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Mısır (Zea mays) Bitkisinin Çimlenmesi Sırasında Fungal Patojen Fusarium verticillioides’in Patojenitesini Azaltmak için Fungal CYP51 Geninin Agroenfeksiyon Aracılığı ile Virüs Kaynaklı Gen Susturulması

Year 2025, , 306 - 319, 27.03.2025

Yaprak Narmanlı , Buse Didem Perendi , Fatma Aydinoglu

https://doi.org/10.18016/ksutarimdoga.vi.1573791

Abstract

Fusarium verticillioides, tahıllarda özellikle mısır (Zea mays) bitkisinde yıkıcı hastalıklara neden olan ve tarımda büyük ekonomik kayıplara yol açan bir fungal patojendir. Günümüzde, F. verticillioides enfeksiyonları ile mücadelede başlıca, sitokrom P450 lanosterol C-14α-demetilazı (CYP51) hedef alan azol içerikli kimyasal fungisitler kullanılmaktadır. Buna alternatif olarak, bu çalışmada, patojenin kontrolü için endojen RNA interferans (RNAi) mekanizmasına dayanan ve agroenfeksiyon aracılıklı Virüs Kaynaklı Gen Susturma (VIGS) yaklaşımının potansiyeli değerlendirilmiştir. Bu amaçla, Agrobacterium tumefaciens, F. verticillioides’in üç CYP51 geninde ortak olan 313 nükleotidlik fragmenti taşıyan Brome Mozaik Virüs (BMV3) türevi bir vektörle transforme edildikten sonra fungal patojen ile birlikte kültüre alınmıştır. Sonuçlar, benzer şekilde fakat boş vektörle muamele edilmiş kontrol grubuyla morfolojik, biyokimyasal ve transkripsiyonel olarak karşılaştırmalı olarak analiz edilmiştir. Buna göre, CYP51A, CYP51B ve CYP51C genlerinin ifade seviyelerinde, sırasıyla, %49, %65 ve %51 oranında azalış saptanmıştır. Fungal büyüme oranının %13 oranında azaldığı; çimlenme oranının %20 ve ROS miktarının %56 oranında kontrole göre daha yüksek olduğu tespit edilmiştir. Bu bulgular, CYP51'in susturulmasının hücrelerde ROS birikimine neden olarak F. verticillioides'in patojenitesinin inhibisyonuna yol açtığını göstermektedir. Bu çalışma, F. verticillioides kaynaklı bitki hastalıklarını kontrol etmek için tarımsal olarak sürdürülebilir ve çevre dostu bir alternatif yöntem olarak agroenfeksiyon aracılı VIGS uygulamasıyla CYP51 geninin hedeflenmesinin potansiyelini ortaya koymuştur.

Keywords

RNA interferans Brome mozaik virusü Agrobacterium tumefaciens Sitokrom P450 lanosterol C-14α-demetilaz VIGS

Project Number

TÜBİTAK 221O321 ve COST CA20110

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Agroinfection-Mediated Virus Induced Gene Silencing of Fungal Pathogen Fusarium verticillioides CYP51 Gene to Reduce its Pathogenicity during Maize (Zea mays) Germination

Year 2025, , 306 - 319, 27.03.2025

Yaprak Narmanlı , Buse Didem Perendi , Fatma Aydinoglu

https://doi.org/10.18016/ksutarimdoga.vi.1573791

Abstract

Fusarium verticillioides is a fungal pathogen, resulting in devastating diseases in cereals, especially in maize (Zea mays) and causing massive economic losses in agriculture. F. verticillioides infections in the field are managed by chemical fungicides, mainly azoles which target cytochrome P450 lanosterol C-14α-demethylase (CYP51). Alternative to chemical fungicide, this study evaluated the potential of agroinfection-mediated Virus Induced Gene Silencing (VIGS) approach in controlling F. verticillioides pathogenicity, which is based on naturally occurring RNA interference (RNAi) mechanism. For this purpose, F. verticillioides was co-cultivated with Agrobacterium tumefaciens transformed with a Brome Mosaic Virus (BMV3) derived vector carrying a 313 nucleotide length fragment common to the three existing F. verticillioides CYP51 genes and comparatively evaluated with the mock group treated similarly but with empty vector at morphologically, biochemically and transcriptionally. It was detected that agroinfection-mediated VIGS treatment of F. verticillioides reduced expression levels of CYP51A by 49%, CYP51B by 65% and CYP51C by 51% and the growth rate by 13% while the germination rate was 20% and ROS amount 56% higher compared to mock-treated F. verticillioides. These findings pointed out that silencing of CYP51 caused ROS accumulation in cells leading to inhibition of the pathogenicity of F. verticillioides. This study represented the potential of targeting CYP51 gene by agroinfection-mediated VIGS treatment as an agriculturally sustainable and environmentally friendly alternative method to control F. verticillioides-caused plant diseases.

Keywords

RNA interference Brome mosaic virus Agrobacterium tumefaciens Cytochrome P450 lanosterol C-14α-demethylase VIGS

Ethical Statement

Etik kurulu onay belgesine gerek yoktur.

Supporting Institution

TÜBİTAK 221O321 ve COST CA20110

Project Number

TÜBİTAK 221O321 ve COST CA20110

Thanks

This study was supported by COST 2519 program of Scientific and Technological Research Council of Turkey (TUBITAK) (Grant number 221O321) and rely upon COST (European Cooperation in Science and Technology) Action CA20110-RNA communication across kingdoms: new mechanisms and strategies in pathogen control (exRNA-PATH). Coauthors deeply appreciate TUBITAK Science Fellowships and Grant Programme Directorate (BIDEB) for their supports.

References

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Details

Primary Language English
Subjects Plant Pathology, Microbial Genetics
Journal Section RESEARCH ARTICLE
Authors

Yaprak Narmanlı 0000-0002-1820-0460

Buse Didem Perendi 0009-0008-9789-9361

Fatma Aydinoglu 0000-0002-9974-045X

Project Number TÜBİTAK 221O321 ve COST CA20110
Early Pub Date March 20, 2025
Publication Date March 27, 2025
Submission Date October 25, 2024
Acceptance Date February 26, 2025
Published in Issue Year 2025

Cite

APA Narmanlı, Y., Perendi, B. D., & Aydinoglu, F. (2025). Agroinfection-Mediated Virus Induced Gene Silencing of Fungal Pathogen Fusarium verticillioides CYP51 Gene to Reduce its Pathogenicity during Maize (Zea mays) Germination. Kahramanmaraş Sütçü İmam Üniversitesi Tarım Ve Doğa Dergisi, 28(2), 306-319. https://doi.org/10.18016/ksutarimdoga.vi.1573791

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