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Projects / Programmes source: ARIS

Protein complexes from the fungal genus Pleurotus, new bio-pesticides for controlling Colorado potato beetle and western corn rootworm

Research activity

Code Science Field Subfield
4.03.00  Biotechnical sciences  Plant production   

Code Science Field
B390  Biomedical sciences  Phytotechny, horticulture, crop protection, phytopathology 

Code Science Field
4.01  Agricultural and Veterinary Sciences  Agriculture, Forestry and Fisheries 
Keywords
aegerolysin protein family, MACPF protein family, pesticide, protein, membrane, lipid, genetically modified plants, Colorado potato beetle.
Evaluation (rules)
source: COBISS
Researchers (29)
no. Code Name and surname Research area Role Period No. of publicationsNo. of publications
1.  15595  Katarina Babnik    Technical associate  2019 - 2022 
2.  24290  PhD Matej Butala  Biochemistry and molecular biology  Researcher  2019 - 2022  236 
3.  34130  PhD Anna Coll Rius  Biochemistry and molecular biology  Researcher  2019 - 2022  164 
4.  18122  Boštjan Drolc  Veterinarian medicine  Technical associate  2019 - 2022  10 
5.  12449  PhD Robert Frangež  Veterinarian medicine  Researcher  2019 - 2022  280 
6.  18324  PhD Gordana Glavan  Neurobiology  Researcher  2019 - 2022  146 
7.  12688  PhD Kristina Gruden  Biotechnology  Researcher  2019 - 2022  990 
8.  32099  PhD Maja Grundner  Biochemistry and molecular biology  Researcher  2020 - 2022  35 
9.  24447  PhD Anita Jemec Kokalj  Biology  Researcher  2019 - 2022  296 
10.  52386  Eva Kočar  Biochemistry and molecular biology  Technical associate  2019 - 2020  29 
11.  00412  PhD Igor Križaj  Biochemistry and molecular biology  Researcher  2019 - 2022  729 
12.  37409  PhD Maja Križnik  Biotechnology  Researcher  2019 - 2022  42 
13.  53699  Amela Kujović  Biochemistry and molecular biology  Technical associate  2019  25 
14.  18802  PhD Adrijana Leonardi  Biochemistry and molecular biology  Researcher  2019 - 2022  157 
15.  36323  PhD Tjaša Lukan  Biology  Researcher  2019 - 2021  127 
16.  25805  MSc Špela Modic  Plant production  Researcher  2019 - 2022  383 
17.  39099  PhD Katja Molan  Biochemistry and molecular biology  Junior researcher  2019  41 
18.  55449  Martina Mravinec  Biochemistry and molecular biology  Researcher  2021  11 
19.  33683  Nina Orehar    Technical associate  2019 
20.  39090  PhD Anastasija Panevska  Biochemistry and molecular biology  Researcher  2019 - 2022  56 
21.  29617  PhD Marko Petek  Biotechnology  Researcher  2019 - 2022  172 
22.  04570  PhD Jože Pungerčar  Biochemistry and molecular biology  Researcher  2019 - 2022  320 
23.  26091  PhD Jaka Razinger  Plant production  Researcher  2019 - 2022  451 
24.  15328  PhD Kristina Sepčić  Biochemistry and molecular biology  Head  2019 - 2022  731 
25.  33137  PhD Matej Skočaj  Biochemistry and molecular biology  Researcher  2019 - 2022  107 
26.  21553  PhD Jernej Šribar  Biochemistry and molecular biology  Researcher  2019 - 2022  108 
27.  06905  PhD Tom Turk  Biochemistry and molecular biology  Researcher  2019 - 2022  620 
28.  13334  PhD Milka Vrecl Fazarinc  Veterinarian medicine  Researcher  2019 - 2022  269 
29.  22588  PhD Monika Cecilija Žužek  Veterinarian medicine  Researcher  2019 - 2022  67 
Organisations (5)
no. Code Research organisation City Registration number No. of publicationsNo. of publications
1.  0481  University of Ljubljana, Biotechnical Faculty  Ljubljana  1626914  66,817 
2.  0105  National Institute of Biology  Ljubljana  5055784  13,304 
3.  0106  Jožef Stefan Institute  Ljubljana  5051606000  91,035 
4.  0401  Agricultural institute of Slovenia  Ljubljana  5055431  20,103 
5.  0406  University of Ljubljana, Veterinary Faculty  Ljubljana  1627139  10,832 
Abstract
To meet increasing food demands and to comply with stricter environmental demands, agriculture must increase food production and quality while decreasing its detrimental ecological impact. These strategies can also be directed towards efficient control of pests, by replacing the use of first-generation chemical pesticides with novel, safer, and environmentally friendly biopesticides. Ostreolysin A6 (OlyA6), pleurotolysin A2 (PlyA2), and erylysin A (EryA) are proteins from the mushroom genus Pleurotus which belong to the aegerolysin family. OlyA6, PlyA2 and EryA have recently been found to specifically bind ceramide phosphoethanolamine, a membrane sphingolipid that is specific to invertebrates, especially insects. In concert with pleurotolysin B (PlyB) with the membrane-attack-complex/perforin domain which is also produced by Pleurotus mushrooms, these aegerolysins can form multimeric bicomponent pore complexes in artificial and biological membranes containing physiologically relevant concentrations of ceramide phosphoethanolamine. These characteristics make these aegerolysin/PlyB protein complexes interesting as tools for for selective suppression of organisms whose cell membranes contain the aegerolysin molecular lipid target: the ceramide phsphoethanolamine. The proposed project is based on encouraging results that were obtained within the recently concluded national research Project J4-7162, in which we demonstrated, by feeding assays, the potential of the OlyA6/PlyB, PlyA2/PlyB and EryA/PlyB protein complexes to selectively kill larvae and adults of two economically most important coleopteran pests, the western corn rootworm (Diabrotica v. virgifera) and the Colorado potato beetle (Leptinotarsa decemlineata). The toxicities of these protein complexes are comparable to, and in the case of CPB even greater than, that of the proteinaceous Cry toxins from Bacillus thuringiensis, which have been introduced into genetically modified crops to control western corn rootworm, Colorado potato beetle, and other insect pests. Cry toxins exert their insecticidal action through binding to (glyco)protein receptors in the insect midgut, and as a consequence, insects are continuously developing resistance against these toxins. Due to the interactions of OlyA6/PlyB, PlyA2/PlyB and EryA/PlyB with their specific insect membrane lipid receptor and not with pest proteins that can be prone to mutation, the chances of evolving resistance to these insecticidal aegerolysin-based protein complexes should be extremely small.  Within the proposed project, we aim to further evaluate the potential of OlyA6/PlyB, PlyA2/PlyB and EryA/PlyB to act as biopesticides against Colorado potato beetle, through the development of genetically modified potatoes that can produce these insecticidal complexes. We will also further elucidate the molecular mechanisms of insecticidal activity of these protein complexes, by exploring the mechanism of the pore formation on ceramide phosphoethanolamine – containing membranes, and by searching for putative additional protein and lipid receptor(s) in the insect midgut. Finally, we will further explore the toxicity of aegerolysin/PlyB complexes against additional agriculturally important pests, as well as against non-target insects (e.g. the bees), and rodents. The proposed study will complete the missing parts in the molecular mechanisms of action of these novel lipid-binding biopesticides, and will pave the way for the development of new environment-friendly and long-term effective insecticides.
Significance for science
The research approach of this project aims to provide solutions for problems that conventional pest control faces nowadays, including insecticide resistance and inadequate pest control, along with the often-associated environmental and human health concerns. A combination of biochemical and ‘-omics’ approaches offers the possibility to define the mechanisms of action and to predict potential obstacles in the introduction of the proposed new insecticides. The proposed study will complete the missing parts in the molecular mechanisms of these novel lipid-binding biopesticides, and will pave the way for the development of new environment-friendly and long-term effective insecticides.    The Project Group is composed of experienced senior scientists in such a way that it enables multidisciplinary approaches to the successful completion of the project. This is important nowadays, as publications in high-impact journals in the life sciences demand multidisciplinarity and many different experimental approaches. The Project Goup has experience with publications in high-impact journals, and we anticipate that the outcomes of this project will be some papers in high-ranking scientific journals, along with some patents. This will have a positive impact on the promotion groups and affiliated institutions that are involved in the project, and on Slovenian science and Slovenia in general. In particular, the results will represent support to the local food industry in assuring food security. The project will also provide good training opportunities for young scientists. They will be included in all stages of the project, which will represent the main theme of their BSc, MSc or PhD studies. Foreign students from Croatia will also be included into the project through project work within the ERASMUS exchange programme. These students will have access to the modern equipment at all of the participating institutions. Together with the senior researchers included in the project, this will provide a stimulating environment for good training in modern life science disciplines, such as biochemistry, molecular biology, microbiology, toxicity testing, and biophysics. Finally, the outcome of the project will have direct economic benefits to the proposers and to the Slovenia economy: we see the possibility of marketing the developed innovative low-risk pest-management solutions through selected European biotech/ IPM companies (e.g., e-nema GmbH, Germany; Globachem NV, Belgium; CBC Europe S.r.l., Italy), and through international companies that have strong presence in the field of biotechnological solutions for plant protection (e.g., Pioneer, BASF, DOW Agrosciences, Bayer Crop Science, Du Pont). Our aim is to develop protein biopesticides and their applications to TRL 5 or higher, and to find licensees who can bring the solution onto the market to influence and change the existing agronomic practices.
Significance for the country
The research approach of this project aims to provide solutions for problems that conventional pest control faces nowadays, including insecticide resistance and inadequate pest control, along with the often-associated environmental and human health concerns. A combination of biochemical and ‘-omics’ approaches offers the possibility to define the mechanisms of action and to predict potential obstacles in the introduction of the proposed new insecticides. The proposed study will complete the missing parts in the molecular mechanisms of these novel lipid-binding biopesticides, and will pave the way for the development of new environment-friendly and long-term effective insecticides.    The Project Group is composed of experienced senior scientists in such a way that it enables multidisciplinary approaches to the successful completion of the project. This is important nowadays, as publications in high-impact journals in the life sciences demand multidisciplinarity and many different experimental approaches. The Project Goup has experience with publications in high-impact journals, and we anticipate that the outcomes of this project will be some papers in high-ranking scientific journals, along with some patents. This will have a positive impact on the promotion groups and affiliated institutions that are involved in the project, and on Slovenian science and Slovenia in general. In particular, the results will represent support to the local food industry in assuring food security. The project will also provide good training opportunities for young scientists. They will be included in all stages of the project, which will represent the main theme of their BSc, MSc or PhD studies. Foreign students from Croatia will also be included into the project through project work within the ERASMUS exchange programme. These students will have access to the modern equipment at all of the participating institutions. Together with the senior researchers included in the project, this will provide a stimulating environment for good training in modern life science disciplines, such as biochemistry, molecular biology, microbiology, toxicity testing, and biophysics. Finally, the outcome of the project will have direct economic benefits to the proposers and to the Slovenia economy: we see the possibility of marketing the developed innovative low-risk pest-management solutions through selected European biotech/ IPM companies (e.g., e-nema GmbH, Germany; Globachem NV, Belgium; CBC Europe S.r.l., Italy), and through international companies that have strong presence in the field of biotechnological solutions for plant protection (e.g., Pioneer, BASF, DOW Agrosciences, Bayer Crop Science, Du Pont). Our aim is to develop protein biopesticides and their applications to TRL 5 or higher, and to find licensees who can bring the solution onto the market to influence and change the existing agronomic practices.
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