Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1.

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field

Endophyte-mediated resistance against white pine blister rust in Pinus monticola

Abstract Induced resistance responses, including fungal endophyte-mediated resistance, have been well studied in both agricultural crops and grass systems. Yet, the effect of these processes and symbionts in forest trees is poorly known. Fungal endophytes have been found in all conifer forest systems examined to date and have been hypothesised to be involved in resistance-mediated responses. However, in the absence of functional studies the influence of these endophytes on the extended phenotype of the host plant is unclear. In this study we demonstrate that fungal endophytes from Pinus monticola were effective at increasing survival in host plants against the exotic pathogen Cronartium ribicola, which is responsible for the devastating disease called white pine blister rust. Seedlings previously inoculated with fungal endophytes lived longer than endophyte-free seedlings and also showed some reduction in white pine blister rust disease severity. This endophyte-mediated resistance was found to be effective over time, indicating persistence, and is hypothesised to be a form of induced resistance. Overall, this suggests fungal endophytes may play a determinative role in the structure of biological communities and could provide a useful alternative or ancillary management tool for combating pests and diseases.

Attracting and retaining women in forest entomology and forest pathology

Inclusion of women in the forest entomology and forest pathology workforce has been a difficult journey. While policies and resources exist for organizations and departments to increase diversity and retain women, there still exist large gaps in gender parity at forest research institutions globally. It is imperative that we better understand the barriers that exist for women in forest entomology and pathology so that more inclusive environments can be created that are welcoming towards women and other underrepresented groups. To assess these barriers and subsequent opportunities for improvement, we surveyed forest entomology and pathology professionals globally to ask about their experiences in the workforce. We also provide examples of trends in gender representation at relevant institutions. Barriers to success for women and men were very different; women experienced more barriers related to family caretaking while men experienced more barriers related to funding opportunities. These differences show where opportunities lie to better attract, support, and retain women in forest entomology and forest pathology. Although some trends in gender representation are promising, large gaps continue to exist which need to be addressed

Role of insect vectors in epidemiology and invasion risk of Fusarium circinatum, and risk assessment of biological control of invasive Pinus contorta

Pitch canker, caused by the pathogen Fusarium circinatum, is a serious disease of pines, Pinus species. It is a threat to natural and planted pine forests, and to date it has invaded countries across five continents. Pine-feeding insects can play a key role in the epidemiology of the disease, as wounding agents allowing pathogen access or as vectors transmitting the pathogen from infected to healthy trees. We reviewed the role of insects in the epidemiology of pitch canker worldwide and assessed which insects are present in New Zealand that may act as wounding agents or vectors to determine whether pathogen invasion could adversely affect Pinus radiata plantation forests and urban trees. We also evaluated whether cone or seed insects of pines could be introduced as biological control agents of invasive Pinus contorta and how this may affect the impact of a potential F. circinatum invasion. As there are no native pines or other Pinaceae in New Zealand, there are only a few pine insects, mainly accidental introductions. None of the insects recorded on pines in New Zealand is likely to be a vector, suggesting low disease risk. Of six potentially suitable biocontrol candidates, the European pine cone weevil Pissodes validirostris is the most promising regarding host specificity and impact on seed production, but there is uncertainty about its ability to act as a vector of F. circinatum. Our methodology to review and evaluate the vector potential of pine associates can be used as a generic framework to assess the potential impacts of F. circinatum invasion

Dothistroma pini, a Forest Pathogen, Contains Homologs of Aflatoxin Biosynthetic Pathway Genes

Homologs of aflatoxin biosynthetic genes have been identified in the pine needle pathogen Dothistroma pini. D. pini produces dothistromin, a difuranoanthraquinone toxin with structural similarity to the aflatoxin precursor versicolorin B. Previous studies with purified dothistromin suggest a possible role for this toxin in pathogenicity. By using an aflatoxin gene as a hybridization probe, a genomic D. pini clone was identified that contained four dot genes with similarity to genes in aflatoxin and sterigmatocystin gene clusters with predicted activities of a ketoreductase (dotA), oxidase (dotB), major facilitator superfamily transporter (dotC), and thioesterase (dotD). A D. pini dotA mutant was made by targeted gene replacement and shown to be severely impaired in dothistromin production, confirming that dotA is involved in dothistromin biosynthesis. Accumulation of versicolorin A (a precursor of aflatoxin) by the dotA mutant confirms that the dotA gene product is involved in an aflatoxin-like biosynthetic pathway. Since toxin genes have been found to be clustered in fungi in every case analyzed so far, it is speculated that the four dot genes may comprise part of a dothistromin biosynthetic gene cluster. A fifth gene, ddhA, is not a homolog of aflatoxin genes and could be at one end of the dothistromin cluster. These genes will allow comparative biochemical and genetic studies of the aflatoxin and dothistromin biosynthetic pathways and may also lead to new ways to control Dothistroma needle blight