The global population is steadily increasing, leading to greater demands for high-quality protein in our diets to sustain growth and various bodily functions, especially considering the aging demographic (Partridge et al., 2018). However, challenges like environmental pollution and limited agricultural space hinder the expansion of animal husbandry. Sole reliance on animal protein falls short of meeting the growing protein needs (Sanchez, 2010). Urgent measures are required to explore suitable alternatives to bridge this protein supply gap. There’s a notable surge in interest surrounding new sources of protein such as plant-based, algae, fungal, and other innovative protein sources, accompanied by advancements in their respective processing technologies (Ulhas et al., 2023).

Edible fungi boast a unique umami taste and are abundant in high-quality protein (ranging from 15–34.7%), vitamins, minerals, and chitin, among other essential nutrients (Ranogajec et al., 2010, Assemie and Abaya, 2022). Moreover, their easy cultivation, simple processing, and excellent preservation properties allow for consistent annual production, potentially addressing protein supply shortages (Zhang et al., 2013). However, the growth rate of edible fungi can be slow, and during cultivation, strain degradation may occur, leading to reduced protein content. Such occurrences pose unpredictable economic losses for farmers (Dong et al., 2022). Therefore, the pivotal step toward establishing large-scale production of edible fungal protein lies in the careful selection of strains that yield high mycelial protein.

In the selection of edible fungi, when meeting the demand for protein yield becomes challenging, accelerating the mutation process through physical and biological mutagenesis methods becomes essential (Zhang et al., 2023a). Subsequently, screening for beneficial mutations becomes imperative to fulfill these requirements. One promising method gaining traction in biotechnology is Atmospheric and Room Temperature Plasma (ARTP). This technique, unlike traditional mutagenesis approaches, employs plasma that encompasses ultraviolet radiation, heat, electromagnetic fields, charged particles, and reactive oxygen species (Cheng et al., 2016). ARTP minimizes thermal damage to microorganisms during mutagenesis, exerting a profound effect on them. The highly active and evenly distributed particles of ARTP instantly target the DNA chain, inducing an incomplete gene repair process that leads to gene mutation (Zhang et al., 2019a). Screening these mutations yields new strains with enhanced yield, quality, or novel traits. Moreover, in terms of biological induction, sexual reproduction occurs through mycelial mating, culminating in the formation of a new sclerotium (Lin et al., 2021). This sexual reproduction method boosts genetic variation in mushrooms, enhancing their adaptability and biodiversity. Subsequent selection allows us to obtain anticipated yields from the desired strains of edible fungi (Liu et al., 2020a).

The selection and breeding of edible fungi stand as crucial pathways to attain high-yield and premium-quality produce, significantly impacting production cycles, costs, and overall quality (Zhang et al., 2023b). Despite the burgeoning research and development in edible fungi protein, there remains a dearth of targeted breeding efforts aimed at maximizing mycelial protein yield. Pleurotus djamor, an edible mushroom with a nutrient-rich fruit body, represents a low-fat, high-protein healthy food option. This study focused on the selection of Pleurotus djamor spores and the subsequent generation of multiple strains through a combination of ARTP technology-induced mutagenesis and mycelia hybridization. Following this, a rigorous screening process was conducted to identify multiple Pleurotus djamor strains characterized by high mycelial protein production. This selection process involved assessing both protein content and mycelial growth rates.