Protein Research Department, Genetic Engineering and Biotechnology Research Institute, City of Scientific Research and Technological Applications, Alexandria, Egypt.
Pharmaceutical Bio-Products Research Department, Genetic Engineering and Biotechnology Research Institute, City of Scientific Research and Technological Applications, Alexandria, Egypt.
ScientificWorldJournal. 2021 Jan 17;2021:6639850. doi: 10.1155/2021/6639850. eCollection 2021.
After introducing the idea of using concentrations equal to or less than the minimum inhibition concentration (MIC) of some active chemical compounds for evacuating microbial cells, different types of microbes were evacuated. The original protocol was given the name sponge-like protocol and then was reduced and modified from a microorganism to another to prepare microbial ghosts for various applications such as immunological applications, drug delivery, and isolation of DNA and protein. Fungal pathogens that infect plants critically affect cost effectiveness, quality, and quantity of their production. They kill plant cells and/or cause plant stress. Plant fungal infections can originate from many sources such as infected soil, seeds, or crop debris causing diseases and quality losses around the world with billions of US dollars annually as costs of the associated productivity loss. This study focused on the application of the sponge-like protocol in protecting tissue cultures of plants against fungal pathogens. This can be useful for research purposes or may be developed to be introduced in field applications. and infection in tissue culture of jojoba ( (Link) Schn.) was used as a model to establish the employment of this protocol to control plant fungal diseases. The best conditions for and ghosts production previously mapped by randomization experimental design (reduced Plackett-Burman experimental design) were used to prepare fungal ghosts. SDS, NaOH, NaHCO, and HO were used in their MIC (+1 level) or minimum growth concentration (MGC, -1 level) according to the determined optimal experimental design. The release of both of DNA and protein from the fungal cells was evaluated spectrophotometrically at 260 and 280, respectively, as an indicator for cell loss of their cytoplasm. Fungal ghost cells were also examined by transmission electron microscopy. After confirming the preparation of high-quality fungal ghost cells, the same conditions were mimicked to control plant fungal infection. Jojoba grown in tissue culture was sprayed with fungal cells (about 10 CFU) as a control experiment or fungal cells followed by treatment with solution (a) represents the fungal ghost cells formation calculated critical concentration (FGCCC) of SDS, NaOH, and NaHCO and then treatment with solution (b) represents HO FGCCC. The plant was examined on day 0 (plant grown before any infection or infection followed by treatment), day 5 (plant at day 5 after infection or infection followed by treatment), and day 10 (plant at day 10 after infection or infection followed by treatment). We observed fungal growth in case of control experiments at days 5 and 10 on the tissue culture medium, as well as plant, and the absence of any fungal growth in case of plant treated with FGCCC even after day 10. We recommend using this FGCCC in the form of chemical spraying formulation to treat the plants aiming to control different plant fungal infections in tissue culture systems or applied in field.
在介绍使用等于或低于某些活性化学化合物的最小抑制浓度 (MIC) 的浓度来清除微生物细胞的想法后，清除了不同类型的微生物。最初的方案被命名为海绵状方案，然后从一种微生物减少和修改到另一种微生物，以制备用于各种应用的微生物幽灵，如免疫应用、药物输送以及 DNA 和蛋白质的分离。感染植物的真菌病原体严重影响其生产的成本效益、质量和数量。它们会杀死植物细胞并/或导致植物压力。植物真菌感染可能来自许多来源，例如受感染的土壤、种子或作物残渣，导致世界各地的疾病和质量损失，每年损失数十亿美元的相关生产力。本研究专注于海绵状方案在保护植物组织培养物免受真菌病原体侵害中的应用。这对于研究目的可能很有用，或者可以开发为引入田间应用。将 jojoba（(Link) Schn.）组织培养物中的 和 感染用作模型，以建立使用该方案控制植物真菌病的方法。先前通过随机化实验设计（简化 Plackett-Burman 实验设计）映射的 和 幽灵生产的最佳条件用于制备真菌幽灵。根据确定的最佳实验设计，SDS、NaOH、NaHCO 和 HO 分别在其 MIC（+1 级）或最小生长浓度（MGC，-1 级）下使用。通过分光光度法分别在 260 和 280 处评估 DNA 和蛋白质从真菌细胞中的释放，作为细胞质细胞损失的指标。通过透射电子显微镜检查真菌幽灵细胞。在确认制备高质量真菌幽灵细胞后，模拟相同条件以控制植物真菌感染。在组织培养中生长的霍霍巴用真菌细胞（约 10 CFU）喷洒作为对照实验，或用真菌细胞处理后用溶液（a）代表 SDS、NaOH 和 NaHCO 的计算临界浓度（FGCCC），然后用溶液（b）代表 HO FGCCC。在第 0 天（植物在任何感染之前或感染后处理）、第 5 天（感染后第 5 天或感染后处理）和第 10 天（感染后第 10 天或感染后处理）检查植物。我们在组织培养培养基、植物上的对照实验中观察到第 5 天和第 10 天的真菌生长，而在用 FGCCC 处理的植物中即使在第 10 天后也没有任何真菌生长。我们建议以化学喷雾制剂的形式使用这种 FGCCC 来处理植物，旨在控制组织培养系统或田间应用中的不同植物真菌感染。
