Abstract
Plasmodiophora brassicae is a Plasmodiophorid and obligate biotrophic protist in the eukaryotic kingdom of Rhizaria. It causes clubroot disease of cruciferous crops by producing resting spores that can survive in the soil for more than ten years. Although up to date there is no effective chemical control but now a strain of the bacterium Bacillus subtilis XF-1, isolated from China, has been found to inhibit P. brassicae and the mycelial growth of other plant-pathogenic fungi, such as Magnaporthe oryzae, Curvularia lunata,Fusarium oxysporum and so on. The antimicrobial activity of this biocontrol bacterium lies in its water-soluble protein (PBT1) which was isolated and purified by means of molecular sieve and affinity chromatography. The single band of the purified protein has a molecular mass of 25 kDa, estimated by a sodium dodecyl sulfate- polyacrylamide gel electrophoresis and it was heat resistant. The result of the sequencing proved PBT1 to be a new protein. Recombinant plasmid pQE81L-PBT1 was constructed using the coding gene of Pbt1 and plasmid PQE81-L, and expressed in Escherichia coli BL21. The fusion protein of BL21 exhibited significant detrimental effects on P. brassicae spores. In addition, pQE81L-PBT1 was successfully inserted into Pseudomonas fluorescens strain 2P24 which developed antagonistic effect to the clubroot pathogen that was not suppressed by 2P24 alone. The results showed that PBT1 protein might play a key role in the biocontrol of clubroot disease of crucifers.
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Acknowledgments
This research was supported by the Ministry of Agriculture of China, for Special Fund for the Agro-scientific Research in the Public Interest (2010029030), the Natural Science Foundation of Yunnan Province (2008CC024), the National Natural Science Foundation of China (20972166, 31070288 & 31161140345), and the Ministry of Education of China through its 211 and 985 programs (B08044, MUC 985-6 & MUC 98506-01000101).
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Zhao, J., Wu, YX., Ho, HH. et al. PBT1, a novel antimicrobial protein from the biocontrol agent Bacillus subtilis XF-1 against Plasmodiophora brassicae . Eur J Plant Pathol 145, 583–590 (2016). https://doi.org/10.1007/s10658-016-0905-y
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DOI: https://doi.org/10.1007/s10658-016-0905-y