Bioscience Evidence 2026, Vol.16, No.4, 316-328 http://bioscipublisher.com/index.php/be 322 Figure 3 a Substrate blocks at stage 1: mycelial run. b Substrate blocks at stage 2: coat formation. c Substrate blocks at stage 3: bump formation. d Substrate blocks at stage 4: pigmentation. e Substrate blocks at stage 5: coat hardening. f Substrate blocks at pin head formation stage. g Substrate blocks at fruit development stage (Adopted from Sassine et al., 2024) 4.2 Effects of substrate on biological efficiency and yield stability Biological efficiency (BE) is an important indicator for evaluating the conversion capacity of shiitake cultivation substrates. It reflects the ability of a substrate to produce fresh mushrooms per unit weight of dry substrate. The type of substrate materials, nutrient supplementation strategies, and carbon-to-nitrogen balance are major factors determining BE. Studies evaluating different substrate components have shown that oak sawdust substrates achieved a BE of 92.35%, together with higher mushroom numbers and a shorter mycelial growth period (Ranjbar and Olfati, 2017). High-quality woody materials can provide suitable physical structure and continuous nutrient supply, and therefore still have important value in some commercial production systems. However, for agricultural by-product-based substrates, nutrient supplementation strategies often determine substrate conversion efficiency. Improving the utilization efficiency of agricultural residues requires not only replacing sawdust but also adjusting nutrient balance through appropriate nitrogen supplementation. The performance across multiple flushes often provides a better indication of substrate quality than the highest yield from a single flush. Under a fixed substrate system, optimization of inoculum composition can improve overall production performance across two flushes, suggesting that stable shiitake production is influenced not only by substrate composition but also by the establishment process of mycelial growth. Increasing nutrient supplementation does not always improve yield. Different nitrogen addition levels can significantly affect fresh mushroom production, and moderate nitrogen supplementation generally performs better
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