Biology subjects

Sorg, M.

Publications and source records attributed to Sorg, M..

12 recordsLinked to original sources

bioRxiv2026

Charting the insect biodiversity of Crete: insights from a pilot metabarcoding survey

Among eukaryotes, insects are by far the most diverse organisms on Earth, yet their global decline threatens ecosystem stability. Understanding local and regional biodiversity patterns is critical for conservation planning, ecosystem management, and predicting responses to environmental change, but traditional surveys for assessing insect diversity (e.g., manual collection, morphological identification, and counting) are highly labor-intensive, time-consuming, and often require rare or simply unavailable dedicated taxonomic expertise. DNA metabarcoding offers an efficient, high-resolution alternative to assess insect communities. Here, we report on the first insect metabarcoding survey on Crete that spans two years of sample collection between 2021 and 2023 from a small area in Southern Central Crete in the context of a citizen science project. A total of 29 samples yielded 10,865 Exact Sequence Variants (ESVs), 10,516 of which were assigned to insects, covering 988 species, 900 genera, and 227 families across 14 orders. A comparison with the existing observation records reveals 406 potential newly-observed species and an estimated 690 unclassified species, indicating substantial cryptic diversity. Our results demonstrate that even small-scale sampling can unravel substantial insect diversity and highlight critical gaps in barcode reference databases. Our study demonstrates how DNA metabarcoding can accelerate biodiversity discovery and monitoring in understudied regions.

ecology↗

bioRxiv2026

Nucleoid-binding protein RicO anchors replication origins to the membrane to ensure correct chromosome segregation in Staphylococcus aureus

The mechanisms underlying chromosome segregation in bacteria, particularly in non-canonical models, remain incompletely understood. The coccoid bacterium Staphylococcus aureus encodes a limited set of conserved proteins involved in chromosome segregation, including the Structural Maintenance of Chromosomes (SMC) complex and an incomplete partition system. Here, we identified a protein of previously unknown function that interacts with the nucleoid and ensures accurate chromosome segregation in S. aureus. RicO (Regulator and Insulator of Chromosomal Origins) contains a conserved N-terminal DNA-binding domain that recognizes a specific sequence motif in the origin-proximal region of the chromosome, as well as a C-terminal amphipathic helix that interacts with the cell membrane, thereby bridging DNA and the membrane. RicO localizes in membrane-proximal foci at the tip of each hemisphere (or cell poles). Cells lacking RicO fail to correctly position replication origins at the cell poles and have weakened genomic interactions in the origin-proximal region, resulting in chromosome segregation defects. The combined absence of RicO and a functional SMC complex leads to nearly half of the population appearing as anucleate cells, indicating that these two systems are key contributors to chromosome segregation in S. aureus. Collectively, our data support a model in which RicO anchors chromosomal origins at the cell periphery to ensure genome maintenance in this spherical bacterial pathogen.

microbiology↗

bioRxiv2025

Chromosome segregation dynamics during the cell cycle of Staphylococcus aureus

Research on chromosome organization and cell cycle progression in spherical bacteria, particularly Staphylococcus aureus, remains limited and fragmented. In this study, we established a working model to investigate chromosome dynamics in S. aureus using a Fluorescent Repressor-Operator System (FROS), which enabled precise localization of specific chromosomal loci. This approach revealed that the S. aureus cell cycle and chromosome replication cycle are not coupled, with cells exhibiting two segregated origins of replication at the start of the cell cycle. The chromosome has a specific origin-terminus-origin conformation, with origins localizing near the membrane, towards the tip of each hemisphere, or the "cell poles". We further used this system to assess the role of various proteins with a role in S. aureus chromosome biology, focusing on the ParB-parS and SMC-ScpAB systems. Our results demonstrate that ParB binds five parS chromosomal sequences and the resulting complexes influence chromosome conformation, but play a minor role in chromosome compaction and segregation. In contrast, the SMC-ScpAB complex plays a key role in S. aureus chromosome biology, contributing to chromosome compaction, segregation and spatial organization. Additionally, we systematically assessed and compared the impact of proteins linking chromosome segregation to cell division--Noc, FtsK, SpoIIIE and XerC--on origin and terminus number and positioning. This work provides a comprehensive study of the factors governing chromosome dynamics and organization in S. aureus, contributing to our knowledge on chromosome biology of spherical bacteria.

microbiology↗

bioRxiv2023

Insect biomass of protected habitats under the impact of arable farming in Germany

Five years after the well-known study on insect biomass decline in nature protected habitats in Germany over three decades, the project DINA (Diversity of Insects in Nature protected Areas) has investigated the status of insects in 21 selected nature reserves across Germany in the years 2020 and 2021. We used the same methods and protocols for trapping and measuring the biomass of flying insects as in the mentioned study. Across two seasons, we accumulated a comprehensive data set of 1621 data points of two-week emptying intervals. The measured overall insect biomass remained at low levels and corresponds to the published latest figures from the years 2007-2016. There were no significant regional differences, but biomass was negatively correlated with agricultural production area within 2 km of nature reserves. Differences between the two consecutive years were very likely due to well-known natural fluctuations of insect populations, changes in agricultural cultivation and local weather events. The results show that protected habitats are essential for insects, but not sufficient in their function, and that further steps need to be taken for a better protection and sustainment of insects, which fulfil key functions in many ecosystems.

ecology↗

bioRxiv2023

German-wide Malaise trap metabarcoding estimates over 33,000 insect species

Mitigating ongoing losses of insects and their key functions (e.g., pollination) requires accurately tracking large-scale and long-term community changes. However, doing so has been notoriously hindered by uniquely high insect species diversity that requires prohibitively high investments of time, funding, and taxonomic expertise. Here, we show that these concerns can be addressed through a comprehensive, scalable and cost-efficient DNA metabarcoding workflow. We use 1,815 samples from 75 Malaise traps across Germany from 2019 and 2020 to demonstrate how metabarcoding can be incorporated into large-scale insect monitoring networks for less than 50 {euro} per sample, including supplies, labor and maintenance. With on average 1.4M sequence reads per sample we uncovered 10,803 validated insect species, of which 83.9% were represented by a single OTU. We estimated another 21,403 plausible species, which likely either lack a reference barcode or are undescribed. The total of 31,846 species is similar to the number of insect species known for Germany ([~]35,500). Because Malaise traps capture only a subset of insects, our approach identified many species likely unknown from Germany or new to science. Our reproducible workflow ([~]80% OTU-similarity among years) provides a blueprint for large-scale biodiversity monitoring of insects and other biodiversity components in near real time.

ecology↗

bioRxiv2022

Spatial, temporal and taxonomic patterns of insect extinction in Germany

Red lists represent an important instrument for evaluating the decline of species in space and time, for improving decisionmaking and for guiding conservation planning. However, globally, only a fraction of species has been categorized according to a red list, even in countries where insects are relatively well-studied. Such large knowledge gaps hinder conservation planning and ultimately jeopardize the maintenance of ecosystem functions. Given the recent reports on severe insect decline, it is now more than ever of great importance to obtain a reliable complete picture of the state of insects. We here derive an estimate of extinction rates and of the proportion of threatened species for the total insect community in Germany, and asses spatial and temporal of extinction patterns. We found a regional extinction rate of 4.5% (1773-1937 species) for the area of Germany. Among extant insect species, 6% are classified as critically endangered (1856-2024 species), while among remaining species, a staggering 36.1% (10758-11086 species) is classified as threatened. Higher trophic levels of zoophagous insects are often more sensitive to negative environmental changes due to their position in the food web, and at the same time are underrepresented in Red Lists. They are therefore disproportionately affected by these knowledge gaps. This concerns particularly parasitoids which are taxa of regulatory importance and often higher extinction risk levels due to their trophic position. Exemplary examination of the spatial scaling of red list categories indicate a far higher rate and risk and exemplary over ten times higher regional extinction rate when the reference area is gradually scaled down. This illustrates the actual situation regarding the magnitude of regional species extinction events and extirpation risks that we have to assume for certain parts of the reference areas. For a given region, the loss of the gene pool of populations specially adapted to a given region usually represents an irreversible biodiversity loss. In order to avoid further irreparable damage, the species threatened with extinction must be preserved with top priority. There is thus a considerable need for research in order to assess the conservation status of more than 56% of the insect species diversity in Germany and to immediately achieve a more balanced trait group representation in red lists.

ecology↗

bioRxiv2022

Development of an insect sample fractionizer for biodiversity research

We describe a new mechanical tool for dividing mixed insects and other invertebrate samples into subsamples. The device enables the division to equal parts by means of a movable hemispherical bowl and a separating disc. Due to the complete stainless steel manufacturing, the sample divider is sterilizable by using chemicals or heating and thus suitable for DNA-based methods. The production of equally sized subsamples is of particular importance for biodiversity studies today, especially when using metabarcoding combined with insect homogenisation for species determination of mixed insect samples. The device allows sub-samples to be analyzed separately using the same or different methods, or getting archived for museal preservation and future research.

ecology↗

bioRxiv2022

A CRISPRi-based genetic resource to study essential Staphylococcus aureus genes

We have optimized a CRISPR interference system to facilitate gene knockdown in the gram-positive bacterial pathogen Staphylococcus aureus. Our approach used a CRISPRi system derived from Streptococcus pyogenes, which involves the co-expression of the dcas9 gene encoding a catalytically inactive Cas9 protein and a customizable single guide RNA (sgRNA). In our system, dcas9 is expressed from a single copy in the chromosome of methicillin resistant S. aureus (MRSA) strains COL or JE2, under the control of a tightly regulated promoter, inducible by anhydrotetracycline. The sgRNAs are expressed from a replicative plasmid under the control of a constitutively active promoter. This system enables efficient, inducible, knockdown of both essential and non-essential genes. Using this approach, we constructed the Lisbon CRISPRi Mutant Library (LCML) comprising 261 strains, in the JE2 background, containing sgRNAs targeting 200 essential genes/operons. This library facilitates the study of the function of essential S. aureus genes and is complementary to the Nebraska Transposon Mutant Library which consists of nearly 2000 strains, each carrying a transposon insertion within a non-essential gene. The availability of these two libraries will facilitate the study of S. aureus pathogenesis and biology. Abstract ImportanceStaphylococcus aureus is an important clinical pathogen that causes a high number of antibiotic resistant infections. The study of S. aureus biology, and particularly of the function of essential proteins, is of particular importance to develop new approaches to combat this pathogen. We have optimized a CRISPRi system that allows efficient targeting of essential S. aureus genes. Furthermore, we have used that system to construct a library of 261 strains which allow the depletion of essential proteins encoded in 200 genes/operons. This library, which we have named Lisbon CRISPRi Mutant Library (LCML), should facilitate the study of S. aureus pathogenesis and biology.

microbiology↗

bioRxiv2022

A universal insect trait tool (ITT, v1.0) for statistical analysis and evaluation of biodiversity research data

We present a unique data set of trait information for 586 insect families in Central Europe, covering the largest known part of described species (over 34,000 species). Life history information and major functional traits were evaluated with fuzzy coding and weighted according to the number of known species in Germany. An overall analysis of the German insect fauna is given and the data set is exemplarily applied to metabarcoding results of malaise trap samples. Due to the high functional and taxonomic diversity in insects, further developments and refinements of traits to be included will be an ongoing process with advancements of upcoming database versions to be subsequently published.

ecology↗

bioRxiv2022

Recommendations for tissue homogenisation and extraction in DNA metabarcoding of Malaise trap samples

With increased application of DNA metabarcoding in fast and high-resolution biodiversity assessment, various laboratory protocols have been optimised in recent years and their further evaluation is subject of current research. Homogenisation of bulk samples and subsequent DNA extraction from destructed tissue is one way of starting the metabarcoding process. This essential step in the protocol can either be conducted from wet sample material (e.g. bulk insect samples) soaked in fixative or from completely dried individuals. While the latter method appears to produce more consistent results, it is time consuming and more prone to cross-contamination. We tested both homogenisation approaches with regard to time efficiency and biodiversity assessment of complex arthropod bulk samples, in particular how the amount of processed tissue affects taxon recovery. Both approaches reveal similar taxa compositions and detect a similar total OTU diversity in a single extraction reaction. Increased amounts of tissue used in DNA extraction improved OTU diversity detection and recovered particularly specific low-biomass taxa, making this approach valuable for samples with high biomass and/or diversity. Due to less handling time and lower vulnerability for cross-contamination we recommend the processing of wet material when sample homogenisation is applied.

molecular biology↗

bioRxiv2021

Metabarcoding malaise trap plant components enables monitoring the diversity of plant-insect interactions

The declines observed in insect abundance and diversity in the past decades has also been observed in plants, and these events are most certainly correlated. Rapid largescale biomonitoring of both plants and insects can help monitor these changes and inform decisions for land management and species protection. Malaise traps have been used for nearly 80 years for passive insect sampling of primarily flying insects, and when they enter these traps, they carry the fragments of the plants they have visited, either as plant fragments and pollen on the body surface, or as digested food material in gut contents. DNA metabarcoding is a potential method to identify these plant traces in the ethanol of the malaise bottles, which is not possible with traditional microscopy. Metabarcoding could offer more insight into what plants insects are directly interacting with at a given time, and allow for the detection of rare plants, and neophyte species visited by insects. This study, to our knowledge, is the first examination of DNA metabarcoding plant traces from Malaise trap samples, we examine 105 samples from 21 sites throughout Germany collected in a 2-week period in May of 2020. Here we report on the feasibility of sequencing these sample types, analysis of the resulting taxa, the usage of cultivated plants by insects near nature conservancy areas, and the detection of rare and neophyte species.

molecular biology↗

bioRxiv2020

Pooling size sorted malaise trap fractions to maximise taxon recovery with metabarcoding

O_LISmall and rare specimens can remain undetected when metabarcoding bulk samples with a high size heterogeneity of specimens. This is especially critical for malaise trap samples, where most of the biodiversity is often contributed by small specimens. How to size sort and in which proportions to pool these samples has not been widely explored. We set out to find a size sorting strategy that maximizes taxonomic recovery but remains highly scalable and time efficient. C_LIO_LIThree 3 malaise trap samples where size sorted into 4 size classes using dry sieving. Each fraction was homogenized and lysed. The corresponding lysates were pooled to simulate samples never sorted, pooled in equal proportions and in 4 different proportions favoring the small size fractions. DNA from the pooled fractions as well as the individual size classes were extracted and metabarcoded using the FwhF2 and Fol-degen-rev primer set. Additionally wet sieving strategies were explored. C_LIO_LIThe small size fractions harbored the highest diversity, and were best represented when pooling in favor of small specimens. Not size sorting a sample leads to a 45-77% decrease in taxon recovery compared to size sorted samples. A size separation into only 2 fractions (below 4 mm and above) can already double taxon recovery compared to not sorting. However, increasing the sequencing depth 3-4 fold can also increase taxon recovery to comparable levels, but remains biased toward biomass rich taxa in the sample. C_LIO_LIWe demonstrate that size fractionizing bulk malaise samples can increase taxon recovery. The most practical approach is wet sieving into two size fractions, and proportional pooling of the lysates in favor of the small size fraction (80-90% volume). However, in large projects with time constraints, increasing sequencing depth can also be an alternative solution. C_LI

molecular biology↗