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<p><i>Ralstonia solanacearum</i>, the causative agent of bacterial wilt disease, poses a significant threat to agriculture. The demand for safe and high-quality food has increased interest in biological control agents (BCAs), despite challenges related to stability and cost. Garbage enzymes (GEs) are a promising alternative, rich in beneficial microbes and organic compounds. In this study, four types of GEs (onion, garlic, ginger, and mixed fruit) were individually tested against <i>R. solanacearum </i>in vitro, with only onion GE showing significant inhibition. Filtered onion GE, which lacked any microbes, also showed inhibition; however, its heat-treated form exhibited a reduced inhibitory effect, indicating the role of heat-sensitive compounds in inhibiting <i>R. solanacearum</i>. An antagonistic bacterial strain A1, isolated from onion GE, inhibited pathogen growth by up to 75% through volatile compounds. Cell-free culture filtrate of the strain A1 also inhibited <i>R. solanacearum </i>in vitro. Strain A1 exhibited nitrogen fixation, siderophore production, indole acetic acid (IAA) synthesis, and extracellular enzyme production, positioning it as a potent biocontrol agent. The genome analysis of the strain A1 revealed the presence of several plant growth-promoting genes. In vivo studies with GE, GE-filtered, and strain A1 demonstrated significant pathogen inhibition and promoted tobacco plant growth. Disease incidence was reduced to 26.6% with GE, 46.67% with microbe-free GE, and 40% with strain A1. Overall, these treatments positively impacted plant root and shoot lengths as well as both fresh and dry weights. Our findings highlight onion GE as a potential, environmentally friendly method for controlling bacterial wilt and enhancing plant development, offering an alternative approach to traditional chemical controls in agriculture.</p> Graphical Abstract <p>(1) In vitro testing of 4 kinds of GEs against <i>R. solanacearum</i> strain YNQK. (2) Only the GE composed of onion waste showed inhibition of YNQK on the agar plates. (3) Three different kinds of bacteria were isolated from the zone of clearance and were tested against the pathogen in vitro; only A1 showed inhibition against the pathogen. (4) Upon sequencing of 16S rRNA&#xa0;and <i>gyrB</i> gene and constructing a phylogenetic tree, the A1 was identified as belonging to the <i>Bacillus</i> genus. (5) The A1 bacteria and GE were tested for their inhibitory action against <i>R. solanacearum</i> strain YNQK through the pot experiment.</p>

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Eco-friendly Biocontrol of Ralstonia solanacearum and Plant Growth Promotion in Tobacco Using Garbage Enzyme and Bacillus velezensis A1

  • 期刊论文

  • 发表日期：

    2025年4月3日

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  • Syed Sib Tul Hassan Shah,
  • Wangjie Shan,
  • Ying Wang,
  • Zhisheng Zheng,
  • Shuo Feng,
  • Lingxiao Wang,
  • Xiufang Hu,
  • Lin Li

摘要

Ralstonia solanacearum, the causative agent of bacterial wilt disease, poses a significant threat to agriculture. The demand for safe and high-quality food has increased interest in biological control agents (BCAs), despite challenges related to stability and cost. Garbage enzymes (GEs) are a promising alternative, rich in beneficial microbes and organic compounds. In this study, four types of GEs (onion, garlic, ginger, and mixed fruit) were individually tested against R. solanacearum in vitro, with only onion GE showing significant inhibition. Filtered onion GE, which lacked any microbes, also showed inhibition; however, its heat-treated form exhibited a reduced inhibitory effect, indicating the role of heat-sensitive compounds in inhibiting R. solanacearum. An antagonistic bacterial strain A1, isolated from onion GE, inhibited pathogen growth by up to 75% through volatile compounds. Cell-free culture filtrate of the strain A1 also inhibited R. solanacearum in vitro. Strain A1 exhibited nitrogen fixation, siderophore production, indole acetic acid (IAA) synthesis, and extracellular enzyme production, positioning it as a potent biocontrol agent. The genome analysis of the strain A1 revealed the presence of several plant growth-promoting genes. In vivo studies with GE, GE-filtered, and strain A1 demonstrated significant pathogen inhibition and promoted tobacco plant growth. Disease incidence was reduced to 26.6% with GE, 46.67% with microbe-free GE, and 40% with strain A1. Overall, these treatments positively impacted plant root and shoot lengths as well as both fresh and dry weights. Our findings highlight onion GE as a potential, environmentally friendly method for controlling bacterial wilt and enhancing plant development, offering an alternative approach to traditional chemical controls in agriculture.

Graphical Abstract

(1) In vitro testing of 4 kinds of GEs against R. solanacearum strain YNQK. (2) Only the GE composed of onion waste showed inhibition of YNQK on the agar plates. (3) Three different kinds of bacteria were isolated from the zone of clearance and were tested against the pathogen in vitro; only A1 showed inhibition against the pathogen. (4) Upon sequencing of 16S rRNA and gyrB gene and constructing a phylogenetic tree, the A1 was identified as belonging to the Bacillus genus. (5) The A1 bacteria and GE were tested for their inhibitory action against R. solanacearum strain YNQK through the pot experiment.