Trichoderma harzianum Cellobiohydrolase Thph2 Induces Reactive Oxygen Species-Mediated Resistance Against Southern Corn Leaf Blight in Maize

The pathogenic plant fungus <i>Bipolaris maydis</i> is responsible for southern corn leaf blight (SCLB), a widespread agricultural disease that significantly reduces maize yield in various agroecological zones. The present research focuses on characterizing the role of <i>Trichoderma harzianum</i> cellobiohydrolase (CBH) Thph2 in induced maize resistance to SCLB by triggering the production of reactive oxygen species (ROS) in leaves. First of all, we demonstrated the potential activities of Thph2 in triggering ROS burst and PDC in a model plant, <i>Nicotiana benthamiana</i>. Cell death, ROS burst, and programmed cell death (PCD) were observed in <i>N. benthamiana</i> leaves following transient expression of Thph2, indicating its defensive role against <i>Sclerotinia sclerotiorum</i> infection. The removal of the signal peptide from Thph2 resulted in the complete loss of the cell death phenotype and the accumulation of reactive oxygen species (ROS), confirming that Thph2 functions as a microbial elicitor that primes host plant immunity through ROS-mediated signaling, thereby inducing systemic resistance (ISR). Furthermore, the Thph2 protein conferred resistance against <i>B. maydis</i> in maize, significantly increasing reactive oxygen species (ROS) accumulation (1.5-fold compared to the control) at 48 h post-inoculation (hpi),and leading to the reduction in the lesion area of SCLB by 15.9% at 2 days post-inoculation (dpi). Our results demonstrated that the Thph2 protein markedly enhanced the expression of <i>lox5</i>, <i>aos</i>, and <i>hpl</i> in maize leaves, thereby confirming its function in triggering plant defense mechanisms primarily via the jasmonic acid signaling pathway. This research reveals new molecular mechanisms by which <i>T. harzianum</i> enhances plant defense and showcases the biocontrol efficacy of Thph2 against southern corn leaf blight (SCLB).