Institut für Biologie

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Industrial chicory genome gives insights into the molecular timetable of anther development and male sterility
(2023) Waegneer, Evelien; Rombauts, Stephane; Baert, Joost; Dauchot, Nicolas; De Keyser, Annick; Eeckhaut, Tom; Haegeman, Annelies; Liu, Chang; Maudoux, Olivier; Notté, Christine; Staelens, Ariane; van der Veken, Jeroen; van Laere, Katrijn; Ruttink, Tom
Industrial chicory (Cichorium intybus var. sativum) is a biannual crop mostly cultivated for extraction of inulin, a fructose polymer used as a dietary fiber. F1 hybrid breeding is a promising breeding strategy in chicory but relies on stable male sterile lines to prevent self-pollination. Here, we report the assembly and annotation of a new industrial chicory reference genome. Additionally, we performed RNA-Seq on subsequent stages of flower bud development of a fertile line and two cytoplasmic male sterile (CMS) clones. Comparison of fertile and CMS flower bud transcriptomes combined with morphological microscopic analysis of anthers, provided a molecular understanding of anther development and identified key genes in a range of underlying processes, including tapetum development, sink establishment, pollen wall development and anther dehiscence. We also described the role of phytohormones in the regulation of these processes under normal fertile flower bud development. In parallel, we evaluated which processes are disturbed in CMS clones and could contribute to the male sterile phenotype. Taken together, this study provides a state-of-the-art industrial chicory reference genome, an annotated and curated candidate gene set related to anther development and male sterility as well as a detailed molecular timetable of flower bud development in fertile and CMS lines.
Limitations of soil-applied non-microbial and microbial biostimulants in enhancing soil P turnover and recycled P fertilizer utilization: A study with and without plants
(2024) Herrmann, Michelle Natalie; Griffin, Lydia Grace; John, Rebecca; Mosquera-Rodríguez, Sergio F.; Nkebiwe, Peteh Mehdi; Chen, Xinping; Yang, Huaiyu; Müller, Torsten
Introduction: Phosphorus recovery from waste streams is a global concern due to open nutrient cycles. However, the reliability and efficiency of recycled P fertilizers are often low. Biostimulants (BS), as a potential enhancer of P availability in soil, could help to overcome current barriers using recycled P fertilizers. For this, a deeper understanding of the influence of BSs on soil P turnover and the interaction of BSs with plants is needed. Methods: We conducted an incubation and a pot trial with maize in which we testednon-microbial (humic acids and plant extracts) and microbial BSs (microbial consortia) in combination with two recycled fertilizers for their impact on soil P turnover, plant available P, and plant growth. Results and discussion: BSs could not stimulate P turnover processes (phosphatase activity, microbial biomass P) and had a minor impact on calcium acetate-lactate extractable P (CAL-P) in the incubation trial. Even though stimulation of microbial P turnover by the microbial consortium and humic acids in combination with the sewage sludge ash could be identified in the plant trial with maize, this was not reflected in the plant performance and soil P turnover processes. Concerning the recycled P fertilizers, the CAL-P content in soil was not a reliable predictor of plant performance with both products resulting in competitive plant growth and P uptake. While this study questions the reliability of BSs, it also highlights the necessity toimprove our understanding and distinguish the mechanisms of P mobilization in soil and the stimulation of plant P acquisition to optimize future usage.
Low-input high-molecular-weight DNA extraction for long-read sequencing from plants of diverse families
(2022) Russo, Alessia; Mayjonade, Baptiste; Frei, Daniel; Potente, Giacomo; Kellenberger, Roman T.; Frachon, Léa; Copetti, Dario; Studer, Bruno; Frey, Jürg E.; Grossniklaus, Ueli; Schlüter, Philipp M.
Long-read DNA sequencing technologies require high molecular weight (HMW) DNA of adequate purity and integrity, which can be difficult to isolate from plant material. Plant leaves usually contain high levels of carbohydrates and secondary metabolites that can impact DNA purity, affecting downstream applications. Several protocols and kits are available for HMW DNA extraction, but they usually require a high amount of input material and often lead to substantial DNA fragmentation, making sequencing suboptimal in terms of read length and data yield. We here describe a protocol for plant HMW DNA extraction from low input material (0.1 g) which is easy to follow and quick (2.5 h). This method successfully enabled us to extract HMW from four species from different families (Orchidaceae, Poaceae, Brassicaceae, Asteraceae). In the case of recalcitrant species, we show that an additional purification step is sufficient to deliver a clean DNA sample. We demonstrate the suitability of our protocol for long-read sequencing on the Oxford Nanopore Technologies PromethION® platform, with and without the use of a short fragment depletion kit.
Plastid phylogenomics reveals evolutionary relationships in the mycoheterotrophic orchid genus Dipodium and provides insights into plastid gene degeneration
(2024) Goedderz, Stephanie; Clements, Mark A.; Bent, Stephen J.; Nicholls, James A.; Patel, Vidushi S.; Crayn, Darren M.; Schlüter, Philipp M.; Nargar, Katharina; Goedderz, Stephanie; Australian Tropical Herbarium, James Cook University, Cairns, QLD, Australia; Clements, Mark A.; Centre for Australian National Biodiversity Research (joint venture between Parks Australia and CSIRO), Canberra, ACT, Australia; Bent, Stephen J.; Data61, Commonwealth Industrial and Scientific Research Organisation (CSIRO), Brisbane, QLD, Australia; Nicholls, James A.; Australian National Insect Collection, Commonwealth Industrial and Scientific Research Organisation (CSIRO), Canberra, ACT, Australia; Patel, Vidushi S.; National Research Collections Australia, Commonwealth Industrial and Scientific Research Organisation (CSIRO), Canberra, ACT, Australia; Crayn, Darren M.; Australian Tropical Herbarium, James Cook University, Cairns, QLD, Australia; Schlüter, Philipp M.; Department of Plant Evolutionary Biology, Institute of Biology, University of Hohenheim, Stuttgart, Germany; Nargar, Katharina; Australian Tropical Herbarium, James Cook University, Cairns, QLD, Australia
The orchid genus Dipodium R.Br. (Epidendroideae) comprises leafy autotrophic and leafless mycoheterotrophic species, with the latter confined to sect. Dipodium . This study examined plastome degeneration in Dipodium in a phylogenomic and temporal context. Whole plastomes were reconstructed and annotated for 24 Dipodium samples representing 14 species and two putatively new species, encompassing over 80% of species diversity in sect. Dipodium . Phylogenomic analysis based on 68 plastid loci including a broad outgroup sampling across Orchidaceae found that sect. Leopardanthus is the sister lineage to sect. Dipodium. Dipodium ensifolium , the only leafy autotrophic species in sect. Dipodium , was found to be a sister to all leafless, mycoheterotrophic species, supporting a single evolutionary origin of mycoheterotrophy in the genus. Divergence-time estimations found that Dipodium arose ca. 33.3 Ma near the lower boundary of the Oligocene and that crown diversification commenced in the late Miocene, ca. 11.3 Ma. Mycoheterotrophy in the genus was estimated to have evolved in the late Miocene, ca. 7.3 Ma, in sect. Dipodium . The comparative assessment of plastome structure and gene degradation in Dipodium revealed that plastid ndh genes were pseudogenised or physically lost in all Dipodium species, including in leafy autotrophic species of both Dipodium sections. Levels of plastid ndh gene degradation were found to vary among species as well as within species, providing evidence of relaxed selection for retention of the NADH dehydrogenase complex within the genus. Dipodium exhibits an early stage of plastid genome degradation, as all species were found to have retained a full set of functional photosynthesis-related genes and housekeeping genes. This study provides important insights into plastid genome degradation along the transition from autotrophy to mycoheterotrophy in a phylogenomic and temporal context.
Regulatory modules of metabolites and protein phosphorylation in arabidopsis genotypes with altered sucrose allocation
(2022) Stefan, Thorsten; Wu, Xu Na; Zhang, Youjun; Fernie, Alisdair; Schulze, Waltraud X.
Multi-omics data sets are increasingly being used for the interpretation of cellular processes in response to environmental cues. Especially, the posttranslational modification of proteins by phosphorylation is an important regulatory process affecting protein activity and/or localization, which, in turn, can have effects on metabolic processes and metabolite levels. Despite this importance, relationships between protein phosphorylation status and metabolite abundance remain largely underexplored. Here, we used a phosphoproteomics–metabolomics data set collected at the end of day and night in shoots and roots of Arabidopsis to propose regulatory relationships between protein phosphorylation and accumulation or allocation of metabolites. For this purpose, we introduced a novel, robust co-expression measure suited to the structure of our data sets, and we used this measure to construct metabolite-phosphopeptide networks. These networks were compared between wild type and plants with perturbations in key processes of sugar metabolism, namely, sucrose export (sweet11/12 mutant) and starch synthesis (pgm mutant). The phosphopeptide–metabolite network turned out to be highly sensitive to perturbations in sugar metabolism. Specifically, KING1, the regulatory subunit of SnRK1, was identified as a primary candidate connecting protein phosphorylation status with metabolism. We additionally identified strong changes in the fatty acid network of the sweet11/12 mutant, potentially resulting from a combination of fatty acid signaling and metabolic overflow reactions in response to high internal sucrose concentrations. Our results further suggest novel protein-metabolite relationships as candidates for future targeted research.

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