Characterization of Antimicrobial-Producing Beneficial Bacteria Isolated from Huanglongbing Escape Citrus Trees

Abstract

The microbiome associated with crop plants has a strong impact on their health and productivity. <i>Candidatus</i> Liberibacter asiaticus (Las), the bacterial pathogen responsible for Huanglongbing (HLB) disease, lives inside the phloem of citrus plants including the root system. It has been suggested that Las negatively affects citrus microbiome. On the other hand, members of citrus microbiome also influence the interaction between Las and citrus. Here, we report the isolation and characterization of multiple putative beneficial bacteria from healthy citrus rhizosphere. Firstly, six bacterial strains showing antibacterial activity against two bacteria closely related to Las: <i>Agrobacterium tumefaciens</i> and <i>Sinorhizobium meliloti</i> were selected. Among them, <i>Burkholderia metallica</i> strain A53 and <i>Burkholderia territorii</i> strain A63 are within the β-proteobacteria class, whereas <i>Pseudomonas granadensis</i> strain 100 and <i>Pseudomonas geniculata</i> strain 95 are within the γ-proteobacteria class. Additionally, two gram-positive bacteria <i>Rhodococcus jialingiae</i> strain 108 and <i>Bacillus pumilus</i> strain 104 were also identified. Secondly, antimicrobial activity against three fungal pathogens: <i>Alternaria alternata</i>, <i>Colletotrichum acutatum</i>, <i>Phyllosticta citricarpa</i>, and two oomycetes: <i>Phytophthora nicotianae</i> and <i>Phytophthora palmivora.</i> Four bacterial strains <i>Burkholderia territorii</i> A63, <i>Burkholderia metallica</i> A53, <i>Pseudomonas geniculata</i> 95, and <i>Bacillus pumilus</i> 104 were shown to have antagonistic activity against the citrus root pathogen <i>Phytophthora nicotianae</i> based on dual culture antagonist assays and compartmentalized petri dish assays. The four selected bacteria were sequenced. Genes involved in phosphate solubilization, siderophore production and iron acquisition, volatile organic compound production, osmoprotection and osmotic tolerance, phytohormone production, antagonism, and nutrient competition were predicted and discussed related to the beneficial traits.