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Full Research Article

Susceptibility of Durum Wheat (Triticum durum Desf.) to Fusarium Head Blight Caused by Fusarium culmorum under Field Conditions

S
Saliha Rahmani1,2,*
A
Abdelkrim Mebarkia1
A
Ahmad Boulal2
S
Said Boudeffeur3
1Department of Agronomic Sciences, Faculty of Nature and Life Sciences, Laboratory of Applied Microbiology, Ferhat ABBAS University - Setif 1, 19000, Algeria.
2Department of biology, Faculty of Nature and Life Sciences, Ahmed Draïa University, Adrar, 01000, Algeria.
3National Institute of Agronomic Research of Adrar, 01000, Algeria.

ABSTRACT

Background: Durum wheat (Triticum durum Desf.) represents a key cereal species cultivated across Mediterranean environments. Despite its agronomic importance, this crop remains highly vulnerable to Fusarium head blight (FHB), a disease mainly attributed to Fusarium culmorum, which can severely losses yields and adversely affect grain quality.

Methods: This study evaluated the susceptibility of five durum wheat varieties (Boutaleb, Mohamed Ben Bachir, Vitron, Oued-elbared and Aoures) to Fusarium culmorum under field conditions in southwestern Algeria during the 2020/2021 cropping season. Inoculations were performed at the beginning of flowering (Zadoks GS 60) using three spore concentrations (102, 104 and 106  spores mL-1). Disease severity, percentage of Fusarium-damaged grains and yield-related parameters were assessed.

Result: Results showed significant differences among wheat varieties and inoculum concentrations. Disease severity and grain damage increased with increasing spore concentration. The highest severity (74%) and grain damage were recorded in the variety Mohamed Ben Bachir, while Vitron showed comparatively lower susceptibility with a severity of 34% and only 10% damaged grains at the highest inoculum level. The study highlights considerable variability in susceptibility among durum wheat genotypes. The variety Vitron showed relatively better tolerance to Fusarium culmorum, suggesting its potential use in breeding strategies focused on enhancing resistance to Fusarium head blight.

INTRODUCTION

Durum wheat (Triticum durum Desf.) is widely recognized as a major cereal crop of global economic importance, playing a central role in global food security and agro-industrial systems in the Mediterranean and North African countries (Royo et al., 2009; Tidiane et al., 2019). However, wheat cultivation in Algeria is frequently challenged by numerous biotic stresses, among which fungal pathogens are considered as the main cause for the yield and quality loss.
       
Among these diseases, Fusarium head blight (FHB) is identified as the most devastating diseases of wheat in the world (Dweba et al., 2017). This disease affects wheat spikes during the flowering stage and can cause severe yield losses as well as significant deterioration in grain quality (Moreno-Amores et al., 2020; Kaur et al., 2026).
       
FHB is a destructive disease associated with several species of the genus Fusarium (Malik and Verma, 2026). Among them, Fusarium culmorum and Fusarium graminearum are the most commonly reported pathogens in many wheat-growing regions (Bouanaka et al., 2021; Saharan, 2020). These fungi infect wheat spikes at anthesis, resulting in premature bleaching of spikelets, shriveled kernels and significant reductions in thousand-grain weight (Bilska et al., 2018). Moreover, Fusarium culmorum produces type B trichothecene mycotoxins, including deoxynivalenol (DON), which pose a health risk to humans and animals (Yekkour et al., 2015; Bilska et al., 2018).
       
In Algeria, different studies have consistently reported Fusarium culmorum as the dominant species associated with FHB on durum wheat (Yekkour et al., 2015; Touati-Hattab et al., 2016; Laraba et al., 2017). Field-based investigations have revealed substantial variability in both the aggressiveness of Fusarium culmorum isolates and the susceptibility of durum wheat genotypes under local agro-climatic conditions (Hadjout et al., 2022).
       
The management of FHB relies on integrated strategies combining agronomic practices, chemical control and host resistance. Among these approaches, breeding and cultivating resistant wheat varieties are considered the most efficient, cost-effective and environmentally friendly technique to mitigate disease development and reduce mycotoxin accumulation in the grains (Saharan, 2020; Ghimire et al., 2020). However, the success of resistance breeding programs depends largely on accurate and reliable phenotyping under field conditions, where environmental factors strongly influence disease development and expression. Hence, resistance to FHB is regarded to be quantitative trait governed by both genetic and environmental conditions, making its evaluation under field conditions particularly relevant.
       
In this context, the present study aims to assess the susceptibility of five durum wheat genotypes to Fusarium culmorum under field conditions in southwestern Algeria. By using artificial inoculation and evaluating disease severity and yield-related parameters, this work seeks to provide reliable data on genotype performance under realistic agronomic conditions. The results are expected to enhance current knowledge on FHB epidemiology and to support breeding and crop protection strategies for durum wheat in Algeria and similar Mediterranean regions.

MATERIALS AND METHODS

Fungal material
 
Isolation of Fusarium culmorum
 
Symptomatic wheat samples were randomly collected during the 2019/2020 agricultural season from several farms located in southwest Algeria. Contaminated stem, leaf and seed fragments underwent surface disinfection using 2% sodium hypochlorite solution for five minutes, followed by three rinses with sterile distilled water. The sterilized tissues were then dried at room temperature on sterile Whatman paper. Five fragments or seeds were inoculated per Petri dish containing Potato Dextrose Agar (PDA) medium and incubated at 28±2oC for seven days. Pure fungal isolates were obtained through repeated subculturing on PDA.
 
Morphological identification
 
Isolates were identified through macroscopic observation of colony appearance and microscopic examination of mycelium and spore morphology. Identification was conducted utilizing the taxonomic keys provided by Leslie and Summerell (2006) and Xia et al., (2019).
 
Molecular characterization
 
The identification was confirmed through the use of molecular identification. Genomic DNA was extracted using the commercial NucleoSpin Plant II kit (Macherey-Nagel, Germany). The primers used were ITS1 and ITS4. PCR products were separated by electrophoresis in a 1.5% agarose gel, using a 10 μL deposit of the amplification products. Gel migration was followed by staining in an ethidium bromide bath (0.5 μg/mL) and DNA bands were visualized and captured using the Gel Doc system (Bio-Rad, USA). After purification, PCR products were sequenced according to the Sanger procedure (Sanger et al., 1977) using the BigDye v3.1 kit (Applied Biosystems) and the original primers. The resulting sequences were edited and cleaned using CHROMAS PRO software. The products were then compared using the BLAST tool.
 
Plant material
 
Five durum wheat (Triticum durum Desf.) genotypes widely cultivated in Algeria were used in this study: Boutaleb, Mohamed Ben Bachir (MBB), Vitron, Oued-elbared and Aoures. All genotypes were provided by the Technical institute of field crops (TIFC), Algeria. The name, pedigree and origin of durum wheat genotypes are presented in Table 1.

Table 1: Name, pedigree and origin of durum wheat varieties.


 
Study site and crop management
 
The study was carried out during the 2020 / 2021 cropping season at the INRA research station in Adrar, Algeria (27o50’N, 0o18’W). The soil profile is medium-deep with a sandy-clay-loam texture. The region features arid climate with a mean annual rainfall of approximately 15 mm. Temperature data indicates a mean minimum of 12oC in winter (December-January) and a maximum of 40oC in summer (July) (INRA Adrar, 2019). Sowing was performed manually during the autumn season (November-December) at a seeding rate of 250 seeds m-2, with a row spacing of 25 cm. Weed management was carried out manually at the herbaceous tillering stage. Nitrogen fertilization consisted of 46% urea applied at a rate of 150 kg ha-1, split into two equal doses at the tillering and stem elongation stages. Flood irrigation was used on the crop, twice to three times a week.
 
Inoculum preparation
 
The inoculum was prepared following to the protocol of Siou et al., (2014), with slight modifications. A preserved isolate of Fusarium culmorum was sub-cultured on PDA plates and incubated at 28±2oC for seven days to obtain actively growing cultures. Three agar discs were excised from the margins of colony and transferred into 100 mL of liquid medium consisting of potato extract and dextrose. The cultures were maintained under continuous agitation for five days to enhance sporulation. The conidial suspension was then filtered using fine cheesecloth to remove mycelial fragments. Spore concentration was determined using a Malassez hemocytometer and adjusted with sterile distilled water to final concentrations of 102, 104  and 106 spores mL-1. The prepared conidial inoculum was stored at 4oC for inoculation in the field.
 
Experimental design and field inoculation
 
A randomized complete block design was used with three replications. Each cultivar was subjected to four treatments: Three inoculum concentrations (10², 104 and 106 spores mL-1) and a non-inoculated control. Artificial inoculation was carried out at the flowering stage (Zadoks, 65). Spore suspensions of each concentration were applied by spraying directly onto both sides of 10 plots of each wheat variety using a hand-held sprayer. Control plots were not inoculated. To promote infection, the spikes were covered with transparent polyethylene bag for 72 h and inoculated plants were irrigated immediately after inoculation to maintain high humidity levels, favorable for fungal development.
 
Disease assessment and yield analysis
 
FHB severity was evaluated 21 days after inoculation. Disease severity was expressed as the percentage of infected spikelets per spike. In addition, the rate of Fusarium-damaged grains (FDG %) was assessed after harvest by visually separating shriveled, discolored, or lightweight kernels from healthy ones. Yield-related parameters were also measured, including the number of grains per spike (NGS), grain weight per spike [GWS (g)] and thousand grain weight [TGW (g)].
 
Statistical analysis
 
The experimental design utilized a randomized complete block design (RCBD) with three replications. The analyzed variables were: genotype (five durum wheat varieties), treatment (0 (control), 10², 104 and 106  spores mL-1) and their interaction. Two-way analysis of variance (ANOVA) was utilized in determining the main effects of genotype and inoculum concentration, as well as their interaction, on disease severity, FDG percentage and yield components. The significance of effects was checked at a probability level of p≤0.05. In cases where significant differences existed, the means were compared using the Tukey’s honestly significant difference (HSD) test at the 5% significance level. The statistical analysis was done using SPSS software.

RESULTS AND DISCUSSION

Identification
 
Isolates identified as Fusarium culmorum typically show rapid colony growth, with aerial mycelium often starting whitish and developing grayish-rose to reddish/burgundy pigmentation and the formation of reddish-brown sporodochia. Microscopic examination generally highlights abundant macroconidia (commonly 3-5 septa).
 
Evaluation of FHB on wheat
 
Disease severity
 
Disease severity was significantly affected by inoculum concentration (p<0.001) and cultivar (p = 0.001), while the interaction was not significant (p = 0.438). At the main-effect level, disease severity increased across inoculum levels from 2.00% (control) to 55.53% (106), demonstrating a clear dose-response relationship between inoculum concentration and disease development (Table 2). Across varieties, the severity ranged from 17.75% (Vitron) to 39.50% (MBB) (Table 3). At 106 spores mL-1, MBB shows the highest severity, followed by Oued-elbared, then Boutaleb and Aoures, while Vitron remains the lowest (Fig 1).

Table 2: Severity (%), FDG (%), NGS, GWS (g), TGW (g) of different treatment (0 (control), 10², 104 and 106 spores mL-1).



Table 3: Severity (%), FDG (%), NGS, GWS (g), TGW (g) of five durum wheat genotypes.



Fig 1: FHB severity % (A), FDG % (B) of five durum wheat genotypes inoculated with different concentrations of Fusarium culmorum under field conditions.


 
Fusarium-damaged grains (FDG)
 
The percentage of FDG increased significantly with increasing inoculum concentration for all genotypes (p≤0.05). At the highest concentration (106 spores mL-1), severe kernel discoloration and shriveling were observed 52.15% (Table 2). Among the tested genotypes, Vitron consistently exhibited the lowest FDG percentages across all inoculum levels, whereas MBB showed the highest kernel damage, particularly at the highest inoculum concentration. The remaining genotypes displayed intermediate responses (Table 3; Fig 1).
 
Yield- component
 
Inoculation with Fusarium culmorum significantly affected all measured yield-related parameters, including number of grains per spike, grain weight per spike and thousand grains weight (TGW) (p≤0.05) (Table 2). At 10v  spores mL-1, a pronounced reduction in TGW was recorded for all genotypes, with losses exceeding 50% in the most susceptible cultivars. In contrast, Vitron exhibited the lowest reduction in TGW, indicating a higher level of tolerance to FHB infection. MBB showed the highest TGW loss at the highest inoculum concentration, reflecting its greater sensitivity to the pathogen (Fig 2).

Fig 2: TGW loss (%) of five durum wheat genotypes inoculated with different concentrations of Fusarium culmorum under field conditions.


       
Analysis of the isolates showed a high prevalence of Fusarium culmorum, which may be explained by several environmental and agronomic factors specific to the region, including climatic conditions that favor the development of this species, particularly elevated temperatures and sufficiently high relative humidity (Kherroubi and Medjdoub-Bensaad, 2025). Indeed, F. culmorum typically thrives under warm and humid conditions, which enhances its ability to cause Fusarium head blight (FHB) (Dill-Macky and Jones, 2000). Most of the isolates characterized in this study belonged to Fusarium culmorum, in agreement with previous investigations conducted in Algeria reporting the predominance of this species in durum wheat grains exhibiting FHB symptoms (Yekkour et al., 2015; Touati-Hattab et al., 2016; Laraba et al., 2017; Abdallah-Nekache et al., 2019). Kant et al., (2011) considered F. culmorum to be one of the most important wheat pathogens due to its capacity to infect multiple cereal tissues. Globally, this pathogen has been reported as predominant in Canada and in northern, central and western Europe (Demeke et al., 2005; Wagacha and Muthomi, 2007), as well as in western Germany and the Rhineland region (Muthomi et al., 2000; Lieneman, 2002).
       
The present study provides a field-based evaluation of the susceptibility of selected durum wheat genotypes to Fusarium culmorum under the agro-climatic conditions of southwestern Algeria. The results clearly demonstrated that disease severity, FDG and yield losses increased significantly with increasing inoculum concentration, confirming the strong pathogenic potential of F. culmorum and its dose-dependent effect on FHB development. The observed increase in FHB severity with higher inoculum concentrations is consistent with previous studies reporting a close relationship between pathogen pressure and disease expression (De Wolf et al., 2003; Stein et al., 2009; Xu and Nicholson, 2009; Francesconi et al., 2019; Gyorgy et al., 2020). Artificial inoculation at anthesis proved to be effective in inducing disease symptoms, highlighting the vulnerability of durum wheat during the flowering stage, which is generally recognized as the key stage for FHB infection (Scherm et al., 2012; Alisaac and Mahlein, 2023). These findings support the relevance of field inoculation assays for evaluating varietal response under realistic agronomic conditions. Significant differences among genotypes across all inoculum concentrations indicate the existence of genetic variability in susceptibility to F. culmorum. Among the tested cultivars, Vitron consistently exhibited lower disease severity, reduced kernel damage and smaller TGW losses, suggesting a relatively higher level of tolerance. In contrast, Oued-elbared and MBB showed higher susceptibility, particularly at the highest inoculum concentration, as reflected by elevated FHB severity, increased FDG percentages and substantial reductions in TGW.
       
Yield-related traits were significantly affected by F. culmorum infection, with reductions in grain number, grain weight per spike and TGW becoming more pronounced at higher inoculum concentrations. These reductions can be attributed to impaired grain filling and premature kernel abortion caused by fungal colonization of spike tissues (Scherm et al., 2012; Alisaac and Mahlein, 2023; Ma et al., 2025). The lower TGW losses recorded for Vitron further support its classification as a more tolerant genotype, whereas the severe TGW losses observed in MBB reflect its greater sensitivity to FHB. The significant genotype ´  inoculum concentration interaction highlights the importance of evaluating varietal performance under different levels of pathogen pressure. This interaction suggests that cultivar ranking may change depending on disease intensity, emphasizing the need for multi-level inoculation assays in resistance screening programs.
       
Overall, this study confirms the high aggressiveness of Fusarium culmorum under field conditions and underscores the necessity of integrating varietal resistance into FHB management strategies. Further research involving multi-location trials and the assessment of mycotoxin contamination would be essential to validate the stability of genotype responses and to fully investigate the effects of FHB on grain quality and safety.

CONCLUSION

Among the assessment cultivars, Vitron showed a comparatively higher level of tolerance, whereas MBB was identified as highly susceptible. These findings emphasize the importance of varietal choice as a key component of integrated FHB management in Algeria. The results provide valuable information for breeding programs and support the deployment of tolerant genotypes as a practical and sustainable strategy to mitigate Fusarium head blight and its associated yield and quality losses in Algerian durum wheat production systems.

ACKNOWLEDGEMENT

The present study was supported by Laboratory of Applied Microbiology, Ferhat ABBAS University - Setif 1, Algeria and National Institute of Agronomic Research of Adrar, Algeria.
 
Disclaimers
 
The views and conclusions expressed in this article are solely those of the authors and do not necessarily represent the views of their affiliated institutions. The authors are responsible for the accuracy and completeness of the information provided, but do not accept any liability for any direct or indirect losses resulting from the use of this content.

CONFLICT OF INTEREST

The authors declare that there are no conflicts of interest regarding the publication of this article. No funding or sponsorship influenced the design of the study, data collection, analysis, decision to publish, or preparation of the manuscript.

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    Full Research Article

    Susceptibility of Durum Wheat (Triticum durum Desf.) to Fusarium Head Blight Caused by Fusarium culmorum under Field Conditions

    S
    Saliha Rahmani1,2,*
    A
    Abdelkrim Mebarkia1
    A
    Ahmad Boulal2
    S
    Said Boudeffeur3
    1Department of Agronomic Sciences, Faculty of Nature and Life Sciences, Laboratory of Applied Microbiology, Ferhat ABBAS University - Setif 1, 19000, Algeria.
    2Department of biology, Faculty of Nature and Life Sciences, Ahmed Draïa University, Adrar, 01000, Algeria.
    3National Institute of Agronomic Research of Adrar, 01000, Algeria.

    ABSTRACT

    Background: Durum wheat (Triticum durum Desf.) represents a key cereal species cultivated across Mediterranean environments. Despite its agronomic importance, this crop remains highly vulnerable to Fusarium head blight (FHB), a disease mainly attributed to Fusarium culmorum, which can severely losses yields and adversely affect grain quality.

    Methods: This study evaluated the susceptibility of five durum wheat varieties (Boutaleb, Mohamed Ben Bachir, Vitron, Oued-elbared and Aoures) to Fusarium culmorum under field conditions in southwestern Algeria during the 2020/2021 cropping season. Inoculations were performed at the beginning of flowering (Zadoks GS 60) using three spore concentrations (102, 104 and 106  spores mL-1). Disease severity, percentage of Fusarium-damaged grains and yield-related parameters were assessed.

    Result: Results showed significant differences among wheat varieties and inoculum concentrations. Disease severity and grain damage increased with increasing spore concentration. The highest severity (74%) and grain damage were recorded in the variety Mohamed Ben Bachir, while Vitron showed comparatively lower susceptibility with a severity of 34% and only 10% damaged grains at the highest inoculum level. The study highlights considerable variability in susceptibility among durum wheat genotypes. The variety Vitron showed relatively better tolerance to Fusarium culmorum, suggesting its potential use in breeding strategies focused on enhancing resistance to Fusarium head blight.

    INTRODUCTION

    Durum wheat (Triticum durum Desf.) is widely recognized as a major cereal crop of global economic importance, playing a central role in global food security and agro-industrial systems in the Mediterranean and North African countries (Royo et al., 2009; Tidiane et al., 2019). However, wheat cultivation in Algeria is frequently challenged by numerous biotic stresses, among which fungal pathogens are considered as the main cause for the yield and quality loss.
           
    Among these diseases, Fusarium head blight (FHB) is identified as the most devastating diseases of wheat in the world (Dweba et al., 2017). This disease affects wheat spikes during the flowering stage and can cause severe yield losses as well as significant deterioration in grain quality (Moreno-Amores et al., 2020; Kaur et al., 2026).
           
    FHB is a destructive disease associated with several species of the genus Fusarium (Malik and Verma, 2026). Among them, Fusarium culmorum and Fusarium graminearum are the most commonly reported pathogens in many wheat-growing regions (Bouanaka et al., 2021; Saharan, 2020). These fungi infect wheat spikes at anthesis, resulting in premature bleaching of spikelets, shriveled kernels and significant reductions in thousand-grain weight (Bilska et al., 2018). Moreover, Fusarium culmorum produces type B trichothecene mycotoxins, including deoxynivalenol (DON), which pose a health risk to humans and animals (Yekkour et al., 2015; Bilska et al., 2018).
           
    In Algeria, different studies have consistently reported Fusarium culmorum as the dominant species associated with FHB on durum wheat (Yekkour et al., 2015; Touati-Hattab et al., 2016; Laraba et al., 2017). Field-based investigations have revealed substantial variability in both the aggressiveness of Fusarium culmorum isolates and the susceptibility of durum wheat genotypes under local agro-climatic conditions (Hadjout et al., 2022).
           
    The management of FHB relies on integrated strategies combining agronomic practices, chemical control and host resistance. Among these approaches, breeding and cultivating resistant wheat varieties are considered the most efficient, cost-effective and environmentally friendly technique to mitigate disease development and reduce mycotoxin accumulation in the grains (Saharan, 2020; Ghimire et al., 2020). However, the success of resistance breeding programs depends largely on accurate and reliable phenotyping under field conditions, where environmental factors strongly influence disease development and expression. Hence, resistance to FHB is regarded to be quantitative trait governed by both genetic and environmental conditions, making its evaluation under field conditions particularly relevant.
           
    In this context, the present study aims to assess the susceptibility of five durum wheat genotypes to Fusarium culmorum under field conditions in southwestern Algeria. By using artificial inoculation and evaluating disease severity and yield-related parameters, this work seeks to provide reliable data on genotype performance under realistic agronomic conditions. The results are expected to enhance current knowledge on FHB epidemiology and to support breeding and crop protection strategies for durum wheat in Algeria and similar Mediterranean regions.

    MATERIALS AND METHODS

    Fungal material
     
    Isolation of Fusarium culmorum
     
    Symptomatic wheat samples were randomly collected during the 2019/2020 agricultural season from several farms located in southwest Algeria. Contaminated stem, leaf and seed fragments underwent surface disinfection using 2% sodium hypochlorite solution for five minutes, followed by three rinses with sterile distilled water. The sterilized tissues were then dried at room temperature on sterile Whatman paper. Five fragments or seeds were inoculated per Petri dish containing Potato Dextrose Agar (PDA) medium and incubated at 28±2oC for seven days. Pure fungal isolates were obtained through repeated subculturing on PDA.
     
    Morphological identification
     
    Isolates were identified through macroscopic observation of colony appearance and microscopic examination of mycelium and spore morphology. Identification was conducted utilizing the taxonomic keys provided by Leslie and Summerell (2006) and Xia et al., (2019).
     
    Molecular characterization
     
    The identification was confirmed through the use of molecular identification. Genomic DNA was extracted using the commercial NucleoSpin Plant II kit (Macherey-Nagel, Germany). The primers used were ITS1 and ITS4. PCR products were separated by electrophoresis in a 1.5% agarose gel, using a 10 μL deposit of the amplification products. Gel migration was followed by staining in an ethidium bromide bath (0.5 μg/mL) and DNA bands were visualized and captured using the Gel Doc system (Bio-Rad, USA). After purification, PCR products were sequenced according to the Sanger procedure (Sanger et al., 1977) using the BigDye v3.1 kit (Applied Biosystems) and the original primers. The resulting sequences were edited and cleaned using CHROMAS PRO software. The products were then compared using the BLAST tool.
     
    Plant material
     
    Five durum wheat (Triticum durum Desf.) genotypes widely cultivated in Algeria were used in this study: Boutaleb, Mohamed Ben Bachir (MBB), Vitron, Oued-elbared and Aoures. All genotypes were provided by the Technical institute of field crops (TIFC), Algeria. The name, pedigree and origin of durum wheat genotypes are presented in Table 1.

    Table 1: Name, pedigree and origin of durum wheat varieties.


     
    Study site and crop management
     
    The study was carried out during the 2020 / 2021 cropping season at the INRA research station in Adrar, Algeria (27o50’N, 0o18’W). The soil profile is medium-deep with a sandy-clay-loam texture. The region features arid climate with a mean annual rainfall of approximately 15 mm. Temperature data indicates a mean minimum of 12oC in winter (December-January) and a maximum of 40oC in summer (July) (INRA Adrar, 2019). Sowing was performed manually during the autumn season (November-December) at a seeding rate of 250 seeds m-2, with a row spacing of 25 cm. Weed management was carried out manually at the herbaceous tillering stage. Nitrogen fertilization consisted of 46% urea applied at a rate of 150 kg ha-1, split into two equal doses at the tillering and stem elongation stages. Flood irrigation was used on the crop, twice to three times a week.
     
    Inoculum preparation
     
    The inoculum was prepared following to the protocol of Siou et al., (2014), with slight modifications. A preserved isolate of Fusarium culmorum was sub-cultured on PDA plates and incubated at 28±2oC for seven days to obtain actively growing cultures. Three agar discs were excised from the margins of colony and transferred into 100 mL of liquid medium consisting of potato extract and dextrose. The cultures were maintained under continuous agitation for five days to enhance sporulation. The conidial suspension was then filtered using fine cheesecloth to remove mycelial fragments. Spore concentration was determined using a Malassez hemocytometer and adjusted with sterile distilled water to final concentrations of 102, 104  and 106 spores mL-1. The prepared conidial inoculum was stored at 4oC for inoculation in the field.
     
    Experimental design and field inoculation
     
    A randomized complete block design was used with three replications. Each cultivar was subjected to four treatments: Three inoculum concentrations (10², 104 and 106 spores mL-1) and a non-inoculated control. Artificial inoculation was carried out at the flowering stage (Zadoks, 65). Spore suspensions of each concentration were applied by spraying directly onto both sides of 10 plots of each wheat variety using a hand-held sprayer. Control plots were not inoculated. To promote infection, the spikes were covered with transparent polyethylene bag for 72 h and inoculated plants were irrigated immediately after inoculation to maintain high humidity levels, favorable for fungal development.
     
    Disease assessment and yield analysis
     
    FHB severity was evaluated 21 days after inoculation. Disease severity was expressed as the percentage of infected spikelets per spike. In addition, the rate of Fusarium-damaged grains (FDG %) was assessed after harvest by visually separating shriveled, discolored, or lightweight kernels from healthy ones. Yield-related parameters were also measured, including the number of grains per spike (NGS), grain weight per spike [GWS (g)] and thousand grain weight [TGW (g)].
     
    Statistical analysis
     
    The experimental design utilized a randomized complete block design (RCBD) with three replications. The analyzed variables were: genotype (five durum wheat varieties), treatment (0 (control), 10², 104 and 106  spores mL-1) and their interaction. Two-way analysis of variance (ANOVA) was utilized in determining the main effects of genotype and inoculum concentration, as well as their interaction, on disease severity, FDG percentage and yield components. The significance of effects was checked at a probability level of p≤0.05. In cases where significant differences existed, the means were compared using the Tukey’s honestly significant difference (HSD) test at the 5% significance level. The statistical analysis was done using SPSS software.

    RESULTS AND DISCUSSION

    Identification
     
    Isolates identified as Fusarium culmorum typically show rapid colony growth, with aerial mycelium often starting whitish and developing grayish-rose to reddish/burgundy pigmentation and the formation of reddish-brown sporodochia. Microscopic examination generally highlights abundant macroconidia (commonly 3-5 septa).
     
    Evaluation of FHB on wheat
     
    Disease severity
     
    Disease severity was significantly affected by inoculum concentration (p<0.001) and cultivar (p = 0.001), while the interaction was not significant (p = 0.438). At the main-effect level, disease severity increased across inoculum levels from 2.00% (control) to 55.53% (106), demonstrating a clear dose-response relationship between inoculum concentration and disease development (Table 2). Across varieties, the severity ranged from 17.75% (Vitron) to 39.50% (MBB) (Table 3). At 106 spores mL-1, MBB shows the highest severity, followed by Oued-elbared, then Boutaleb and Aoures, while Vitron remains the lowest (Fig 1).

    Table 2: Severity (%), FDG (%), NGS, GWS (g), TGW (g) of different treatment (0 (control), 10², 104 and 106 spores mL-1).



    Table 3: Severity (%), FDG (%), NGS, GWS (g), TGW (g) of five durum wheat genotypes.



    Fig 1: FHB severity % (A), FDG % (B) of five durum wheat genotypes inoculated with different concentrations of Fusarium culmorum under field conditions.


     
    Fusarium-damaged grains (FDG)
     
    The percentage of FDG increased significantly with increasing inoculum concentration for all genotypes (p≤0.05). At the highest concentration (106 spores mL-1), severe kernel discoloration and shriveling were observed 52.15% (Table 2). Among the tested genotypes, Vitron consistently exhibited the lowest FDG percentages across all inoculum levels, whereas MBB showed the highest kernel damage, particularly at the highest inoculum concentration. The remaining genotypes displayed intermediate responses (Table 3; Fig 1).
     
    Yield- component
     
    Inoculation with Fusarium culmorum significantly affected all measured yield-related parameters, including number of grains per spike, grain weight per spike and thousand grains weight (TGW) (p≤0.05) (Table 2). At 10v  spores mL-1, a pronounced reduction in TGW was recorded for all genotypes, with losses exceeding 50% in the most susceptible cultivars. In contrast, Vitron exhibited the lowest reduction in TGW, indicating a higher level of tolerance to FHB infection. MBB showed the highest TGW loss at the highest inoculum concentration, reflecting its greater sensitivity to the pathogen (Fig 2).

    Fig 2: TGW loss (%) of five durum wheat genotypes inoculated with different concentrations of Fusarium culmorum under field conditions.


           
    Analysis of the isolates showed a high prevalence of Fusarium culmorum, which may be explained by several environmental and agronomic factors specific to the region, including climatic conditions that favor the development of this species, particularly elevated temperatures and sufficiently high relative humidity (Kherroubi and Medjdoub-Bensaad, 2025). Indeed, F. culmorum typically thrives under warm and humid conditions, which enhances its ability to cause Fusarium head blight (FHB) (Dill-Macky and Jones, 2000). Most of the isolates characterized in this study belonged to Fusarium culmorum, in agreement with previous investigations conducted in Algeria reporting the predominance of this species in durum wheat grains exhibiting FHB symptoms (Yekkour et al., 2015; Touati-Hattab et al., 2016; Laraba et al., 2017; Abdallah-Nekache et al., 2019). Kant et al., (2011) considered F. culmorum to be one of the most important wheat pathogens due to its capacity to infect multiple cereal tissues. Globally, this pathogen has been reported as predominant in Canada and in northern, central and western Europe (Demeke et al., 2005; Wagacha and Muthomi, 2007), as well as in western Germany and the Rhineland region (Muthomi et al., 2000; Lieneman, 2002).
           
    The present study provides a field-based evaluation of the susceptibility of selected durum wheat genotypes to Fusarium culmorum under the agro-climatic conditions of southwestern Algeria. The results clearly demonstrated that disease severity, FDG and yield losses increased significantly with increasing inoculum concentration, confirming the strong pathogenic potential of F. culmorum and its dose-dependent effect on FHB development. The observed increase in FHB severity with higher inoculum concentrations is consistent with previous studies reporting a close relationship between pathogen pressure and disease expression (De Wolf et al., 2003; Stein et al., 2009; Xu and Nicholson, 2009; Francesconi et al., 2019; Gyorgy et al., 2020). Artificial inoculation at anthesis proved to be effective in inducing disease symptoms, highlighting the vulnerability of durum wheat during the flowering stage, which is generally recognized as the key stage for FHB infection (Scherm et al., 2012; Alisaac and Mahlein, 2023). These findings support the relevance of field inoculation assays for evaluating varietal response under realistic agronomic conditions. Significant differences among genotypes across all inoculum concentrations indicate the existence of genetic variability in susceptibility to F. culmorum. Among the tested cultivars, Vitron consistently exhibited lower disease severity, reduced kernel damage and smaller TGW losses, suggesting a relatively higher level of tolerance. In contrast, Oued-elbared and MBB showed higher susceptibility, particularly at the highest inoculum concentration, as reflected by elevated FHB severity, increased FDG percentages and substantial reductions in TGW.
           
    Yield-related traits were significantly affected by F. culmorum infection, with reductions in grain number, grain weight per spike and TGW becoming more pronounced at higher inoculum concentrations. These reductions can be attributed to impaired grain filling and premature kernel abortion caused by fungal colonization of spike tissues (Scherm et al., 2012; Alisaac and Mahlein, 2023; Ma et al., 2025). The lower TGW losses recorded for Vitron further support its classification as a more tolerant genotype, whereas the severe TGW losses observed in MBB reflect its greater sensitivity to FHB. The significant genotype ´  inoculum concentration interaction highlights the importance of evaluating varietal performance under different levels of pathogen pressure. This interaction suggests that cultivar ranking may change depending on disease intensity, emphasizing the need for multi-level inoculation assays in resistance screening programs.
           
    Overall, this study confirms the high aggressiveness of Fusarium culmorum under field conditions and underscores the necessity of integrating varietal resistance into FHB management strategies. Further research involving multi-location trials and the assessment of mycotoxin contamination would be essential to validate the stability of genotype responses and to fully investigate the effects of FHB on grain quality and safety.

    CONCLUSION

    Among the assessment cultivars, Vitron showed a comparatively higher level of tolerance, whereas MBB was identified as highly susceptible. These findings emphasize the importance of varietal choice as a key component of integrated FHB management in Algeria. The results provide valuable information for breeding programs and support the deployment of tolerant genotypes as a practical and sustainable strategy to mitigate Fusarium head blight and its associated yield and quality losses in Algerian durum wheat production systems.

    ACKNOWLEDGEMENT

    The present study was supported by Laboratory of Applied Microbiology, Ferhat ABBAS University - Setif 1, Algeria and National Institute of Agronomic Research of Adrar, Algeria.
     
    Disclaimers
     
    The views and conclusions expressed in this article are solely those of the authors and do not necessarily represent the views of their affiliated institutions. The authors are responsible for the accuracy and completeness of the information provided, but do not accept any liability for any direct or indirect losses resulting from the use of this content.

    CONFLICT OF INTEREST

    The authors declare that there are no conflicts of interest regarding the publication of this article. No funding or sponsorship influenced the design of the study, data collection, analysis, decision to publish, or preparation of the manuscript.

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