Resistance of the Wheat Cultivar ‘Renan’ to Septoria Leaf Blotch Explained by a Combination of Strain Specific and Strain Non-Specific QTL Mapped on an Ultra-Dense Genetic Map

Quantitative resistance is considered more durable than qualitative resistance as it does not involve major resistance genes that can be easily overcome by pathogen populations, but rather a combination of genes with a lower individual effect. This durability means that quantitative resistance could be an interesting tool for breeding crops that would not systematically require phytosanitary products. Quantitative resistance has yet to reveal all of its intricacies. Here, we delve into the case of the wheat/Septoria tritici blotch (STB) pathosystem. Using a population resulting from a cross between French cultivar Renan, generally resistant to STB, and Chinese Spring, a cultivar susceptible to the disease, we built an ultra-dense genetic map that carries 148,820 single nucleotide polymorphism (SNP) markers. Phenotyping the interaction was done with two different Zymoseptoria tritici strains with contrasted pathogenicities on Renan. A linkage analysis led to the detection of three quantitative trait loci (QTL) related to resistance in Renan. These QTL, on chromosomes 7B, 1D, and 5D, present with an interesting diversity as that on 7B was detected with both fungal strains, while those on 1D and 5D were strain-specific. The resistance on 7B was located in the region of Stb8 and the resistance on 1D colocalized with Stb19. However, the resistance on 5D was new, so further designated Stb20q. Several wall-associated kinases (WAK), nucleotide-binding and leucine-rich repeats (NB-LRR) type, and kinase domain carrying genes were present in the QTL regions, and some of them were expressed during the infection. These results advocate for a role of Stb genes in quantitative resistance and for resistance in the wheat/STB pathosystem being as a whole quantitative and polygenic.

Isolation and Identification of Lipopeptide-Producing Bacillus velezensis Strains from Wheat Phyllosphere with Antifungal Activity against the Wheat Pathogen Zymoseptoria tritici

Septoria tritici blotch, caused by the fungal pathogen Zymoseptoria tritici, is a highly significant disease on wheat crops worldwide. The objective of the present study was to find out new bacterial strains with bio-antimicrobial activity against Z. tritici. Two phyllospheric bacteria (S1 and S6) were isolated from wheat ears and identified as Bacillus velezensis strains according to 16S rRNA Sanger sequencing. Antagonistic assays performed with either living strains or cell-free culture filtrates showed significant in vitro antifungal activities against Z. tritici. For the culture filtrates, the half-maximal inhibitory dilution and the minimal inhibitory dilution were 1.4% and 3.7% for the strain S1, and 7.4% and 15% for the strain S6, respectively. MALDI—ToF analysis revealed that both strains synthesize cyclic lipopeptides but from different families. Interestingly, only strain S1 produces putative bacillomycin D. Such differential lipopeptide production patterns might explain the difference observed between the antifungal activity of the culture filtrates of the two strains. This study allows the identification of new lipopeptide-producing strains of B. velezensis with a high potential of application for the biocontrol of Z. tritici.

Two Novel Dimorphism-Related Virulence Factors of Zymoseptoria tritici Identified Using Agrobacterium-Mediated Insertional Mutagenesis

Diseases caused by dimorphic phytopathogenic and systemic dimorphic fungi have markedly increased in prevalence in the last decades, and understanding the morphogenic transition to the virulent state might yield novel means of controlling dimorphic fungi. The dimorphic fungus Z. tritici causes significant economic impact on wheat production, and yet the regulation of the dimorphic switch, a key first step in successful plant colonization, is still largely unexplored in this fungus. The fungus is amenable to suppression by fungicides at this switch point, and the identification of the factors controlling the dimorphic switch provides a potential source of novel targets to control Septoria tritici blotch (STB). Inhibition of the dimorphic switch can potentially prevent penetration and avoid any damage to the host plant. The aim of the current work was to unveil genetic determinants of the dimorphic transition in Z. tritici by using a forward genetics strategy. Using this approach, we unveiled two novel factors involved in the switch to the pathogenic state and used reverse genetics and complementation to confirm the role of the novel virulence factors and further gained insight into the role of these genes, using transcriptome analysis via RNA-Seq. The transcriptomes generated potentially contain key determinants of the dimorphic transition.

SNP-based Genetic Structure of Tunisian Durum Wheat Landraces and Distribution of the Resistance to Septoria Tritici Blotch

Background: Septoria tritici blotch (STB) has marked durum wheat production worldwide. This fungal disease is until today a challenge for farmers, researchers and breeders all united in the aim of reducing its damage and improving wheat resistance. Tunisian durum wheat landraces were reported to be valuable genetic resources for resistance to biotic and abiotic stresses and are therefore prominently deployed in breeding programs to develop new varieties adapted to fungal diseases as STB and to climate change constraints overall.Results: A total of 366 local durum wheat accessions were assessed for resistance to two virulent Tunisian isolates of Zymoseptoria tritici Tun06 and TM220 under field conditions. Population structure analysis of the durum wheat accessions, performed with 286 polymorphic SNPs (PIC >0.3) covering the entire genome, identified three genetic subpopulations (GS1, GS2 and GS3) with 22% of admixed genotypes. Interestingly, all of the resistant genotypes were among GS2 or admixed with GS2. Conclusions: This study revealed the population structure and the genetic distribution of the resistance to Z. tritici in the Tunisian durum wheat landraces. The grouping pattern of accessions appear to be associated, to some extent, with the geographical pattern of the landraces. We suggested that GS2 accessions were mostly introduced from eastern Mediterranean populations, unlike GS1 and GS3 that originated from the west. Resistant GS2 accessions belonged to landraces Taganrog, Sbei glabre, Richi, Mekki, Badri, Jneh Khotifa and Azizi. Furthermore, we suggested that admixture contributed to transmit STB resistance from GS2 resistant landraces to initially susceptible landraces such as Mahmoudi (GS1), but also resulted in the loss of resistance in the case of GS2 suscpetible Azizi and Jneh Khotifa accessions.

Management of septoria tritici blotch using cultivar mixtures

Septoria tritici blotch (STB) is among the most devastating diseases in European wheat production. In recent years, there has been increased interest in using cultivar mixtures as part of an integrated control strategy against diseases. This study investigated different cultivar mixtures for their ability to control STB across three years and at seven trial sites in Denmark with a range of fungicide strategies, yielding a total of 194 individual cultivar mixture combinations. The mixtures were composed of two, three or four cultivars which were either similar or contrasting in their susceptibility to STB. Across all trials, the cultivar mixtures reduced disease severity significantly, by 14% compared to the component cultivars grown in monoculture. The reductions were larger when the disease pressure was high and when the mixtures included more cultivars. Mixtures composed of four cultivars reduced disease severity significantly, by 24%. Across all trials, cultivar mixtures significantly increased yield by 2% compared to the component cultivars grown in monoculture. The yield increase was significant for plots treated with one or two fungicide applications, and cultivar mixtures increased yield significantly, by 4.4% in untreated plots. The yield increase was smaller for mixtures with a high proportion of resistant cultivars. Based on the results from this study, cultivar mixtures can contribute positively to an IPM strategy, by reducing disease severity for STB and increasing yield. The most pronounced benefits from cultivar mixtures were found in fields with moderate to low fungicide input, under conditions with high disease pressure, when combining four cultivars with varying susceptibilities.

SNP-Based Genetic Structure of Tunisian Durum Wheat Landraces and Distribution of the Resistance to Septoria Tritici Blotch

Background: Septoria tritici blotch (STB) has marked durum wheat production worldwide. This fungal disease is until today a challenge for farmers, researchers and breeders all united in the aim of reducing its damage and improving wheat resistance. Tunisian durum wheat landraces were reported to be valuable genetic resources for resistance to biotic and abiotic stresses and are therefore prominently deployed in breeding programs to develop new varieties adapted to fungal diseases as STB and to climate change constraints overall.Results: A total of 366 local durum wheat accessions were assessed for resistance to two virulent Tunisian isolates of Zymoseptoria tritici Tun06 and TM220 under field conditions. Population structure analysis of the durum wheat accessions, performed with 286 polymorphic SNPs (PIC >0.3) covering the entire genome, identified three genetic subpopulations (GS1, GS2 and GS3) with 22% of admixed genotypes. Interestingly, all of the resistant genotypes were among GS2 or admixed with GS2. Conclusions: This study revealed the population structure and the genetic distribution of the resistance to Z. tritici in the Tunisian durum wheat landraces. The grouping pattern of accessions appear to be associated, to some extent, with the geographical pattern of the landraces. We suggested that GS2 accessions were mostly introduced from eastern Mediterranean populations, unlike GS1 and GS3 that originated from the west. Resistant GS2 accessions belonged to landraces Taganrog, Sbei glabre, Richi, Mekki, Badri, Jneh Khotifa and Azizi. Furthermore, we suggested that admixture contributed to transmit STB resistance from GS2 resistant landraces to initially susceptible landraces such as Mahmoudi (GS1), but also resulted in the loss of resistance in the case of GS2 suscpetible Azizi and Jneh Khotifa accessions.

Advances in breeding techniques for durable Septoria tritici blotch (STB) resistance in cereals

Septoria tritici blotch (STB), caused by the hemibiotrophic fungus Zymoseptoria tritici, is one of the most important foliar diseases of winter cereal crops. Recent advances are helping to understand the genetic basis and architecture of resistance to STB. To date, at least 22 genes for qualitative resistance and over 200 quantitative trait loci (QTL) for quantitative resistance have been identified in cereals. This knowledge is enabling cereal breeding programs to develop varieties with more durable resistance to STB. This chapter reviews recent research on genetic resistance loci and breeding strategies based on both conventional and biotechnology-based breeding approaches (molecular marker/genomic-assisted breeding, genetic transformation, and gene-editing) to achieve achieving durable resistance to STB infection and minimise grain yield losses.

A Microbial Fermentation Mixture Reduces Fusarium Head Blight and Promotes Grain Weight but does not impact Septoria tritici blotch

Fusarium graminearum and Zymoseptoria tritici cause economically important diseases of wheat. F. graminearum is one of the primary causal agents of Fusarium head blight (FHB) and Z. tritici is the causal agent of Septoria tritici blotch (STB). Alternative control methods are required in the face of fungicide resistance and EU legislation which seek to cut pesticide use by 2030. Both fungal pathogens have been described as either hemibiotrophs or necrotrophs. A microbial fermentation-based product (MFP) was previously demonstrated to control the biotrophic pathogen powdery mildew, on wheat. Here we investigated if MFP would be effective against the non-biotrophic fungal pathogens of wheat, F. graminearum and Z. tritici. We assessed the impact of MFP on fungal growth, disease control and also evaluated the individual constituent parts of MFP. Antifungal activity towards both pathogens was found in vitro but MFP only significantly decreased disease symptoms of FHB in planta. In addition, MFP was found to improve the grain number and weight, of uninfected and F. graminearum infected wheat heads.

Understanding plant-pathogen interactions in Septoria tritici blotch infection of cereals

Zymoseptoria is a major fungal pathogen of wheat, responsible for the Septoria Tritici Blotch (STB) disease. Recently, STB has been the subject of intensive molecular studies. Notably, massive transcriptomic analyses have helped to explore this particular bi-phasic (asymptomatic/necrotrophic) infection process. Cytological analyses have also improved our understanding of the asymptomatic phase. These advances suggest that Zymoseptoria behaves as a hemi-biotrophic fungus, acting like an endophyte during its asymptomatic phase. STB is still difficult to control. The emergence of fungicide-resistant isolates has reduced the efficacy of many fungicides requiring the development of novel fungicides and methods to counteract/reduce fungicide resistance. Likewise, because Stb-resistant wheat cultivars have all been successively defeated by virulent isolates, there is a need to identify new resistance genes in wheat, and to develop better disease resistance management methods (pyramiding, mixture/alternation) to sustainably control this pathogen.