AUTHOR=Duan Yanan , Chen Ran , Zhang Rong , Jiang Weitao , Chen Xuesen , Yin Chengmiao , Mao Zhiquan 

TITLE=Isolation, Identification, and Antibacterial Mechanisms of Bacillus amyloliquefaciens QSB-6 and Its Effect on Plant Roots

JOURNAL=Frontiers in Microbiology

VOLUME=12

YEAR=2021

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2021.746799

DOI=10.3389/fmicb.2021.746799

ISSN=1664-302X

ABSTRACT=<p>Apple replant disease (ARD) is a common problem in major apple planting areas, and biological factors play a leading role in its etiology. Here, we isolated the bacterial strain QSB-6 from the rhizosphere soil of healthy apple trees in a replanted orchard using the serial dilution method. Strain QSB-6 was provisionally identified as <italic>Bacillus amyloliquefaciens</italic> based on its morphology, physiological and biochemical characteristics, carbon source utilization, and chemical sensitivity. Maximum likelihood analysis based on four gene sequences [16S ribosomal RNA gene (16S rDNA), DNA gyrase subunit A (gyrA), DNA gyrase subunit B (gyrB), and RNA polymerase subunit B (rpoB)] from QSB-6 and other strains indicated that it had 100% homology with <italic>B. amyloliquefaciens</italic>, thereby confirming its identification. Flat standoff tests showed that strain QSB-6 had a strong inhibitory effect on <italic>Fusarium proliferatum</italic>, <italic>Fusarium solani</italic>, <italic>Fusarium verticillioides</italic>, <italic>Fusarium oxysporum</italic>, <italic>Alternaria alternata</italic>, <italic>Aspergillus flavus</italic>, <italic>Phoma</italic> sp., <italic>Valsa mali</italic>, <italic>Rhizoctonia solani</italic>, <italic>Penicillium brasilianum</italic>, and <italic>Albifimbria verrucaria</italic>, and it had broad-spectrum antibacterial characteristics. Extracellular metabolites from strain QSB-6 showed a strong inhibitory effect on <italic>Fusarium</italic> hyphal growth and spore germination, causing irregular swelling, atrophy, rupture, and cytoplasmic leakage of fungal hyphae. Analysis of its metabolites showed that 1,2-benzenedicarboxylic acid and benzeneacetic acid, 3- hydroxy-, methyl ester had good inhibitory effects on <italic>Fusarium</italic>, and increased the length of primary roots and the number of lateral roots of <italic>Arabidopsis thaliana</italic> plantlet. Pot experiments demonstrated that a QSB-6 bacterial fertilizer treatment (T2) significantly improved the growth of <italic>Malus hupehensis</italic> Rehd. seedlings. It increased root length, surface area, tips, and forks, respiration rate, protective enzyme activities, and the number of soil bacteria while reducing the number of soil fungi. Fermentation broth from strain QSB-6 effectively prevented root damage from <italic>Fusarium</italic>. terminal restriction fragment length polymorphism (T-RFLP) and quantitative PCR (qPCR) assays showed that the T2 treatment significantly reduced the abundance of <italic>Fusarium</italic> in the soil and altered the soil fungal community structure. In summary, <italic>B. amyloliquefaciens</italic> QSB-6 has a good inhibitory effect on <italic>Fusarium</italic> in the soil and can significantly promote plant root growth. It has great potential as a biological control agent against ARD.</p>