| Summary: | <i>Fusarium pseudograminearum</i> causes crown rot in wheat. This study aimed to assess the effects of the bacterial strain QTH8 isolated from <i>Cotinus coggygria</i> rhizosphere soil against <i>F. pseudograminearum</i>. Bacterial strain QTH8 was identified as <i>Bacillus halotolerans</i> in accordance with the phenotypic traits and the phylogenetic analysis of 16S rDNA and <i>gyrB</i> gene sequence. Culture filtrates of bacterial strain QTH8 inhibited the mycelial growth of <i>F. pseudograminearum</i> and resulted in mycelial malformation such as tumor formation, protoplast condensation, and mycelial fracture. In addition, bacterial strain QTH8 also inhibited the mycelial growth of <i>Hainesia lythri</i>, <i>Pestalotiopsis</i> sp., <i>Botrytis cinerea</i>, <i>Curvularia lunata</i>, <i>Phyllosticta theaefolia</i>, <i>Fusarium graminearum</i>, <i>Phytophthora nicotianae</i>, and <i>Sclerotinia sclerotiorum</i>. The active compounds produced by bacterial strain QTH8 were resistant to pH, ultraviolet irradiation, and low temperature, and were relatively sensitive to high temperature. After 4 h exposure, culture filtrates of bacterial strain QTH8—when applied at 5%, 10%, 15%, 20%, 25%, and 30%—significantly reduced conidial germination of <i>F. pseudograminearum</i>. The coleoptile infection assay proved that bacterial strain QTH8 reduced the disease index of wheat crown rot. In vivo application of QTH8 to wheat seedlings decreased the disease index of wheat crown rot and increased root length, plant height, and fresh weight. Iturin, surfactin, and fengycin were detected in the culture extract of bacterial strain QTH8 by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Bacterial strain QTH8 was identified for the presence of the <i>ituC</i>, <i>bacA</i>, <i>bmyB</i>, <i>spaS</i>, <i>srfAB</i>, <i>fend</i>, and <i>srfAA</i> genes using the specific polymerase chain reaction primers. <i>B. halotolerans</i> QTH8 has a vital potential for the sustainable biocontrol of wheat crown rot.
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