Fusarium proliferatum: isolation, identification and evaluation of its association with the model insect Galleria mellonella (L.) (Lepidoptera: Pyralidae)
Abstract
Although the genus Fusarium is widely recognized for including significant plant pathogens and mycotoxin producers, many Fusarium species have also demonstrated promising potential for insect control. These species exhibit characteristics such as high mortality rates, rapid action, and prolific sporulation, making them attractive candidates for agricultural pest management. In this study, the micro and macromorphology, DNA sequencing, and the effects on the model insect Galleria mellonella of Fusarium proliferatum isolated from soil using the trap method were investigated. Morphological analyses revealed detailed colony and spore structures on various agar media. Species identification was confirmed through DNA analysis using ITS and CaM gene sequences. F. proliferatum was observed to prolong the larval stage of model ınsect G. mellonella while reducing pupal duration, weight, and total egg production. Our findings indicate that the fungus exerts a dose-dependent impact on the development and reproduction of G. mellonella. Future studies should focus on identifying secondary metabolites responsible for the observed biological effects, evaluating possible effects on non-target organisms, and conducting field trials under different environmental conditions.
Keywords
Fusarium proliferatum Galleria mellonella developmental biology fungal pathogenicity identification isolation
Supporting Institution
TÜBİTAK
Project Number
TUBITAK-1001 (122O398) and TUBITAK 2209-A Research Project Support Program for Undergraduate Students
Thanks
Our work was financially supported by TUBITAK-1001, The Scientific and Technological Research Projects Funding Program (122O398) and TUBITAK 2209-A Research Project Support Program for Undergraduate Students.
References
- Aoki, T., Smith, J.A., Kasson, M.T., Freeman, S., Geiser, D.M., Geering, A.D., & O’Donnell, K. (2019). Three novel Ambrosia Fusarium Clade species producing clavate macroconidia known (F. floridanum and F. obliquiseptatum) or predicted (F. tuaranense) to be farmed by Euwallacea spp. (Coleoptera: Scolytinae) on woody hosts. Mycologia, 111(6), 919-935. DOI: 10.1080/00275514.2019.1647074
- Azor, M., Gené, J., Cano, J., Manikandan, P., Venkatapathy, N., & Guarro, J. (2009). Less- frequent Fusarium species of clinical interest: correlation between morphological and molecular identification and antifungal susceptibility. Journal of Clinical Microbiology, 47(5), 1463- 1468. DOI: 10.1128/JCM.02467-08
- da Silva Santos, A.C., Diniz, A.G., Tiago, P.V., & de Oliveira, N.T. (2020). Entomopathogenic Fusarium species: a review of their potential for the biological control of insects, implications and prospects. Fungal Biology Reviews, 34(1), 41-57. DOI: 10.1016/j.fbr.2019.12.002
- De Silva, D.D., Crous, P.W., Ades, P.K., Hyde, K.D., & Taylor, P.W. (2017). Life styles of Colletotrichum species and implications for plant biosecurity. Fungal Biology Reviews, 31(3), 155- 168. DOI: 10.1016/j.fbr.2017.05.001
- Fan, J.H., Xie, Y.P., Xue, J.L., Xiong, Q., Jiang, W.J., Zhang, Y., & Ren, Z. (2014). The strain HEB01 of Fusarium sp., a new pathogen that infects brown soft scale. Annals of Microbiology, 64, 333-341. DOI: 10.1007/s13213-013-0668-z
- Fancelli, M., Dias, A.B., Delalibera, I.J., Cerqueira de Jesus, S., Souza do Nascimento, A., & Oliveira e Silva, S. (2013). Beauveria bassiana Strains for Biological Control of Cosmopolites sordidus (Germ.) (Coleoptera: Curculionidae) in Plantain. BioMed Research International, 2013(1), 184756. DOI: 10.1155/2013/184756
- Fujita, S. (2013). Simple modified method for fungal slide preparation. Journal of Medical Mycology, 54(2), 141-146. DOI: 10.3314/mmj.54.141
- Ganassi, S., Moretti, A., Stornelli, C., Fratello, B., Bonvicini Pagliai, A.M., Logrieco, A., & Sabatini, M.A. (2001). Effect of Fusarium, Paecilomyces and Trichoderma formulations against aphid Schizaphis graminum. Mycopathologia, 151, 131-138. DOI: 10.1023/a:1017940604692
- Geiser, D.M., del Mar Jiménez-Gasco, M., Kang, S., Makalowska, I., Veeraraghavan, N., Ward, T.J., & O'donnell, K. (2004). FUSARIUM-ID v. 1.0: A DNA sequence database for identifying Fusarium. European Journal of Plant Pathology, 110, 473-479. DOI: 10.1023/B:EJPP.0000032386.75915.a0
- Goettel, M.S., Eilenberg, J., & Glare, T. (2010). Entomopathogenic fungi and their role in regulation of insect populations. In: Gilbert, L.I. & Gill, S.S. (Eds.), Insect Control: Biological and Synthetic Agents, 387-432p, Academic Press., Cambridge.
- Gradmann, C. (2008). A matter of methods: the historicity of Koch's postulates. Medical Journal, 43(2), 121-148. https://www.jstor.org/stable/25805450
- Gurjar, G., Barve, M., Giri, A., & Gupta, V. (2009). Identification of Indian pathogenic races of Fusarium oxysporum f. sp. Ciceris with gene specific, ITS and random markers. Mycologia, 101, 484-495. DOI: 10.3852/08-085
- Hall, T.A. (1999). BioEdit: a user-friendly biological sequence alignment editor and analysis program for windows 95/98/NT. In Nucleic acids symposium series, 41, 95-98.
- Kuruvilla, S., & Jacob, A. (1979). Comparative susceptibility of nymphs and adults of Nilaparvata lugens to Fusarium oxysporum and its use in microbial control. Agricultural Research Journal Kerala, 17, 287-288.
- Kuruvilla, S., & Jacob, A. (1980). Studies on Fusarium oxysporum Schlecht infecting rice brown plant hopper. Agricultural Research Journal Kerala, 18, 51-54.
- Larone, D.H. (1995). Medically Important Fungi, A Guide to İdentification (3rd ed.). American Society for Microbiology, Washington. Laurence, M.H., Walsh, J.L., Shuttleworth, L.A., Robinson, D.M., Johansen, R.M., Petrovic, T., Vu, T.H., Burgess, L.W., Summerell, B.A., & Liew, E.C.Y. (2015). Six novel species of Fusarium from natural ecosystems in Australia. Fungal Diversity, 77, 349-366. DOI: 10.1007/s13225-015-0337-6
- Leger, R.J.S., Wang, C., & Fang, W. (2011). New perspectives on insect pathogens. Fungal Biology Reviews, 25, 84-88. DOI: 10.1016/j.fbr.2011.04.005
- Nilsson, R.H., Larsson, K.H., Taylor, A.F.S., Bengtsson-Palme, J., Jeppesen, T.S., Schigel, D., & Abarenkov, K. (2019). The UNITE database for molecular identification of fungi: handling dark taxa and parallel taxonomic classifications. Nucleic Acids Research, 47(D1), D259-D264. DOI: 10.1093/nar/gky1022
- O’Donnell, K., Humber, R.A., Geiser, D.M., Kang, S., Park, B., Robert, V.A.R.G., Crous, P.W., Johnston, P.R., Aoki, T., Rooney, A.P. & Rehner, S.A. (2012). Phylogenetic diversity of insecticolous fusaria inferred from multilocus DNA sequence data and their molecular identification via FUSARIUM-ID and Fusarium MLST. Mycologia, 104, 427-445. DOI: 10.3852/11-179
- O’Donnell, K., Sutton, D.A., Rinaldi, M.G., Sarver, B.A.J., Balajee, S.A., Schroers, H., Summerbell, R.C., Robert, V.A.R.G., Crous, P.W., Zhang, N., Aoki, T., Jung, K., Park, J., Lee, Y., Kang, S., Park, B., & Geiser, D.M. (2010). Internet-accessible DNA sequence database for identifying fusaria from human and animal infections. Journal of Clinical Microbiology, 48, 3708-3718. DOI: 10.1128/JCM.00989-10
- O'Donnell, K., Cigelnik, E. & Casper, H.H. (1998). Molecular phylogenetic, morphological, and mycotoxin data support reidentification of the Quorn mycoprotein fungus as Fusarium venenatum. Fungal Genetic Biology, 23, 57-67. DOI: 10.1006/fgbi.1997.1018
- Padmaja G.K.V. (2001). Use of the fungus Beauveria bassiana (Bals.) Vuill (Moniliales: Deuteromycetes) for controlling termites. Current Science, 81(6), 647-647.
- Sak, O., & Uçkan, F. (2009). Cypermethrinin Galleria mellonella L. (Lepidoptera: Pyralidae)’nın Puplaşma ve Ölüm Oranlarına Etkisi. Uludag Bee Journal, 9(3), 88-96.
- Sanchez-Pena, S.R., Lara, J.S.J., & Medina, R.F. (2011). Occurrence of entomopathogenic fungi from agricultural and natural ecosystems in Saltillo, México, and their virulence towards thrips and whiteflies. Journal of Insect Science, 11, 1. DOI: 10.1673/031.011.0101
- Sang, T., Crawford, D.J., & Stuessy, T.F. (1995). Documentation of reticulate evolution in peonies (Paeonia) using internal transcribed spacer sequences of nuclear ribosomal DNA: Implications for biogeography and concerted evolution. Proceedings of the National Academy of Sciences, 92, 6813-6817. DOI: 10.1073/pnas.92.15.681
- Sharma, L., & Marques, G. (2018). Fusarium, an entomopathogen-a myth or reality? Pathogens, 7(4). DOI: 10.3390/pathogens7040093
- Summerell, B.A., & Leslie, J.F. (2011). Fifty years of Fusarium: how could nine species have ever been enough? Fungal Diversity, 50, 135-144. DOI: 10.1007/s13225-011-0132-y
- Thangam, S.D., Selvakumar, G., Verghese, A., Kamala, K., & Jayanthi, P.D. (2014). Natural mycosis of mango leafhoppers (Cicadellidae: Hemiptera) by Fusarium sp. Biocontrol Science and Technology, 24(2), 229-232. DOI: 10.1080/09583157.2013.851171
- Torbati, M., Arzanlou, M., Sandoval-Denis, M., & Crous, P.W. (2018). Multigene phylogeny reveals new fungicolous species in the Fusarium tricinctum species complex and novel hosts in the genus Fusarium from Iran. Mycological Progress, 18, 119-133. DOI: 10.1007/s11557-018-1422-5
- Tosi, L., Beccari, G., Rondoni, G., Covarelli, L., & Ricci, C. (2015). Natural occurrence of Fusarium proliferatum on chestnut in Italy and its potential entomopathogenicity against the Asian chestnut gall wasp Dryocosmus kuriphilus. Journal of Pest Science, 88, 369-381. DOI: 10.1007/s10340-014- 0624-0
- Van Diepeningen, A. D. & de Hoog, G.S. (2016). Challenges in Fusarium, a Trans-Kingdom Pathogen. Mycopathologia, 181, 161-163. DOI: 10.1007/s11046-016-9993-7
- Vannini, A., Vettrainoa, A., Martignonia, D., Morales Rodriguezc, C., Contarini, M., Caccia Paparatti, B., & Speranza, S. (2017). Does Gnomoniopsis castanea contribute to the natural biological control of chestnut gall wasp? Fungal Biology, 121, 44-52. DOI: 10.1016/j.funbio.2016.08.013
- White, T.J., Bruns, T., Lee, S., & Taylor, J.W. (1990). Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis, M.A., Gelfand, D.H., Sninsky, J. J. & White T. J. (eds.) PCR Proto-cols: A Guide to Methods and Applications, 315-322p. Academic Press, Inc., New York.
Fusarium proliferatum: izolasyonu, tanımlaması ve model Böcek Galleria mellonella (L.) (Lepidoptera: Pyralidae) ile ilişkisinin değerlendirilmesi
Abstract
Fusarium cinsi, önemli bitki patojenleri ve mikotoksin üreticileri içermesiyle dikkat çekmesine rağmen birçok Fusarium türünün böcekleri kontrol etmede etkili olduğu ve yüksek ölüm oranlarına neden olma, hızlı etki ve bol sporlanma gibi tarımsal zararlı kontrolü için umut verici özellikler sergilediği bilinmektedir. Bu çalışmada, topraktan tuzak metodu ile izole edilen Fusarium proliferatum'un mikro-makro morfolojisi, DNA dizilimi ve model böcek Galleria mellonella üzerindeki etkisi araştırılmıştır. Morfolojik analizler, farklı agarlarda ayrıntılı koloni ve spor yapıları ortaya koymuştur. DNA analizi ile ITS ve CaM gen dizileri kullanılarak tür tanımlamasını doğrulanmıştır. F. proliferatum'un model böcek G. mellonella 'nın larval evresi uzattığı ve pupa süresini, ağırlığını ve toplam yumurta sayısını azalttığı gözlenmiştir. Çalışmamız fungusun G. mellonella 'nın gelişme ve üremesi üzerinde doza bağlı bir etkiye sahip olduğunu göstermektedir. Gelecekteki çalışmalar, gözlenen biyolojik etkilerden sorumlu ikincil metabolitlerin tanımlanmasına, hedef dışı organizmalar üzerindeki olası etkilerin değerlendirilmesi ve farklı çevre koşulları altında saha denemelerinin yürütülmesine odaklanmalıdır.
Keywords
Fusarium proliferatum fungal patojenite Galleria mellonella gelişim biyolojisi izolasyon tanımlama
Project Number
TUBITAK-1001 (122O398) and TUBITAK 2209-A Research Project Support Program for Undergraduate Students
References
- Aoki, T., Smith, J.A., Kasson, M.T., Freeman, S., Geiser, D.M., Geering, A.D., & O’Donnell, K. (2019). Three novel Ambrosia Fusarium Clade species producing clavate macroconidia known (F. floridanum and F. obliquiseptatum) or predicted (F. tuaranense) to be farmed by Euwallacea spp. (Coleoptera: Scolytinae) on woody hosts. Mycologia, 111(6), 919-935. DOI: 10.1080/00275514.2019.1647074
- Azor, M., Gené, J., Cano, J., Manikandan, P., Venkatapathy, N., & Guarro, J. (2009). Less- frequent Fusarium species of clinical interest: correlation between morphological and molecular identification and antifungal susceptibility. Journal of Clinical Microbiology, 47(5), 1463- 1468. DOI: 10.1128/JCM.02467-08
- da Silva Santos, A.C., Diniz, A.G., Tiago, P.V., & de Oliveira, N.T. (2020). Entomopathogenic Fusarium species: a review of their potential for the biological control of insects, implications and prospects. Fungal Biology Reviews, 34(1), 41-57. DOI: 10.1016/j.fbr.2019.12.002
- De Silva, D.D., Crous, P.W., Ades, P.K., Hyde, K.D., & Taylor, P.W. (2017). Life styles of Colletotrichum species and implications for plant biosecurity. Fungal Biology Reviews, 31(3), 155- 168. DOI: 10.1016/j.fbr.2017.05.001
- Fan, J.H., Xie, Y.P., Xue, J.L., Xiong, Q., Jiang, W.J., Zhang, Y., & Ren, Z. (2014). The strain HEB01 of Fusarium sp., a new pathogen that infects brown soft scale. Annals of Microbiology, 64, 333-341. DOI: 10.1007/s13213-013-0668-z
- Fancelli, M., Dias, A.B., Delalibera, I.J., Cerqueira de Jesus, S., Souza do Nascimento, A., & Oliveira e Silva, S. (2013). Beauveria bassiana Strains for Biological Control of Cosmopolites sordidus (Germ.) (Coleoptera: Curculionidae) in Plantain. BioMed Research International, 2013(1), 184756. DOI: 10.1155/2013/184756
- Fujita, S. (2013). Simple modified method for fungal slide preparation. Journal of Medical Mycology, 54(2), 141-146. DOI: 10.3314/mmj.54.141
- Ganassi, S., Moretti, A., Stornelli, C., Fratello, B., Bonvicini Pagliai, A.M., Logrieco, A., & Sabatini, M.A. (2001). Effect of Fusarium, Paecilomyces and Trichoderma formulations against aphid Schizaphis graminum. Mycopathologia, 151, 131-138. DOI: 10.1023/a:1017940604692
- Geiser, D.M., del Mar Jiménez-Gasco, M., Kang, S., Makalowska, I., Veeraraghavan, N., Ward, T.J., & O'donnell, K. (2004). FUSARIUM-ID v. 1.0: A DNA sequence database for identifying Fusarium. European Journal of Plant Pathology, 110, 473-479. DOI: 10.1023/B:EJPP.0000032386.75915.a0
- Goettel, M.S., Eilenberg, J., & Glare, T. (2010). Entomopathogenic fungi and their role in regulation of insect populations. In: Gilbert, L.I. & Gill, S.S. (Eds.), Insect Control: Biological and Synthetic Agents, 387-432p, Academic Press., Cambridge.
- Gradmann, C. (2008). A matter of methods: the historicity of Koch's postulates. Medical Journal, 43(2), 121-148. https://www.jstor.org/stable/25805450
- Gurjar, G., Barve, M., Giri, A., & Gupta, V. (2009). Identification of Indian pathogenic races of Fusarium oxysporum f. sp. Ciceris with gene specific, ITS and random markers. Mycologia, 101, 484-495. DOI: 10.3852/08-085
- Hall, T.A. (1999). BioEdit: a user-friendly biological sequence alignment editor and analysis program for windows 95/98/NT. In Nucleic acids symposium series, 41, 95-98.
- Kuruvilla, S., & Jacob, A. (1979). Comparative susceptibility of nymphs and adults of Nilaparvata lugens to Fusarium oxysporum and its use in microbial control. Agricultural Research Journal Kerala, 17, 287-288.
- Kuruvilla, S., & Jacob, A. (1980). Studies on Fusarium oxysporum Schlecht infecting rice brown plant hopper. Agricultural Research Journal Kerala, 18, 51-54.
- Larone, D.H. (1995). Medically Important Fungi, A Guide to İdentification (3rd ed.). American Society for Microbiology, Washington. Laurence, M.H., Walsh, J.L., Shuttleworth, L.A., Robinson, D.M., Johansen, R.M., Petrovic, T., Vu, T.H., Burgess, L.W., Summerell, B.A., & Liew, E.C.Y. (2015). Six novel species of Fusarium from natural ecosystems in Australia. Fungal Diversity, 77, 349-366. DOI: 10.1007/s13225-015-0337-6
- Leger, R.J.S., Wang, C., & Fang, W. (2011). New perspectives on insect pathogens. Fungal Biology Reviews, 25, 84-88. DOI: 10.1016/j.fbr.2011.04.005
- Nilsson, R.H., Larsson, K.H., Taylor, A.F.S., Bengtsson-Palme, J., Jeppesen, T.S., Schigel, D., & Abarenkov, K. (2019). The UNITE database for molecular identification of fungi: handling dark taxa and parallel taxonomic classifications. Nucleic Acids Research, 47(D1), D259-D264. DOI: 10.1093/nar/gky1022
- O’Donnell, K., Humber, R.A., Geiser, D.M., Kang, S., Park, B., Robert, V.A.R.G., Crous, P.W., Johnston, P.R., Aoki, T., Rooney, A.P. & Rehner, S.A. (2012). Phylogenetic diversity of insecticolous fusaria inferred from multilocus DNA sequence data and their molecular identification via FUSARIUM-ID and Fusarium MLST. Mycologia, 104, 427-445. DOI: 10.3852/11-179
- O’Donnell, K., Sutton, D.A., Rinaldi, M.G., Sarver, B.A.J., Balajee, S.A., Schroers, H., Summerbell, R.C., Robert, V.A.R.G., Crous, P.W., Zhang, N., Aoki, T., Jung, K., Park, J., Lee, Y., Kang, S., Park, B., & Geiser, D.M. (2010). Internet-accessible DNA sequence database for identifying fusaria from human and animal infections. Journal of Clinical Microbiology, 48, 3708-3718. DOI: 10.1128/JCM.00989-10
- O'Donnell, K., Cigelnik, E. & Casper, H.H. (1998). Molecular phylogenetic, morphological, and mycotoxin data support reidentification of the Quorn mycoprotein fungus as Fusarium venenatum. Fungal Genetic Biology, 23, 57-67. DOI: 10.1006/fgbi.1997.1018
- Padmaja G.K.V. (2001). Use of the fungus Beauveria bassiana (Bals.) Vuill (Moniliales: Deuteromycetes) for controlling termites. Current Science, 81(6), 647-647.
- Sak, O., & Uçkan, F. (2009). Cypermethrinin Galleria mellonella L. (Lepidoptera: Pyralidae)’nın Puplaşma ve Ölüm Oranlarına Etkisi. Uludag Bee Journal, 9(3), 88-96.
- Sanchez-Pena, S.R., Lara, J.S.J., & Medina, R.F. (2011). Occurrence of entomopathogenic fungi from agricultural and natural ecosystems in Saltillo, México, and their virulence towards thrips and whiteflies. Journal of Insect Science, 11, 1. DOI: 10.1673/031.011.0101
- Sang, T., Crawford, D.J., & Stuessy, T.F. (1995). Documentation of reticulate evolution in peonies (Paeonia) using internal transcribed spacer sequences of nuclear ribosomal DNA: Implications for biogeography and concerted evolution. Proceedings of the National Academy of Sciences, 92, 6813-6817. DOI: 10.1073/pnas.92.15.681
- Sharma, L., & Marques, G. (2018). Fusarium, an entomopathogen-a myth or reality? Pathogens, 7(4). DOI: 10.3390/pathogens7040093
- Summerell, B.A., & Leslie, J.F. (2011). Fifty years of Fusarium: how could nine species have ever been enough? Fungal Diversity, 50, 135-144. DOI: 10.1007/s13225-011-0132-y
- Thangam, S.D., Selvakumar, G., Verghese, A., Kamala, K., & Jayanthi, P.D. (2014). Natural mycosis of mango leafhoppers (Cicadellidae: Hemiptera) by Fusarium sp. Biocontrol Science and Technology, 24(2), 229-232. DOI: 10.1080/09583157.2013.851171
- Torbati, M., Arzanlou, M., Sandoval-Denis, M., & Crous, P.W. (2018). Multigene phylogeny reveals new fungicolous species in the Fusarium tricinctum species complex and novel hosts in the genus Fusarium from Iran. Mycological Progress, 18, 119-133. DOI: 10.1007/s11557-018-1422-5
- Tosi, L., Beccari, G., Rondoni, G., Covarelli, L., & Ricci, C. (2015). Natural occurrence of Fusarium proliferatum on chestnut in Italy and its potential entomopathogenicity against the Asian chestnut gall wasp Dryocosmus kuriphilus. Journal of Pest Science, 88, 369-381. DOI: 10.1007/s10340-014- 0624-0
- Van Diepeningen, A. D. & de Hoog, G.S. (2016). Challenges in Fusarium, a Trans-Kingdom Pathogen. Mycopathologia, 181, 161-163. DOI: 10.1007/s11046-016-9993-7
- Vannini, A., Vettrainoa, A., Martignonia, D., Morales Rodriguezc, C., Contarini, M., Caccia Paparatti, B., & Speranza, S. (2017). Does Gnomoniopsis castanea contribute to the natural biological control of chestnut gall wasp? Fungal Biology, 121, 44-52. DOI: 10.1016/j.funbio.2016.08.013
- White, T.J., Bruns, T., Lee, S., & Taylor, J.W. (1990). Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis, M.A., Gelfand, D.H., Sninsky, J. J. & White T. J. (eds.) PCR Proto-cols: A Guide to Methods and Applications, 315-322p. Academic Press, Inc., New York.
Details
| Primary Language | English |
|---|---|
| Subjects | Animal Physiology-Ecophysiology |
| Journal Section | Articles |
| Authors | |
| Project Number | TUBITAK-1001 (122O398) and TUBITAK 2209-A Research Project Support Program for Undergraduate Students |
| Early Pub Date | July 15, 2025 |
| Publication Date | |
| Submission Date | April 23, 2025 |
| Acceptance Date | June 27, 2025 |
| Published in Issue | Year 2025 Volume: 10 Issue: 4 |
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