zotero.bib

@article{kiss_modified_2024,
	title = {A modified {CTAB} method for the extraction of high-quality {RNA} from mono-and dicotyledonous plants rich in secondary metabolites},
	volume = {20},
	issn = {1746-4811},
	url = {https://plantmethods.biomedcentral.com/articles/10.1186/s13007-024-01198-z},
	doi = {10.1186/s13007-024-01198-z},
	abstract = {Abstract
            
              Background
              High-quality RNA extraction from woody plants is difficult because of the presence of polysaccharides and polyphenolics that bind or co-precipitate with the RNA. The CTAB (cetyl trimethylammonium bromide) based method is widely used for the isolation of nucleic acids from polysaccharide-rich plants. Despite the widespread use of the CTAB method, it is necessary to adapt it to particular plant species, tissues and organs. Here we described a simple and generalized method for RNA isolation from mature leaf tissues of several economically important woody (17) and herbaceous plants (2) rich in secondary metabolites. High yields were achieved from small amount (up to 50 mg) of plant material. Two main modifications were applied to the basic protocol: an increase in β-mercaptoethanol concentration (to 10\%v/v) and the use of an effective DNase treatment. As opposed to similar studies, we tried to describe a more detailed protocol for isolating RNA, including the exact quantity and concentration of the reagents were used.
            
            
              Results
              
                Our modified CTAB method is proved to be efficient in extracting the total RNA from a broad range of woody and herbaceous species. The RNA yield was ranged from 2.37 to 91.33 µg/µl. The A
                260
                :A
                280
                and A
                260
                :A
                230
                absorbance ratios were measured from 1.77 to 2.13 and from 1.81 to 2.22. The RIN value (RNA Integrity Number) of the samples fell between 7.1 and 8.1, which indicated that a small degree of RNA degradation occurred during extraction. The presence of a single peak in the melt curve analyses and low standard errors of the Ct values of replicated measurements indicated the specificity of the primers to bind to the cDNA.
              
            
            
              Conclusions
              
                Our RNA isolation method, with fine-tuned and detailed instructions, can produce high quality RNA from a small amount of starting plant material that is suitable for use in downstream transcriptional analyses. The use of an increased concentration of the reducing agent β-mercaptoethanol in the extraction buffer, as well as the application of DNaseI-treatment resulted in a method suitable for a wide range of plants without the need of further optimalization, especially in
                Rhus typhina
                (Staghorn sumac), for which molecular-genetic studies have not yet been sufficiently explored.},
	language = {en},
	number = {1},
	urldate = {2024-11-04},
	journal = {Plant Methods},
	author = {Kiss, Tibor and Karácsony, Zoltán and Gomba-Tóth, Adrienn and Szabadi, Kriszta Lilla and Spitzmüller, Zsolt and Hegyi-Kaló, Júlia and Cels, Thomas and Otto, Margot and Golen, Richárd and Hegyi, Ádám István and Geml, József and Váczy, Kálmán Zoltán},
	month = may,
	year = {2024},
	pages = {62},
}

@article{di_pietro_endopolygalacturonase_1996,
	title = {Endopolygalacturonase from {Fusarium} oxysporum f. sp. lycopersici: {Purification}, {Characterization}, and {Production} {During} {Infection} of {Tomato} {Plants}},
	volume = {86},
	url = {https://www.apsnet.org/publications/phytopathology/backissues/Documents/1996Abstracts/Phyto_86_1324.htm},
	number = {12},
	journal = {Phytopathology},
	author = {Di Pietro, Antonio and G. Roncero, M. Isabel},
	year = {1996},
}

@article{segorbe_three_2017,
	title = {Three \textit{{Fusarium} oxysporum} mitogen‐activated protein kinases ({MAPKs}) have distinct and complementary roles in stress adaptation and cross‐kingdom pathogenicity},
	volume = {18},
	issn = {1464-6722, 1364-3703},
	url = {https://bsppjournals.onlinelibrary.wiley.com/doi/10.1111/mpp.12446},
	doi = {10.1111/mpp.12446},
	abstract = {Summary
            
              Mitogen‐activated protein kinase (MAPK) cascades mediate cellular responses to environmental signals. Previous studies in the fungal pathogen
              Fusarium oxysporum
              have revealed a crucial role of Fmk1, the MAPK orthologous to
              Saccharomyces cerevisiae
              Fus3/Kss1, in vegetative hyphal fusion and plant infection. Here, we genetically dissected the individual and combined contributions of the three MAPKs Fmk1, Mpk1 and Hog1 in the regulation of development, stress response and virulence of
              F. oxysporum
              on plant and animal hosts. Mutants lacking Fmk1 or Mpk1 were affected in reactive oxygen species (ROS) homeostasis and impaired in hyphal fusion and aggregation. Loss of Mpk1 also led to increased sensitivity to cell wall and heat stress, which was exacerbated by simultaneous inactivation of Fmk1, suggesting that both MAPKs contribute to cellular adaptation to high temperature, a prerequisite for mammalian pathogens. Deletion of Hog1 caused increased sensitivity to hyperosmotic stress and resulted in partial rescue of the restricted colony growth phenotype of the
              mpk1
              Δ mutant. Infection assays on tomato plants and the invertebrate animal host
              Galleria mellonella
              revealed distinct and additive contributions of the different MAPKs to virulence. Our results indicate that positive and negative cross‐talk between the three MAPK pathways regulates stress adaptation, development and virulence in the cross‐kingdom pathogen
              F. oxysporum
              .},
	language = {en},
	number = {7},
	urldate = {2024-10-07},
	journal = {Molecular Plant Pathology},
	author = {Segorbe, David and Di Pietro, Antonio and Pérez‐Nadales, Elena and Turrà, David},
	month = sep,
	year = {2017},
	pages = {912--924},
}

@article{andersen_mapping_2012,
	title = {Mapping the polysaccharide degradation potential of {Aspergillus} niger},
	volume = {13},
	issn = {1471-2164},
	url = {https://bmcgenomics.biomedcentral.com/articles/10.1186/1471-2164-13-313},
	doi = {10.1186/1471-2164-13-313},
	abstract = {Abstract
            
              Background
              The degradation of plant materials by enzymes is an industry of increasing importance. For sustainable production of second generation biofuels and other products of industrial biotechnology, efficient degradation of non-edible plant polysaccharides such as hemicellulose is required. For each type of hemicellulose, a complex mixture of enzymes is required for complete conversion to fermentable monosaccharides. In plant-biomass degrading fungi, these enzymes are regulated and released by complex regulatory structures. In this study, we present a methodology for evaluating the potential of a given fungus for polysaccharide degradation.
            
            
              Results
              
                Through the compilation of information from 203 articles, we have systematized knowledge on the structure and degradation of 16 major types of plant polysaccharides to form a graphical overview. As a case example, we have combined this with a list of 188 genes coding for carbohydrate-active enzymes from
                Aspergillus niger
                , thus forming an analysis framework, which can be queried. Combination of this information network with gene expression analysis on mono- and polysaccharide substrates has allowed elucidation of concerted gene expression from this organism. One such example is the identification of a full set of extracellular polysaccharide-acting genes for the degradation of oat spelt xylan.
              
            
            
              Conclusions
              
                The mapping of plant polysaccharide structures along with the corresponding enzymatic activities is a powerful framework for expression analysis of carbohydrate-active enzymes. Applying this network-based approach, we provide the first genome-scale characterization of all genes coding for carbohydrate-active enzymes identified in
                A. niger
                .},
	language = {en},
	number = {1},
	urldate = {2024-10-02},
	journal = {BMC Genomics},
	author = {Andersen, Mikael R and Giese, Malene and De Vries, Ronald P and Nielsen, Jens},
	month = dec,
	year = {2012},
	pages = {313},
}

@article{de_vries_aspergillus_2001,
	title = {\textit{{Aspergillus}} {Enzymes} {Involved} in {Degradation} of {Plant} {Cell} {Wall} {Polysaccharides}},
	volume = {65},
	issn = {1092-2172, 1098-5557},
	url = {https://journals.asm.org/doi/10.1128/MMBR.65.4.497-522.2001},
	doi = {10.1128/MMBR.65.4.497-522.2001},
	abstract = {SUMMARY
              Degradation of plant cell wall polysaccharides is of major importance in the food and feed, beverage, textile, and paper and pulp industries, as well as in several other industrial production processes. Enzymatic degradation of these polymers has received attention for many years and is becoming a more and more attractive alternative to chemical and mechanical processes. Over the past 15 years, much progress has been made in elucidating the structural characteristics of these polysaccharides and in characterizing the enzymes involved in their degradation and the genes of biotechnologically relevant microorganisms encoding these enzymes. The members of the fungal genus Aspergillus are commonly used for the production of polysaccharide-degrading enzymes. This genus produces a wide spectrum of cell wall-degrading enzymes, allowing not only complete degradation of the polysaccharides but also tailored modifications by using specific enzymes purified from these fungi. This review summarizes our current knowledge of the cell wall polysaccharide-degrading enzymes from aspergilli and the genes by which they are encoded.},
	language = {en},
	number = {4},
	urldate = {2024-10-02},
	journal = {Microbiology and Molecular Biology Reviews},
	author = {De Vries, Ronald P. and Visser, Jaap},
	month = dec,
	year = {2001},
	pages = {497--522},
}

@article{roncero_fusarium_2003,
	title = {Fusarium as a model for studying virulence in soilborne plant pathogens},
	volume = {62},
	copyright = {https://www.elsevier.com/tdm/userlicense/1.0/},
	issn = {08855765},
	url = {https://linkinghub.elsevier.com/retrieve/pii/S0885576503000432},
	doi = {10.1016/S0885-5765(03)00043-2},
	language = {en},
	number = {2},
	urldate = {2024-10-02},
	journal = {Physiological and Molecular Plant Pathology},
	author = {Roncero, M},
	month = feb,
	year = {2003},
	pages = {87--98},
}

@article{rispail_fusarium_2009,
	title = {\textit{{Fusarium} oxysporum} {Ste12} {Controls} {Invasive} {Growth} and {Virulence} {Downstream} of the {Fmk1} {MAPK} {Cascade}},
	volume = {22},
	issn = {0894-0282, 1943-7706},
	url = {https://apsjournals.apsnet.org/doi/10.1094/MPMI-22-7-0830},
	doi = {10.1094/MPMI-22-7-0830},
	abstract = {A conserved mitogen-activated protein kinase (MAPK) cascade homologous to the yeast Fus3/Kss1 mating/filamentation pathway regulates virulence in fungal plant pathogens. In the soilborne fungus Fusarium oxysporum, the MAPK Fmk1 is required for infection and development of vascular wilt disease on tomato plants. Knockout mutants lacking Fmk1 are deficient in multiple virulence-related functions, including root adhesion and penetration, invasive growth, secretion of pectinolytic enzymes, and vegetative hyphal fusion. The transcription factors mediating these different outputs downstream of the MAPK cascade are currently unknown. In this study, we have analyzed the role of ste12 which encodes an orthologue of the yeast homeodomain transcription factor Ste12p. F. oxysporum mutants lacking the ste12 gene were impaired in invasive growth on tomato and apple fruit tissue and in penetration of cellophane membranes. However, ste12 was not required for adhesion to tomato roots, secretion of pectinolytic enzymes, and vegetative hyphal fusion, suggesting that these Fmk1-dependent functions are mediated by other downstream MAPK targets. The Δste12 strains displayed dramatically reduced virulence on tomato plants, similar to the Δfmk1 mutant. These results indicate that invasive growth is the major virulence function controlled by the Fmk1 MAPK cascade and depends critically on the transcription factor Ste12.},
	language = {en},
	number = {7},
	urldate = {2024-10-02},
	journal = {Molecular Plant-Microbe Interactions®},
	author = {Rispail, Nicolas and Di Pietro, Antonio},
	month = jul,
	year = {2009},
	pages = {830--839},
}

@unpublished{noci_effects_nodate,
	title = {Effects of the lack of regulation during growth and infection in the pathogenic fungus {Fusarium} oxysporum},
	language = {es},
	author = {Noci, Alfonso José Cañuelo},
}

@article{benocci_regulators_2017,
	title = {Regulators of plant biomass degradation in ascomycetous fungi},
	volume = {10},
	issn = {1754-6834},
	url = {http://biotechnologyforbiofuels.biomedcentral.com/articles/10.1186/s13068-017-0841-x},
	doi = {10.1186/s13068-017-0841-x},
	language = {en},
	number = {1},
	urldate = {2024-10-01},
	journal = {Biotechnology for Biofuels},
	author = {Benocci, Tiziano and Aguilar-Pontes, Maria Victoria and Zhou, Miaomiao and Seiboth, Bernhard and De Vries, Ronald P.},
	month = dec,
	year = {2017},
	pages = {152},
}

@article{chroumpi_identification_2020,
	title = {Identification of a gene encoding the last step of the {L}-rhamnose catabolic pathway in {Aspergillus} niger revealed the inducer of the pathway regulator},
	volume = {234},
	issn = {09445013},
	url = {https://linkinghub.elsevier.com/retrieve/pii/S0944501319312790},
	doi = {10.1016/j.micres.2020.126426},
	language = {en},
	urldate = {2024-10-01},
	journal = {Microbiological Research},
	author = {Chroumpi, Tania and Aguilar-Pontes, Maria Victoria and Peng, Mao and Wang, Mei and Lipzen, Anna and Ng, Vivian and Grigoriev, Igor V. and Mäkelä, Miia R. and De Vries, Ronald P.},
	month = apr,
	year = {2020},
	pages = {126426},
}

@article{khosravi_vivo_2017,
	title = {In vivo functional analysis of {L}-rhamnose metabolic pathway in {Aspergillus} niger: a tool to identify the potential inducer of {RhaR}},
	volume = {17},
	issn = {1471-2180},
	shorttitle = {In vivo functional analysis of {L}-rhamnose metabolic pathway in {Aspergillus} niger},
	url = {http://bmcmicrobiol.biomedcentral.com/articles/10.1186/s12866-017-1118-z},
	doi = {10.1186/s12866-017-1118-z},
	language = {en},
	number = {1},
	urldate = {2024-10-01},
	journal = {BMC Microbiology},
	author = {Khosravi, Claire and Kun, Roland Sándor and Visser, Jaap and Aguilar-Pontes, María Victoria and De Vries, Ronald P. and Battaglia, Evy},
	month = dec,
	year = {2017},
	pages = {214},
}

@misc{kowalczyk_molecular_2017,
	type = {Dissertation},
	title = {Molecular mechanisms shaping the plant biomass utilization potential in {Aspergillus}},
	copyright = {Open Access (free)},
	url = {https://dspace.library.uu.nl/handle/1874/356771},
	abstract = {Coordination of the induction of genes encoding extracellular enzymes and sugar uptake systems in fungi are often mediated by transcription factors (TFs). The aim of this thesis was to study the transcriptional regulatory network in more detail by analysis of sugar-specific TFs in A. niger and A. nidulans. This research was focused on interactive aspects of regulation such as (1) combinatorial control of gene expression and (2) identification of novel transcriptional network motifs linking the TFs. In addition, a few previously characterized TFs involved in plant biomass degradation were re-evaluated in a broader context. Chapter 2 describes genetic interactions between A. nidulans AraR, XlnR and GalR in regulation of pentose and D-galactose catabolism. AraR and XlnR were previously shown to regulate genes encoding enzymes involved in L-arabinose and D-xylose conversion in both A. nidulans and A. niger, while GalR was linked to regulation of the Leloir D-galactose pathway. Our data showed that GalR is involved in the oxido-reductive D-galactose catabolic pathway in A. nidulans, but not in the pentose catabolic pathway. In contrast, AraR and XlnR not only control the pentose catabolic pathways, but also genes of the oxido-reductive D-galactose catabolic pathway. Chapter 3 describes characterization of D-galacturonic acid responsive TF GaaR in A. niger by studying the transcriptome response of the reference strain and ΔgaaR grown on D-galacturonic acid and pectin. GaaR was shown to mainly control expression of genes encoding enzymes involved in degradation of the homogalacturonan part of pectin as well as D-galacturonic acid transport and metabolism. Complete degradation of pectin might require co-operative action of several TFs in A. niger. Previous studies identified two TFs involved in degradation of pectin, AraR, responding to L-arabinose, and RhaR, responding to L-rhamnose. Chapter 4 describes combinatorial control of gene expression by AraR, RhaR and the recently characterized GaaR in A. niger grown on sugar beet pectin. The results of this study demonstrate that GaaR, AraR and RhaR are all involved in regulation of pectinolytic gene expression during growth on a complex carbon source, which is rich in pectin. Moreover, the study reveals genes under combined control of two or three TFs. Transcriptional regulatory networks are based on interactions between TFs and their target genes. In some cases, expression of a gene relies on two TFs, of which the ‘primary’ TF regulates the ‘modulating’ TF. The primary TF is activated by the presence of an inducer, while the modulating TF is transcriptionally activated by the primary TF. Chapter 5 describes the identification of two putative XlnR and AmyR-affected modulating TFs and asses their involvement in plant biomass degradation in A. niger. In Chapter 6, we studied the physiological role of XlnR and AraR in colonization of unprocessed wheat bran by high-resolution microscopy and exo-proteomics. Deletion of xlnR in A. niger reduced secretion of hemicellulolytic enzymes and resulted in reduced biofilm formation especially on the smooth surface of the wheat bran flake. This is the first in vivo demonstration of the physiological importance of these regulators. The results are summarized and discussed in Chapter 7.},
	language = {en},
	urldate = {2024-10-01},
	author = {Kowalczyk, Joanna E.},
	month = oct,
	year = {2017},
	note = {Accepted: 2017-11-17T17:46:53Z
ISBN: 9789462957367
Publisher: Utrecht University},
}

@phdthesis{palos-fernandez_papel_2023,
	title = {Papel de la homeostasis del cobre en la patogénesis fúngica},
	copyright = {https://creativecommons.org/licenses/by-nc-nd/4.0/},
	url = {http://helvia.uco.es/xmlui/handle/10396/26386},
	abstract = {In fungi, copper acquisition is mainly carried out by specific high-affinity systems consisting of Fre metalloreductases and Ctr copper transporters, which are transcriptionally activated under copper limiting conditions by the copper-sensing transcription factor Mac1. Mac1 is required for full virulence in several animal pathogenic fungi, but its role in phytopathogenic fungi has not been described so far. In this work we show that Mac1 controls the transcriptional response of the vascular wilt pathogen Fusarium oxysporum f. sp. lycopersici to copper limitation and that it is essential for growth under copper limiting conditions and for virulence on tomato plants. Importantly, simultaneous overexpression of the high-affinity copper transporter ctr3 and the metalloreductase fre9 in a mac1Δ background restored growth under copper limitation and pathogenicity, indicating that the main role of Mac1 during infection is to ensure efficient copper acquisition. Because inactivation of Mac1 in F. oxysporum leads to increased sensitivity to oxidative stress and reduced virulence, we studied the role of superoxide dismutases (SODs), a class of copper-dependent enzymes that are relevant for pathogenicity in other fungi. We found that the two SODs of F. oxysporum that use copper as a cofactor, Sod1 and Sod5, play a role in the pathogenicity. Although the activity of Sod5 could not be detected in our in vitro assays, the enzymatic activity of Sod1 was reduced in mac1Δunder copper-limiting conditions, suggesting that failure to activate Mac1-dependent high-affinity copper acquisition could lead to reduced activity of copper-dependent SODs and explain the virulence phenotype of the mac1Δ mutant. On the other hand, the response to oxidative stress in F. oxysporum was previously shown to be controlled by the Mpk1 mitogen-activated protein kinase (MAPK) signalling pathway. This MAPK cascade regulates cell wall integrity and is involved in virulence-related processes that are also controlled in parallel by the Fmk1 MAPK pathway, but the downstream components that mediate these diverse functions are largely unknown. Here we find that expression of the GATA-type transcription factor Pro1 is regulated at least in part, by the Mpk1 and Fmk1 MAPK pathways, as well as by other signalling components such as the Velvet regulatory complex and the signaling protein Soft (Fso1). We further show that Pro1 controls a subset of functions downstream of Mpk1 including quorum sensing, hyphal fusion and chemotropism, whereas it is dispensable for other processes such as invasive hyphal growth, response to cell wall stress or virulence.},
	language = {spa},
	urldate = {2024-10-01},
	author = {Palos-Fernandez, Rafael},
	year = {2023},
	note = {Accepted: 2023-12-19T09:29:33Z
Publisher: Universidad de Córdoba, UCOPress},
}

@phdthesis{gomez_gil_structure_2022,
	title = {Structure and dynamics of chromosomes: role in the genomic and pathogenic plasticity of {Fusarium} oxysporum},
	shorttitle = {Structure and dynamics of chromosomes},
	url = {http://helvia.uco.es/xmlui/handle/10396/23702},
	language = {eng},
	urldate = {2024-10-01},
	author = {Gómez Gil, Lucía},
	year = {2022},
	note = {Publisher: Universidad de Córdoba, UCOPress},
}

@article{ma_fusarium_2013,
	title = {\textit{{Fusarium}} {Pathogenomics}},
	volume = {67},
	issn = {0066-4227, 1545-3251},
	url = {https://www.annualreviews.org/doi/10.1146/annurev-micro-092412-155650},
	doi = {10.1146/annurev-micro-092412-155650},
	abstract = {Fusarium is a genus of filamentous fungi that contains many agronomically important plant pathogens, mycotoxin producers, and opportunistic human pathogens. Comparative analyses have revealed that the Fusarium genome is compartmentalized into regions responsible for primary metabolism and reproduction (core genome), and pathogen virulence, host specialization, and possibly other functions (adaptive genome). Genes involved in virulence and host specialization are located on pathogenicity chromosomes within strains pathogenic to tomato (Fusarium oxysporum f. sp. lycopersici) and pea (Fusarium ‘solani’ f. sp. pisi). The experimental transfer of pathogenicity chromosomes from F. oxysporum f. sp. lycopersici into a nonpathogen transformed the latter into a tomato pathogen. Thus, horizontal transfer may explain the polyphyletic origins of host specificity within the genus. Additional genome-scale comparative and functional studies are needed to elucidate the evolution and diversity of pathogenicity mechanisms, which may help inform novel disease management strategies against fusarial pathogens.},
	language = {en},
	number = {1},
	urldate = {2024-10-01},
	journal = {Annual Review of Microbiology},
	author = {Ma, Li-Jun and Geiser, David M. and Proctor, Robert H. and Rooney, Alejandro P. and O'Donnell, Kerry and Trail, Frances and Gardiner, Donald M. and Manners, John M. and Kazan, Kemal},
	month = sep,
	year = {2013},
	pages = {399--416},
}

@article{navarro-velasco_galleria_2011,
	title = {Galleria mellonella as model host for the trans-kingdom pathogen {Fusarium} oxysporum},
	volume = {48},
	copyright = {https://www.elsevier.com/tdm/userlicense/1.0/},
	issn = {10871845},
	url = {https://linkinghub.elsevier.com/retrieve/pii/S1087184511001587},
	doi = {10.1016/j.fgb.2011.08.004},
	language = {en},
	number = {12},
	urldate = {2024-10-01},
	journal = {Fungal Genetics and Biology},
	author = {Navarro-Velasco, Gesabel Y. and Prados-Rosales, Rafael C. and Ortíz-Urquiza, Almudena and Quesada-Moraga, Enrique and Di Pietro, Antonio},
	month = dec,
	year = {2011},
	pages = {1124--1129},
}

@incollection{brand_galleria_2012,
	address = {Totowa, NJ},
	title = {Galleria mellonella as a {Model} for {Fungal} {Pathogenicity} {Testing}},
	volume = {845},
	isbn = {978-1-61779-538-1 978-1-61779-539-8},
	url = {http://link.springer.com/10.1007/978-1-61779-539-8_33},
	language = {en},
	urldate = {2024-10-01},
	booktitle = {Host-{Fungus} {Interactions}},
	publisher = {Humana Press},
	author = {Fallon, John and Kelly, Judy and Kavanagh, Kevin},
	editor = {Brand, Alexandra C. and MacCallum, Donna M.},
	year = {2012},
	doi = {10.1007/978-1-61779-539-8_33},
	note = {Series Title: Methods in Molecular Biology},
	pages = {469--485},
}

@article{curtis_galleria_2022,
	title = {Galleria mellonella {Larvae} as a {Model} for {Investigating} {Fungal}—{Host} {Interactions}},
	volume = {3},
	issn = {2673-6128},
	url = {https://www.frontiersin.org/articles/10.3389/ffunb.2022.893494/full},
	doi = {10.3389/ffunb.2022.893494},
	abstract = {Galleria mellonella
              larvae have become a widely accepted and utilised infection model due to the functional homology displayed between their immune response to infection and that observed in the mammalian innate immune response. Due to these similarities, comparable results to murine studies can be obtained using
              G. mellonella
              larvae in assessing the virulence of fungal pathogens and the
              in vivo
              toxicity or efficacy of anti-fungal agents. This coupled with their low cost, rapid generation of results, and lack of ethical/legal considerations make this model very attractive for analysis of host-pathogen interactions. The larvae of
              G. mellonella
              have successfully been utilised to analyse various fungal virulence factors including toxin and enzyme production
              in vivo
              providing in depth analysis of the processes involved in the establishment and progression of fungal pathogens (e.g.,
              Candida spps, Aspergillus spp., Madurella mycetomatis, Mucormycetes
              , and
              Cryptococcus neoformans
              ). A variety of experimental endpoints can be employed including analysis of fungal burdens, alterations in haemocyte density or sub-populations, melanisation, and characterisation of infection progression using proteomic, histological or imaging techniques. Proteomic analysis can provide insights into both sides of the host-pathogen interaction with each respective proteome being analysed independently following infection and extraction of haemolymph from the larvae.
              G. mellonella
              can also be employed for assessing the efficacy and toxicity of antifungal strategies at concentrations comparable to those used in mammals allowing for early stage investigation of novel compounds and combinations of established therapeutic agents. These numerous applications validate the model for examination of fungal infection and development of therapeutic approaches
              in vivo
              in compliance with the need to reduce animal models in biological research.},
	urldate = {2024-10-01},
	journal = {Frontiers in Fungal Biology},
	author = {Curtis, Aaron and Binder, Ulrike and Kavanagh, Kevin},
	month = apr,
	year = {2022},
	pages = {893494},
}

@article{admella_straightforward_2022,
	title = {A {Straightforward} {Method} for the {Isolation} and {Cultivation} of {Galleria} mellonella {Hemocytes}},
	volume = {23},
	copyright = {https://creativecommons.org/licenses/by/4.0/},
	issn = {1422-0067},
	url = {https://www.mdpi.com/1422-0067/23/21/13483},
	doi = {10.3390/ijms232113483},
	abstract = {Galleria mellonella is an alternative animal model of infection. The use of this species presents a wide range of advantages, as its maintenance and rearing are both easy and inexpensive. Moreover, its use is considered to be more ethically acceptable than other models, it is conveniently sized for manipulation, and its immune system has multiple similarities with mammalian immune systems. Hemocytes are immune cells that help encapsulate and eliminate pathogens and foreign particles. All of these reasons make this insect a promising animal model. However, cultivating G. mellonella hemocytes in vitro is not straightforward and it has many difficult challenges. Here, we present a methodologically optimized protocol to establish and maintain a G. mellonella hemocyte primary culture. These improvements open the door to easily and quickly study the toxicity of nanoparticles and the interactions of particles and materials in an in vitro environment.},
	language = {en},
	number = {21},
	urldate = {2024-10-01},
	journal = {International Journal of Molecular Sciences},
	author = {Admella, Joana and Torrents, Eduard},
	month = nov,
	year = {2022},
	pages = {13483},
}