The Development of a Phytopathogenic Fungi Control Trial: Aspergillus flavus and Aspergillus niger Infection in Jojoba Tissue Culture as a Model

Abstract

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 in vitro tissue cultures of plants against fungal pathogens. This can be useful for research purposes or may be developed to be introduced in field applications. Aspergillus flavus and Aspergillus niger infection in tissue culture of jojoba (Simmondsia chinensis (Link) Schn.) was used as a model to establish the employment of this protocol to control plant fungal diseases. The best conditions for A. flavus and A. niger ghosts production previously mapped by randomization experimental design (reduced Plackett–Burman experimental design) were used to prepare fungal ghosts. SDS, NaOH, NaHCO3, and H2O2 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 260nm and 280nm, 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 103 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 NaHCO3 and then treatment with solution (b) represents H2O2 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 in vitro tissue culture systems or applied in field.

Impact of Enterococcus italicus ONU547 on the growth and acclimatization of micropropagated Rubus fruticosus L. and Paulownia tomentosa Steud. plants to ex vitro conditions

“…Mechnikov National University (Odesa, Ukraine). These selected species are commonly associated with plant diseases in field conditions and are also frequent contaminants in plant tissue culture (Omamor et al, 2007 ; El-Baky et al, 2021 ; Liu et al, 2021; Wang et al, 2021 ). The fungi were cultured on potato dextrose agar (PDA) at 28°C.…”

Recent Approaches towards Control of Fungal Diseases in Plants: An Updated Review

“…Our research group has recently developed a phytopathogenic fungi control trial involving a model of Aspergillus flavus and Aspergillus niger infection in the tissue culture of the jojoba plant [12]. In this study, we applied the sponge-like protocol for evacuating microbial cells [111][112][113][114][115][116][117] in protecting in vitro tissue cultures of plants against fungal pathogens to establish the use of this protocol and ghost techniques in controlling plant fungal diseases.…”

“…The sponge-like protocol was principally created to employ cheap and safe chemical compounds (NaOH, SDS, NaHCO 3 , and H 2 O 2 ) for microbial ghost cell preparation for various applications [112]. Ghosts from various microbes were produced by this gentle chemical protocol that was applied to induce evacuation-pores at the microbial cell wall [12,[111][112][113][114][115][116][117]. This protocol is unique as it combines the use of minimum inhibition concentration (MIC, which kills microbial cells under minimal killing conditions) and minimum growth concentration (MGC, which allows cells to escape and live but still affects their cell wall) of active chemicals responsible for killing microbes according to the optimal experimental design to prepare microbial ghosts mapped by the full or reduced Plackett-Burman experimental design.…”

“…Accordingly, development of healthy, non-toxic, and eco-friendly alternate approaches (green strategies of fungal control) to chemical and synthetic fungicides is very helpful in the control of plant fungal infections [3][4][5][6]. These safe and effective alternative control means against plant fungal diseases include biological control of phytopathogenic fungi [7], microbial fungicides [7,8], botanical fungicides [9], agronanotechnology [10,11], and fungal cell deactivation and evacuation using ghost techniques [12].…”

Development of rapid DNA extraction and PCR amplification methods for fungi and parasitic plants

“…Phytopathogenic fungi and parasitic plants are the leading cause of devastating agricultural losses or death in crops. Phytopathogenic fungi damage plants by killing their cells and causing biotic stress (El-Baky et al, 2021), while parasitic plants penetrate their host tissues and form vascular connections to redirect the host's nutrients via the haustoria in their stems or roots (Těšitel, 2016). To characterise the phytopathogens, besides morphological characterisation, one of the reliable and trending methods is molecular identification (López et al, 2009).…”