Begin with a culture dish containing a rice leaf with small wounds, exposing the underlying epidermis.
Apply a moistening solution containing Tween-20, creating a conducive environment for fungal growth.
Place a mycelial plug of Magnaporthe grisea, consisting of conidiophores with conidia — reproductive fungal spores, over the wounded site.
Incubate the leaf in a humid chamber under photoperiod conditions to promote fungal growth.
During incubation, conidiophores release spores that adhere to the exposed epidermis, developing germ tubes.
The germ tube differentiates into a dome-shaped appressorium, which generates high turgor pressure, forming a sharp hypha — or penetration peg.
The penetration peg secretes cell wall-degrading enzymes, facilitating its entry into the epidermal cell and differentiation into invasive hyphae for nutrient acquisition.
These hyphae grow and spread within the epidermal cells through plasmodesmata — connecting channels between cells, and establishing the infection.
The fungus secretes toxins, causing cell necrosis and lesion formation near the wounded site on the leaves, indicating fungal infection.
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