Mitigation of Powdery Mildew Disease by Integrating Biocontrol Agents and Shikimic Acid with Modulation of Antioxidant Defense System, Anatomical Characterization, and Improvement of Squash Plant Productivity

Squash (Cucurbita pepo L.) is a globally important vegetable, the production of which is severely constrained by powdery mildew caused by Podosphaera xanthii. In this study, we examined the effects of Trichoderma asperellum (MW965676), Streptomyces rochei (MN700192), and a mixture of the two foliar sprays with or without shikimic acid seed priming treatment on powdery mildew severity, plant growth, and total yield during the 2020–2021 and 2021–2022 growing seasons. We also studied their immune eliciting properties by examining their enzymatic, phenolic, and hormonal functions. The combination of Trichoderma asperellum, Streptomyces rochei, and shikimic acid triggered plant defense responses, which elicited enzyme activities such as peroxidase (POD), superoxide dismutase (SOD), and catalase (CAT), phenolic compound accumulation, and increased salicylic acid (SA) and jasmonic acid (JA) content. This approach yielded high-quality results in the control of powdery mildew during the two growing seasons under greenhouse conditions. Additionally, relatively large statistical differences in plant growth, total yield, mineral components, and physiological traits were observed. A GC–MS analysis of Trichoderma asperellum (MW965676) showed hemin cation as a major component, while Streptomyces rochei (MN700192) contained 2,4-di-tert-butyl phenol and the hexadecenoic acid methyl ester. With respect to the morphological changes induced by powdery mildew and the treatments, plants treated with a mixture of Trichoderma asperellum, Streptomyces rochei and shikimic acid showed an improvement in the thickness of the midvein, increased dimensions of the main midvein bundle, a larger number of xylem rows in the main midvein bundle, greater mean diameters of vessels and of parenchyma cells in the ground tissues, as well as increased thickness of the upper and lower epidermis, lamina, palisade tissue and spongy tissue. This extensive, new study is the first step toward a more profound understanding of the use of Trichoderma asperellum and Streptomyces rochei with shikimic acid-primed seeds as a potential alternative technique for attenuating powdery mildew infection in squash.