Kathie T. Hodge

Living tissues are still far from being used as practical components in biohybrid robots because of limitations in life span, sensitivity to environmental factors, and stringent culture procedures. Here, we introduce fungal mycelia as an easy-to-use and robust living component in biohybrid robots. We constructed two biohybrid robots that use the electrophysiological activity of living mycelia to control their artificial actuators. The mycelia sense their environment and issue action potential–like spiking voltages as control signals to the motors and valves of the robots that we designed and built. The paper highlights two key innovations: first, a vibration- and electromagnetic interference–shielded mycelium electrical interface that allows for stable, long-term electrophysiological bioelectric recordings during untethered, mobile operation; second, a control architecture for robots inspired by neural central pattern generators, incorporating rhythmic patterns of positive and negative spikes from the living mycelia. We used these signals to control a walking soft robot as well as a wheeled hard one. We also demonstrated the use of mycelia to respond to environmental cues by using ultraviolet light stimulation to augment the robots’ gaits.

The postharvest fruit pathogen Paecilomyces niveus produces ascospores that can survive some pasteurization temperatures, spoil fruit products, and contaminate them with patulin, an FDA-regulated mycotoxin. Preventing P. niveus from entering food systems requires a robust detection method to effectively determine sources of P. niveus spoilage and disease inoculum. We designed a new robust and culture-independent detection method using species-specific primers (PnPATf/r) based on the patK gene, encoding a 6-methylsalicylic acid synthase, in P. niveus , for use in a rapid qPCR assay. Primer specificity was validated using 24 different P. niveus isolates and 16 other important food spoilage and fruit pathogenic fungi. The threshold for detection of qPCR was 18 genome equivalents. To further validate our new detection method, we demonstrate its use in detecting P. niveus in infected fruits, infested soils and ciders, and in fruit arising from apple blossoms sprayed with a P. niveus spore suspension. Results from this study may help fruit producers address spoilage and patulin contamination by this food spoiling fungus. Highlights: New primers specific to Paecilomyces niveus (PnPATf/r) were developed based on the patK gene A qPCR assay to detect P. niveus was validated, and shown to be able to detect quantities of P. niveus DNA as low as 18 genome equivalents The new qPCR assay was used to investigate the ability of P. niveus ascospores to infect strawberry fruits and enter apple fruits through apple blossom infestation

Hard apple cider is considered to be a low-risk product for food spoilage and mycotoxin contamination due to its alcoholic nature and associated food sanitation measures. However, the thermotolerant mycotoxin-producing fungus Paecilomyces niveus may pose a significant threat to hard cider producers. Pa. niveus is known to infect apples ( Malus x domestica ), and previous research indicates that it can survive thermal processing and contaminate finished apple juice with the mycotoxin patulin. To determine if hard apple cider is susceptible to a similar spoilage phenomenon, cider apples were infected with Pa. niveus or one of three patulin-producing Penicillium species and the infected fruits underwent benchtop fermentation. Cider was made with lab inoculated Dabinett and Medaille d’Or apple cultivars, and patulin was quantified before and after fermentation. Results show that all four fungi can infect cider apples and produce patulin, some of which is lost during fermentation. Only Pa. niveus was able to actively grow throughout the fermentation process. To determine if apple cider can be treated to hinder Pa. niveus growth, selected industry-grade sanitation measures were tested, including chemical preservatives and pasteurization. High concentrations of preservatives inhibited Pa. niveus growth, but apple cider flash pasteurization was not found to significantly impact spore germination. This study confirms that hard apple cider is susceptible to fungal-mediated spoilage and patulin contamination. Pa. niveus should be of great concern to hard apple cider producers due to its demonstrated thermotolerance, survival in fermentative environments, and resistance to sanitation measures. Highlights: Apple fruits of traditional cider cultivars Dabinett and Medaille d’Or were found to be susceptible to infection by three patulin-producing Penicillium spp. and Paecilomyces niveus Pa. niveus can grow in finished fermented hard cider at 5.22% ethanol Patulin levels in cider were reduced by fermentation but still exceeded 50 µg/kg, a maximum limit set by various regulatory agencies Pa. niveus was observed to be able to grow in low concentrations of three preservatives: potassium sorbate, sulfur dioxide, and sodium benzoate

Fungi of the Conidiobolus group belong to the family Ancylistaceae (Entomophthorales, Entomophthoromycotina, Zoopagomycota) and include over 70 predominantly saprotrophic species in four similar and closely related genera, that were separated phylogenetically recently. Entomopathogenic fungi of the genus Batkoa are very close morphologically to the Conidiobolus species. Their thalli share similar morphology, and they produce ballistic conidia like closely related entomopathogenic Entomophthoraceae. Ballistic conidia are traditionally considered as an efficient tool in the pathogenic process and an important adaptation to the parasitic lifestyle. Our study aims to reconstruct the phylogeny of this fungal group using molecular and genomic data, ancestral lifestyle and morphological features of the conidiobolus-like group and the direction of their evolution. Based on phylogenetic analysis, some species previously in the family Conidiobolaceae are placed in the new families Capillidiaceae and Neoconidiobolaceae, which each include one genus, and the Conidiobolaceae now includes three genera. Intermediate between the conidiobolus-like groups and Entomophthoraceae, species in the distinct Batkoa clade now belong in the family Batkoaceae. Parasitism evolved several times in the Conidiobolus group and Ancestral State Reconstruction suggests that the evolution of ballistic conidia preceded the evolution of the parasitic lifestyle.

Paecilomyces rot of apples is a postharvest disease caused by Paecilomyces niveus , a problematic spoiling agent of fruit juices and derivatives. The fungus produces ascospores that can survive food processing and germinate in finished fruit products. Processing apple fruits infected with Paecilomyces rot can lead to P. niveus contaminated juices. Because the fungus produces the mycotoxin patulin, juice spoilage by P. niveus is an important health hazard. Little is known about the disease biology and control mechanisms of this recently described postharvest disease. Following Koch’s postulates, we determined that a range of previously untested rosaceous fruits and popular apple cultivars are susceptible to Paecilomyces rot infection. We also observed that two closely related food spoiling fungi, Paecilomyces fulvus and Paecilomyces variotti , were unable to infect, cause symptoms in, or reproduce in wounded fruits. Our results highlight the unique abilities of Paecilomyces niveus to infect a variety of fruits, produce patulin, and form highly-resistant spores capable of spoiling normally shelf-stable products.

Within is a comprehensive review of current apple powdery mildew literature, with the taxonomy, morphology, and life cycle of the causal phytopathogen Podosphaera leucotricha described. A discussion of economic impact, environmental aspects contributing to apple powdery mildew epidemiology, options for cultural, chemical, and biopesticide disease control, and fungicide resistance in P. leucotricha is also detailed. The present status of breeding for host resistance and apple cultivar susceptibility is discussed. Finally, emerging research into the biology and management of apple powdery mildew is discussed along with how they may aid future research endeavors.

The fungal genus Massospora (Zoopagomycota: Entomophthorales) includes more than a dozen obligate, sexually transmissible pathogenic species that infect cicadas (Hemiptera) worldwide. At least two species are known to produce psychoactive compounds during infection, which has garnered considerable interest for this enigmatic genus. As with many Entomophthorales, the evolutionary relationships and host associations of Massospora spp. are not well understood. The acquisition of M. diceroproctae from Arizona, M. tettigatis from Chile, and M. platypediae from California and Colorado provided an opportunity to conduct molecular phylogenetic analyses and morphological studies to investigate whether these fungi represent a monophyletic group and delimit species boundaries. In a three-locus phylogenetic analysis including the D1–D2 domains of the nuclear 28S rRNA gene (28S), elongation factor 1 alpha-like (EFL), and beta-tubulin (BTUB), Massospora was resolved in a strongly supported monophyletic group containing four well-supported genealogically exclusive lineages, based on two of three methods of phylogenetic inference. There was incongruence among the single-gene trees: two methods of phylogenetic inference recovered trees with either the same topology as the three-gene concatenated tree (EFL) or a basal polytomy (28S, BTUB). Massospora levispora and M. platypediae isolates formed a single lineage in all analyses and are synonymized here as M. levispora. Massospora diceroproctae was sister to M. cicadina in all three single-gene trees and on an extremely long branch relative to the other Massospora, and even the outgroup taxa, which may reflect an accelerated rate of molecular evolution and/or incomplete taxon sampling. The results of the morphological study presented here indicate that spore measurements may not be phylogenetically or diagnostically informative. Despite recent advances in understanding the ecology of Massospora, much about its host range and diversity remains unexplored. The emerging phylogenetic framework can provide a foundation for exploring coevolutionary relationships with cicada hosts and the evolution of behavior-altering compounds.

The fungal genus Massospora (Zoopagomycota: Entomophthorales) includes more than a dozen obligate, sexually transmissible pathogenic species that infect cicadas (Hemiptera) worldwide. At least two species are known to produce psychoactive compounds during infection, which has garnered considerable interest for this enigmatic genus. As with many Entomophthorales, the evolutionary relationships and host associations of Massospora spp. are not well understood. The acquisition of M. diceroproctae from Arizona, M. tettigatis from Chile, and M. platypediae from California and Colorado provided an opportunity to conduct molecular phylogenetic analyses and morphological studies to investigate if these fungi represent a monophyletic group and delimit species boundaries. In a three-locus phylogenetic analysis including the D1–D2 domains of the nuclear 28S rRNA gene (28S), elongation factor 1 alpha-like (EFL), and beta-tubulin (BTUB), Massospora was resolved in a strongly supported monophyletic group containing four well-supported genealogically exclusive lineages, based on two of three methods of phylogenetic inference. There was incongruence among the single-gene trees: two methods of phylogenetic inference recovered trees with either the same topology as the 3-gene concatenated tree (EFL), or a basal polytomy (28S, BTUB). Massospora levispora and M. platypediae isolates formed a single lineage in all analyses and are synonymized here as M. levispora . Massospora diceroproctae was sister to M. cicadina in all three single-gene trees and on an extremely long branch relative to the other Massospora , and even the outgroup taxa, which may reflect an accelerated rate of molecular evolution and/or incomplete taxa sampling. The results of the morphological study presented here indicate that spore measurements may not be phylogenetically or diagnostically informative. Despite recent advances in understanding the ecology of Massospora , much about its host range and diversity remains unexplored. The emerging phylogenetic framework can provide a foundation for exploring co-evolutionary relationships with cicada hosts and the evolution of behavior-altering compounds.

Paecilomyces niveus is an important food spoilage fungus that survives thermal processing in fruit products, where it produces the mycotoxin patulin. Spoilage of products has been attributed to soil contamination; however, little is known about the ecology of this organism. In this study, orchard soils and culled apple fruit were surveyed and the ability of P. niveus to infect apple was tested on two popular apple varieties. P. niveus was found in 34% of sampled orchard soils from across New York. Completing Koch’s postulates, P. niveus was demonstrated to cause postharvest disease in Gala and Golden Delicious apple. Symptoms of this disease, named Paecilomyces rot, resemble several other apple diseases, including black rot, bitter rot, and bull’s-eye rot. External symptoms of Paecilomyces rot include brown, circular, concentrically ringed lesions, with an internal rot that is firm and cone-shaped. Both Gala and Golden Delicious apple fruit inoculated with P. niveus developed lesions ≥43 mm in size at 22 days after inoculation. There is some evidence that the size of lesions and rate of infection differ between Gala and Golden Delicious, which may indicate differing resistance to P. niveus. This work shows that P. niveus is common in New York orchard soil and can cause a novel postharvest fruit disease. Whether infected fruit can serve as an overlooked source of inoculum in heat-processed apple products requires further study.

Paecilomyces niveus is an extremotolerant fungus with surprising powers to survive high temperatures and infect apples and aphids. These abilities make it a formidable enemy in food and agricultural environments. In addition, it produces patulin, the most significant mycotoxin in apples.