The Effect of Banana Rhizosphere Chemotaxis and Chemoattractants on Bacillus velezensis LG14-3 Root Colonization and Suppression of Banana Fusarium Wilt Disease

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Abstract

Fusarium oxysporum f. sp. cubense (Foc) causes banana Fusarium wilt disease, which is a destructive soil-borne disease. Many plants can recruit rhizosphere microorganisms using their root exudates, thereby shaping the rhizosphere microbiome to resist pathogen infection. Therefore, this study was conducted to explore the role of root exudates in the process of biocontrol strain colonization and resistance to pathogens. In this study, the banana root exudates used as chemoattractants were obtained by hydroponics. Bacillus velezensis strain LG14-3 was isolated from the infected area of the root system of banana and showed significant chemotaxis to banana root exudates and strong inhibition of Fusarium oxysporum f. sp. cubense. Further analysis found that LG14-3 showed chemotaxis toward the components of banana root exudates, such as citric acid, succinic acid, glycine, D-galactose and D-maltose, and glycine and citric acid, which resulted in more significant chemotaxis of LG14-3. Moreover, banana root exudates enhanced the swarming motility and biofilm formation of LG14-3. Pot experiments showed that glycine and citric acid enhanced the colonization ability of Bacillus velezensis LG14-3 in the banana rhizosphere and reduced the disease severity index of banana fusarium wilt. Glycine and citric acid enhanced the growth-promoting ability of LG14-3 under pathogen stress. Our results showed that the addition of chemotactic substances enhanced the biocontrol potential of Bacillus velezensis LG14-3 to prevent banana Fusarium wilt.

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A universal method to detect and determine siderophores was developed by using their high affinity for iron(III). The ternary complex chrome azurol S/iron(III)/hexadecyltrimethylammonium bromide, with an extinction coefficient of approximately 100,000 M-1 cm-1 at 630 nm, serves as an indicator. When a strong chelator removes the iron from the dye, its color turns from blue to orange. Because of the high sensitivity, determination of siderophores in solution and their characterization by paper electrophoresis chromatography can be performed directly on supernatants of culture fluids. The method is also applicable to agar plates. Orange halos around the colonies on blue agar are indicative of siderophore excretion. It was demonstrated with Escherichia coli strains that biosynthetic, transport, and regulatory mutations in the enterobactin system are clearly distinguishable. The method was successfully used to screen mutants in the iron uptake system of two Rhizobium meliloti strains, DM5 and 1021.

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Disease-suppressive soils are exceptional ecosystems in which crop plants suffer less from specific soil-borne pathogens than expected owing to the activities of other soil microorganisms. For most disease-suppressive soils, the microbes and mechanisms involved in pathogen control are unknown. By coupling PhyloChip-based metagenomics of the rhizosphere microbiome with culture-dependent functional analyses, we identified key bacterial taxa and genes involved in suppression of a fungal root pathogen. More than 33,000 bacterial and archaeal species were detected, with Proteobacteria, Firmicutes, and Actinobacteria consistently associated with disease suppression. Members of the γ-Proteobacteria were shown to have disease-suppressive activity governed by nonribosomal peptide synthetases. Our data indicate that upon attack by a fungal root pathogen, plants can exploit microbial consortia from soil for protection against infections.