New finds of the click beetle Agriotes sordidus (Illiger, 1807) and an overview on its current distribution in Germany

Die Schnellkäfergattung Agriotes beinhaltet einige der in Ackerbaukulturen schädlichsten Drahtwürmer. Von den fünf in Deutschland als wirtschaftlich bedeutend eingestuften Agriotes-Arten ist eine, Agriotes sordidus, als Neuankömmling mit rezent erfolgter Nordausbreitung zu betrachten. Diese ursprünglich westmediterrane, als sehr schädigend angesehene Art wird auch im bundesweiten Schnellkäfermonitoring erfasst. Sie hat inzwischen ein geschlossenes Gebiet im Oberrheingraben besiedelt, sich entlang einiger Rheinnebenflüsse weiter ausgebreitet und ist über den Mittelrhein bis an den südlichsten Rand der Kölner Bucht vorgestoßen. Dieser Standort Westerhausen stellt wahrscheinlich den ersten Fund für Nordrhein-Westfalen dar. Ein weiterer Fund bei Zusmars­hausen stellt den wohl ersten Fund für Bayern abseits des an den Oberrhein anknüpfenden Vorkommens entlang des Mains dar. Überraschend sind Funde in Küstennähe in Niedersachsen und Schleswig-Holstein weitab vom geschlossenen Verbreitungsgebiet im Südwesten Deutschlands. Norddeutsche Tiere unterschieden sich zwar optisch von den Tieren aus dem südlichen Verbreitungs­gebiet, aber die morphologische Bestimmung wurde mittels PCR bestätigt. A. sordidus ist demnach in Deutschland weiter verbreitet als angenommen. In den meisten Gebieten fehlt er aber noch, so z.B. im Osten Deutschlands. Die Habitatpräferenzen sowie das Schadpotential in den neu besiedelten Gebieten sind damit aber nicht geklärt. Die Ausbreitung dieser Art sollte weiter überwacht werden.    Some of the most damaging wireworms in middle European arable land belong to the genus Agriotes. One of the five species of this genus which are of economic importance in Germany, is a newcomer. The species in question, Agriotes sordidus, is essentially western Mediterranean in its distribution, however, showing a range ex­tension to the north in Western Europe in recent decades. In Germany A. sordidus now shows a continuous distribution along the Upper Rhine valley and its tributaries (e.g. Neckar, Nahe, Main), as was shown recently. The species is also recorded in the countrywide click beetle monitoring with additional finds at the southern edge of the Cologne lowland (first find in North Rhine-Westphalia), the region of Zusmarshausen in western Bavaria, and most surprising, two locations close to the North Sea coast in Lower Saxony and Schleswig-Holstein (first find in this federal state). The latter two sites and also the Bavarian site near Zusmarshausen are far from the area of continuous distribution in southwestern Germany. As northern German specimens differed visually from the southern German finds the morphological determination was verified by PCR. Therefore A. sordidus is already more widespread than previously thought, but still missing in most regions. No records exist from eastern Ger­many yet. However, the habitat preferences and the damage potential of this species are still not fully understood. The spread of A. sordidus should be monitored further.   &nbsp

Identification of Agriotes wireworms – Are they always what they appear to be?

Die Schnellkäfergattung Agriotes beinhaltet einige der in Ackerbaukulturen schädlichsten Drahtwürmer. Von den fünf in Deutschland als wirtschaftlich bedeutend eingestuften Agriotes-Arten sind insbesondere die drei Arten A. lineatus, A. obscurus, A. sputator im Larvalstadium schwierig zu unterscheiden. Eine genaue Identifikation ist jedoch entscheidend, um mögliche Unterschiede, zum Beispiel in der Präferenz für Bodenarten, für Bodenfeuchte oder in der Nahrungspräferenz, überhaupt zu erkennen. Aber wichtige morphologische Unterschiede, die in allgemein genutzten Bestimmungsschlüsseln für Drahtwürmer angegeben werden, ermöglichten nicht immer eine korrekte Bestimmung. Ebenso war es nicht in allen Fällen möglich, die Tiere über eine PCR zu identifizieren, da teils Kreuzreaktionen auftraten. Daher entwickelten wir neue Primer für die Arten. Nachfolgend wurden Populationen von verschiedenen Standorten untersucht, um zu überprüfen, ob die Anteile aberranter Exemplare sich zwischen den Standorten unterschieden. DOI: 10.5073/JfK.2015.04.03, https://doi.org/10.5073/JfK.2015.04.03Wireworms are internationally recognized as economically important pests with the most damaging wireworms in middle European arable land belonging to the genus Agriotes. From the five species of greater importance in Germany, three (A. lineatus, A. obscurus, A. sputator) are very difficult to identify in their larval stage. Precise identification is crucial when trying to determine whether there are eco­logical differences between these three widespread species, e.g. in the reactions to different soil types, to soil moisture or in food choice. We report on the differentiation of A. lineatus, A. obscurus and A. sputator wireworms by comparing the identification by morphological traits of the larvae with the identification by PCR using a part of the mitochondrial DNA. Morphological traits given in commonly used identification keys for wireworms were not always reliable. Likewise the PCR did not always produce results, in part due to cross reactions. Therefore we developed new primers for reducing cross-reactions previously encountered. In the following, samples from different populations from the three species were compared to investigate if the proportion of aberrant specimens differed at different locations. DOI: 10.5073/JfK.2015.04.03, https://doi.org/10.5073/JfK.2015.04.0

Detection of the methylated Trichothecenegenes Tri5 and Tri14 in Fusarium sporotrichioides using the endonuclease MspJI during host plant infections

Bei Fusarium sporotrichioides wurde die Methylierung von DNA der beiden Mykotoxin-Gene Tri5 und Tri14 bei verschiedenen Temperaturen und bei Infektionsbedingungen untersucht. Dazu wurde das Restriktionsenzym MspJI verwendet, das DNA immer nur dann schneidet, wenn diese auch bei der Nukleotid-Folge Guanosin/Cytosin (G/C) methyliert ist. Unmethylierte G/C-haltige DNA wird von diesem Enzym nicht geschnitten, weshalb hier in einfacher Weise eine Unterscheidung in der Methylierung möglich ist. Eine Restriktion oder Nichtrestriktion wurde mit PCR-Primern nachgewiesen, die die Sequenzen beider Gene abdeckten. So war eine Methylierung der für die Mykotoxin-Gene Tri5 und Tri14 codierende DNA mit MspJI nur nachweisbar, wenn F. sporotrichioides seine Wirtspflanzen Gerste und Mais erfolgreich infi­zierte. Alle anderen Bedingungen hatten keinen Einfluss auf die Methylierung der DNA beider Gene.    A survey was made for Fusarium sporotrichioides on its methylation of DNA by choosing the mycotoxin genes Tri5 and Tri14 and applying different temperature and infection conditions. The restriction endonuclease MspJI was used which only digests DNA when the nucleotide sequence Guanosin/Cytosine (G/C) is methylated. Unmethylated G/C-DNA is not restricted by this enzyme and therefore a differentiation is possible in a simple way. A successful restriction and non-restriction was proved by using PCR-primer, covering both genes. Methylation of the coding DNA for the two mycotoxin genes Tri5 und Tri14 was detected by MspJI, only when F. sporotrichioides infected its host plants barley and maize. All other conditions did not have any influence on the methylation of the two genes.   &nbsp

Use of inert cultivation material for semi-sterile amplification of potato wart for lab-experiments applying the Glynne-Lemmerzahl method

Um semi-sterile Kartoffelkrebswucherungen für die unterschiedlichsten Untersuchungszwecke im Labor zu erhalten, wurde der methodische Schritt der Glynne-Lemmerzahl-Methode an derjenigen Stelle modifiziert, an der frisch infizierte Kartoffelkeime zur Gewinnung von Kartoffelkrebswucherungen mit Kompost überschichtet werden. Insgesamt kamen anstelle des Komposts drei verschiedene Materialien zum Einsatz: Perlit, Vliestücher und Sägemehl. Es zeigte sich, dass mit anfälligen Kartoffelsorten im Sägemehl, unabhängig vom Baumtyp, ähnlich große Wucherungen produzierbar waren wie im Kompost, während die anderen beiden Substanzen schlechtere Wucherungen hervorbrachten. Bei der Verwendung von Sägemehl waren die Kartoffelkrebswucherungen deutlich geringer kontaminiert als bei den Kompost-Varianten.In order to obtain semi-sterile potato wart pathotypes for different kinds of lab-research, parts of the methodological approach of the Glynne-Lemmerzahl method were modified at the step where the wart-infected sprouts were covered with compost to generate new infective warts. Three different subsidising substances were applied: Perlit, fleece cloths and saw dust. Only with saw dust the galls obtained were of a similar size as those from the previously used compost, independently of the kind of tree the saw dust came from, whereas the other two substances resulted in minor gall formations. The use of saw dust produced far less contaminated galls of potato wart than the compost version

Epigenetics – a new way to understand reactions of plant pathogenic fungi

Fusarium sporotrichioides-Isolat 62423 aus der JKI-(Julius Kühn-Institut)-Stammsammlung wurde verschiedenen Umweltbedingungen ausgesetzt, was eine Veränderung des Phänotyps bewirkte. Die chromosomale DNA wurde mit den isoschizomeren Endonukleasen MspI und HpaII komplett im Palindrom CCGG verdaut, was je nach Methylierungsgrad der DNA unterschiedlich stark erfolgte und mittels PCR und Primern des Tri14-Gens nachweisbar war. So korrelierte die phänotypische Veränderung des Isolates 62423, die sich in Form von farblich unterschiedlichen Ringen auf Kartoffeldextrose-(PDA)-Agarplatten unter Blaulicht zeigte, mit einem veränderten DNA-Methylierungsgrad im Vergleich zum unter „normalen“ Tageslicht-Bedingungen gewachsenen Isolat, was mit der jeweiligen Restriktionsintensität und den amplifizierten PCR-Fragmenten sichtbar wurde.    Fusarium sporotrichioides isolate 62423 from the JKI-strain collection was incubated under different environmental conditions resulting in a change of its phenotype. The chromosomal DNA was allowed to be restricted completely by using the isoschizomere endonucleases MspI and HpaII both recognizing the palindrome CCGG, however, occurring differently according to their degree of methylation. This could be shown by PCR amplification using primers of the Tri14 gene. The phenotypic change of isolate 62423 was manifested when growing on PDA by forming colored rings when dark blue light was applied. There was a correlation of the methylation intensity when the DNA of the fungus was compared with the same isolate incubated at normal daylight resulting in different amounts of amplified DNA.   &nbsp

Newly characterized pathogenicity-correlated genes are suitable in a PCR-test for a further differentiation of Fusarium oxysporum f. sp. lycopersici-isolates from the JKI strain collection

Am Beispiel von Fusarium oxysporum forma specialis (f. sp.) lycopersici aus der JKI-Stammsammlung wurde das Vorkommen des Pathogenitäts-korrelierten, bedingt überzähligen (conditional dispensable, CD) Chromosoms mittels PCR untersucht. So ließ sich nur bei F. oxysporum f. sp. lycopersici das Vorhandensein des CD-Chromosoms mit der PCR nachweisen, während bei der nahe verwandten f. sp. radicis-lycopersici dieses Chromosom fehlte. Die These einer stattgefundenen Übertragung des CD-Chromosoms auf verschiedene klonale Linien von F. oxysporum wurde durch die Verwendung von teilspezifischen Primern zur Untersuchung des chromosomalen „Hintergrunds“ der einzelnen Fusarium oxysporum-Isolate getestet. Bei zehn in der JKI-Stammsammlung vorhandenen F. oxysporum f. sp. lycopersici-Isolaten ließen sich deutlich zwei verschiedene chromosomale „Hintergründe“ mit insgesamt drei verwendeten teilspezifischen Primerpaaren nachweisen. Mit diesen Versuchen konnte gezeigt werden, dass eine Polyphyletie bei F. oxysporum f. sp. lycopersici vorliegt, es also zwei verschiedene Fusarium oxysporum f. sp. lycopersici-Linien in der JKI-Stammsammlung gibt, die dieses CD-Chromosom besitzen. Polyphyletische Studien könnten zukünftig als zusätzliches Kriterium zur Einordnung von Fusarium-Isolaten verwendet werden.Fusarium oxysporum forma specialis (f. sp.) lycopersici was evaluated as an example from the JKI strain col­lection to detect the occurrence of the pathogenicity-correlated conditional dispensable (CD) chromosome via PCR. Only in Fusarium oxysporum f. sp. lycopersici the CD-chromosome could be detected, while it was not detected in the closely related F. sp. radicis-lycopersici. The theory of a transfer of the CD-chromosome into different clonal lines of F. oxysporum was tested by the application of partly-specific primers to investigate the chromosomal background of each isolate. Ten conserved Fusarium oxysporum f. sp. lycopersici isolates of the JKI strain collection were clearly differentiated into two chromosomal categories, when three partly-specific primer pairs were applied. With these experiments it could be shown that Fusarium oxysporum f. sp. lycopersici was polyphyletic, i. e. there are two different Fusarium oxysporum f. sp. lycopersici lines in the JKI strain collection possessing the CD chromosome. In future, polyphyletic studies could be used as an additional criterion for the classification of Fusarium isolates

Application of the SNP-analysis for characterisation of Phytophthora infestans-isolates

Using the single nucleotid polymorphism (SNP) method P. infestans could be easily characterised. P. infestans-isolates collected in the period from 2000 to 2004 did show a continuous change of the chromosomal area used for the SNP analysis. At the end of the survey only 25% of the isolates corresponded with the original DNA sequence (230 bp fragment). Since a non-coding DNA region was used for the SNP analysis, genetic changes could easier be found within the surveyed period. For monitoring the genetic changes of P. infestans-isolates a host change was performed (potato-tomato respectively black nightshade). Parts of the surveyed P. infestans isolates showed a stable SNP-fragment of the proper size, being independently from the host change. The other part of the isolates did not only show visual microscopic differences in the sporangia but also in the detection of the 230 bp SNP-fragment. Since it is not plausible that in a highly variable gene a fast change via point mutation can occur that fast, it is assumed that in that case a population mixture was surveyed. Therefore, a stress in the form of a host change might favour the one or other P. infestans-isolate exhibiting a change in the SNP-fragment. When the P. infestans isolates grown on different hosts were re-inoculated onto their potato hosts, the acquired change in the SNP-pattern was kept.vokMyynti MTT Tietopalvelut 31600 Jokioine

Developing a systems thinking approach when it comes to developing possible solutions to climate change

Few issues facing society are more urgent than reducing our vulnerability to climate impacts, preparing for the staggering transition to a low-carbon economy, and building resilient communities. Yet K-12 schools, higher education, and free-choice learning institutions are often not prepared or focused on building awareness and inspiring action to care for our communities and our planet. Through activation of nations and communities extensive education systems with comprehensive climate change education, communities can more quickly embrace a low-carbon future, inspire future leaders, showcase their cities’ adaptability, and create stronger communities. The need for comprehensive, interdisciplinary climate change education is more important now than ever before. Since 1988, the Intergovernmental Panel on Climate Change (IPCC), an intergovernmental body of the United Nations, has provided the world with an objective, scientific view of climate change and its political and economic impacts. On October 8, 2018, the IPCC released the most important climate report to date. This IPCC special report, Global Warming of 1.5° C, provides insight into the collective global greenhouse gas emission choices that led to a warming of 1.5° C or higher above pre-industrial levels and serves as an urgent call to rapidly transition our global communities to low-carbon economies. It is also the first international climate report to provide a viable way to reach the goals set forth in the landmark Paris Climate Agreement to combat climate change, and accelerate and intensify the actions and investments needed for a sustainable low-carbon future. The Climate Science and Education stated in the 2009 "Climate Literacy: The Essential Principles of Climate Science guide for Individuals and Communities” (USGCRP, 2009) why climate is key scientific issue that students need to address in their learning;  “To protect fragile ecosystems and to build sustainable communities that are resilient to climate change— including extreme weather and climate events—a climate-literate citizenry is essential. This climate science literacy guide identifies the essential principles and fundamental concepts that individuals and communities should understand about Earth’s climate system. Such understanding improves our ability to make decisions about activities that increase vulnerability to the impacts of climate change and to take precautionary steps in our lives and livelihoods that would reduce those vulnerabilities.” The Climate Literacy Guide was used to create the climate related standards supporting the learning of more than three-quarters (84%) of U.S. students live in states that have education standards influenced by the Framework for K-12 Science Education and/or the Next Generation Science Standards. To strengthen the learning related to climate change, we need to recognize and identify learning pathways that involves the complex, dynamic systems that demand a systems thinking approach when it comes to developing possible solutions. A systems thinking approach is increasingly recognized as a critical approach for education to address climate change. Climate change epitomizes a problem that demands a systems thinking and system dynamics approach: it is dynamic, complex, and crosses disciplines and societal sectors. Addressing the impacts and societal problems resulting from climate change requires an unprecedented level of integration and education across scientific, social science, civic/government, and humanities fields. Systems thinking offers an opportunity to integrate knowledge across disciplines and move society’s capabilities to rapidly transition to a low-carbon economy and address the impacts of climate change.

PCR - detection of Synchytrium endobioticum (Schilb.) Perc.

The ITS-DNA region of Synchytrium endobioticum (Schilb.) Perc, the causal agent of po-tato wart disease, was used to generate specific primers for a PCR. DNA was extracted from wart galls of all S. endobioticum pathotypes (1, 2, 6, 18) currently occurring and used for official testing purposes in Germany. Using the universal ITS primer # 4 and the S. endobioticum specific primer Kbr1, a PCR fragment of 525 bp was obtained from all four fungal pathotypes. DNA for PCR based diagnosis of the fungus could be extracted easily from summer sori, however, we did not succeed to extract amplifiable DNA from resting sori found in contaminated soils. To circumvent this problem, zoospores emerging from resting sori were used for DNA extraction and indirect detection. PCR was also useful to discriminate between of weakly resistant and moderately susceptible response of potato varieties in addition to the routine visual inspection, since only alive summer sori of the weakly susceptible reacting potato varieties released quantities of zoospores sufficient to be detected. No PCR signal was obtained from weakly resistant or resistant potato varie-ties, since no zoospores were released from the dead sori, due to the resistance response of the potato.vokMyynti MTT Tietopalvelut 31600 Jokioine