grown on 55%, 15%,
15%, 10% and 5% of the fields respectively. Before planting, all fields were thoroughly
ploughed and levelled by the owner of the land. Accordingly, planting started on September 6
and was completed within three consecutive days for 18 farms whereas two farms (farms 19
and 20) were planted on September 15. In all cases, there was frequent rainfall during the
early plant stages, which guaranteed good initial plant establishment. All fields were weeded
twice during the growing period. During our frequent field supervision, no severe disease
symptoms were observed on the farms. However, on several farms there was occurrence of
boll worm (Helicoverpa) during flowering and pod setting. As a control measure, Lambdacyhalothrin chemical was sprayed for each plot. Nevertheless, yield loss due to this problem
was inevitable as some pods were holed and devoid of seeds during harvesting. Harvesting
started around mid-December and finished on January 19 as the farms differed with respect to
time to maturity.
26

2.9. Observations and Data collection

2.9.1. Crop phenology

Days to emergence was recorded for each plot when more than 50% of the plants emerged.
Around three weeks after planting, the total number of plants emerged was counted from the
middle 13 rows to conduct a stand count at emergence. Similarly, days to flowering and
maturity were recorded when more than 50% of the plants in each plot attained flowering and
physiological maturity respectively. At the time of flowering, six plants were randomly
selected from the 13 middle rows and their height from the ground to the tip measured using
a ruler. From the six randomly uprooted sample plants at maturity (see 2.9.4), the number of
branches (both primary and secondary) was counted and averaged to give number of
branches plant-1 (NBPP).
2.9.2. Nodulation

Nodulation assessment was conducted at three physiological stages. For all treatments, at 23
days after sowing (DAS), six plants were selected from the two border rows, 3 from each,
and gently uprooted. The root was washed with tap water to remove the adhering soil.
The number of nodules per plant was counted and the values averaged to give the number of
nodules per plant (NNPP). The same plants were used for nodule scoring per plant (NSPP) on
0-5 scale according to a protocol developed by Bala et al. (2010) where plants with nodule
number of zero, <5, 5-10, 11-20, 21-50, >50 were scored 0,1,2,3,4 and 5 respectively. These
scores refer to absent, rare, few, moderate, abundant and super nodulation of roots in
ascending order. At 45 DAS, which is around flowering time, again six plants were randomly
selected and uprooted from the interior rows and these plants were separated in to shoot and
root and NNPP and NSPP were recorded. In addition, the nodules were severed from the
roots, oven dried at 70C for 48 hours and their dry weight recorded to give nodule dry
weight per plant (NDWPP) at 45 DAS. At harvesting time, six plants were randomly marked,
watered a day ahead, and gently uprooted the next day. The roots of these plants were washed
with tap water and after removing all the adhering soil and debris, NNPP and NSPP were
recorded.
2.9.3. Shoot weight

The shoot part of the plants that were sampled at 45 DAS were cleaned and weighed, and the
values were averaged to constitute shoot fresh weight per plant. The same plants were oven
27

dried at 70C for 48 hours to a constant weight and weighed again to determine dry shoot
weight per plant.
2.9.4. Grain yield, yield related traits and total biomass production

Just before harvesting, the total number of plants was counted from the interior 13 rows of
each plot to give stand count at harvest. During harvesting time, data was recorded for yield
attributing parameters such as number of pods per plant, number of seeds per pod, and
thousand seed weight for each plot. For this purpose, six plants were randomly selected from
the interior 13 rows and dug up.
All of the plants, apart from the two external rows, were manually harvested and brought
together on a plastic sheet for each plot and measured to give weight of total biomass per
plot. Then the plants were threshed separately for each plot and the seeds blown to air to
separate seeds from debris/husks. The grain was put in a cloth sack and weighed to give yield
per plot. On the same date, seeds were sampled and taken to the laboratory to measure their
moisture content using a seed moisture meter (Model HOH- Express He 50). To keep
uniformity, the final grain yield of each plot was adjusted to 12% seed moisture content.
2.9.5. Data on household characteristics

After harvest, farmers were interviewed to get their opinion about the treatments as well as their
household characteristics. For this purpose, a questionnaire or field evaluation book designed by
the N2Africa project was used and farmers were interviewed at their homestead. Data was
collected of farmers household characteristics, previous management practices and their
perception on fertility of their farm. The household level data were explored to explain the
differences in treatment performance between farms.

2.10. Statistical analysis

After checking the compliance of the data with the assumptions of the statistical test, analysis
of variance was conducted using a split plot design where soil type was applied to the whole
plot and the four treatments were applied to the subplot. Each farm was considered as a
block. A threshold P value of <0.05 was used to declare effects and interactions to be
significant. When the effects were found to be significant (P<0.05), a LSD test ( = 0.05) was
used to study which means differed significantly and the comparison was made by employing
Duncan multiple range test (DMRT). In addition, the relationship among the studied
parameters was assessed using Pearsons simple correlation analysis.
28

3. Results
3.1. Soil characteristics of the study farms
3.1.1. Physicochemical properties

The soil textural class of fourteen of the experimental fields was loam with average
proportions of 24% clay, 40% silt and 36% sand. Of the remaining fields, soil texture was silt
loam, clay and clay loam on two, one and three farms respectively (Appendix 1). This result
is in agreement with that of Kassa (2009) who reported the soil texture of Taba kebele to be
loam. The pH value of the soil ranged from 5.8 to 7.3. Of the twenty soil samples, pH values
of soils from fourteen farms ranged between 6 and 7. Generally, the average pH value was
6.5 which is only slightly acidic and within the optimum range for crop production (Havlin et
al. 1999), as well as for chickpea infective strains of rhizobia (Rodrigues et al. 2006).
According to Havlin et al. (1999), total nitrogen content (TN) of a soil can be classified as
very low (<0.1%), low (0.1-0.15%), medium (0.15-0.25%), and high (>0.25%). According to
this classification, the total nitrogen content of the soils from the study farms was found to be
very low, low, and medium for 6, 12 and 2 farms respectively. The average total nitrogen for
the twenty farms was 0.11%, which is within the bottom range of the low total nitrogen class.
Indicative ranges of available phosphorus have been established by Olsen et al. (1954),
including < 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). Based on this criterion, the available phosphorus content was found to be
very low, low, medium and high for 1, 9, 6 and 4 farms respectively. As far as soil organic
carbon (OC) is concerned, the value ranged from 0.55 to 1.62%, which is within the range of
low organic carbon content (Landon 1991). The cation exchange capacity (CEC) is referred
to be low (5-15 cmol kg-1), medium (15-25 cmol kg-1), or high (25-40 cmol kg-1). For this
parameter, soils of nineteen farms had a medium CEC and only one farm was characterized
by low CEC.
The two soil types showed differences in their initial content of major nutrients and
characteristics, with the black soil somewhat more fertile than the red soil. The black soil
contained significantly (P=0.013) higher TN (0.12 %) and highly significantly (P<0.001)
higher OC (1.4 %) than the red soil with TN 0.091% and OC 0.96%. For P, CEC and pH
values no significant difference was observed between the soil types.

29

Nonetheless, the black soil contained higher mean P (19 mg kg-1) and CEC (17.9 cmol kg-1)
than the red soil whose mean P and CEC were 13 mg kg-1 and 17.5 cmol kg-1 respectively.
Opposite to this, the pH was higher for the red soil (6.7) than for the black soil (6.4).
Generally, the soils of the experimental fields were deficient in total nitrogen content,
available phosphorus, and organic carbon, whereas for almost all farms, the CEC was in the
medium category (Appendix 1). Since these are the major elements of soil fertility, the result
indicates poor fertility status of the soil that would in turn limit crops to achieve their
potential productivity.
3.1.2. Native rhizobia populations
The MPN test revealed that the population of indigenous rhizobia of the soils in the study
area was very low, ranging from none to <10 rhizobia cells gram-1 of soil. In 50% of the soil
samples tested, there was no viable and infective rhizobia recovered and in the remaining
50% of the samples, the rhizobia population was <10 rhizobia cell gram-1 of soil (Table 2).
This indicates that the population is not abundant enough to initiate optimum nodulation. In
75% of the farms on black soils, there was no measurable population of rhizobia, whereas on
the red soil type, 25% of the farms had no measurable rhizobia.
Table 2. Most probable number test result of soil samples for some farms of the study area
(n=8)
Soil type
Farm number
rhizobia population g-1 of soil
Black
2
none
Black
7
none
Black
19
none
Black
20
< 10
Red
4
< 10
Red
9
none
Red
13
< 10
17
< 10
Red

30

3.2. Rainfall data

212 and 214 mm of rainfall was received during the whole growing period (September to
January) on black and red soils respectively (Appendix 2). In both cases, the highest amount
of rain (100 mm) was recorded in the month of September. Generally, rainfall amount
declined from planting time to harvest with less than 5mm in December (Appendix 2). Both
the daily and cumulative rainfall remained almost similar for the two soil types (Fig. 3).

Cummulativerainfall(mm)

250

200
BS

150

RS

100

50

Dateofrainfall
Figure3:Cumulativerainfallofthetwosoiltypesforthegrowingperiod.BS=blacksoil,RS=redsoil.

31

3.3. Effects of soil types on nodulation, growth, pod production and days to
maturity of chickpea (Cicer arietinum L.)
The effect of soil type was studied for all the variables investigated in this study. However,
significant effects of soil type were only observed for the six variables presented in Table 3.
At 23 DAS a higher mean nodule number plant-1 was produced by plants grown on red soil
(26) than plants grown on black soil (19) (Table 3). Similarly, nodule score plant-1 at 23 DAS
was significantly (P=0.036) higher for plants raised on red soil with mean value of 3.6
whereas mean NSPP was 3.1 for plants on black soil type. Chickpea plants grown on black
soil were significantly (P<0.001) taller (45 cm) than those on red soil (38 cm) (Table 3).
Likewise, a significant (P=0.015) effect of soil type was observed for number of branches
plant-1 (NBPP). A higher NBPP was recorded for plants on black soil (11) than on of red soil
(9) (Table 3). In addition, a significantly (P=0.018) higher number of pods plant-1 (NPPP)
was produced by plants raised on black soil (56 pods) compared with plants on red soil (44
pods). The effect of soil type was also significant (P=0.046) for days to maturity with plants
on red soil requiring 98 days, whereas 108 days were required for plants on black soil to
reach maturity (Table 3).
Table 3. Effects of soil types on nodulation, growth, pod production and days to maturity of
chickpea(CicerarietinumL.)(n=40)
NBPP
NPPP
Daysto
Soiltype
NNPPat
NSPPat
Plant
maturity
23DAS
23DAS
height
(cm)
a
a
Blacksoil
19
3.1
46b
11b
56b
108b
Redsoil
26b
3.6b
38a
9a
44a
98a
Notes: NNPP = nodule number per plant, DAS = days after sowing, NSPP = nodule score per plant, NBPP =
number of branches per plant, NPPP = number of pods per plant. Means followed by the same letter(s) with
in a column are not significantly different at P = 0.05 (Duncans Multiple Range Test).

32

3.4. Effects of Rhizobium inoculation and / or Phosphorus fertilization on

nodulation of chickpea (Cicer arietinum L.)
3.4.1. Nodule number plant 1

The assessment of nodule number plant-1 (NNPP) was carried out at three different
physiological stages of the crop and except for NNPP at 23DAS, a significant and positive
effect of the soil fertility treatments was observed. At the early crop stages, nodule number
plant-1 of chickpea was improved only by combined application of inoculation and P
fertilizer. In later stages, however, application of all the three soil fertility treatments resulted
in increased NNPP. At all the three stages, maximum improvement of NNPP was achieved
by the combined application of inoculation and P fertilizer. In addition, NNPP of chickpea
was highest at 45 DAS and lowest at maturity (Fig. 4).
Similarly, a highly significant (P<0.001) effect of the soil fertility treatments was observed
for NNPP at 45 DAS. For this parameter, the maximum nodule number (48) was obtained
from application of the I+P treatment which was 138% higher than the NNPP of the control
treatment. Also at maturity the effect of the soil fertility treatments on NNPP remained highly
significant (P<0.001), with the maximum NNPP (13) obtained from the application of I+P
(Fig. 4).

33

50

+I

Numberofnodulesplant1

c
b

40

+P
+P+I

30

ab

20

b
a

10

0
23DAS

45DAS
Timeofnodulecount

Maturity

Figure4:Nodulenumbersplant1atthreephysiologicalstages(DAS=daysaftersowing)forfourtreatments(C=control,
+I = inoculation, +P = phosphorus fertilized, +I+P = both inoculation and phosphorus application). Treatments are
significantlydifferentiftheyhavenoletterincommonP=0.03at23DAS,P<0.001at45DASandmaturity.

3.4.2. Nodule score plant1

Nodule score plant-1 (NSPP) of chickpea followed an almost identical trend with NNPP and
was enhanced by application of inoculation and/ or P fertilization. At 23 DAS NSPP was
significantly (P=0.026) improved only by the combined application of inoculation and P,
resulting in the maximum (3.6) NSPP. On the other hand, NSPP at 45 DAS was observed to
be significantly (P<0.001) and positively enhanced by all the treatments, with a more
pronounced effect when inoculation and P were applied in combination (Table 4). The effect
of the treatments on NSPP remained highly significant (P<.001) at maturity. Accordingly, the
highest (2.7) NSPP was recorded for plants that received the treatment I + P. There was no
significant difference between inoculation and P treatments, but both gave higher NSPP than
the control treatment (Table 4).
3.4.3. Nodule dry weight plant 1 at 45 DAS, (NDWPP at 45DAS)

The application of inoculation and / or P significantly (P<.001) enhanced nodule dry weight
plant-1 of chickpea at 45 days after sowing Although all the three treatments gave
significantly higher NDWPP over the control treatment, the maximum value (78 mg plant-1)
was recorded from plants that received the I+P treatment (Table 4).

34

There was no significant difference between inoculation and P fertilization, but both gave
higher NDWPP than the control (55 mg plant-1) (Table 4).
Table4.EffectsofRhizobiuminoculationand/orphosphorusfertilizationonnodulationofchickpea
(CicerarietinumL.)(n=20)
Treatments
NDWPPat45DAS NSPPat23DAS
NSPPat45DAS
NSPPatMaturity
mg/plant
Control
55a
3.3a
3.4a
2.3a
b
ab
c
Inoculation
70
3.4
4.3
2.6b
3.3a
4.1b
2.6b
Phosphorus
65b
c
b
d
Inoculation+P
78
3.6
4.5
2.7c
Notes: DAS = days after sowing, NDWPP = nodule dry weight per plant in milligram, NSPP = nodule score per
plant. Means followed by the same letter(s) with in a column are not significantly different at P = 0.05 (Duncans
Multiple Range Test).

35

3.5. Effects of Rhizobium inoculation and / or phosphorus fertilization on

phenological and agronomic traits of chickpea (Cicer arietinum L.)
3.5.1. Shoot fresh weight plant 1

Shoot development of chickpea was enhanced by P fertilization and Rhizobium inoculation

with an even more pronounced improvement under combined application of inoculation and
P (I+P). Though no statistically significant difference was observed between inoculation and
P, and between I+P and P treatments, separate application of inoculation and P resulted in
significantly higher SFWPP than the control treatment (Table 5). The maximum SFWPP (9.0
g plant-1) was recorded from plants that received a treatment in which inoculation and P were
combined.
3.5.2. Days to maturity

Days to maturity was significantly (P<0.001) extended by the applied soil fertility treatments
with the shortest maturity time observed from the control treatment (Table 5). Inoculation,
both alone or in combination with P, resulted in the longest (104 days) growing period of
chickpea. Likewise, a separate application of P also exerted a highly significant effect on
days to maturity with mean maturity days of 103, which was significantly shorter than that of
I+P and I treatments but longer than that of the control treatment (Table 5). Nevertheless,
days to maturity did not change considerably with the fertility treatments, ranging only from
101 to 104 days.
3.5.3. Number of branches plant 1

Number of branches plant-1 (NBPP) of chickpea increased significantly (P<.001) due to the
applied soil fertility treatments. The maximum NBPP (11) was produced by plants that
received the I+P treatment whereas the plants in the control produced the lowest number of
branches (8). Likewise, compared to the control higher NBPP was produced by plants that
received a separate application of inoculation and P (Table 5). However, non-significant
differences in NBPP between inoculated and P fertilized plants, as well as between I+P and P
treated plants were observed (Table 5).
3.5.4. Plant height (cm)

The application of inoculation and / or phosphorus resulted in significantly (P<.001) taller

chickpea plants over the control treatment.

36

Though all the treatment effects remained on par, plant height was significantly improved as
compared to the control, with the combined application of inoculation and P resulting in the
tallest plants (43 cm) (Table 5).
Table 5. Effects of Rhizobium inoculation and / or phosphorus fertilization on phenological and
morphologicalcharacteristicsofChickpea(CicerarietinumL.)(n=20)
Treatments
SFWPPat45DAS Daystomaturity NBPP
Plantheight
(g/plant)
(cm)
a
a
a
Control
7.0
101
8
40a
Inoculation
8.4b
104c
10b
43b
Phosphorus
8.5bc
103b
10bc
42b
Inoculation+P
9.0c
104c
11c
43b
Notes: SFWPP = shoot fresh weight per plant, DAS = days after sowing, NBPP = number of branches per
plant. Means followed by the same letter(s) with in a column are not significantly different at P = 0.05
(Duncans Multiple Range Test).

37

3.6. Effects of Rhizobium inoculation and / or Phosphorus fertilization on

yield and yield related traits of chickpea (Cicer arietinum L.)
3.6.1. Number of pods plant1 (NPPP) and number of seeds pod1 (NSPP)

Inoculation and/ or P significantly increased the number of pods plant-1 compared to the
control treatment. Accordingly, the lowest NPPP (45) was recorded in the control treatment
whereas the highest (54) NPPP was recorded for plants receiving I+P treatment followed by
51 and 50 NPPP for the inoculation and P treatments respectively (Table 6).
Similarly, inoculation, either alone or in combination with P, significantly (P=0.006)
increased number of seeds pod-1 (NoSPP). Nonetheless, a separate application of P gave
NoSPP which was statistically not different to that of the control. The maximum mean
NoSPP (1.6) was recorded from the inoculated plants whereas the lowest mean (1.4) NoSPP
was recorded from the plants in the control treatment (Table 6).
3.6.2. Total biomass production (ton ha1)

Analysis of variance indicated that total biomass of chickpea was significantly (P<.001)
improved by inoculation and / or phosphorus fertilization treatments. Application of each of
the fertility treatments had a positive and highly significant effect on total biomass of
chickpea as compared to the control. Consequently, the maximum mean total biomass (4.5
ton ha-1) resulted from application of the I+P treatment, which was followed by 4.3 ton ha-1
from the inoculation treatment. The lowest mean total biomass (3.5 ton ha-1) was recorded
from the control treatment (Table 6). For this parameter, there were no statistically significant
differences observed between inoculation and P treatments, and between sole P and I+P
treatments.
3.6.3. Grain yield (ton ha1)

Analysis of variance revealed a highly significant (P<.001) effect of the treatments on the
grain yield of chickpea. The maximum mean grain yield (2.1 ton ha -1) was obtained from the
I+P treatment which, in fact, was statistically on par with that of the inoculation treatment
(2.0 ton ha-1) and slightly greater than the treatment with P only (Table 6).
3.6.4. Thousand Seeds weight (gram)

The weight of thousand seeds of chickpea responded positively and significantly (P=0.002)
to the application of the fertility treatments with heavier seeds compared with the control.
38

The maximum mean weight of thousand seeds was recorded in the I+P treatment (308 g),
whose effect was statistically on par with that of sole P (307 g) which, in turn, remained
statistically similar to that of sole inoculation (294 g) (Table 6).
Table 6. Effects of Rhizobium inoculation and / or phosphorus fertilization on yield and yield
relatedtraitsofChickpea(CicerarietinumL.)(n=20)
Treatments
NPPP
NSPP
Totalbiomass Grainyield
1000SW
1
1
(tonha )
(tonha )
(g)
a
a
a
a
Control
45
1.4
3.5
1.6
285a
Inoculation
51bc
1.6b
4.3bc
2.0bc
294ab
Phosphorus
50b
1.5ab
4.0b
1.9b
307bc
Inoculation+P 54c
1.6b
4.5c
2.1c
308c
Notes: NPPP = number of pods per plant, NSPP = number of seeds per pod, 1000 SW= thousand seeds
weight. Means followed by the same letter(s) with in a column are not significantly different at P= 0.05
(Duncans Multiple Range Test).

39

3.7. Effects of Rhizobium inoculation and/ or phosphorus fertilization on

straw total nitrogen (STN, %) and phosphorus content (STP, mg kg1) and
total N uptake of chickpea
The application of all three fertility treatments significantly (P <0.001) improved the total
nitrogen content of chickpea straw over that of the control treatment. STN of the inoculation
treatment (0.84%) remained statistically on par with the STN of both the P fertilization
treatment (0.72%) and the I+P treatment (0.98%) (Table 7). Furthermore, inoculation, either
alone or in combination with P increased the total N uptake compared with the control. The
least total N uptake (28 kg ha-1) was recorded from the control treatment whereas the
maximum total N uptake (65 kg ha-1) was recorded from the I+P treatment that was
statistically alike with the total N uptake of the I treatment (53 kg ha-1) (Table 7).
Similarly, also for total phosphorus content of chickpea straw the effect of the treatments was
highly significant (P< 0.001) All the fertility treatments, except when only inoculation is
used, were able to improve the STP over the control treatment. The highest STP (367 mg kg1

) was recorded from plants receiving P fertilization, followed by that of I+P treatment (334

mg kg-1), with which it was statistically on par (Table 7).

Table 7. Effects of Rhizobium inoculation and/or phosphorus fertilization on straw total
nitrogen and phosphorus content, and total N uptake of chickpea (CicerarietinumL.)(n=6)
Total N uptake
Treatments
STN %
(kg ha-1)
STP (mg kg -1)
a
28
Control
0.54a
261a
b
bc
53
Inoculation
0.84
273a
42ab
Phosphorus
0.72b
367 b
65b
Inoculation + Phosphorus
0.98c
334 b
Notes: STN % = straw total nitrogen %, STP = straw total phosphorus (mg kg -1). Means
followed by the same letter(s) with in a column are not significantly different at P = 0.05
(Duncans Multiple Range Test).

40

3.8. Interaction effects between soil fertility treatments and soil types on
growth and performance of chickpea
Interaction effects of Rhizobium inoculation and/or P fertilization and the soil types were
studied for all the variables investigated. However, the analysis of variance showed that only
three variables among all were significantly influenced by the interaction of the soil fertility
treatments and the soil types. Only these three variables (NNPP at 45 DAS, NSPP at 45 DAS
and days to maturity) are presented in Table 8 and the below text.
3.8.1. Nodule number and score plant1 at 45 days after sowing (45 DAS)

There was a highly significant (P<0.001) effect exerted on NNPP at 45 DAS by the
interaction between the treatments and the two soil types. There was large variation observed
among various combinations of factors for this variable. The number of nodules per plant
ranged from 20 in black soil of the control treatment to 52 in red soil of the I+P treatment
(Table 8). For control and P fertilization treatments, NNPP remained alike in both soil types.
However, for inoculation and I+P treatments nodule numbers were higher in the red soil.
The interaction effect between the treatments and the soil types was also significant
(P=0.003) for nodule score plant-1 studied at 45 days after sowing. In general, the highest
(4.6) score was recorded for plants received I+P treatment on red soil whereas the lowest
(3.4) score was recorded from control treatment under both soil types (Table 8).
3.8.2. Days to maturity

The interaction between soil types and the treatments had a significant (P=0.007) effect on
days to maturity of chickpea. With the inoculation and control treatments, chickpea had a
shorted maturity period when grown on red soil, whereas no effect of soil type was observed
for the sole P and I+P treatments. Of all treatment combinations, the shortest (96 days)
maturity period was recorded for the control treatment on the red soil type. In contrast to this,
application of I+P on the black soil resulted in the longest (109 days) growing period.
Generally, the soil fertility treatments advanced the maturity of chickpea on the black soil
whereas the treatments delayed maturity on the red soil (Table 8).

41


Table8.Effectsofinteractionbetweensoilfertilitytreatmentsandsoiltypeonnodulationand
daystomaturityofchickpea(CicerarietinumL)(n=20)
Treatments
Control
Inoculation
Phosphorus
Inoc+P

Soiltype
Black
Red
Black
Red
Black
Red
Black
Red

NNPPat45DAS
20a
20a
39b
48cd
40b
39b
43bc
52d

NSPPat45DAS
3.4a
3.4a
4.2bc
4.4cd
4.2bc
4.1b
4.3bc
4.6d

Daystomaturity
110d
96a
105c
98ab
108bcd
99ab
109cd
99abc

Notes: NNPP = nodule number per plant, DAS = days after sowing, NSPP = nodule score per plant. Means
followed by the same letter(s) with in a column are not significantly different at P = 0.05 (Duncans
Multiple Range Test).

42

3.9. Correlations among the symbiotic and agronomic variables of chickpea

Positive and significant correlation among related traits was found whereas the opposite was
observed between divergent traits. Grain yield of chickpea was positively and significantly
correlated with number of seeds pod-1 (r = 0.358***), number of pods plant-1 (r = 0.418***),
number of nodules plant-1 at 45 DAS (r = 0.308**), days to maturity (r = 0.322**), plant
height (r = 0.397***) and total biomass (r = 0.855***) (Appendix 8). Similarly, positive and
significant correlation was observed for total biomass production with NNPP at 45 DAS (r =
0.273*), initial soil organic carbon (r = 0.229*), initial soil available P (r = 0.305**), plant
height (r = 0.351**), number of branches plant-1 (r = 0.342**) and shoot fresh weight at 45
DAS (r = 0.253*) (Appendix 8). On the other hand, negative correlation (r = -0.295**) was
observed between the number of nodule plant-1 at 23 DAS. Similarly, days to maturity was
negatively correlated with NNPP at 23 DAS (r = -0.306**), stand count at harvest (r = 0.363***), shoot fresh weight plant-1 at 45 DAS (r = -0.554***), and shoot dry weight plant-1 at
45 DAS (r = -0.576***) (Appendix 8).

43

3.10. Across farm variability in response to the soil fertility treatments

3.10.1. Yield and total biomass production

Across the twenty farms, large variability in growth and stand of chickpea plants was
observed for each treatment (Fig. 6). Nevertheless, a positive response to the soil fertility
treatments was recorded on more than 85% of the farms, the least yield being recorded from
the control treatment. Overall, the yield ranged from 0.5 ton ha-1 on a control plot to 3.0 ton
ha-1 on a P treated plot (Fig. 5A). Regardless of the huge variability, on the majority of farms
yield response was most pronounced under I+P treatment application. For all four treatments
the lowest yield, including a weak response, was produced on farm 20 and this might be
related to poorer plant establishment (16%, 14%, 17% and 22% plant emergence recorded for
C, I, P and I+P treatments respectively) due to waterlogging, which also affected other crops
on this farm.
Almost similar response to the soil fertility treatments was recorded for the total chickpea
biomass. A positive response of the soil fertility treatments was recorded on more than 70%
of the farms, in which the control treatment produced the lowest total biomass and the I+P
treatment the highest total biomass. As such, the total biomass production ranged from 1.4
ton ha-1 on a control plot to 7.1 ton ha-1 on a plot that received the I+P treatment (Fig. 5B).
For both yield and biomass, positive responses were observed across the range in control
yields/biomass (Figs. 5A and 5B).
8

A
TBMinPand/orItreatments(tonha1)

YieldinPand/orItreatments(ton ha1)

3.5
3.0
2.5
2.0
1.5
+I
1.0

+P

0.5

+P+I

0.0
0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

7
6
5
4
3
2

+I
+P
+P+I

1
0
0
2
4
6
TBMincontroltreatment(tonha1)

Yieldincontroltreatment(tonha1)

Figure 5: Response of chickpea grain yield (A) and total biomass (B) (ton ha1) to inoculation and/or P treatments.
Notes:+I=inoculation,+P=phosphorusfertilized,I+P=bothinoculatedandPfertilized,TBM=totalbiomass.

44

Figure6:Betweenfarmdifferenceofchickpeagrowthinresponsetothetreatments

3.10.2. Nodule number plant1 and shoot fresh weight plant1 at 45 DAS

The trend of response and variability was almost alike for most of the variables studied. Here
only NNPP at 45 DAS and SFWPP are presented, as they are clearly linked to yield and give
an insight of the variability in almost all other parameters. Both nodule number plant-1
(NNPP) at 45 DAS and SFWPP positively responded to the P and/or I treatments on almost
all farms studied (Figs 7A and 7B). On the majority of farms, the lowest NNPP was recorded
for the plants from the control treatment whereas the highest NNPP was counted from the
plants receiving the I+P treatment. Overall, the NNPP ranged from 13 in the control
treatment of farm 20 to 57 in the I+P treatment of farm 10.
The SFWPP response to the treatments also exhibited large across farm variability. For the
control treatment SFWPP ranged from 2.6 g plant-1 to 14.7 g plant-1 on farm 17 and 9
respectively whereas for I+P treatment alone the SFWPP ranged from 4 g plant-1 to 19 g
plant-1 on farm 4 and 9 respectively. Nonetheless, for the majority of the farms the lowest
SFWPP was recorded on the control (Fig. 7B).

45

Similar to yield and biomass, positive responses were observed across the range in control
NNPP and SFWPP (Figs 7A and 7B), with still moderate across farm variability in response
to the treatments.

25

SFWPP(gp1)inIand/orPtreatments

NNPPat45DASinPand/Itreatments

60
50
40
+I

30

+P
+P+I

20
10

B
20

15
+I
+P

10

+P+I

0
0

20
40
NNPPat45DASinthecontrol

60

5
10
15
20
SFWPP(gp1)inthecontrol

25

Figure7:Responsetoinoculationand/orphosphorusfertilizationofnodulenumberplant1(A)andshootfreshweightplant1 (g
1
plant )(B)at45daysaftersowingofchickpea.Notes:+I=inoculation,+P=phosphorusfertilized,+I+P=bothinoculationand
phosphorusadded,DAS=daysaftersowing,NNPP=nodulenumbersplant1,SFWPP=shootfreshweightplant1.

3.11. Across farm variability in nodulation, biomass, yield and their

response to the soil fertility treatments under the two soil types
3.11.1. Nodule number plant1
For both soil types, moderate across farm variability was exhibited for NNPP at 45 DAS.

However, the variability appears to be smaller on the red soil than the black soil (Figs 8A and
8B).

46

50
40
+I

30

+P
+P+I

20
10

50
40
30

+I
+P

20

+P+I
10
0

0
0

60

A
NNPPinPand/orItreatments

NNPPinPand/orItreatments

60

20
40
NNPPat45DASinthecontrol

60

20
40
NNPPat45DASinthecontrol

60

Figure 8: Nodule number plant1 at 45 days after sowing response of chickpea to inoculation and/or phosphorus
1
fertilization
on the black soil (A) and on the red soil (B). NNPP = nodule number plant , DAS = days after sowing, I =

inoculation,P=phosphorusfertilization,I+P=inoculationandphosphorusfertilization.

3.11.2. Yield and biomass

Moderate to large across farm variability in yield and biomass production of the treatments
occurred under both soil types. Nonetheless, both soil types resulted in positive response to
the soil treatments. For both yield and biomass, the variability was smaller on the red soil
(Figs 9C and 9D) than on the black soil (Figs 9A and 9B).

47


3.5

3.5

YieldinPand/orI(t/ha)

3.0
YieldinPand/orI(t/ha)

2.5
2.0
+I
1.5

+P
+P+I

1.0
0.5

2.5
2

+I
+P

1.5

+P+I

1
0.5

0.0

0
0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

B
BiomassinPand/orI(t/ha)

BiomassyieldinPand/orI(t/ha)

7
6
5
4

+I

+P
+P+I

0.5

1.5

2.5

3.5

Yieldinthecontrol(t/ha)

Yieldinthecontrol(t/ha)

7
6
5
4

+I

+P

+P+I

0
0

Biomassinthecontrol(t/ha)

Biomassinthecontrol(t/ha)

Figure9:Yieldandbiomassresponseofchickpeatoinoculationand/orPfertilizationontheblacksoil(A&B)andthe
redsoil(C&D).+I=inoculation,+P=phosphorusfertilized,andP+I=bothPfertilizationandinoculation.

3.11.3. Yield response to the soil fertility treatments of the two soil types

Although positive response to the soil fertility treatments was recorded on both soil types,
larger magnitude of response to the applied soil fertility treatments was recorded on the red
than the black soil. Application on the red soil of I, P, and I+P treatments resulted in 0.27 ton
ha-1, 0.21 ton ha-1 and 0.2 ton ha-1 more average grain yield than their application on the black
soil respectively (Fig 10).

48

0.7

IC

PC

(I+P)C

Yieldresponse(tonha1)

0.6

0.6
0.5

0.5
0.4

0.4

0.4
0.3

0.3

0.2

0.2

0.1
0.0
Blacksoil

Soiltype

Redsoil

Figure10:Yieldresponsetoinoculationand/orphosphorusfertilizationofchickpeaonblackandredsoil
types.IC=yieldresponseduetoinoculation(I),PC=yieldresponseduetophosphorusfertilization(P),
(I+P)C=yieldresponseduetoapplicationofbothinoculationandphosphorusfertilization(I+P),C=yield
inthecontroltreatment.

3.12. The effect of initial soil fertility on yield and magnitude of yield
response to the soil fertility treatments
A highly significant and positive correlation was found only between the available P content
of the soil and yield in +I (r = 0.874) and I+P (r = 0.882) treatments (Appendix 9).
Nonetheless, for the control and I+P treatments yield was observed to increase with increase
in OC%, TN%, and Av. P content of the soil (Fig. 11). The yield increment seems to be more
pronounced under the control treatment compared to the I+P treatment (Fig. 11).
For the magnitude of the yield response (I-C, I-P, and (I+P)-C) to the soil fertility treatments,
significant correlations were found between OC and I-C (r = -0.58) and P-C (r = -0.357)
(Appendix 9). Nonetheless, except for TN and (I+P) - C, negative correlations were observed
between initial soil fertility and yield response to the applied soil fertility treatments
(Appendix 9 and Fig. 12).

49

YieldinI+P(tonha1)

YieldinC(tonha1)

3.0
2.5
2.0
1.5
1.0
0.5
0.0

2.5
2.0
1.5
1.0
0.5
0.0

0.1
TN%

2.0

3.0
2.5
2.0
1.5
1.0
0.5

0.2

3.0

3.5

2.5

3.0

2.0
1.5
1.0
0.5
0.0
0

20
Av.P(mgkg1)

40

0.2

2.5
2.0
1.5
1.0
0.5
0.0
0

20
Av.P(mgkg1)

40

Figure11:Scatterplotsofchickpeayieldincontrol(left)andI+P(right)treatmentsagainstinitialsoil
fertility variables, with regression line plotted. Notes: OC = organic carbon content, TN = total
nitrogencontentandAv.P=availablephosphorus,C=control,andI+P=inoculation+phosphorus.

50

1.0
OC%

0.0
0.1
TN%

0.0
3.5

YieldinI+P(tonha1)

Yieldincontrol(tonha1)

2.0

3.0

1.0
OC%

YieldinI+P(tonha1)

Yieldincontrol(tonha1)

0.0

3.5
3.0
2.5
2.0
1.5
1.0
0.5
0.0

1.2
Yieldresponse(tonha1)

1.0

(I+P)C

1.4

IC

1.2

IC

PC

1.0

PC

0.8
0.6
0.4
0.2
0.0

0.2

10

20

30

40

Yieldresponse(tonha1)

1.4

0.6
0.4
0.2
0.0
0.0

0.5

1.0

1.5

2.0

0.4

0.6

Availablephosphorus(mgkg1)

1.4

0.6

(I+P)C

1.2

PC

0.8

Organiccarbon(%)

Figure 12: The relation between initial soil fertility

(availableP,totalnitrogenandorganiccarboncontent)
and the magnitude of the yield response to the soil
fertilitytreatments,whereIC=inoculationcontrol,P
C = phosphorus fertilized control, and (I+P)C =
inoculation and P treatment the control. In all cases
the solid upper line, the middle dotted line, and the
lower line refer to the (I+P)C, IC, and PC yield
responses

IC

1.0
Yieldresponse(tonha1)

0.8

0.2

0.4

(I+P)C

0.6
0.4
0.2
0.0

0.2

0.05

0.1

0.15

0.2

0.4
0.6
Totalnitrogen(%)

3.13. Household characteristics

Family size ranged from 3 to 8 with average family size of 6 per household. In addition, the
livestock holdings of the households ranged from 0.13 to 3.74 tropical livestock unit (TLU)
per household with an average of 1.9 TLU per household. This is consistent with the previous
report of Amede et al. (2001) that indicated the average livestock holding of households in
Wolaita area to be 1-2 heads. The average landholding of the households in the study area
was 0.6 ha, with a range from 0.25 to 2 hectares. Per capita landholding of the households
also ranged from 0.06 0.25 ha with the average of 0.11ha (Appendix 3).

51

The statistical analysis showed that both the initial fertility of the soil, except for P, and yield
of the farms were not significantly affected by the livestock herd size. However, positive
relation was observed between livestock herd size and OC and TN. This might have resulted
from the small herd sizes across the board. Lack of significant effect of the livestock herd
size on initial fertility and yield of the plots could also be attributed to the competing uses for
crop residues and manure as livestock feed and fuel respectively. In conditions when crop
residues are used as livestock feed and fuel instead of soil amendments, more livestock leads
to larger decrease in soil fertility serving as driver of the competing use of the crop residue.
The landholding of the households was negatively and highly significantly correlated to Av.
P of the soil (r = -0.189) and P-C yield response (-0.292) (Appendix 9). Also, positive and
significant (r = 0.49) correlations were recorded between TLU and Av.P of the soil. Although
statistically not significant, yield of the control treatment and per capita landholding of the
households were also positively correlated (Appendix 9).

52

4. Discussion
In these experiments, positive effects of inoculation and/or P fertilization on nodulation,
growth and yield of chickpea were found.

4.1. Nodulation
As expected from the well-established knowledge on the phenomenon of symbiosis between
legumes and rhizobia, the inoculation treatments (I and I+P) resulted in increased nodulation
of chickpea that in turn, led to enhanced growth and yield through increased supply of
nitrogen. Unlike at 23 DAS where only the I+P treatment gave higher nodules than the
control, nodulation of chickpea was improved by all the three soil fertility treatments at 45
DAS and maturity, with the most pronounced improvement at 45 DAS. Of the three stages,
minimum nodulation was recorded at maturity which can be attributed to nodule degradation
(Shukla and Yadav 1982).
Inoculation did not significantly improve nodulation of chickpea at 23 DAS. This could be
due to the relatively long time required for BNF to begin, which, usually takes place between
2-5 weeks after planting (Werner and Newton 2005). In contrast, inoculation improved
nodulation significantly at 45 DAS. Similar results were reported by Kassa (2009), who
indicated that in the same location, inoculation of chickpea seed improved NNPP by 60%
over that of the control. Moreover, a positive and significant response of legume nodulation
to inoculation was also reported by others (Otieno et al. 2009; Namvar and Sharifi 2011;
Verma et al. 2013) in Kenya, Iran and India respectively. Similar to the effect of inoculation,
P application did not improve nodulation of chickpea in the early stage of plant growth, i.e. at
23 DAS. This could be related to the fact that P dissolution takes about 3-6 weeks.
Nonetheless, the application of P resulted in a highly significant improvement of nodulation
in the later stages. This is in agreement with the findings of a previous study in which on
farm application of 46 kg P ha-1 increased nodule numbers and NDWPP of chickpea by 157%
and 114% over the control treatment respectively (Kassa 2009). Such positive effects of P on
legume nodulation have also been reported by several other studies conducted on chickpea,
haricot bean and faba bean in Ethiopia, Tanzania and Pakistan (Yoseph 2011; Amijee and
Giller 1998; Shukla and Yadav 1982; Ali et al. 2004; Ghizaw et al. 1999). From an
experiment that included five levels of P, Idris et al. (1989) also reported that phosphorus
application significantly improved both number and dry weight of nodule in chickpea. Also
for soybean, the alleviation of phosphorus deficiency resulted in highly significant increase in
53

nodule mass, nodule number, and nodule dry weight plant-1 (Israel (1987). Generally, the
observed improvement is explained by the positive effect of P on root growth, nodule
initiation and functioning.
Nodulation was significantly affected by soil type only at 23 DAS, when the red soil had
higher NNPP and NSPP than chickpea on the black soil. This might have resulted from the
relatively higher resident rhizobia found in the red soil compared with the black soil (Table
2). The result of interaction between the treatments and the soil types showed that
inoculation, either alone or in combination with P, resulted in higher nodulation when applied
on red soil compared to the black soil. In this experiment nodulation was assessed with two
methods, i.e. counting all the nodules, resulting in NNPP and visually scoring the nodules,
resulting in NSPP. Though both methods assess the number of nodules per plant, the latter
employs a 1-5 scale, which eases the work as counting all nodules is not required. Unlike
NNPP, the method also takes into account the distribution of nodules. However, the lack of
accuracy and the need for adjustment for each crop and location can be mentioned as
disadvantages of NSPP. For small sample size using NNPP might enable accurate
measurement and show all the differences existing between treatments. In this study, both
methods were used and mostly found to follow the same trend for all the treatments.
The MPN test result showed that the majority of the soils in the study area are inhabited by
very small populations of rhizobia. Similarly, Rupela et al. (1987b) found fields with low
native rhizobial populations in regions where chickpea had been grown for centuries. As
Rhizobium concentrations of <102 g-1 of soil are insufficient to establish effective symbiosis
and initiate nodulation (Rupela et al. (1987b), BNF in the soils of the study area was severely
constrained. Absence of viable native rhizobia in these soils might have resulted in the
observed positive response of chickpea plants to inoculation. Thus, artificial inoculation is
strongly recommended to improve the productivity of chickpea in this area.

4.2. Morphological and phenological traits of chickpea

Both separate and combined application of inoculation and P significantly improved shoot
fresh weight plant-1 (SFWPP), number of branches plant-1 (NBPP) and plant height of
chickpea. The role of inoculation in enhancing nitrogen fixation and thereby increasing N
uptake by plants contributed to improved vegetative growth of chickpea.
On the other hand, the poor shoot weight recorded from the control treatment might be due to
the low population of native rhizobia that ranged from none to <10 cells gram-1 of soil.
54

This result might be explained further by the below optimum available P content of these
soils (Appendix 1). Such positive response of chickpea shoot development to inoculation and
P fertilization was also reported elsewhere (Elkoca et al. 2007; Verma et al. 2013).
Application of the soil fertility treatments extended the growing period of chickpea compared
with the control. The longer maturity periods might have been caused by the promoted
vegetative growth due to enhanced supply of N and P. Eventually, late maturing crops were
observed to give higher yield compared to the early maturing ones. A similar effect was
reported earlier where inoculation delayed maturity time of chickpea (Gan et al. 2009b) and
white bean (Buttery et al. 1987). Also the delaying effect of P and K fertilizer was reported
for white bean (Buttery et al. 1987). Of the two soil types, plants growing on black soils took
longest to mature. The higher water holding capacity of the black soil might have favoured
vegetative growth of the plants due to sufficient water availability even at a later stage.
Similarly, Gan et al. (2009b) has reported that maturity of chickpea varied from 91 to 136
days mainly due to differences in soil environment.
With respect to the morphological traits, our findings are in accordance with those of Ahmed
et al. (2010) and Dutta and Bandyopadhyay (2009) confirming positive and significant effects
of inoculation and P on number of branches and plant height of chickpea respectively. Also
numerous other studies (Ghizaw et al. 1999; Elkoca et al. 2007; Togay et al. 2008; Ahmed et
al. 2010; Namvar and Sharifi 2011; Yoseph 2011) reported enhanced morphological growth
due to inoculation and/or P due to the increased supply of nitrogen, through BNF, and direct
application of phosphorus. Again, the taller plants and higher number of branches on the
black soil compared to the red soil might have resulted from the higher moisture holding
capacity of the former

4.3. Yield and yieldrelated traits of chickpea

Positive effects of the soil fertility treatments were also observed for NPPP and NoSPP of
chickpea. The results of the analysis of variance showed that NPPP was significantly superior
in all the three treatments compared to the control whereas NoSPP was improved by the
application of inoculation and I+P treatments only. Our results are in accordance with the
findings of several other studies (Kassa 2009; Ahmed et al. 2010; Yoseph 2011; Namvar and
Sharifi 2011; Tellawi et al. 1986; Togay et al. 2008; Dutta and Bandyopadhyay 2009) and
explained by the increased supply of nitrogen through BNF and the direct supplementation of
phosphorus that in turn play important roles in enhanced growth and assimilate accumulation,
55

thereby improving the reproductive performance of the plants. NPPP was higher on the black
soil than on the red soil, which might be due to higher moisture availability in the black soil
during pod setting and flowering of the plants.
The total biomass of chickpea is of great importance in farm level nutrient cycling due its role
as either animal feed or organic matter to be returned to the soil. In this study, total biomass
of chickpea was highly significantly improved by the soil fertility treatments due to the
positive effects of nitrogen and phosphorus on vegetative growth, and the synergy of these
two nutrients on plant growth. This result is in agreement with some previous findings
reported on chickpea (Ahmed et al. 2010; Ali et al. 2004; Elkoca et al. 2007). Moreover, from
an experiment conducted under similar condition, Kassa (2009) reported that inoculated
chickpea produced 22% more straw than the control. Also, the positive effect of P on total
biomass was in agreement with earlier studies (Kassa 2009; Gidago et al. 2012; Idris et al.
1989; Togay et al. 2008), as well as the strongest effect by the combined application of
inoculation and P (Ali et al. 2004).
Inoculation of chickpea seed significantly improves its grain yield (Rokhzadi and Toashih
2011; Togay et al. 2008; Ahmed et al. 2010; Namvar and Sharifi 2011; Verma et al. 2013). In
this study, the soil fertility treatments resulted in a significantly higher yield compared to the
control (1.6 ton ha-1), with the highest (2.1 ton ha-1) grain yield recorded in the I+P treatment.
Similarly, inoculation of two different strains increased chickpea yield in the same study area
by 45% and 32% compared to the control (Kassa 2009). Also, Gan et al. (2009a) reported
that inoculated chickpea produced a 37% higher seed yield over the control, which was
similar to that obtained from a 112 kg N ha-1 fertilized plot. In our study, application of 46 kg
ha-1 of P fertilizer has also resulted in 19% higher chickpea grain yield over the control.
Similar effect of P on grain yield of legumes was found by previous researchers (Idris et al.
1989; Dutta and Bandyopadhyay 2009; Kassa 2009) and attributed to the role of P in the
meristematic activity of plant tissue, and in the synthesis of other growth components from
carbohydrates (Ahmed and Badr 2009). The effect was even more pronounced when
inoculation and P were applied in combination resulting in a yield advantage of 33% over the
control. This might have resulted from the positive effects of P on the process of N2 fixation
where the increased supply of N through inoculation result in enhanced plant growth that
eventually leads to P deficiency.

56

Thus, application of P again improves the growth of the plants further. In line with this,
Fatima et al. (2007) reported that combined application of inoculation and P increased
cowpea grain yield by 63% over that of the control. The observed yield improvements are
due to the increased N and P availability and are in line with the improvements observed for
the yield related traits discussed above. In general, the observed improvement in chickpea
grain yield due to the soil fertility treatments is attributed to the enhanced supply of nitrogen
and P elements whose role in both vegetative growth and grain filling are of paramount
importance.
Unlike most of the parameters studied in this experiment, thousand seeds weight of chickpea
was not significantly increased by inoculation. Similar result was reported by Elkoca et al.
(2007) and Kassa (2009) who found no significant effect of inoculation on thousand seeds
weight of chickpea. However, contrary to these, some studies found a positive response of
seed weight to inoculation of annual legumes (Namvar and Sharifi 2011; Ali et al. 2004;
Yoseph 2011). Nonetheless, P, either alone or in combination with inoculation, resulted in a
significantly larger weight of thousand seeds of chickpea, which is in good agreement with
the findings of some other studies (Ali et al. 2004; Yoseph 2011). The reason that the
maximum 1000 seeds weight was recorded in the I+P treatment could be due to enhanced
growth and development of plants that resulted from phosphorus supply and its positive
effect on nitrogen fixation. The resulting increased N availability might have promoted the
supply of assimilates to seed thereby enabling them to gain more weight.

4.4. Straw quality and total N uptake

Positive effects of inoculation and P fertilization were also recorded for total N and total P
content of chickpea straw. As such, separate application of inoculation resulted in 56% higher
STN% compared to the control, which is clearly related to the enhanced availability of
nitrogen through fixation of atmospheric N2. This result is in good agreement with the
findings of Gan et al. (2010) who reported that straw of inoculated chickpea gave 1.3 mg g-1
higher N concentration than the non- inoculated crop. In line with this result, enhanced straw
N concentration of chickpea due to inoculation was also reported by some other studies
(Kassa 2009; Zaidi et al. 2003; Rokhzadi and Toashih 2011; Alagawadi and Gaur 1988).
Total N uptake of legumes can serve as good indication of N2 fixation. The statistical
analysis of variance showed that the total N uptake was increased by application of the soil
fertility treatments. Inoculation, either alone or in combination with P, significantly improved
57

the total N uptake with yet a more pronounced improvement under the combined application
of inoculation and P. Given the very low native rhizobial populations of the study soil, this
improvement in total N uptake due to inoculation could be due to the positive role of
inoculation on the process of N2 fixation.
Likewise, the separate application of P was also able to result in significant improvement of
STN content of chickpea over the control. A similar positive effect of P on TN concentration
of chickpea straw was reported (Kassa 2009). Moreover, this finding is supported by the
result of some other authors (Fatima et al. 2007; Zaidi et al. 2003; Rokhzadi and Toashih
2011).
However, the effects were even more pronounced when the I+P treatment was applied,
resulting in the highest STN of about 0.98%, exceeding the control treatment by 82%. This
result is in agreement with the findings of Fatima et al. (2007) who reported the maximum N
concentration in soybean straw under the combined application of inoculation and P. The
resulting highest N shoot uptake was attributed to the increased supply of P having a positive
role in N2 fixation by rhizobia, thereby enhancing nitrogen uptake by the plant shoots. The
observed positive effect of P, either alone or in combination with inoculation, on STP content
of chickpea might have resulted from increased availability of P in the soil and its subsequent
vital role in physiological and developmental processes of plants and its favourable effect on
growth and nutrient uptake. Generally, the results of this study are in line with the previous
conclusions that in addition to its role in host plant growth, phosphorus has specific roles in
nodule initiation, growth, and functioning (Israel 1987).

4.5. Across farm variability

For all treatments and variables, a consistent pattern of high variability was observed across
farms. Nonetheless, on the majority of farms, more than 85% in case of yield, positive effects
of the soil fertility treatments were recorded. In line with Tittonell et al. (2010), the soil test
result showed wide differences between farms for initial soil fertility. However, soil fertility
was poor across all farms. The magnitude of response to the soil fertility treatments was also
different across the farms with better yield response observed on relatively less fertile soils.
Although the red soil contained relatively higher number of native rhizobia than the black
soil, for all treatments including the control higher yield and yield related traits were recorded
on the black soil than the red soil which could be due to the better holding capacity of the
black soil.
58

Contrary, the magnitude of response to the treatments was higher on the red soil than the
black soil. Such positive but variable on-farm yield response to inoculation and P treatments
was also reported for cowpea, groundnut, and soybean in Ghana (Bressers 2012).
Mostly regional or national recommendations are made based on on-station trials where
management and inputs are optimized. In the present studys area, there were few replicated
on-farm trials. Usually, experiments are conducted only on a single farm but meant to
represent all farms located in the kebele. Given the huge across farm variability observed in
response to the soil fertility treatments, it is important to test technology packages on several
farms thereby identifying how often and to which extent do the technologies perform.
Nonetheless, according to the results of this study especially farmers with low fertile soil
seem to benefit from application of P and I. In this regard, the information obtained from the
current study fills an important knowledge gap.

4.6. Cost benefit analysis

As indicated above, the average landholding of the households in the area is about 0.6 ha and
farmers usually plant chickpea on small part of their land. Assuming chickpea is at least
planted on a quarter of their land, the use of these technologies will enable each household to
get a yield increment of 60 kg, 45 kg and 75 kg per household from inoculation, P
fertilization, and P+I treatments respectively. Converting this into monetary value and the
cost of these inputs considered, the net benefit that each household would get by using these
technologies would be 63, 33, and 63 USD for using inoculation, P fertilization, and P+I
treatments respectively with no difference in return due to inoculation and P+I treatments.
While making the analysis, the costs of P fertilizer, inoculants and chickpea grain were
assumed to be 100 USD ha-1, 8 USD ha-1 and 1 USD kg-1 respectively. As such, a yield
improvement of 1 kg, 15 kg and 16 kg is required to compensate the cost of inputs for
inoculation, P fertilization, and I+P treatments respectively, using the application rates as in
the experiment. There seem to be equal benefit obtained from the use of inoculation and I+P
treatments. Given the high cost of P fertilizer, the use of inoculation appears to be a priority
for smallholder farmers growing chickpea in this area. Likewise, it can be inferred that
inoculation is a means to increase the economic benefits of P fertilizer use.
The vast majority of the households in Taba area are resource poor farmers who have been
experiencing shortage of food fuelled by the low agricultural input usage on the already
degraded land. At the same time, in spite of using low yielding local varieties and lack of
59

fertilizer input, these farmers have been practising crop rotation through inclusion of food
grain legumes in cereal dominated farming. Given the small landholdings, the observed yield
improvement seems to be small. However, the observed yield improvements due to the
treatments can still somehow increase the harvest of each household which can be either used
for consumption or sold and serve as cash income. On the other hand, the increased total
biomass production can increase the fertility status of the soil for the next cereal crop, if left
on the farm or serve as good source of livestock feed.

60

5. Conclusions
In this study the response of chickpea to inoculation and/or P fertilization was evaluated on
farmers fields in Taba kebele of Wolaita area. The results of the soil test clearly showed that
regardless of the fact that in Taba chickpea has been cultivated for a long time, the soils of
the area are inhabited by small populations of rhizobia. Furthermore, soils of all the study
farms were deficient in major plant nutrients. As was expected in such soils, artificial
inoculation of chickpea seed improved the growth and yield of the crop. For all treatments,
including the control, larger yields and higher values for yield related traits were recorded on
the black soil than on the red soil. On the other hand, the response to the soil fertility
treatments was larger on the red soil than the black soil.
Both nodulation and phenological traits of chickpea were improved by inoculation and P
fertilization, consequently resulting in improved yield and yield related traits. For all plant
variables studied, the improvement was more pronounced for the combined application of
phosphorus and inoculation than for their separate application. This underlines the positive
effect of P on biological nitrogen fixation. For all the variables studied, moderate to large
across farm variability was exhibited, with relatively smaller variability on the red soil.
Assuming farmers would sow chickpea on a quarter of their cropland (average 0.6 ha),
households would get an average extra income of 63, 33, and 63 USD by applying
inoculation, P fertilization, and P+I treatments respectively.
However, most of the farmers in this area cannot afford P fertilizers, so credit facilities or
direct supply of fertilizer would be necessary. Generally, depending on the content of native
rhizobia of a soil and the availability of elite strain, Rhizobium inoculation of chickpea seed
may either reduce or substitute the need for costly N fertilizers for smallholder farmers.
Although there are limited number of inoculant producing companies in the country, the on
market unavailability of this product may hinder the likely absorption of the technology.
Moreover, the small landholding of the households might negatively affect the adoption of
these technologies as the improvement might be considered insignificant by the farmers.

61

62

6. Implications of the results for policy and research

The results of this study suggest that increasing chickpea productivity on smallholder farms is
possible and that the use of Rhizobium inoculants is an appealing strategy to achieve this.
However, the experiment only included one strain of bacteria and a single variety of Desi
type chickpea. Basically, there is huge variability among the regions of Ethiopia for rainfall,
soil type, and other soil characteristics. Hence, though positive and appealing yield and
growth improvement were observed with the strain and chickpea variety used, I would
suggest that more strains (domestic or imported) and more varieties (including Kabuli type)
would be tested in order to ascertain the best alternative. Based on visual assessment, the
performance of chickpea plants in the control treatment was better than the plants sown by
the owners of the farms, which could be due to the differences in time and method of sowing.
Most of the farmers normally broadcast chickpea and, compared with the row planted plots,
the stand in the broadcasted fields was poor. Moreover, most of the farmers in the area were
sowing chickpea late September whereas for this experiment plating was done early
September. Given the observed shortage of rainfall towards maturity, once the land is
available it is strongly advisable to sow chickpea as early as possible starting from midAugust. This study was conducted only in the southern region of Ethiopia, whose share of
Ethiopian chickpea production is only about 2%. Therefore, regarding promotion of
technologies related to chickpea, priority should be given to farmers in Amhara and Oromia
regions of Ethiopia whose chickpea share is about 61% and 32% of the country respectively.
In addition, it is important to note that P fertilizers, except the commonly known DAP, are
not available on market across the country except what is imported for research purpose by
agricultural research organizations. Moreover, the very limited numbers of inoculant
producing companies in the country need to be further strengthen in order to ensure active
involvement of private companies as well as governmental organizations. In addition,
attempts should be made in creating farmers awareness concerning the potential benefits of
these technologies and this needs to be followed by establishing a link between the actual
production and input supply to the farmers.

63

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68

8. Appendices
Appendix1.Physicochemicalpropertiesofsoilsfromthetwentystudiedfarms
CEC & Exchangeable Bases (cmol(+)/kg Soil)

Texture

EC
Soil
pHH2O OC
TN
Av. P
%
%
%
(mS/cm) Clay Silt
colour 1:2.5 (%)
(%)
(mg/Kg) CEC
Ca
Mg
Na
K
Sand Soil class
Black
5.8
1.5
0.17
12
20
11.6
6.6
0.22
0.82
0.08
25
50
25
Silt- loam
Black
7.2
1.8
0.17
30
23
39.9
9.3
0.18
2.45
0.18
23
50
27
Loam
Black
5.9
1.4
0.12
21
0.1
18.4
10.9
2.62
0.26
0.84
24
50
26
Loam
Red
6.4
0.9
0.07
12
17
16
6.3
0.18
0.86
0.07
25
34
41
Loam
Black
6.7
1.6
0.13
22
9.4
3.3
0.1
2.8
0.29
24
44
31
Loam
Red
6.8
0.9
0.08
11
15
10.5
3.2
0.16
1.16
0.06
23
32
45
Loam
Black
6.3
1.2
0.1
18
12
14.6
4.8
0.24
1.02
0.08
18
46
36
Loam
Red
7.1
1
0.1
11
20
9.1
6
0.14
2.55
0.15
20
34
46
Loam
Red
7.2
1.3
0.13
25
19
12.7
3.6
0.12
3.51
0.16
20
35
45
Loam
Red
7.3
1.1
0.11
16
17
12.8
4.8
0.14
3.23
0.11
35
37
29
Clay- loam
Red
6.5
0.7
0.09
14
18
12.5
6.8
0.17
1.32
0.08
43
34
24
clay
Red
6.6
0.8
0.07
10
16
14
4.5
0.14
0.91
0.06
28
30
42
Clay loam
Red
6.4
1.2
0.1
19
18
9.1
7.1
0.19
1.27
0.15
24
37
38
Loam
Black
6.0
1.4
0.1
33
17
9.2
5.4
0.21
0.86
0.08
19
50
31
Loam
Red
6.1
1.1
0.09
12
19
11.3
5.9
0.19
1.08
0.1
24
36
40
Loam
Black
6.2
1.1
0.1
16
17
10.4
3.3
0.24
0.87
0.11
16
38
46
Loam
Red
6.6
0.6
0.07
4
15
10.8
5.5
0.17
1.26
0.04
37
26
37
Clay- loam
Black
6.4
1.4
0.12
27
18
2.3
12.8
0.14
1.58
0.07
19
45
36
Loam
Black
6.5
1.3
0.1
5
16
7.1
4.5
0.18
1.49
0.12
19
42
38
Loam
Black
6.7
1.4
0.1
9
16
12.7
4.9
0.36
1.16
0.09
17
50
33
Silt- loam
RS
6.7
0.96
0.09
13
17
11.9
5.4
0.2
1.7
0.1
28
34
39
Clay loam
Average BS
6.4
1.4
0.12
19
18
13.6
6.6
0.4
1.3
0.2
20
47
33
Loam
OA
6.5
1.2
0.11
16
17
12.7
6.0
0.30
1.52
0.40
24
40
36
Loam
Notes: OC = organic carbon, TN = total nitrogen, Av. P = available phosphorus, EC = electrical conductivity, RS = red soil, BS = black soil, OA = over all. NB:
The result for Av. P of farm 5 was found to be an outlier and excluded.
Farm
No.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20

69

Appendix2.Rainfalldataforthewholegrowingperiodofthetwosoiltypes
Date
Amountofrainfall(mm)recorded
Blacksoil
Redsoil
Sept.4/2012
10.4
9.6
Sept.7/2012
11.9
12.2
Sept.9/2012
0.5
0.7
Sept.11/2013
5.6
5.6
Sept.13/2012
3.6
4.6
Sept.14/2012
0.6
1.1
Sept.19/2012
4.0
4.2
Sept.20/2012
19.1
21.5
Sept.23/2012
4.7
2.6
Sept.25/2012
3.3
4.0
Sept.27/2012
19.1
18.6
Sept.28/2012
13.2
12.3
Sept.29/2012
3.5
3.2
Sum(Sept.)
100
100
Oct.1/2012
25.7
28.0
Oct.2/2012
28.0
32.2
Oct.22/2012
6.3
4.8
Oct.29/2012
2.2
1.4
Oct.30/2012
1.3
8.1
Sum(Oct.)
63.4
74.4
Nov.5/2012
5.8
3.9
Nov.26/2012
3.0
0.5
Nov.28/2012
2.2
0.0
Nov.30/2012
16.7
19.6
Sum(Nov.)
27.6
24
Dec.18/2012
1.6
2.1
Dec.19/2012
3.2
1.6
Sum(Dec.)
4.8
3.7
Jan.7/2013
1.4
2.7
Jan.9/2013
2.7
3.5
Jan.10/2013
4.4
2.8
Jan.11/2013
Sum(Jan)
Total

8.5

3.2

17

212.3

12.2
214.3

70


Appendix3.Householdcharacteristicsofthefarmsunderthepresentstudy
TLU
Percapita Cattle Sheep/ Poultry Donkey
Farm Family
Land
goat

No
size
holding landholding
(ha)
(ha)
1
0
7
1
1
6
0.5
0.08
0.97
3
0
0
0
2
7
0.8
0.11
2.1
3
2
0
0
3
5
0.5
0.10
2.3
2
0
5
0
4
7
0.6
0.09
1.45
2
0
0
0
5
4
0.6
0.16
1.4
4
0
0
0
6
5
0.5
0.10
2.8
2
1
9
0
7
6
1.0
0.17
1.59
2
1
3
0
8
6
0.5
0.08
1.53
4
0
2
1
9
8
2.0
0.25
3.02
2
3
0
0
10
5
0.3
0.05
1.7
3
0
0
0
11
7
0.5
0.07
2.1
1
0
0
0
12
7
0.5
0.07
0.7
2
0
0
0
13
6
0.4
0.06
1.4
5
0
4
1
14
6
0.8
0.13
3.74
2
2
0
0
15
6
0.5
0.08
1.6
4
0
0
1
16
6
0.8
0.13
3
1
2
0
0
17
7
0.5
0.07
0.9
4
4
10
1
18
3
0.5
0.17
3.5
0
1
3
0
19
4
0.3
0.06
0.13
3
1
0
0
20
5
1.0
0.20
2.2
2.5
Mean
5.8
0.6
0.11
0.85
2.15
0.25
1.9
Notes:TLU=tropicallivestockunit,where1TLU=0.7cattle,=0.2pigs,0.1sheep/goat,and0.01
=poultry.

71

Appendix 4. F values for chickpea nodulation traits

Source of
DF
NNPP at
NNPP at
NNPPM
NDWPP at
NSPP at
NSPP at
NSPPM
variation
23 DAS
45 DAS
45 DAS
23 DAS
45 DAS
*
***
*
Soil type
1
890
372.67
35.113
0.3
5.1681
0.23472
0.2
Error a
18
159.69
84.75
57.163
1191.5
1.0004
0.14444
0.59178
Treatments
3
78.34*
2991.35***
38.193***
1886.1
0.5194*
4.2912***
0.61733***
***
**
Soil type *Trt
3
12.66
157.09
0.909
114.8
0.0949
0.25231
0.00933
Error b
54
24.35
17.1
2.046
137.7
0.1565
0.04774
0.03444
Note: *** = significant at 0.1%, ** = significant at 1% and, * = significant at 5%. DF = degrees of freedom, NNPP = number of nodule plant-1, DAS =
days after sowing, NNPPM = number of nodule plant-1 at maturity, NDWPP = nodule dry weight plant-1, NSPP = nodule score plant-1, NSPPM =
nodule score plant-1 at maturity.

72

Appendix 5. F values for morphological and phenological traits of chickpea

Source of variation
DF SFWPP at 45 Days to
NBPP
Plant height
DAS
maturity
Soil type
1 43.5199
2268.45*
72.178*
1156.467***
Error a
18 43.7288
496.033
10.034
73.842
***
***
***
Treatments
3 15.3695
75.517
27.786
29.228***
**
Soil type *Treatment 3 1.8452
8.45
0.541
1.893
Error b
54 0.9912
1.9
1.805
3.492
Note: *** = significant at 0.1%, ** = significant at 1% and, * = significant at 5%. DF =
degrees of freedom, SFWPP = shoot fresh weight plant-1, NBPP = number of branches plant1
.

73

Appendix 6. F values for yield and yield related traits of chickpea

Source of
DF NPPP
NSPP
TBM
Grain yield 1000 SW
-1
variation
(ton ha )
(ton ha-1)
(g)
*
Soil type
1
2814.47
0.49568
2.3033
0.25385
5837.1
Error a
18 416.5
0.14096
5.2254
1.23023
3517.3
Treatments
3
294.75***
0.10738**
3.9199***
0.96371*** 2436.1**
Soil type *Trt
3
16.65
0.0406
0.4303
0.07741
446
Error b
54 27.39
0.02337
0.3211
0.03936
435.3
Note: *** = significant at 0.1%, ** = significant at 1% and, * = significant at 5%. DF =
degrees of freedom, NPPP = number of pods plant-1, NSPP = number of seeds pod-1, total
biomass.

74

Appendix 7. F values for total nitrogen and phosphorus in chickpea straw

Source of variation
DF
STN (%)
STP(mg kg-1)
Soil type
1
0.07707
13151
Error a
18
0.01258
39048
***
Treatments
3
0.20036
15037***
Soil type * Treatment 3
0.00581
728
Error b
54
0.01797
1190
Note: *** = significant at 0.1%, DF = degrees of freedom, STN = straw total
nitrogen, STP = straw total phosphorus.

75

Appendix 8. Pearsons correlation coefficients among the variables (n=80)

NSPPat
23DAS

NSPPAt
45DAS

1
0.14
0.293**
0.102

1
0.26*
0.956***

1
0.244*

TBM(tonha1)

0.33**
0.959***
0.262*
0.26*
0.253*
0.388***
0.356***
0.209
0.453***

0.299**
0.127
0.243*
0.278*
0.086
0.285*
0.042
0.278*
0.069

Yield(tonha1)
1000SW(g)

0.53***
0.202

0.032
0.054

NSPPM
NSPPM
NSPPat23DAS
NSPPAt45DAS
NNPPAt23DAS
NNPPAt45DAS
NNPPM
SFWPPat45DAS
Plht(cm)
NBPP
NPPP
NoSPP
DM

NNPPAt
23DAS

0.96*** 0.28*
0.349**
0.078
**
0.311
0.261*
0.099
0.337
0.198
0.16
0.035
0.37***
0.022
0.123
0.071
0.306**
0.279*
0.047
0.282*
0.253*

0.017
0.048

NNPPAt NNPPM
45DAS

SFWPPat
45DAS
Plht(cm)

1
1

0.38***
*
**
0.29

1
0.349
0.228*
0.238*
0.123
0.284*
0.247*
0.18
0.05
0.404***
0.05
0.338** 0.146
0.025
0.186
0.554***
0.056
0.44***
0.253*
0.273*
0.518***
0.044
0.308**
0.204

0.203

0.023

NBPP

NPPP

1
0.578***
0.779***
0.602***
0.697***
0.351**

1
0.63***
0.392***
0.247*
0.342**

1
0.584***
0.589***
0.346**

0.397***
0.175

0.217
0.241*

0.418***
0.216

Notes: ***, ** and * = significant at 0.001, 0.01 and 0.05 respectively. DAS = days after sowing, NSPPM = nodule score plant-1 at maturity, NSPP = nodule
score plant-1, Pl ht = plant height, SFWPP = shoot fresh weight plant-1, TBM = total biomass, NoSPP = number of seeds pod-1, NPPP = number of pods plant-1,
NNPP = number of nodules plant-1, NNPPM = number of nodules plant-1 at maturity, NBPP = number of branches plant-1, DM = days to maturity, 1000SW =
thousand seeds weight.

76

Appendix 8. Pearsons correlation coefficients among the variables (Continued)

NoSPP

DM

TBM(ton
ha1)

TBM(tonha1)

1
0.507***
0.182

1
0.11

Yield(tonha1)

0.358***

0.322**

0.855***

NSPP
DM

0.138

1000SW(g)
*** **

0.144

0.251

Yield(ton
ha1)

1000SW(g)

0.113

Notes: , and = significant at 0.001, 0.01 and 0.05 respectively. NoSPP = number of seeds
pod-1, DM = days to maturity, TBM = total biomass, 1000SW = thousand seeds weight.

77


Appendix 9. Pearsons correlation coefficients of soil fertility variables, yield of the treatments, yield response and household characteristics (n=20)
Yield in
Yield in
Yield in
Yield in
(+I)-C
(+P)-C
(I+P)-C
TLU
PCLH
Landholding
OC%
TN
Av. P
(C)
(+I)
(+P)
(I+P)
OC%
1
TN
0.841
1
*
Av. P
0.02
-0.08
1
Yield in (C)
0.178
0.025 0.301
1
Yield in (+I)
0.06
-0.059 0.874***
1
0.888***
***
Yield in( +P)
0.02
0.08 0.086
0.74
0.874*** 1
***
Yield in (I+P)
0.122
0.005 0.882
0.921***
0.781***
0.882***
1
**
(+I)-C
-0.58
-0.24
0.126
0.32
0.272
0.126
0.17
1
(+P)-C
-0.357* -0.147 0.311
0.046
0.107
0.572*
0.228
0.311
1
(I+P)-C
-0.236
0.021 0.192
0.087
0.32
-0.359
0.471*
1
0.231
0.192
*
TLU
0.449
0.38
0.49
0.227
0.154
-0.074
-0.128
-0.119
1
0.171
0.006
*
PCLH
0.553
0.443 -0.174
-0.052
-0.117
-0.434
-0.277
0.471
0.615
1
0.084
0.174
***
***
Landholding
0.35
0.239 -0.189
-0.078
-0.148
-0.359
-0.292
-0.148
0.449
0.861
1
0.042
0.189
Notes: ***, ** and * = significant at 0.001, 0.01 and 0.05 respectively. OC = soil organic carbon, TN = soil total nitrogen, Av. P = soil available phosphorus, Yield in C= yield in
control treatment, +I = yield in inoculated treatment, +P = yield in phosphorus fertilized treatment, (I+P) = yield in inoculated and phosphorus fertilized treatment, (I-C) = yield of
inoculated treatment yield of the control treatment, (P-C) = yield of phosphorus fertilized treatment yield of the control treatment, (I+P)-C = yield of inoculated and P fertilized
treatment yield of the control treatment, TLU, tropical livestock unit, PCLH = per capita land holding.

78