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Qualitative and Quantitative Analyses of the Colonization Characteristics of Bacillus subtilis Strain NCD-2 on Cotton Root

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Abstract

Bacillus subtilis strain NCD-2 is an excellent biocontrol agent against plant soil-borne diseases. With the purpose of understanding the colonization characteristics of strain NCD-2, firstly, a constitutive expression promoter was cloned from strain NCD-2 and was used to construct GFP-labeled strain NCD-2. The GFP-labeled strain NCD-2 showed strong green fluorescence under planktonic cells and biofilm formation. The colonization characteristics of strain NCD-2 on different parts of cotton root were qualitatively observed by confocal laser scanning microscopy (CLSM). Results showed that strain NCD-2 mainly colonized on the zone of differentiation and elongation. Rhizosphere populations of B. subtilis strain NCD-2 on different cotton root were quantitatively evaluated by traditional plating count and quantitative PCR (qPCR) analysis in both autoclaved soil and non-autoclaved soil, respectively. Results showed that both traditional plating count and qPCR analysis showed similar trend for colonization characteristics of strain NCD-2. The greatest strain NCD-2 populations were in the root tip, at 9.19 × 107 CFU g−1 root and 6.75 × 107 CFU g−1 root as estimated by qPCR in non-autoclaved and autoclaved soil, respectively. This study provides a clearer understanding of the interactions between biocontrol agent and plant, as well as with the indigenous microorganisms in the soil.

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Acknowledgements

This work was funded by the National Key Research and Development Program of China (2017YFD0200400), the earmarked fund for China Agriculture Research System (CARS-15–17), the Chinese National Natural Science Foundation (31572051 and 31601680), and the Natural Science Foundation of Hebei Province (C2016301069 and C2017301069).

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  1. Institute of Plant Protection, Hebei Academy of Agricultural and Forestry Sciences, Integrated Pest Management Centre of Hebei Province, 437# Dongguan street, Baoding, 071000, Hebei, China

    Lihong Dong, Qinggang Guo, Peipei Wang, Xiaoyun Zhang, Zhenhe Su, Weisong Zhao, Xiuyun Lu, Shezeng Li & Ping Ma

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284_2020_1971_MOESM1_ESM.tif

Supplementary file1 (TIF 188 kb) Fig. S1 The biofilm formation of wild type strain NCD-2 and GFP-labeled strain NCD-2. The error bars indicate the standard deviations of the means calculated from three independent samples.

284_2020_1971_MOESM2_ESM.eps

Supplementary file2 (EPS 111363 kb) Fig. S2 Growth curve of the wild-type strain NCD-2 and the GFP-labeled strain NCD-2.

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Dong, L., Guo, Q., Wang, P. et al. Qualitative and Quantitative Analyses of the Colonization Characteristics of Bacillus subtilis Strain NCD-2 on Cotton Root. Curr Microbiol 77, 1600–1609 (2020). https://doi.org/10.1007/s00284-020-01971-y

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