| Summary: | The marine yeast <i>Scheffersomyeces spartinae</i> W9 is a promising biocontrol agent for gray mold caused by <i>Botrytis cinerea</i> in strawberries. Improving the biocontrol efficacy of <i>S. spartinae</i> W9 is necessary for its commercial application. In this study, different concentrations of β-glucan were added to the culture medium to evaluate its effect on the biocontrol efficacy of <i>S. spartinae</i> W9. The results showed that 0.1% β-glucan could increase the biocontrol effect of <i>S. spartinae</i> W9 against <i>B. cinerea</i> in strawberries and in vitro. We found that adding 0.1% β-glucan to the culture medium promoted the growth of <i>S. spartinae</i> W9 in wounds of strawberries, enhanced biofilm formation ability, and secreted more β-1,3-glucanase. In addition, 0.1% β-glucan increased the survival rate of <i>S. spartinae</i> W9 under oxidative, thermal, osmotic, and plasma membrane stressors. Transcriptome analysis revealed 188 differential expressed genes in <i>S. spartinae</i> W9 cultured with or without 0.1% β-glucan, including 120 upregulated and 68 downregulated genes. The upregulated genes were associated with stress response, cell wall formation, energy production, growth, and reproduction. Thus, culturing with 0.1% β-glucan is an effective way to improve the biocontrol ability of <i>S. spartinae</i> W9 against gray mold in strawberries.
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