RNA interference (RNAi) is a powerful genetic tool to accelerate research in plant biotechnology and control biotic stresses by manipulating target gene expression. However, the potential of RNAi in wheat to efficiently and durably control the devastating stripe rust fungus Puccinia striiformis f. sp. tritici (Pst) remained largely under explored so far. To address this issue, we generated transgenic wheat (Triticum aestivum) lines expressing dsRNA targeting PsFUZ7 transcripts of Pst. We analyzed expression of PsFUZ7 and related genes, and resistance traits of the transgenic wheat lines. We show that PsFUZ7 is an important pathogenicity factor that regulates infection and development of Pst. A PsFUZ7 RNAi construct stably expressed in two independent transgenic wheat lines confers strong resistance to Pst. Pst hyphal development is strongly restricted, and necrosis of plant cells in resistance responses was significantly induced. We conclude that trafficking of RNA molecules from wheat plants to Pst may lead to a complex molecular dialogue between wheat and the rust pathogen. Moreover, we confirm the RNAi-based crop protection approaches can be used, to our knowledge, as a novel control strategy against rust pathogens in wheat.

Summary: Only few fungal effectors have been described to be delivered into the host cell during obligate biotrophic interactions. RTP1p , from the rust fungi U romyces fabae and U . striatus , was the first fungal protein for which localization within the host cytoplasm could be demonstrated directly. We investigated the occurrence of RTP1 homologues in rust fungi and examined the structural and biochemical characteristics of the corresponding gene products. The analysis of 28 homologues showed that members of the RTP family are most likely to occur ubiquitously in rust fungi and to be specific to the order P ucciniales. Sequence analyses indicated that the structure of the RTPp effectors is bipartite, consisting of a variable N ‐terminus and a conserved and structured C ‐terminus. The characterization of Uf ‐RTP1p mutants showed that four conserved cysteine residues sustain structural stability. Furthermore, the C ‐terminal domain exhibits similarities to that of cysteine protease inhibitors, and it was shown that Uf ‐RTP1p and Us ‐RTP1p are able to inhibit proteolytic activity in P ichia pastoris culture supernatants. We conclude that the RTP1p homologues constitute a rust fungi‐specific family of modular effector proteins comprising an unstructured N ‐terminal domain and a structured C ‐terminal domain, which exhibit protease inhibitory activity possibly associated with effector function during biotrophic interactions.

Bacillus strains can produce various lipopeptides, known for their antifungal properties. This makes them attractive metabolites for applications in agriculture. Therefore, identification of productive wild-type strains is essential for the development of biopesticides. Bacillus velezensis FZB42 is a well-established strain for biocontrol of plant pathogens in agriculture. Here, we characterized an alternative strain, B. velezensis UTB96, that can produce higher amounts of all three major lipopeptide families, namely surfactin, fengycin, and iturin. UTB96 produces iturin A. Furthermore, UTB96 showed superior antifungal activity towards the soybean fungal pathogen Diaporthe longicolla compared to FZB42. Moreover, the additional provision of different amino acids for lipopeptide production in UTB96 was investigated. Lysine and alanine had stimulatory effects on the production of all three lipopeptide families, while supplementation of leucine, valine and isoleucine decreased the lipopeptide bioproduction. Using a 45-litre bioreactor system for upscaling in batch culture, lipopeptide titers of about 140 mg/L surfactin, 620 mg/L iturin A, and 45 mg/L fengycin were achieved. In conclusion, it becomes clear that B. velezensis UTB96 is a promising strain for further research application in the field of agricultural biological controls of fungal diseases.

With >7000 species the order of rust fungi has a disproportionately large impact on agriculture, horticulture, forestry and foreign ecosystems. The infectious spores are typically dikaryotic, a feature unique to fungi in which two haploid nuclei reside in the same cell. A key example is Phakopsora pachyrhizi, the causal agent of Asian soybean rust disease, one of the world’s most economically damaging agricultural diseases. Despite P. pachyrhizi’s impact, the exceptional size and complexity of its genome prevented generation of an accurate genome assembly. Here, we sequence three independent P. pachyrhizi genomes and uncover a genome up to 1.25 Gb comprising two haplotypes with a transposable element (TE) content of ~93%. We study the incursion and dominant impact of these TEs on the genome and show how they have a key impact on various processes such as host range adaptation, stress responses and genetic plasticity.

The search for approaches to a holistic sustainable agriculture requires the development of new cropping systems that provide additional ecosystem services beyond biomass supply for food, feed, material, and energy use. The reduction of chemical synthetic plant protection products is a key instrument to protect vulnerable natural resources such as groundwater and biodiversity. Together with an optimal use of mineral fertilizer, agroecological practices, and precision agriculture technologies, a complete elimination of chemical synthetic plant protection in mineral-ecological cropping systems (MECSs) may not only improve the environmental performance of agroecosystems, but also ensure their yield performance. Therefore, the development of MECSs aims to improve the overall ecosystem services of agricultural landscapes by (i) improving the provision of regulating ecosystem services compared to conventional cropping systems and (ii) improving the supply of provisioning ecosystem services compared to organic cropping systems. In the present review, all relevant research levels and aspects of this new farming concept are outlined and discussed based on a comprehensive literature review and the ongoing research project “Agriculture 4.0 without Chemical-Synthetic Plant Protection”.