Isolation and characterization of Bacillus subtilis strain 1-L-29, an endophytic bacteria from Camellia oleifera with antimicrobial activity and efficient plant-root colonization

PLoS One. 2020 Apr 27;15(4):e0232096. doi: 10.1371/journal.pone.0232096. eCollection 2020.

Abstract

Endophytic bacteria, which are common in plant tissues, may help to control plant pathogens and enhance plant growth. Camellia oleifera, an oil-producing plant, is widely grown in warm, subtropical, hilly regions in China. However, C. oleifera is strongly negatively affected by C. oleifera anthracnose, which is caused by Colletetrichum fructicola. To find a suitable biocontrol agent for C. oleifera anthracnose, 41 endophytes were isolated from the stems, leaves, and roots of C. oleifera. Bacterial cultures were identified based on analyses of 16S rDNA sequences; most strains belonged to the genus Bacillus. The antagonistic effects of these strains on C. fructicola were tested in vitro. In total, 16 strains inhibited C. fructicola growth, with B. subtilis strain 1-L-29 being the most efficient. Strain 1-L-29 demonstrated antagonistic activity against C. siamense, C. asianum, Fusarium proliferatum, Agaricodochium camellia, and Pseudomonas syringae. In addition, this strain produced indole acetic acid, solubilized phosphate, grew on N-free media, and produced siderophores. To facilitate further microecological studies of this strain, a rifampicin-resistant, green fluorescent protein (GFP)-labeled strain, 1-L-29gfpr, was created using protoplast transformation. This plasmid had good segregational stability. Strain 1-L-29gfpr was re-introduced into C. oleifera and successfully colonized root, stem, and leaf tissues. This strain remained at a stable concentration in the root more than 20 d after inoculation. Fluorescence microscopic analysis showed that strain 1-L-29gfpr thoroughly colonized the root surfaces of C. fructicola as well as the root vascular tissues of Arabidopsis thaliana.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Bacterial Agents / metabolism*
  • Bacillus subtilis / classification
  • Bacillus subtilis / growth & development
  • Bacillus subtilis / isolation & purification
  • Bacillus subtilis / metabolism*
  • Camellia / growth & development*
  • Camellia / metabolism
  • Camellia / microbiology
  • Endophytes / growth & development
  • Endophytes / isolation & purification
  • Endophytes / metabolism*
  • Pest Control, Biological
  • Plant Development
  • Plant Diseases / microbiology
  • Plant Diseases / prevention & control*
  • Plant Roots / growth & development*
  • Plant Roots / metabolism
  • Plant Roots / microbiology

Substances

  • Anti-Bacterial Agents

Grants and funding

This work was supported by the National Science Foundation for Young Scientists of China (Grant No. 31600515, http://www.nsfc.gov.cn/) and the Natural Science Foundation of Hunan Province, China (Grant No. 2019JJ50999, http://kjt.hunan.gov.cn/zxgz/zkjj/). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.