Fusiform nanoparticle boosts efficient genetic transformation in Sclerotinia sclerotiorum.

IF 10.6 1区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Journal of Nanobiotechnology Pub Date : 2024-08-20 DOI:10.1186/s12951-024-02736-6
Yijuan Ding, Nan Yang, Yi Lu, Jiming Xu, Kusum Rana, Yangui Chen, Zhigang Xu, Wei Qian, Huafang Wan
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Abstract

Background: Sclerotinia sclerotiorum is a highly destructive phytopathogenic fungus that poses a significant threat to a wide array of crops. The current constraints in genetic manipulation techniques impede a thorough comprehension of its pathogenic mechanisms and the development of effective control strategies.

Results: Herein, we present a highly efficient genetic transformation system for S. sclerotiorum, leveraging the use of fusiform nanoparticles, which are synthesized with FeCl3 and 2,6-diaminopyrimidine (DAP). These nanoparticles, with an average longitude length of 59.00 nm and a positively charged surface, facilitate the direct delivery of exogenous DNA into the mycelial cells of S. sclerotiorum, as well as successful integration with stable expression. Notably, this system circumvents fungal protoplast preparation and tedious recovery processes, streamlining the transformation process considerably. Furthermore, we successfully employed this system to generate S. sclerotiorum strains with silenced oxaloacetate acetylhydrolase-encoding gene Ss-oah1.

Conclusions: Our findings demonstrate the feasibility of using nanoparticle-mediated delivery as a rapid and reliable tool for genetic modification in S. sclerotiorum. Given its simplicity and high efficiency, it has the potential to significantly propel genetic research in filamentous fungi, offering new avenues for elucidating the intricacies of pathogenicity and developing innovative disease management strategies.

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纺锤形纳米粒子提高了硬菌的遗传转化效率。
背景:Sclerotinia sclerotiorum 是一种具有高度破坏性的植物病原真菌,对多种作物构成严重威胁。目前遗传操作技术的限制阻碍了对其致病机理的透彻理解和有效控制策略的开发:在此,我们提出了一种针对 S. sclerotiorum 的高效遗传转化系统,该系统利用纺锤形纳米粒子(由 FeCl3 和 2,6-二氨基嘧啶(DAP)合成)。这些纳米颗粒的平均经度长度为 59.00 纳米,表面带正电荷,有助于将外源 DNA 直接输送到硬核菌的菌丝细胞中,并成功整合和稳定表达。值得注意的是,该系统避免了真菌原生质体的制备和繁琐的回收过程,大大简化了转化过程。此外,我们还成功利用该系统生成了沉默草酰乙酸乙酰水解酶编码基因 Ss-oah1 的 S. sclerotiorum 菌株:我们的研究结果表明,利用纳米颗粒介导的递送作为一种快速、可靠的工具对硬菌进行基因改造是可行的。鉴于其简便性和高效性,它有可能极大地推动丝状真菌的遗传研究,为阐明复杂的致病性和开发创新的疾病管理策略提供新的途径。
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来源期刊
Journal of Nanobiotechnology
Journal of Nanobiotechnology BIOTECHNOLOGY & APPLIED MICROBIOLOGY-NANOSCIENCE & NANOTECHNOLOGY
CiteScore
13.90
自引率
4.90%
发文量
493
审稿时长
16 weeks
期刊介绍: Journal of Nanobiotechnology is an open access peer-reviewed journal communicating scientific and technological advances in the fields of medicine and biology, with an emphasis in their interface with nanoscale sciences. The journal provides biomedical scientists and the international biotechnology business community with the latest developments in the growing field of Nanobiotechnology.
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