Development of a Strain-Specific Quantification Method for Monitoring Bacillus amyloliquefaciens TF28 in the Rhizospheri
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ORIGINAL PAPER
Development of a Strain‑Specific Quantification Method for Monitoring Bacillus amyloliquefaciens TF28 in the Rhizospheric Soil of Soybean Shumei Zhang1,2 · Yinpeng Ma2 · Wei Jiang1,2 · Liqiang Meng1,2 · Xu Cao1,2 · Jihua Hu2 · Jingyu Chen2 · Jing Li1,2 Accepted: 14 August 2020 © Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Bacillus amyloliquefaciens TF28 can be used to control soybean root disease. To assess its commercial potential as a biocontrol agent, it is necessary to develop a strain-specific quantification method to monitor its colonization dynamics in the rhizospheric soil of soybean under field conditions. Based on genomic comparison with the same species in NCBI databases, a strain-unique gene ukfpg was used as molecular marker to develop strain-specific PCR assay. Among three primer pairs, only primer pairs (F2/R2) could specifically differentiate TF28 from other strains of B. amyloliquefaciens with the detection limit of 10 fg and 100 CFU/g for DNA extracted from pure culture and dry soil, respectively. Then, a colony count coupled with PCR assay was used to monitor the population of TF28 in the rhizospheric soil of soybean in the field. The results indicated that TF28 successfully colonized in the rhizospheric soil of soybean. The colonization population of TF28 changed dynamically within the 120-day growth period with high population at the branching (V6) and flowering stages (R2). This study provides an efficient method to quantitatively monitor the colonization dynamics of TF28 in the rhizospheric soil of soybean in the field and demonstrates the potential of TF28 as a biocontrol agent for commercial development. Keywords Bacillus amyloliquefaciens TF28 · Monitoring · Quantification
Introduction Agricultural sustainability requires an environment friendly strategy to control plant diseases. Biological control utilizing beneficial microbes is a promising way to decrease plant diseases and the negative effect of chemicals on the environment [1]. Research on these beneficial microbes as biocontrol agents showed that biocontrol efficacy of these microbes to suppress plant diseases depends on their ability to survive and maintain a sufficient population in the rhizospheric soil [2]. Therefore, successful colonization of biocontrol agents in the rhizospheric soil is a prerequisite for the commercial development [3]. Besides that, study on the microbial * Jing Li [email protected] 1
Institute of Microbiology, Heilongjiang Academy of Sciences, 68 Zhaolin Street, Daoli District, Harbin 150010, Heilongjiang, China
Institute of Advanced Technology, Heilongjiang Academy of Sciences, Harbin 150020, China
2
colonization is essential for discovering and evaluating the promising biocontrol strains. Soil is a complex environmental matrix. Plant root exudates, soil physicochemical properties, and indigenous microbes have a great influence on the colonization of biocontrol agents [4]. The abundance of indigenous microorganisms increases the difficulty of acc
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