Institut für Phytomedizin

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Role of benzoic acid and lettucenin A in the defense response of lettuce against soil-borne pathogens
(2021) Windisch, Saskia; Walter, Anja; Moradtalab, Narges; Walker, Frank; Höglinger, Birgit; El-Hasan, Abbas; Ludewig, Uwe; Neumann, Günter; Grosch, Rita
Soil-borne pathogens can severely limit plant productivity. Induced defense responses are plant strategies to counteract pathogen-related damage and yield loss. In this study, we hypothesized that benzoic acid and lettucenin A are involved as defense compounds against Rhizoctonia solani and Olpidium virulentus in lettuce. To address this hypothesis, we conducted growth chamber experiments using hydroponics, peat culture substrate and soil culture in pots and minirhizotrons. Benzoic acid was identified as root exudate released from lettuce plants upon pathogen infection, with pre-accumulation of benzoic acid esters in the root tissue. The amounts were sufficient to inhibit hyphal growth of R. solani in vitro (30%), to mitigate growth retardation (51%) and damage of fine roots (130%) in lettuce plants caused by R. solani, but were not able to overcome plant growth suppression induced by Olpidium infection. Additionally, lettucenin A was identified as major phytoalexin, with local accumulation in affected plant tissues upon infection with pathogens or chemical elicitation (CuSO4) and detected in trace amounts in root exudates. The results suggest a two-stage defense mechanism with pathogen-induced benzoic acid exudation initially located in the rhizosphere followed by accumulation of lettucenin A locally restricted to affected root and leaf tissues.
Assessment of a postharvest treatment with pyrimethanil via thermo-nebulization in controlling storage rots of apples
(2021) Büchele, Felix; Neuwald, Daniel A.; Scheer, Christian; Wood, Rachael M.; Vögele, Ralf T.; Wünsche, Jens N.
Apples are very susceptible to infections from various fungal pathogens during the growing season due to prolonged exposure to environmental influences in the field. Therefore, a strict and targeted fungicide strategy is essential to protect fruit and trees. Increased environmental and health concerns and pathogen resistance have resulted in a rising demand to reduce fungicide usage and residues on marketed fruit. Thus, producers must develop new plant protection strategies to conform to the legal and social demands while still offering high-quality apples. This study assessed the efficacy of a post-harvest fungicide treatment with pyrimethanil via thermo-nebulization for controlling storage rots and compared the results to those of standard pre-harvest fungicide strategies. The results showed that a single post-harvest application of pyrimethanil successfully controlled storage rots and is comparable to strategies using multiple pre-harvest fungicide applications. The control of fungal rot was sustained even after 5 months of storage and 2 weeks of shelf life. Thermo-nebulization into the storage facility allowed for a lower dosage of fungicide to be used compared to pre-harvest applications, while still maintaining optimal rot control. Residue analyses showed that the post-harvest fungicide treatment did not exceed legal or retailer’s standards.
Exploring the effects of different stubble tillage practices and glyphosate application combined with the new soil residual herbicide cinmethylin against Alopecurus myosuroides Huds. in winter wheat
(2022) Messelhäuser, Miriam Hannah; Saile, Marcus; Sievernich, Bernd; Gerhards, Roland
Effective control of Alopecurus myosuroides Huds. (blackgrass) solely with a chemical treatment is not guaranteed anymore because populations exhibit resistance to almost all herbicide modes of action. Integrated weed management (IWM) against blackgrass is necessary to maintain high weed control efficacies in winter cereals. Four field experiments were conducted in Southwest Germany from 2018 to 2020 to control A. myosuroides with a combination of cultural and chemical methods. Stubble treatments, including flat, deep and inversion soil tillage; false seedbed preparation and glyphosate use, were combined with the application of the new pre-emergence herbicide cinmethylin in two rates in winter wheat. Average densities of A. myosuroides in the untreated control plots were up to 505 plants m−2. The combination of different stubble management strategies and the pre-emergence herbicide cinmethylin controlled 86–97% of A. myosuroides plants at the low rate and 95–100% at the high rate until 120 days after sowing. The different stubble tillage practices varied in their efficacy between trials and years. Most effective and consistent were pre-sowing glyphosate application on the stubble and stale seedbed preparation with a disc harrow. Stubble treatments increased winter wheat density in the first year but had no effect on crop density in the second year. Pre-emergence application of cinmethylin did not reduce winter wheat densities. Multiple tactics of weed control, including stubble treatments and pre-emergence application of cinmethylin, provided higher and more consistent control of A. myosuroides. Integration of cultural weed management could prevent the herbicide resistance development.
Agronomic and technical evaluation of herbicide spot spraying in maize based on high-resolution aerial weed maps - an on-farm trial
(2024) Allmendinger, Alicia; Spaeth, Michael; Saile, Marcus; Peteinatos, Gerassimos G.; Gerhards, Roland; Allmendinger, Alicia; Department of Weed Science, Institute for Phytomedicine, University of Hohenheim, 70599 Stuttgart, Germany; (A.A.);; Spaeth, Michael; Department of Weed Science, Institute for Phytomedicine, University of Hohenheim, 70599 Stuttgart, Germany; (A.A.);; Saile, Marcus; Department of Weed Science, Institute for Phytomedicine, University of Hohenheim, 70599 Stuttgart, Germany; (A.A.);; Peteinatos, Gerassimos G.; ELGO-DIMITRA, Leof Dimokratias 61, Agii Anargiri, 135 61 Athens, Greece;; Gerhards, Roland; Department of Weed Science, Institute for Phytomedicine, University of Hohenheim, 70599 Stuttgart, Germany; (A.A.);; Rossi, Vittorio
Spot spraying can significantly reduce herbicide use while maintaining equal weed control efficacy as a broadcast application of herbicides. Several online spot-spraying systems have been developed, with sensors mounted on the sprayer or by recording the RTK-GNSS position of each crop seed. In this study, spot spraying was realized offline based on georeferenced unmanned aerial vehicle (UAV) images with high spatial resolution. Studies were conducted in four maize fields in Southwestern Germany in 2023. A randomized complete block design was used with seven treatments containing broadcast and spot applications of pre-emergence and post-emergence herbicides. Post-emergence herbicides were applied at 2–4-leaf and at 6–8-leaf stages of maize. Weed and crop density, weed control efficacy (WCE), crop losses, accuracy of weed classification in UAV images, herbicide savings and maize yield were measured and analyzed. On average, 94% of all weed plants were correctly identified in the UAV images with the automatic classifier. Spot-spraying achieved up to 86% WCE, which was equal to the broadcast herbicide treatment. Early spot spraying saved 47% of herbicides compared to the broadcast herbicide application. Maize yields in the spot-spraying plots were equal to the broadcast herbicide application plots. This study demonstrates that spot-spraying based on UAV weed maps is feasible and provides a significant reduction in herbicide use.
CRISPR/SpCas9‐mediated double knockout of barley Microrchidia MORC1 and MORC6a reveals their strong involvement in plant immunity, transcriptional gene silencing and plant growth
(2021) Galli, Matteo; Martiny, Engie; Imani, Jafargholi; Kumar, Neelendra; Koch, Aline; Steinbrenner, Jens; Kogel, Karl‐Heinz
The Microrchidia (MORC) family proteins are important nuclear regulators in both animals and plants with critical roles in epigenetic gene silencing and genome stabilization. In the crop plant barley (Hordeum vulgare), seven MORC gene family members have been described. While barley HvMORC1 has been functionally characterized, very little information is available about other HvMORC paralogs. In this study, we elucidate the role of HvMORC6a and its potential interactors in regulating plant immunity via analysis of CRISPR/SpCas9‐mediated single and double knockout (dKO) mutants, hvmorc1 (previously generated and characterized by our group), hvmorc6a, and hvmorc1/6a. For generation of hvmorc1/6a, we utilized two different strategies: (i) successive Agrobacterium‐mediated transformation of homozygous single mutants, hvmorc1 and hvmorc6a, with the respective second construct, and (ii) simultaneous transformation with both hvmorc1 and hvmorc6a CRISPR/SpCas9 constructs. Total mutation efficiency in transformed homozygous single mutants ranged from 80 to 90%, while upon simultaneous transformation, SpCas9‐induced mutation in both HvMORC1 and HvMORC6a genes was observed in 58% of T0 plants. Subsequent infection assays showed that HvMORC6a covers a key role in resistance to biotrophic (Blumeria graminis) and necrotrophic (Fusarium graminearum) plant pathogenic fungi, where the dKO hvmorc1/6a showed the strongest resistant phenotype. Consistent with this, the dKO showed highest levels of basal PR gene expression and derepression of TEs. Finally, we demonstrate that HvMORC1 and HvMORC6a form distinct nucleocytoplasmic homo‐/heteromers with other HvMORCs and interact with components of the RNA‐directed DNA methylation (RdDM) pathway, further substantiating that MORC proteins are involved in the regulation of TEs in barley.
Successful silencing of the mycotoxin synthesis gene TRI5 in fusarium culmorum and observation of reduced virulence in VIGS and SIGS experiments
(2022) Tretiakova, Polina; Voegele, Ralf Thomas; Soloviev, Alexander; Link, Tobias Immanuel
Crops constantly experience various biotic stresses during their life cycle, and Fusarium spp. remain one of the most serious groups of pathogens affecting plants. The ability to manipulate the expression of certain microorganism genes via RNAi creates the opportunity for new-generation dsRNA-based preparations to control a large number of diseases. In this study, we applied virus-induced gene silencing (VIGS), and spray-induced gene silencing (SIGS) to silence the trichothecene-producing gene TRI5 in F. culmorum as a means to reduce its aggressiveness on spring wheat. Treatment of the fungus with dsTRI5RNA in vitro reduced deoxynivalenol (DON) and 3-acetyldeoxynivalenol (3-A-DON) accumulations by 53–85% and 61–87%, respectively, and reduced TRI5 expression by 84–97%. VIGS decreased the proportion of infected wheat spikelets by 73%, but upregulation was observed for TRI5. SIGS on wheat leaves and ears using certain dsTRI5RNA amounts negatively impacted F. culmorum growth. However, when performing in vivo analyses of TRI5 mRNA levels, the upregulation of the gene was determined in the variants where fungal colonization was restricted, suggesting a compensatory reaction of the pathogen to RNAi.
New approaches to manage Asian soybean rust (Phakopsora pachyrhizi) using Trichoderma spp. or their antifungal secondary metabolites
(2022) El-Hasan, Abbas; Walker, Frank; Klaiber, Iris; Schöne, Jochen; Pfannstiel, Jens; Voegele, Ralf T.
Attempts have been made to determine the in vitro and in planta suppressive potential of particular Trichoderma strains (T16 and T23) and their secondary metabolites (SMs) against Asian soybean rust (ASR) incited by Phakopsora pachyrhizi. Aside from the previously identified SMs 6-pentyl-α-pyrone (6PAP) and viridiofungin A (VFA), the chemical structures of harzianic acid (HA), iso-harzianic acid (iso-HA), and harzianolide (HZL) were characterized in this study. Our results indicate that exposure of urediospores to 200 ppm 6PAP completely inhibits germination. A slightly higher dosage (250 ppm) of HZL and VFA reduces germination by 53.7% and 44%, respectively. Germ tube elongation seems more sensitive to 6PAP than urediospore germination. On detached leaves, application of conidia of T16 and T23 results in 81.4% and 74.3% protection, respectively. Likewise, 200 ppm 6PAP recorded the highest ASR suppression (98%), followed by HZL (78%) and HA (69%). Treatment of undetached leaves with 6PAP, HA, or HZL reduces ASR severity by 84.2%, 65.8%, and 50.4%, respectively. Disease reduction on the next, untreated trifoliate by T23 (53%), T16 (41%), HZL (42%), and 6PAP (32%) suggests a translocation or systemic activity of the SMs and their producers. To our knowledge, this study provides the first proof for controlling ASR using antifungal SMs of Trichoderma. Our findings strongly recommend the integration of these innovative metabolites, particularly 6PAP and/or their producers in ASR management strategies.
Effects of a two-factorial dynamic storage system on apple fruit metabolism, quality attributes, disorder incidences and biochemical properties
(2024) Büchele, Felix; Vögele, Ralf; Kittemann, Dominikus
Dynamic Controlled Atmosphere (DCA) technologies are designed to monitor the metabolism of fruit as a function of time and the oxygen partial pressure (pO2) in the storage atmosphere. By identifying signals sent by the fruit in response to low oxygen stress, this concept allows defining the lowest oxygen level tolerated by the fruit material and the specific and transient pO2, which is proposed to slow down the aerobic respiration of the fruit to a minimum and accordingly the ripening-related loss of fruit quality. This work examines a novel storage technology for pome fruit referred to as DCA-CD+, which can be considered the first generation of a two-factorial dynamic storage system. DCA-CD+ aims to define both a transient optimum pO2 and storage temperature in real-time, based on monitoring the carbon dioxide (CO2) production rate of the stored product. The CO2-release rate is proposed as a dependable indicator of low oxygen stress and an accurate depiction of the metabolic intensity of the fruit in response to temperature variations. The idea behind dynamic temperature adjustments is based on the assumption, that increased storage temperatures can reduce the energy usage of the refrigeration systems during long-term storage substantially, while also mitigating oxidative stress in the fruit, thus contributing to reducing the occurrence of storage disorders. The extremely low pO2 levels established in the storage atmosphere are suggested to counteract the ripening-inducing effects of this dynamic temperature approach. The assessment of DCA-CD+ in this work is based on a comparison to other postharvest conditions such as cold storage (RA) or static controlled atmosphere (CA). Furthermore, the interactions between storage atmosphere, temperature, and applications of the ethylene-inhibitor product 1-methylcyclopropene (1-MCP) are investigated. The conducted biochemical analyses highlight that apples stored under DCA-CD+ undergo repeated periods of hypoxia. Fruit adapt to the energy crisis induced by low oxygen stress by increasing their glycolytic flux, which is coupled to the activation of the fermentative pathway. Importantly, none of the examined apple cultivars in any of the experimental seasons exceeded critical thresholds for volatile fermentative products acetaldehyde (AA) and ethanol (EtOH), which could potentially be associated with the development of off-flavors or internal disorder symptoms. Consequently, the atmospheric conditions implemented did not result in any fruit damage associated with low oxygen stress in any of the tested scenarios. These findings suggest that DCA-CD+, specifically the use of carbon dioxide as an input value, is effective in identifying low oxygen stress in stored fruit and defining the pO2 for optimum quality conservation. pO2 setpoint calculation by the DCA-CD+ algorithm showed an interaction effect with the respective storage temperatures. Furthermore, the sensitivity of the stored fruit to temperature variation was found to be cultivar-dependent and transient during the storage period. Depending on the stored apple cultivar and season, DCA-CD+ calculated temperature setpoints reaching up to 3°C to 4°C, from the baseline temperature of 1°C. Temperature peaks were generally followed by a significant decrease in the calculated temperature setpoints, as an increased CO2 production rate signaled an intensifying fruit metabolism due to elevated storage temperature. The analysis of quality-defining parameters and disorder symptoms support the conclusion that DCA-CD+ allows for dynamic temperature adjustments without accelerating fruit ripening and the associated loss of quality. Preliminary findings indicate that this approach can reduce the energy usage of the cooling system in commercial storage rooms, without requiring cost-intensive additional installation of technology or renovations of room structural components. Lower pO2 setpoints were calculated at higher temperatures, suggesting that increasing the storage temperature can contribute to alleviating low oxygen stress in apples. Improved conservation of fruit quality attributes and a reduction in storage disorder incidences of DCA-CD+ in comparison to CA could be demonstrated in some instances, contingent on experimental season and apple cultivar. These benefits presumably become more pronounced with extended storage durations exceeding eight months. Ultimately, it can be argued that the complementary and interactive effects of dynamic temperature and oxygen in DCA-CD+ with 1-MCP application provide the highest potential for fruit quality conservation, limiting storage disorder, and reducing cooling-related energy usage. DCA-CD+ was demonstrated to potentially counteract detrimental effects of 1-MCP applications, e.g. an increased risk of carbon dioxide injuries. This work aimed to contribute to understanding the mechanisms behind the interference of the established temperature and atmospheric conditions in the DCA-CD+ system with the volatile aroma profile of apples. It was demonstrated that the synthesis capacity of volatile organic compounds (VOC) is primarily suppressed on a principal substrate level, and less in the later conversion of aldehydes to alcohols and esters. The pathways for the synthesis of linear volatiles originating from fatty acids were determined more responsive to low oxygen environments, in comparison to the pathways of branched volatiles derived from amino acids. Further insights were gained into the physiological mechanisms underlying the activation of the fermentative pathway, suggesting it functions as an adaptation mechanism not exclusively linked to low oxygen stress. Moreover, efforts were made to establish a connection between ripening and disorder-related modifications in cell structural components and associated alterations in the volatile profile, primarily highlighting the role of the lipoxygenase pathway. Statistical classification demonstrated that the repeated induction of low oxygen stress in DCA-CD+ storage resulted in a distinct volatile profile and a higher association with aroma defining compounds compared to CA storage. The observed increased EtOH accumulation is discussed to mitigate the ripening-inducing effects of the hormone ethylene, while also providing additional substrate for the synthesis of ethyl esters. Preliminary findings of this work indicate that storage temperatures can play a role in the aroma development of stored apples, even when low pO2 conditions are established. In summary, this study created a comprehensive documentation of the commercial viability of two-factorial dynamic storage systems for pome fruit and provided insights into the metabolomic responses of apples to extremely low oxygen levels, particularly in interaction with dynamic storage temperature adjustments.
Metabolization and sequestration of plant specialized metabolites in insect herbivores: Current and emerging approaches
(2022) Jeckel, Adriana Moriguchi; Beran, Franziska; Züst, Tobias; Younkin, Gordon; Petschenka, Georg; Pokharel, Prayan; Dreisbach, Domenic; Ganal-Vonarburg, Stephanie Christine; Robert, Christelle Aurélie Maud
Herbivorous insects encounter diverse plant specialized metabolites (PSMs) in their diet, that have deterrent, anti-nutritional, or toxic properties. Understanding how they cope with PSMs is crucial to understand their biology, population dynamics, and evolution. This review summarizes current and emerging cutting-edge methods that can be used to characterize the metabolic fate of PSMs, from ingestion to excretion or sequestration. It further emphasizes a workflow that enables not only to study PSM metabolism at different scales, but also to tackle and validate the genetic and biochemical mechanisms involved in PSM resistance by herbivores. This review thus aims at facilitating research on PSM-mediated plant-herbivore interactions.
Facilitation of weed seed predation by living mulch and cover crops
(2022) Schumacher, Matthias; Gerhards, Roland
Weed seed predation (WSP) is an important agro‐ecosystem service that naturally suppresses weed populations. Disturbances directly interfere with the activity of the seed predators, for example, harvest and soil tillage during a peak in weed seed availability (around cereal harvest). Plant soil cover and reduced soil tillage are factors that might positively affect WSP. We investigated cover crops and living mulch along with soil disturbance (no‐till and conventional seeding) in comparison to a fallow and repeated stubble tillage with a chisel plough in terms of WSP, activity‐density of carabid beetles and soil cover. A field experiment with a randomised complete block design and four replications was conducted at two sites from 2017 until 2019. WSP rates were assessed with seed cards and exclosure cages from July until November during a period of eight consecutive days each month. Living mulch exhibited the highest WSP rates of 72% with a maximum of 95% immediately following cereal harvest in August and September of 2017 and 2019, while the remainder of the treatments ranged from 2% to 5% WSP. In October and November mustard showed on average twice as high WSP rates compared to the other treatments (50% vs. 10%–25%). Seeding and soil tillage did not significantly influence carabid beetle activity‐density and WSP, except for living mulch. Predator groups (invertebrates and vertebrates) were not influenced by the various treatments, rather year and month determined the predator groups dominance. Besides their weed suppressive effects, living mulches and cover crops also facilitate WSP.
Characterization of Bacillus velezensis UTB96, demonstrating improved lipopeptide production compared to the strain B. velezensis FZB42
(2022) Vahidinasab, Maliheh; Adiek, Isabel; Hosseini, Behnoush; Akintayo, Stephen Olusanmi; Abrishamchi, Bahar; Pfannstiel, Jens; Henkel, Marius; Lilge, Lars; Vögele, Ralf ; Hausmann, Rudolf
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.
Antioxidant availability trades off with warning signals and toxin sequestration in the large milkweed bug (Oncopeltus fasciatus)
(2023) Heyworth, H. Cecilia; Pokharel, Prayan; Blount, Jonathan D.; Mitchell, Christopher; Petschenka, Georg; Rowland, Hannah M.
In some aposematic species the conspicuousness of an individual's warning signal and the concentration of its chemical defense are positively correlated. Several mechanisms have been proposed to explain this phenomenon, including resource allocation trade‐offs where the same limiting resource is needed to produce both the warning signal and chemical defense. Here, the large milkweed bug (Oncopeltus fasciatus: Heteroptera, Lygaeinae) was used to test whether allocation of antioxidants, that can impart color, trade against their availability to prevent self‐damage caused by toxin sequestration. We investigated if (i) the sequestration of cardenolides is associated with costs in the form of changes in oxidative state; and (ii) oxidative state can affect the capacity of individuals to produce warning signals. We reared milkweed bugs on artificial diets with increasing quantities of cardenolides and examined how this affected signal quality (brightness and chroma) across different instars. We then related the expression of warning colors to the quantity of sequestered cardenolides and indicators of oxidative state—oxidative lipid damage (malondialdehyde), and two antioxidants: total superoxide dismutase and total glutathione. Bugs that sequestered more cardenolides had significantly lower levels of the antioxidant glutathione, and bugs with less total glutathione had less luminant orange warning signals and reduced chroma of their black patches compared to bugs with more glutathione. Bugs that sequestered more cardenolides also had reduced red–green chroma of their black patches that was unrelated to oxidative state. Our results give tentative support for a physiological cost of sequestration in milkweed bugs and a mechanistic link between antioxidant availability, sequestration, and warning signals.
Kann die Saatgutbehandlung mit dem Safener Cyprosulfamid die Verträglichkeit von Isoxaflutole in Mais steigern, die Unkrautbekämpfung verbessern und die Wirkstoffmenge verringern?
(2020) Gerhards, Roland; Santel, Hans-Joachim
Herbizide Wirkstoffe werden in kommerziellen Produkten manchmal mit Safenern kombiniert, um die Verträglichkeit des Herbizids für die Kulturpflanze zu gewährleisten. Dabei regen Safener meist die Metabolisierung von Herbiziden in der Kulturpflanze an. Das Herbizid Isoxaflutole (IFT) wird in Mais mit dem Safener Cyprosulfamid (CSA) als Co-Formulierung angewendet. In dieser Arbeit wurde untersucht, ob CSA auch zeitlich getrennt von IFT als Saatgutbehandlung den Mais vor Herbizidschäden durch IFT schützen kann und ob die Unkrautbekämpfung in diesem Fall höher ist als bei der Co-Formulierung, weil die Unkräuter durch die Saatbehandlung nicht mit CSA in Kontakt kommen. Es wurden von 2016–2018 insgesamt fünf Topfversuche als randomisierte, vollständige Blockanlage mit 15 Wiederholungen durchgeführt. Dabei wurde IFT ohne CSA, mit CSA als Co-Formulierung und mit CSA-Beize in der einfachen und doppelten Feldaufwandmenge im 2‑Blatt-Stadium von Mais appliziert und 28 Tage nach der Applikation die Herbizidschäden an Mais und Unkräutern bonitiert und die Sprosslänge und Trockenmasse der Maisstängel gemessen. In allen Versuchen reagierte Mais sensibel mit Bleichungen an den Blättern auf IFT mit bis zu 62 % Schäden. Die Kombination von IFT und CSA reduzierte den vom Herbizid verursachten Schaden signifikant. Die schützende Wirkung von CSA war stärker, wenn der Safener simultan mit dem Herbzid gespritzt wurde (5–10 % Schaden) als bei Beizanwendung (15–25 % Schaden). Im Mittel aller Versuche war die Sprosslänge des mit der der Co-Formulierung von IFT und CSA behandelten Maises auf dem Niveau der unbehandelten Kontrolle und höher als in den IFT-Behandlungen. Die Beizvariante mit 100 g/ha IFT zeigte die längsten Sprosse. Der Safener konnte die herbizidbedingte Reduktion der Trockenmassebildung nur teilweise verhindern. Die Behandlungen mit IFT ohne Zusatz des Safeners zeigten die niedrigsten Sprossmassen der Maisstängel. Die Bekämpfung von Amaranthus retroflexus, Echinochloa crus-galli und Setaria viridis gelang in allen Behandlungen zu 100 %. Nur gegen Stellaria media war die Bekämpfung ohne Safener besser als mit CSA-Beize (92 % Bekämpfungsrate) und Co-Formulierung (79 %). Mit der Saatgutbeizung können ca. 80 % des Safeners gegenüber der Co-Formulierung eingespart werden.
Camera-guided weed hoeing in winter cereals with narrow row distance
(2020) Gerhards, Roland; Kollenda, Benjamin; Machleb, Jannis; Möller, Kurt; Butz, Andreas; Reiser, David; Griegentrog, Hans-Werner
Farmers are facing severe problems with weed competition in cereal crops. Grass-weeds and perennial weed species became more abundant in Europe mainly due to high percentages of cereal crops in cropping systems and reduced tillage practices combined with continuous applications of herbicides with the same mode of action. Several weed populations have evolved resistance to herbicides. Precision weed hoeing may help to overcome these problems. So far, weed hoeing in cereals was restricted to cropping practices with row distances of more than 200 mm. Hoeing in cereals with conventional row distances of 125–170 mm requires the development of automatic steering systems. The objective of this project was to develop a new automatic guidance system for inter-row hoeing using camera-based row detection and automatic side-shift control. Six field studies were conducted in winter wheat to investigate accuracy, weed control efficacy and crop yields of this new hoeing technology. A three-meter prototype and a 6-meter segmented hoe were built and tested at three different speeds in 150 mm seeded winter wheat. The maximum lateral offset from the row center was 22.53 mm for the 3 m wide hoe and 18.42 mm for the 6 m wide hoe. Camera-guided hoeing resulted in 72–96% inter-row and 21–91% intra-row weed control efficacy (WCE). Weed control was 7–15% higher at 8 km h−1 compared to 4 km h−1. WCE could be increased by 14–22% when hoeing was combined with weed harrowing. Grain yields after camera-guided hoeing at 8 km h−1 were 15–76% higher than the untreated control plots and amounted the same level as the weed-free herbicide plots. The study characterizes camera-guided hoeing in cereals as a robust and effective method of weed control.
Weed control in a pesticide‐free farming system with mineral fertilisers
(2023) Saile, Marcus; Spaeth, Michael; Schwarz, Jürgen; Bahrs, Enno; Claß‐Mahler, Ingrid; Gerhards, Roland
Negative impacts of pesticides on the environment and human health, the risk of pesticide residues in the food chain, and the problems with herbicide‐resistant weed biotypes support the need for alternative cropping systems. The objective of this study was to investigate weed populations, weed management and crop yield in a pesticide‐free cropping system with the use of mineral fertilisers. Conventional‐, organic‐ and mineral‐ecological cropping systems (MECS) with 6‐year crop rotations including winter wheat, maize, winter triticale or winter rye, soyabean or spring pea, and spring barley were established in a randomised complete strip plot design with four repetitions. Experiments were conducted at four locations in Germany. Preventive and sensor‐guided mechanical weed management strategies were applied in all crops in the organic system and in MECS. Herbicide were applied in the conventional farming system. Weed densities, weed species composition, weed control efficacy (WCE) and crop yield were analysed over 2 years in 2020 and 2021. Conventional farming had the highest WCE and 1–7 weeds m−2 (2.7% weed coverage) after herbicide application. In the organic cropping system and MECS, up to 27 weeds m−2 were counted after camera‐guided weed hoeing. Weed coverage in MECS (9.7%) was higher than in the organic cropping system with 7.7%. Crop yield in MECS was equal to the conventional farming system and 20% higher yield than in the organic farming system. MECS represents a promising new and productive cropping practice if an effective integrated weed management strategy is applied.
Spatial metabolomics reveal divergent cardenolide processing in the monarch (Danaus plexippus) and the common crow butterfly (Euploea core)
(2023) Dreisbach, Domenic; Bhandari, Dhaka R.; Betz, Anja; Tenbusch, Linda; Vilcinskas, Andreas; Spengler, Bernhard; Petschenka, Georg
Although being famous for sequestering milkweed cardenolides, the mechanism of sequestration and where cardenolides are localized in caterpillars of the monarch butterfly (Danaus plexippus, Lepidoptera: Danaini) is still unknown. While monarchs tolerate cardenolides by a resistant Na+/K+‐ATPase, it is unclear how closely related species such as the nonsequestering common crow butterfly (Euploea core, Lepidoptera: Danaini) cope with these toxins. Using novel atmospheric‐pressure scanning microprobe matrix‐assisted laser/desorption ionization mass spectrometry imaging, we compared the distribution of cardenolides in caterpillars of D. plexippus and E. core. Specifically, we tested at which physiological scale quantitative differences between both species are mediated and how cardenolides distribute across body tissues. Whereas D. plexippus sequestered most cardenolides from milkweed (Asclepias curassavica), no cardenolides were found in the tissues of E. core. Remarkably, quantitative differences already manifest in the gut lumen: while monarchs retain and accumulate cardenolides above plant concentrations, the toxins are degraded in the gut lumen of crows. We visualized cardenolide transport over the monarch midgut epithelium and identified integument cells as the final site of storage where defences might be perceived by predators. Our study provides molecular insight into cardenolide sequestration and highlights the great potential of mass spectrometry imaging for understanding the kinetics of multiple compounds including endogenous metabolites, plant toxins, or insecticides in insects.
Characterization ofantifungal properties of lipopeptide-producing Bacillus velezensis strains and their proteome-based response to the phytopathogens, Diaporthe spp
(2023) Akintayo, Stephen Olusanmi; Hosseini, Behnoush; Vahidinasab, Maliheh; Messmer, Marc; Pfannstiel, Jens; Bertsche, Ute; Hubel, Philipp; Henkel, Marius; Hausmann, Rudolf; Vögele, Ralf; Lilge, Lars
Introduction: B. velezensis strains are of interest in agricultural applications due to their beneficial interactions with plants, notable through their antimicrobial activity. The biocontrol ability of two new lipopeptides-producing B. velezensis strains ES1-02 and EFSO2-04, against fungal phytopathogens of Diaporthe spp., was evaluated and compared with reference strains QST713 and FZB42. All strains were found to be effective against the plant pathogens, with the new strains showing comparable antifungal activity to QST713 and slightly lower activity than FZB42. Methods: Lipopeptides and their isoforms were identified by high-performance thin-layer chromatography (HPTLC) and mass spectrometric measurements. The associated antifungal influences were determined in direct in vitro antagonistic dual culture assays, and the inhibitory growth effects on Diaporthe spp. as representatives of phytopathogenic fungi were determined. The effects on bacterial physiology of selected B. velezensis strains were analyzed by mass spectrometric proteomic analyses using nano-LC-MS/MS. Results and Discussion: Lipopeptide production analysis revealed that all strains produced surfactin, and one lipopeptide of the iturin family, including bacillomycin L by ES1-02 and EFSO2-04, while QST713 and FZB42 produced iturin A and bacillomycin D, respectively. Fengycin production was however only detected in the reference strains. As a result of co-incubation of strain ES1-02 with the antagonistic phytopathogen D. longicolla, an increase in surfactin production of up to 10-fold was observed, making stress induction due to competitors an attractive strategy for surfactin bioproduction. An associated global proteome analysis showed a more detailed overview about the adaptation and response mechanisms of B. velezensis, including an increased abundance of proteins associated with the biosynthesis of antimicrobial compounds. Furthermore, higher abundance was determined for proteins associated with oxidative, nitrosative, and general stress response. In contrast, proteins involved in phosphate uptake, amino acid transport, and translation were decreased in abundance. Altogether, this study provides new insights into the physiological adaptation of lipopeptide-producing B. velezensis strains, which show the potential for use as biocontrol agents with respect to phytopathogenic fungi.
Role of Bacillus spp. plant growth promoting properties in mitigating biotic and abiotic stresses in lowland rice (Oryza sativa L.)
(2023) Weinand, Tanja; El-Hasan, Abbas; Asch, Folkard
The ability of microorganisms to promote plant growth and mitigate abiotic and biotic stresses makes them an interesting tool for sustainable agriculture. Numerous studies aim to identify new, promising bacteria isolates. Traditional culture-based methods, which focus on selecting microorganisms with plant-growth-promoting traits, such as hormone production, nutrient solubilization, and antifungal properties, are widely used. This study aims to investigate the role of plant-growth-promoting properties in bacteria-mediated stress mitigation and the suitability of traditional culture-based methods as a screening tool for the identification of beneficial bacteria. To this end, we tested three endophytic Bacillus isolates, which have previously been shown to affect tolerance against iron toxicity in lowland rice, (a) for their effect on the resistance against brown spot disease, and (b) for plant-growth-promoting traits using common culture-based methods. Both B. pumilus isolates inhibited fungal growth in vitro and reduced brown spot disease in two of three rice cultivars in planta, although they tested negative for all plant-growth-promoting traits. While B. megaterium was negative for ACC deaminase activity and nutrient solubilization, it exhibited auxin production. Nevertheless, B. megaterium did not suppress brown spot disease in any of the three rice cultivars. This study shows that bacteria do not necessarily have to possess classical plant-growth-promoting properties in order to be beneficial to plants, and it emphasizes the limitation of common culture-based methods in effectively identifying beneficial bacteria. Moreover, our results highlight the significance of the interaction between bacteria and plant cultivars in determining the beneficial effects of Bacillus spp. on plants under biotic or abiotic stresses.
A simple artificial diet for feeding and sequestration assays for the milkweed bugs Oncopeltus fasciatus and Spilostethus saxatilis
(2023) Espinosa del Alba, Laura; Petschenka, Georg
Insect artificial diets are not only an important tool for mass rearing, nutritional research, and maintaining laboratory colonies but also for studying insect‐plant interactions. For herbivorous insects able to sequester plant toxins, feeding and sequestration assays based on artificial diet allow for the investigation of physiological, ecological, and evolutionary questions which may be difficult to study using real plants representing complex chemical environments. We developed a simple artificial diet, consisting of sunflower meal pressed into pills, for the milkweed bugs Oncopeltus fasciatus (Dallas) and Spilostethus saxatilis (Scopoli) (Heteroptera: Lygaeidae), which are capable of sequestering cardenolides and colchicum alkaloids, respectively. We assessed insect performance, suitability of the diet for sequestration assays, and its shelf life. Compared to sunflower seeds which are widely used as a laboratory maintenance diet for milkweed bugs, no differences were found in terms of weight development, presence of deformities, speed of development, or mortality. Importantly, after feeding O. fasciatus and S. saxatilis sunflower pills enriched with crystalline ouabain (cardenolide) or colchicine (colchicum alkaloid), respectively, sequestration was observed in both species. Moreover, as a prerequisite to test ecological hypotheses, our method allows for adequate concentration control and homogenous distribution of toxins across the diet. Under relatively warm conditions (27 °C and 60% r.h.), the new diet was stable for up to 10 days when used for feeding assays with adult bugs. Therefore, studies focusing on the role of plant toxins in predator–prey interactions and plant defense, but also insecticide research could benefit from using this approach.
New species based on the biological species concept within the complex of Lariophagus distinguendus (Hymenoptera, Chalcidoidea, Pteromalidae), a parasitoid of household pests
(2023) Pollmann, Marie; Kuhn, Denise; König, Christian; Homolka, Irmela; Paschke, Sina; Reinisch, Ronja; Schmidt, Anna; Schwabe, Noa; Weber, Justus; Gottlieb, Yuval; Steidle, Johannes Luitpold Maria
The pteromalid parasitoid Lariophagus distinguendus (Foerster) belongs to the Hymenoptera, a megadiverse insect order with high cryptic diversity. It attacks stored product pest beetles in human storage facilities. Recently, it has been shown to consist of two separate species. To further study its cryptic diversity, strains were collected to compare their relatedness using barcoding and nuclear genes. Nuclear genes identified two clusters which agree with the known two species, whereas the barcode fragment determined an additional third Clade. Total reproductive isolation (RI) according to the biological species concept (BSC) was investigated in crossing experiments within and between clusters using representative strains. Sexual isolation exists between all studied pairs, increasing from slight to strong with genetic distance. Postzygotic barriers mostly affected hybrid males, pointing to Haldane's rule. Hybrid females were only affected by unidirectional Spiroplasma‐induced cytoplasmic incompatibility and behavioural sterility, each in one specific strain combination. RI was virtually absent between strains separated by up to 2.8% COI difference, but strong or complete in three pairs from one Clade each, separated by at least 7.2%. Apparently, each of these clusters represents one separate species according to the BSC, highlighting cryptic diversity in direct vicinity to humans. In addition, these results challenge the recent ‘turbo‐taxonomy’ practice of using 2% COI differences to delimitate species, especially within parasitic Hymenoptera. The gradual increase in number and strength of reproductive barriers between strains with increasing genetic distance also sheds light on the emergence of barriers during the speciation process in L. distinguendus.

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