Biocontrol Potential of Aspergillus Species Producing Antimicrobial Metabolites

Microbial metabolites have been recognized as an important source for the discovery of new antifungal agents because of their diverse chemical structures with novel modes of action. In the course of our screening for new antifungal agents from microbes, we found that culture filtrates of two fungal species Aspergillus candidus SFC20200425-M11 and Aspergillus montenegroi SFC20200425-M27 have the potentials to reduce the development of fungal plant diseases such as tomato late blight and wheat leaf rust. From these two Aspergillus spp., we isolated a total of seven active compounds, including two new compounds (4 and 6), and identified their chemical structures based on the NMR spectral analyses: sphaeropsidin A (1), (R)-formosusin A (2), (R)-variotin (3), candidusin (4), asperlin (5), montenegrol (6), and protulactone A (7). Based on the results of the in vitro bioassays of 11 plant pathogenic fungi and bacteria, sphaeropsidin A (1), (R)-formosusin A (2), (R)-variotin (3), and asperlin (5) exhibited a wide range of antimicrobial activity. Furthermore, when plants were treated with sphaeropsidin A (1) and (R)-formosusin A (2) at a concentration of 500 μg/ml, sphaeropsidin A (1) exhibited an efficacy disease control value of 96 and 90% compared to non-treated control against tomato late blight and wheat leaf rust, and (R)-formosusin A (2) strongly reduced the development of tomato gray mold by 82%. Asperlin (5) at a concentration of 500 μg/ml effectively controlled the development of tomato late blight and wheat leaf rust with a disease control value of 95%. Given that culture filtrates and active compounds derived from two Aspergillus spp. exhibited disease control efficacies, our results suggest that the Aspergillus-produced antifungal compounds could be useful for the development of new natural fungicides.

Wheat Leaf Antioxidative Status—Variety-Specific Mechanisms of Zinc Tolerance during Biofortification

In this study, we evaluated the leaf antioxidative responses of three wheat varieties (Srpanjka, Divana, and Simonida) treated with two different forms of zinc (Zn), Zn-sulfate and Zn-EDTA, in concentrations commonly used in agronomic biofortification. Zn concentration was significantly higher in the flag leaves of all three wheat varieties treated with Zn-EDTA compared to control and leaves treated with Zn-sulfate. Both forms of Zn increased malondialdehyde level and total phenolics content in varieties Srpanjka and Divana. Total glutathione content was not affected after the Zn treatment. Zn-sulfate increased the activities of glutathione reductase (GR) and guaiacol peroxidase (GPOD) in both Srpanjka and Divana, while glutathione S-transferase (GST) was only induced in var. Srpanjka. Chelate form of Zn increased the activities of GST and GPOD in both Simonida and Divana. Catalase activity was shown to be less sensitive to Zn treatment and was only induced in var. Srpanjka treated with Zn-EDTA where GPOD activity was not induced. Concentrations of Zn used for agronomic biofortification can induce oxidative stress in wheat leaves. The antioxidative status of wheat leaves could be a good indicator of Zn tolerance, whereas wheat genotype and chemical form of Zn are the most critical factors influencing Zn toxicity.

The Emergence of New Aggressive Leaf Rust Races with the Potential to Supplant the Resistance of Wheat Cultivars

Characterization of the genetic structure and the physiological races of Puccinia triticina is a growing necessity to apply host genetic resistance against wheat leaf rust as a successful control strategy. Herein, we collected and identified about 130 isolates of P. triticina from 16 Egyptian commercial wheat cultivars grown at different locations, over two seasons (2019/2020 and 2020/2021). The 130 isolates of P. triticina were segregated into 17 different physiological races. TTTST and TTTKS were the most common virulent races, whereas TTTST and MTTGT were the most frequent races. The races were classified into three groups, based on their distinct DNA band sizes (150 bp, 200 bp, and 300 bp) after RAPD analysis. The new wheat cultivars (Sakha-94, Sakha-95, and Shandweel-1) infected with the most virulent race (TTTST), Gemmeiza-12, and Misr-3 were resistant to all physiological races. The resistance of these cultivars was mostly due to the presence of Lr19- and Lr28-resistant genes. Our results serve as a warning about emerging aggressive races capable of supplanting resistance to leaf rust, and help in the understanding of the pathotype–cultivar–location association and its role in the susceptibility/resistance of new wheat cultivars to P. triticina.

Impact of Crop Rotation and Soil Tillage on the Severity of Winter Wheat Leaf Blotches

Reduced tillage is considered as one of the main tools to save biological diversity; however, it increases pressure of diseases, including wheat leaf blotches. The aim of present study was to clarify the impact of reduced soil tillage on the development of winter wheat leaf blotches in different schemes of crop rotation. The impact of different growing technologies on the severity of winter wheat disease was evaluated in a two-factorial experiment: A – soil tillage system, and B – different combinations of wheat pre-pre-crop and pre-crop (wheat, oilseed rape, barley and faba beans). Diseases were assessed every 10 days approximately and total impact of diseases was evaluated by calculating AUDPC (Area under Diseases Progress Curve). Tan spot, caused by Pyrenophora tritici-repentis was dominated disease over the long period, regardless of meteorological conditions. Development of Septoria leaf blotch was not influenced by neither crop rotation nor soil tillage. The level of tan spot was essentially higher after wheat, regardless of the pre-pre-crop. Reduced soil tillage promoted severity of tan spot. Ploughing mitigated effect of previous crops and differences in tan spot level were insignificant. Despite many positive effects of conservation tillage, increasing of fungicide treatment could be necessary, in conditions, when the tan spot is most devastating and widespread wheat disease.

A Chromosome-Scale Assembly of the Wheat Leaf Rust Pathogen Puccinia triticina Provides Insights Into Structural Variations and Genetic Relationships With Haplotype Resolution

Despite the global economic importance of the wheat leaf rust pathogen Puccinia triticina (Pt), genomic resources for Pt are limited and chromosome-level assemblies of Pt are lacking. Here, we present a complete haplotype-resolved genome assembly at a chromosome-scale for Pt using the Australian pathotype 64-(6),(7),(10),11 (Pt64; North American race LBBQB) built upon the newly developed technologies of PacBio and Hi-C sequencing. PacBio reads with ∼200-fold coverage (29.8 Gb data) were assembled by Falcon and Falcon-unzip and subsequently scaffolded with Hi-C data using Falcon-phase and Proximo. This approach allowed us to construct 18 chromosome pseudomolecules ranging from 3.5 to 12.3 Mb in size for each haplotype of the dikaryotic genome of Pt64. Each haplotype had a total length of ∼147 Mb, scaffold N50 of ∼9.4 Mb, and was ∼93% complete for BUSCOs. Each haplotype had ∼29,800 predicted genes, of which ∼2,000 were predicted as secreted proteins (SPs). The investigation of structural variants (SVs) between haplotypes A and B revealed that 10% of the total genome was spanned by SVs, highlighting variations previously undetected by short-read based assemblies. For the first time, the mating type (MAT) genes on each haplotype of Pt64 were identified, which showed that MAT loci a and b are located on two chromosomes (chromosomes 7 and 14), representing a tetrapolar type. Furthermore, the Pt64 assembly enabled haplotype-based evolutionary analyses for 21 Australian Pt isolates, which highlighted the importance of a haplotype resolved reference when inferring genetic relationships using whole genome SNPs. This Pt64 assembly at chromosome-scale with full phase information provides an invaluable resource for genomic and evolutionary research, which will accelerate the understanding of molecular mechanisms underlying Pt-wheat interactions and facilitate the development of durable resistance to leaf rust in wheat and sustainable control of rust disease.