生防细菌HX-6菌株对青枯病菌的抑制效果及作用机制

    Antagonistic effects and action mechanism of Bacillus velezensis HX-6 against Ralstonia solanacearum

    • 摘要: 本研究探讨了生防细菌HX-6菌株对植物病原细菌茄科雷尔氏菌(Ralstonia solanacearum,简称青枯病菌)的抑制效果及其作用机制。通过平板对峙试验和琼脂扩散试验评估HX-6发酵液及其无细胞上清液 (CFS) 对青枯病菌的抑制作用;采用微量稀释肉汤法测定了CFS的最小抑菌浓度 (MIC) 和最小杀菌浓度 (MBC);通过菌落形态特征结合16S rRNA和gyrA基因测序对HX-6菌株进行鉴定;利用检测性培养基评估其促生能力相关功能;此外,通过OD600 nm值、电导率、核酸泄漏和丙二醛含量的变化,以及扫描电子显微镜观察,进一步评估CFS对青枯病菌的抑制作用和对细胞完整性的影响。结果显示,HX-6菌株发酵液和CFS对青枯病菌均有显著的抑制作用,发酵液的抑菌圈直径为3.03 cm,CFS的抑制率达到70.48%,MIC和MBC分别为12.5%和50.0%。HX-6菌株被鉴定为贝莱斯芽孢杆菌 (Bacillus velezensis),并具备产吲哚-3-乙酸、铁载体、生物膜和溶解磷酸盐等功能。此外,CFS显著抑制了青枯病菌的活性,并破坏其细胞完整性。通过OD600 nm值、电导率、核酸泄漏情况和丙二醛含量的变化进一步验证了上述效果。同时,扫描电子显微镜观察显示,CFS对青枯病菌完整性造成了严重损伤。研究表明HX-6菌株具有较高的抑菌活性,可能成为新型青枯病生物防治剂,值得在农业应用中进一步研究。

       

      Abstract: In this study, we investigated the bacteriostatic activity and underlying mechanisms of the biocontrol bacterial strain HX-6 against Ralstonia solanacearum. The bacteriostatic activity of strain HX-6 fermentation broth and its cell-free supernatant (CFS) on R. solanacearum were evaluated using antagonistic plate assays and agar well diffusion assays. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of CFS were determined using the micro-dilution broth method. Strain HX-6 was identified based on colony morphological characteristics, along with 16S rRNA and gyrA gene sequencing. Its growth-promoting functions were assessed using selective media. Additionally, the inhibitory effects of CFS on R. solanacearum and its impact on cell integrity were further evaluated through changes in OD600 nm, conductivity, nucleic acid leakage, and malondialdehyde content, as well as observations using scanning electron microscopy. The results demonstrated that both the HX-6 fermentation solution and its CFS exhibited significant inhibitory activity on R. solanacearum. The fermentation solution produced an inhibition zone with a diameter of 3.03 cm, while CFS achieved an inhibitory rate of 70.48%. The MIC and MBC of CFS were determined to be 12.5% and 50.0%, respectively. Strain HX-6 was identified as Bacillus velezensis and possesses functions such as indole-3-acetic acid (IAA) production, siderophore synthesis, biofilm formation, and phosphate solubilization. Furthermore, CFS significantly impaired R. solanacearum activity and damaged its cellular integrity. These effects were evidenced by changes in OD600 nm value, conductivity, nucleic acid leakage, and malondialdehyde content. The results of scanning electron microscopy further revealed CFS caused significant damage to the integrity of R. solanacearum. The findings demonstrate that strain HX-6 exhibits strong antibacterial activity and shows promise as a novel biocontrol agent for bacterial wilt, warranting further investigation for agricultural use.

       

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