ADAMA SCIENCE AND TECHNOLOGY UNIVERSITY

School of Applied Natural Science

Department of Applied Biology

Term Paper on Plant Molecular Pathology

Chocolate Spot Disease Resistance Mechanism in Faba Bean (Vicia faba L.)
and Molecular Breeding to Combat the Challenge

By: Kasahun Amare (PhD student)

Submitted To:Dr. Getache Ayana

Date: June, 2021

Adama, Ethiopia
 CONTENTS
1. INTRODUCTION...............................................................................................................................1
2. PLANT DISEASE RESISTANCE MECHANISMS...............................................................................2
2.1. Passive (Constitutive) resitance....................................................................................................2
2.2. Active (post-infectious) or induced, stimulated resistance............................................................3
3. GENETIC BASIS OF PLANT DISEASE RESISTANCE......................................................................4
3.1. Non-Host Resistance........................................................................................................................4
3.2. Host resistance..................................................................................................................................5
3.3. Plant-pathogen interaction models....................................................................................................5
4. RESESTANCE MECHANIS OF FABA BEAN FOR CHOCOLATE SPOT.........................................7
4.1. Chocolate Spot and Ist Lyfe Cycle...................................................................................................7
4.2. Control Options................................................................................................................................8
4.3. Resestance Mechanism.....................................................................................................................9
5. METHODS OF BREEDING FOR DISESEAS RESISTANCE............................................................10
5.1. Molecular Breeding Methods........................................................................................................10
5.2.Breeding for Biotic Resistance.......................................................................................................11
5.3. Faba Bean Germplasms Resistant to Chocolate Spot......................................................................12
6. REFERENCES......................................................................................................................................14

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 1. INTRODUCTION

Faba bean (Vicia faba L.) (also referred to as the broad bean, horse bean or field bean), is a major
food and feed legume. It is mainly harvested in the form of dry seeds for human food due to its
high protein content (24 to 35%) particularly for low income earners (Crépon et al., 2010; Zong
et al., 2019).

However, this crop is subjected to many abiotic and biotic stresses that seriously compromise the
final yields. Among the threatening biotic stresses, fungal diseases can severely damage faba
bean crops, especially in wet (high moisture or humidity >70%) weather conditions. Ascochyta
blight, chocolate spot, and rust are the three main pathogens affecting faba bean crops globally
(Torres et al., 2006; Emeran et al., 2011 ). The Chocolate spot disease caused by Botrytis fabae
Sardina is more likely to occur in bulky crops and the most economically important diseases that
damage the foliage, limit photosynthesis activity, and reduce faba bean production worldwide
(Torres et al., 2006; Yitayih & Azmeraw, 2018).

The severe epidemics of chocolate spots can be devastating resulting in over 90 % yield
reduction. For example, extensive damage has been reported in Australia with yield loss of 90 %,
United Kingdom (59 %), China (>50 %), and Ethiopia (34-61 %) (Dereje, & Yaynu,2001;
Beyene et al., 2016).

Several attempts have been made to find out ways to control or minimize the impact of plant
diseases on the yield. These include breeding for disease resistance (Sillero et al., 2010)
fungicide control (Emeran et al., 2011), and biological treatment (El-Banoby et al., 2013).
Systemic fungicides provide rapid and effective control, but they are expensive and can be
detrimental to the environment (Emeran et al., 2011). Boris (1997) reported finding both
Botrytis fabae and Botrytis cinerea with resistance to the benzimidazoles and the
dicarboximides. At present, although genetic resistance to these pathogens generally provides
partial protection, the use of resistant cultivars remains the major means to reduce yield losses.

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More recently, biotechnology has been used as a tool to increase field crop productivity in
contrast to sustainable agriculture (Cheikh et al., 2000). Inoculation of plants with compatible
and incompatible pathogens triggers a variety of plant defense responses, including the activation
of genes encoding pathogenesis-related proteins (Sudisha et al., 2012). Many of these proteins
have been shown to exhibit antifungal activity in vitro (Ponstein et al., 1994), for example, the
PR2-class (B-1, 3 glucanase) and PR3 class (chitinase) proteins hydrolyzing the important fungal
cell wall components (Attia, et al., 2007). This suggested that PR proteins may play an important
role in pathogen defense.

The use of resistant cultivars and incorporating favorable resistance genes into the locally
adapted susceptible varieties is one of the most effective ways of controlling chocolate spot
disease (Sillero et al. 2012). Therefore, this paper is intended to highlight some plant(faba bean)
disease resistance mechanisms for chocolate spot and molecular breeding advances for faba bean
disease resistance.

2. PLANT DISEASE RESISTANCE MECHANISMS

All higher plants have developed elaborate and sophisticated system of defence mechanisms,
which enable them to resist and survive pathogen attacks, and various environmental stresses.
The mechanisms of defence can be broadly classified into: 1) passive or pre-existing resistance,
which is also known as pre-infectious or constitutional resistance; 2) active, post-infectious or
induced resistance.

2.1. Passive (Constitutive) resitance

Passive (pre-infectious) or constitutional resistance exists regardless of whether the plant was
attacked by pathogens or not. It is not a true genetic resistance, and is often called pseudo-
resistance. The plant does not develop specific defence responses, but has inherent properties or
certain morphological, anatomical or physiological characteristics, which in case of a pathogen
attack can help prevent infection or the spread of the disease itself. It is also called horizontal
resistance, as it is influenced by a large number of minor genes (small effect genes) (Duvnjak, et
al., 2007). Elements of passive or constitutional plant resistance include resistance to penetration
and spread of pathogens. On the plant surface, there are often physical barriers, such as waxy
layers, cuticles, cork layers, cell wall polymers, lenticels, stomas and trichomes, which prevent

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pathogen invasion. For instance, drops of water are essential for germination of phytopathogenic
fungal spores, but it is hard for them to stay on hydrophobic cuticles and epicuticular wax. Hence
these structures inhibit spore germination and prevent successful infection. In addition, plants
have several constitutive antimicrobial components, for instance simple phenols, saponins and
tannins, which have potent antifungal activity, and thus help prevent colonisation of the tissue
(Osbourn et al., 1998).

2.2. Active (post-infectious) or induced, stimulated resistance

Plants and pathogens have coevolved such that pathogens can recognize plants by the sugars, or
other molecules, they produce. Plants, in turn, can recognize pathogens by the molecules they
produce. The ability to recognize pathogens allows plants to activate defense systems that can
prevent widespread infection. Active defence mechanisms require activation of plant
metabolism, as they are a direct response of the host organism to an invading pathogen. This
usually involves accumulation of phytoalexins, phenolics, ethylene, hydrolytic enzymes,
peroxidases, and numerous stress-related proteins. Active or induced defence responses can be
separated into three categories:
1. Primary responses,
2. Secondary responses and
3. Systemically acquired responses (Hutcheson, 1998).
Primary responses are localized to cells that are in direct contact with the pathogen; secondary
responses occur by the spread of signals in cells that are in close proximity to the sites of primary
infection, while systemically acquired responses are hormonally induced throughout the plant.
Active response mechanisms lead to induced resistance a state of enhanced developed resistance,
acquired as a consequence of primary exposure to a pathogen, which helps stem further infection
(Van et al ., 1998). Induced resistance is generally systemic, as the heightened defence capacity
is developed in not only primary infected tissue, but also in non-infected parts of the plant. Thus,
induced resistance is usually referred to as systematically acquired resistance (SAR). Induced
resistance can also be localized to the parts of the tissue exposed to the primary infection, in
which case it is referred to as localized acquired resistance (LAR). Both LAR and SAR are
effective against a wide range of pathogens.

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3. GENETIC BASIS OF PLANT DISEASE RESISTANCE

The genetic basis of plant resistance to pathogens is divided into qualitative resistance
(monogenic resistance) and quantitative resistance (polygenic resistance) (Jiquel et al., 2021).
Qualitative resistance can be explained by the gene-for-gene model proposed in the fifties by
Flor (1971), who determined the basis of inheritance of resistance to flax rust in flax cultivars.
According to the type of interaction with the pathogen, plant responses at the molecular level are
divided into two types: i) non-host response and ii) host response. These two types of responses
which differ mainly by the molecule type (both from the plant and the pathogen), involved in the
process (Legris et al., 2019).

3.1. Non-Host Resistance

Non-host resistance, is defined as the event where a plant species in particular is resistant to
different kind of pathogens (either bacteria, fungi, oomycetes, or viruses) but, these same
pathogens can infect other plant species (Fan and Doerner, 2012; Bellincampi et al., 2014).

This response spectrum is caused by specific recognition processes between pathogen molecules
called MAMPs (microbe-associated molecular patterns), which are recognized by PRR-type
(pattern recognition receptors) membrane receptor proteins. PRR structures are types of receptor-
like kinases (RLKs) that have functional modular domains (Monaghan and Zipfel, 2012).
However, MAMPs are molecules involved not only in the pathogenesis process; most of them, in
fact, have been described as essential components of the cell, such as flg22. There are other types
of MAMPs such as the HAMPs (herbivore-associated molecular patterns) and the DAMPs
(damage-associated molecular patterns; originally called endogenous elicitors), but in general
most literature talk about MAMPs to refer to this kind of molecules (Conrath et al., 2015).

The molecular response triggered by the recognition of MAMPs is known as PTI (PAMP-
triggered immunity) in which some molecular mechanisms associated with PTI include:
production of ROS, Ca+2 cascades, and the activation of MAPK (mitogen-activated protein
kinases) cascades, involving Ca+2-dependent proteins that ultimately lead to transcriptional
reprograming ( Bigeard et al., 2015; Trapet et al., 2015).

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3.2. Host resistance

Molecular events during ETI (effector-triggered immunity) processes overlap with PTI. This
branch of plant immunity occurs within the cell and originates once the host recognizes the
effectors secreted by the pathogen, in which plant resistance (R) proteins can perceive these
effectors initiating a defense response, including oxidative burst, accumulation of hormones such
as salicylic acid (SA) and NOI (nitrogen oxide), MAPK cascades, changes in calcium levels,
transcriptional reprogramming and synthesis of antimicrobial compounds, expression of
pathogeneses related (PR) genes ( Stael et al., 2015).

Depending on the type of pathogen, the plant can induce programmed cell death (PCD)
processes, also termed hypersensitive response (HR). These processes seek to block the advance
of biotrophic pathogens to avoid an infection in different host tissues. In this process,
chloroplasts play a key role in the production of ROS-type molecules and NOI (Künstler et al.,
2016). For necrotrophic pathogens, the plant cell wall is the first line of defense providing a
dynamic interface for interaction with necrotrophic pathogens. The interaction includes serving
as a rich source of carbohydrates for the growth of pathogens; acting as a physical barrier to
restrict the progression of the pathogens, and playing a role as an integrity sensory system that
can activate intracellular signaling cascades in which plant hormones like jasmonic acid (JA) and
ethylene (ET) play a major role in the defense response against these pathogens (Mandal et al.,
2018).

3.3. Plant-pathogen interaction models

Zig-zag model. In the most basic interaction, the zig zag model involves an interaction between
the pathogen and the host. The interaction can be divided in four phases:

 Phase 1: plants detect MAMPs via PRRs to trigger PAMP-triggered immunity (PTI).
 Phase 2: successful pathogens deliver effectors that interfere with PTI, resulting in
effector triggered susceptibility (ETS).
 Phase 3: an effector can be recognized by an NB-LRR protein, activating effector-
triggered immunity (ETI), which after surpassing a defined threshold induces
hypersensitive cell death (HR).

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Phase 4, pathogen strains that have lost certain effector are selected. They might have also
gained a new set of effectors to respond to the plant defense

Invasion model.This model was proposed by Cook et al. (2015), the authors took into
consideration some limitations of the zig zag model such as: the model is restricted in terms of
what microbe-associated molecule patterns (MAMPs) the plants can perceive through pattern
recognition receptors (PRRs).

The invasion model has been explained in a similar way than the the zig-zag model, except in the
aspects related to the definition of the immunogenic molecules. The authors suggest that these
molecules must be represented as a continuum, and they argue that these molecules play other
roles beyond the pathogenicity. Thus, the evolution can affect these molecules and have effects
in an interaction model. In this sense, if a molecule has a role in a different process some
evolutive forces can alter them, producing changes in the interaction process or even in the
fitness of the species.
Multicompent model. The model was proposed by Andolfo et al. (2016). Like in the previous
model, the authors start showing the disadvantages of the zig-zag model such as the fact that the
model only describes two perception layers (PTI and ETI).
The multicomponent model has two components: activation and modulation, and it is divided in
three phases as follows:
1) Interaction: two principal effects are detected:
 modifications of virulence factor targets and
 specific alterations of primary plant metabolism.
2) Activation: modifications of virulence factor targets induce the Nibblers Triggered
Signaling (NTS) or PPRs Triggered Signaling (PTS), mediated by R-genes activation.
Metabolic alterations induce a feedback regulation of primary metabolic pathways
resulting in a Hormone Tempered Resistance (HTR).
3) Modulation or effective resistance stage, the NTS/ PTS, and the HTR converge to confer
a resistance specific to the lifestyle of pathogen (Pathogen lifestyle- Specific Resistance,
PSR)

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In this case, the authors try to emphasize that the model will be determined specifically by the
pathogen´s life cycle. On the other hand, this model is in certain way connected with a new
approach it which R genes and effectors are described like independent molecules, in a way that
better articulate all the plant-pathogen related information for using molecular techniques to open
a new era in crop breeding

4. RESESTANCE MECHANIS OF FABA BEAN FOR CHOCOLATE SPOT

4.1. Chocolate Spot and Ist Lyfe Cycle

Chocolate spot affects both broad and field beans as Botrytis spores on the leaf germinate and
penetrate the leaf surface. Autumn-sown crops can be infected from early spring, particularly
following periods of frost damage and during prolonged wet and overcast weather. Spring-sown
crops can be affected later in the growing season.

Because chocolate spot is encouraged by overcast and wet conditions, there is always a
proportion of the crop at risk from infection. Control of chocolate spot is dependent on the
application of fungicides as protectant treatments, because once the disease is established, there
is little curative control possible. Timing of fungicides should be related to growth stage and
disease risk (Davidson et al., 2007; Teshome & Kora, 2021).

Both Botrytis cinerea and Botrytis fabae can be seed-borne but the primary source of infection is
from airborne spores that settle on the leaves and germinate to penetrate the leaf surface. Both
fungi are very common, especially in areas where beans are grown regularly. Crop debris and
volunteers from previous crops provide material for overwinter survival of the disease. The
fungus overwinters as sclerotia in the soil, which sporulate and produce conidia in wet
conditions. The conidia contaminate the plant organs, causing lesions on the leaves, flowers and
stems. Optimum conditions for the development of the fungus are when temperatures are
between 15°C and 25C, and relative humidity greater than 80% is required for conidial
sporulation. In dry weather, the fungus does not develop as quickly on the plants(Emeran et al.,
2011; Ahmed, 2017).

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 Figure 1: Chocolate spot infection cycle

4.2. Control Options

Control options for disease include choosing more resistant varieties, fungicide applications, and
planting good quality seed with low infection levels. Testing seed for disease levels prior to
planting can help reduce the risk of infection at the seedling stage and early disease onset in a
field, and should be considered when doing germination tests(Teshome & Kora, 2021).

In some environments, ntegrated disease management may require a single component used
alone (usually host plant resistance) or in combination with one other component (such as
fungicide seed treatment) to adequately combat
diseases of faba bean. The components of ntegrated disease management employed in the
production of faba bean are listed as follows:
 Cultural (agronomic) practices (sowing dates, plant population etc.).

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  Host plant resistance,
 Biological control
 Chemical sprays (fungicides, pesticides), Disease modeling (prediction) for avoidance of
high risk or disease pressure,
The combination of genetic resistance, hygiene and monitoring of crops for threshold levels of
infestation, allows the most economic and effective controls with the result that economic yields
can be maximized( Mitiku, 2017)

4.3. Resestance Mechanism

The deployment of resistant faba bean varieties is an efficient strategy for controlling the disease
and promoting the development of sustainable agriculture (Bouhassan et al., 2004; Rhaïem et al.,
2002). However, only limited studies exist on the defense responses of the faba bean × Botrytis
fabae interaction (Sillero et al., 2010).

Plants defend themselves against fungal infection through the activation of complex defense
responses (Dangl and Jones, 2001). One of the earliest these responses is the rapid generation of
reactive oxygen species (ROS), which includes superoxide anion (O2-), hydroxyl radical (OH•)
and hydrogen peroxide (H2O2) (Patykowski and Urbanek).

There are numerous studies demonstrating the production of these free radicals in plants as an
initial response to microorganisms, both pathogenic and non-pathogenic (Bolwell et al., 2002;
Garcia-Limones et al., 2002; Gill and Tuteja, 2010). ROS may have multiple effects on defense
responses, including direct antimicrobial action, lignin biosynthesis, phytoalexin production and
the triggering of systemic acquired resistance (Bradely et al., 1992)

McKeen 1974 suggested that the cuticle on faba bean leaves was disolved by enzyme produced
of the tip of Botrytis cenerea germ tube and not penetrated mechanically. Microscopic studies by
Manisfield and Richardson 1981 supported the McKeen’s findings and showed that the means of
infection by Botrytis fabae was similar to Botrytis cenerea. They nots that after penetration of the
cuticle hyphae expanded within walls of eppidermal cells.

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Host penetration rarly occurs via stomata for Botrytis spp. but wounded and damaged leaves an
important means ofentery(Verhoeff, 1980). this allow faster estabilishment and spread of
Botrytis fabae infections on faba beans of Botrytis cinerea ussually remain limited(Devrall &
Wood, 1961).

The involvement of PR genes in faba bean plants after pathogen attack has been previously
demonstrated (Attia et al., 2007; Cheng et al., 2012; Rauscher et al., 1999). However, no data are
available regarding the differential induction of the above mentioned PR genes in FB cultivars
possessing different different levels of resistance against Botrytis fabae.

5. METHODS OF BREEDING FOR DISESEAS RESISTANCE

The methods of breeding for disease resistances are essenntially same as those used for other
agronomic traits. theses may include :

1. Introduction
2. Selection
3. Hybridization
4. Budding & Grafting
5. Mutation Breeding
6. Biotechnological methods.

5.1. Molecular Breeding Methods

Faba beans are partially allogamous species having an intermediate level of cross-pollination.
Traditional faba bean breeding involves crossing elite parents, multi-stage testing of progeny,
identify progeny that outperform the parents for specific traits and the release of superior
rvarieties. Typically this process could take up to 10 years to release a commercial cultivar.

Development and advancement of new plant molecular biology techniques and genomic tools
could assist conventional breeding to accelerate the release of faba bean cultivars
(Gnanasambandam et al., 2012). The ability to modify plant genomes and introduce genes for a
specific desired trait into a desired plant gave rise to an array of experiments on several
commercially important crops. Several biotech companies developed seeds for plants that are

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disease-resistant, pest-resistant, herbicide-resistant, or that give high yields. Interspecific sources
of resistance cannot be transferred to faba bean by conventional breeding methods, and the
practical significance of induced mutation is relatively low due to low success rates (Van, 1998).

Molecular approaches to analyse and change qualitative and quantitative traits in cultivated
plants are highly effective to improve yield and quality of food and renewable resources, disease
resistance and abiotic stress tolerance. Molecular plant breeding focuses on the application of
molecular markers and genomics to explore natural variation and on the development of
transgene technologies to expand genetic variation.

The breeding objectives of faba bean programs are similar to other crops and include higher
yield, appropriate phenology, plant growth habit, seed quality, traits to enhance crop
management, breeding for resistance/tolerance to biotic and abiotic stresses, adaptation to the
target environment

Nowaday, biotechnology has been used as a potential tools for faba bean cultivar development,
significant efforts have been made in the last two decades to understand the genetics and
genomics of faba bean. The Lack of suitable cultivar can be easily overcome by application of
modern tools and techniques. Several in-vitro techniques would be very useful for faba bean
breeding. New techniques such as protoplast fusion, regeneration, and embryo-rescue assisted
interspecific crossing could probably be introduced to faba bean to improve yield and
quality(Singh et al., 2012). Linkage maps of faba bean have been constructed based on RFLPs
(Restriction Fragment Length Polymorphisms), RAPDs (Random Amplified Polymorphic DNA)
and SSRs (Simple Sequence repeats) (Satovic et al., 1996), and various QTLs (Quantitative Trait
Loci) have been identified (Torres et al.,210).

5.2.Breeding for Biotic Resistance

Faba bean is affected by a range of biotic stresses, including foliar fungal diseases, soil borne
pathogens, viruses, parasitic weeds, nematodes and bruchids. However, many are of minor or
local importance and are not major objectives in breeding programs. Germplasm collections have
been screened for resistance to a range of diseases and pests.

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Breeding is generally considered the most effective means of disease control. However, for a
genetic approach to disease control to be effective,
1. A good understanding of the host × pathogen interaction is needed;
2. A degree of resistance to the pathogen must be identified; and
3. Efficient and effective screening systems should be available that can process large
populations.
The decision of whether to adopt selection for resistance to a particular disease within a breeding
program will depend on the impact of the disease, likelihood of success, cost and availability of
alternative methods of control and the overall priority of resistance to the particular disease
compared to all other breeding aims. The three fungal diseases, chocolate spot (Botrytis fabae),
ascochyta blight (Ascochyta fabae) and rust (Uromyces viciae-fabae) can lead to very significant
loss in yield and are considered to be the major constraints to the crop. Recent dvance in gene
mapping studies have allowed the identification of genes and QTLs controlling resistance to
some of these diseases. In case of broomrape/ Orobanche, three QTLs more than 70% of the
phenotypic variance of the trait. Concerning Ascochyta, two QTLs located in chromosomes 2
and 3 identified with 45% of variation. A second population sharing the susceptible parental line
also revealed two QTLs, one of them likely sharing chromosomal location and jointly
contributing with a similar percentage of the total phenotypic variance(Torres et al., 2006).

5.3. Faba Bean Germplasms Resistant to Chocolate Spot

The success of resistance breeding depends upon the presence of genetic variability and our
ability to detect and utilise it. This variability is captured in the collection of germplasm. The
largest collection of faba bean worldwide is maintained by ICARDA,with over ten thousand
accessions from 71 countries. The second largest reserve with 5,200 accessions is preserved at
the Chinese Academy of Agriculture `, Beijing, China followed by 2,445 collections in the
Australian Grains Genebank, Victoria, Australia. The USA, Russia, Ethiopia, France, Spain,
Germany and Italy also have large numbers of accessions.

No wild species is sufficiently closely related to faba bean that fertile hybrids have been derived,
so genetic diversity is restricted to the primary gene pool of the species itself (Cubero 2011).
Accessions from the Mediterranean region are considered to have the best potential to provide
genetic variation for rust resistance (Maalouf et al. 2010). Genotypes showing significant rust

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 resistance with late necrosis originated from Central Europe (V-300, V-313, V-1271, V-1272),
East Africa (2N-34) and Spain (V-1335) (Sillero et al. 2000).

The most chocolate spot resistant lines from the Andean region are characteristically very late
flowering and maturing when sown in autumn in a Mediterranean-type environment. Examples
of useful sources of resistance that have emerged are resistance to chocolate spot (BPL 410 from
Ecuador), IO rust (BPL 26IJ to Aseocbyta (BPL472 and PBI line 1/3; JKLLIS, LOCKWOOD
andAuBLKY 1984), and to Orobanche (F402 from Egypt).

The Ethiopian Institute of Agricultural Research (EIAR) released several varieties with a high
level of resistance to chocolate spot. EIAR researchers released several high‐yielding faba bean
varieties through direct selection from germplasm supplied by ICARDA or by transferring good
levels of resistance from ICARDA germplasm into locally adapted varieties. Among the faba
bean varieties released (Temesgen etal., 2015) with partial resistance to chocolate spot are ‘Moti’
(ILB 4432 × Kuse‐2‐27‐33), ‘Gebelcho’ (ILB 4726 × ‘Tesfa’), Obsie (ILB 4427 × CS20DK) and
‘Walki’ (ILB 4615 × Bulga 70) (Table-1). Recently, another variety named ‘Gora’ (ILB2717‐
1 × R878‐1) has been released in Ethiopia with higher degree of resistance to chocolate spot and
larger seed size than traditional cultivars (Temesgen et al., 2015).
Although most breeding programmes focus on developing resistant genotypes for a single
disease of economic importance, efforts have recently been directed to develop faba bean lines
with multiple disease resistance lines (Maalouf et al., 2016).

Table 1: some Resistance and susptible variests for chocolate spot

Pedigree Genotypic cluster Resistant level to Source

Parent lines group CH
ILB-4726 ICARDA I Resistant ICARDA
ILB-938 ICARDA I Resistant ICARDA
BPL-710 ICARDA I Resistant ICARDA
Moti ILB-4432 x kuse-2-27-33 II Moderately resistant HARC
Dosha Landrace collection II Moderately resistant HARC
Gebelcho Tesfa x ILB-4726 II Moderately resistant HARC
Kasa Landrace collection Il Susceptible HARC
NC58 National collection III Susceptible HARC
CS-20-DK National collection III Susceptible HARC
Bulga-70 Landrace collection III Susceptible HARC
Source:( Asnakech, 2014 Doctoria desertation)

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