While the need for biodiversity research is growing, paradoxically, global taxonomical expertise is decreasing as a result of the neglected funding for young academics in taxonomy. Non-destructive approaches for DNA barcoding are necessary for a more efficient use of this dwindling expertise to fill gaps, and identify incorrect entries in sequence databases like BOLD or GenBank. They are efficient because morphological re-examination of species vouchers is still possible post-DNA barcoding. Non-destructive approaches for Diptera with a comprehensive species representation or the consideration of diagnostic fragile morphological characters are missing. Additionally, most non-destructive approaches combine a time intensive and non-destructive digestion step with common DNA extraction methods, such as commercial kits or CTAB DNA isolation. We circumvented those approaches and combined a modified non-destructive TE buffer high-speed DNA extraction, with a PCR inhibitor-resistant PCR reaction system, to a non-destructive DNA barcoding procedure for fresh and frozen samples of the Schizophora (Diptera). This method avoids morphological impairment and the application of harmful chemicals, is cost and time effective, restricts the need for laboratory equipment to a minimum, and prevents cross-contamination risk during DNA isolation. Moreover, the study indicates that the presented non-destructive DNA barcoding procedure is transferable to other soft-bodied insects. We suggest that PCR inhibitor-resistant master mixes enable the development of new—and the modification of existing—non-destructive approaches with the avoidance of further DNA template cleaning.

The tomato is a major crop worldwide and is one of the five most consumed vegetables in Germany. Stemphylium species including S. eturmiunum, S. gracilariae, S. lycii, S. lycopersici, S. rombundicum, S. simmonsii, S. solani, and S. vesicarium have been identified as tomato pathogens in various countries. In Germany, multiple instances of S. lycopersici and S. solani affecting tomato plants have been documented; however, only one incident of S. vesicarium has been reported in 1972. S. vesicarium is known to cause yellowish-brown spots on tomato leaves, which can ultimately lead to defoliation and reduced fruit yield. Therefore, it is crucial to identify the S. vesicarium that cause this disease accurately. In this study, S. vesicarium was isolated from necrotic tomato plants in organic farms located in northern and southern regions of Germany. Single spore isolates were generated and identified as S. vesicarium based on morphological characteristic and molecular analyses using nucleotide sequences of the internal transcribed spacer (ITS) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Koch´s postulates were fulfilled and revealed that S. vesicarium is the causal agent of brown spot on the samples adding a new account of the species.

A new disease was observed on chamomile plants (Matricaria recutita) in various commercially cultivated fields in 2016 and 2017 in Germany. From symptomatic stems and leaves, the fungal species described here was isolated repeatedly. Kochʼs postulates using in planta experiments were fulfilled by inoculation of chamomile plants proving the fungus to be the causal agent of the disease. Morphological studies and phylogenetic analyses using internal transcribed spacer, large subunit, and translation elongation factor-1α sequences suggested that the fungus represents a new species within the genus Rhexocercosporidium (Helotiales). The data are presented together with a description of the growth parameters and comprehensive illustrations of the new species, Rhexocercosporidium matricariae. All species so far assigned to Rhexocercosporidium are compared and discussed. The combination Rhexocercosporidium microsporum is validated.

Golovinomyces longipes is a widespread powdery mildew on Solanaceae (Nicotiana, Petunia and Solanum spp.). In the past, it has been reported only once on Verbena (Verbenaceae), a non-solanaceaous host. Recently, this powdery mildew has been found on the composite Matricaria chamomilla. The identification of the powdery mildew species on this unusual host has been proved by morphological studies and DNA sequence analysis. Both datasets coincide with the characteristic data for G. longipes on Solanaceae. First inoculation experiments with further composites resulted in an infection of Brachyscome hybrid ‘Surdaisy’. To our knowledge, this is the first report of G. lonigpes on hosts belonging to the important family of Asteraceae and an additional proof of the broader host range of G. longipes beyond the Solanaceae.