Effects of Coumarin on Biofilm and Phyllosphere Colonization of Bacillus subtilis B2 Strain

doi:10.7668/hbnxb.20194770

Abstract

Abstract:

In order to identify the synergistic mechanism of disease resistance of secondary metabolite coumarin to Bacillus subtilis B2,the biological control bacterium of walnut anthrax,this study measured the effects of different concentrations of coumarin on the growth,biofilm formation and colonization ability of B.subtilis B2.Coumarin was treated with 0.001(XD-1),0.010(XD-2),0.100(XD-3),1.000(XD-4),2.000 mmol/L (XD-5),and the effects of different concentrations on the growth of B.subtilis B2 strain were determined.The biofilm formation was observed and the biofilm formation ability was quantitatively determined by weighing method and crystal violet staining method.Colonization of the fluorescently labeled B.subtilis B2 strain was determined by coating plate method.The results showed that different concentrations of coumarin could promote the growth of the strains,0.100 mmol/L coumarin treatment was significantly different from the control,2.000 mmol/L coumarin treatment was not significantly different from the control.The 0.100 mmol/L coumarin treatment significantly enhanced the folding and stereoscopic degree of the biofilm,and the microscopic results also showed that the biofilm cells were significantly clustered together.The quantitative results of biofilm showed that 0.100 mmol/L coumarin treatment had the strongest biofilm formation ability.In addition,coumarin could improve the interfoliar colonization ability of strain B2,and the colonization amount reached the peak at 7 days after culture,and the 0.100 mmol/L treatment significantly increased the interfoliar colonization number of strain B2.In summary,this study revealed the important role of coumarin in regulating the biocontrol effect of B.subtilis B2 strain on walnut anthracnose,and gave full play to the biocontrol potential of biocontrol bacteria,providing an effective strategy for the biocontrol of walnut anthracnose.

Key words: Bacillus subtilis, Coumarin, Biofilm, Colonization

WANG Ruidong, LI Jing, SONG Fei, LI Huiping, YAN Aihua. Effects of Coumarin on Biofilm and Phyllosphere Colonization of Bacillus subtilis B2 Strain[J]. Acta Agriculturae Boreali-Sinica, 2024, 39(S1): 245-251. doi: 10.7668/hbnxb.20194770.

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Figures/Tables 6

Fig.1 Effects of different concentrations of coumarin on the growth of B.subtilis B2 strain

Fig.2 Observation on morphology of biofilm of B. subtilis strain B2 with different concentrations of coumarin

Fig.3 Microscopic observation of different concentrations of coumarin on B.subtilis B2 strain A.CK;B.XD-1;C.XD-2;D.XD-3;E.XD-4;F.XD-5.

Fig.4 Effect of coumarin at different concentrations on biofilm weight of B. subtilis B2 strain Different lowercase letters indicate significant differences between different treatments(P<0.05).The same as Fig.5.

Fig.5 Quantitative determination of biofilm of B. subtilis B2 with different concentrations of coumarin

Tab.1 Effect of different concentrations of coumarin on colonization of chloramphenicol labeled B.subtilis B2 10³cfu/g

处理Treatment	1 d	4 d	7 d	10 d
CK	0.46±0.07a	1.42±0.07bc	2.23±0.06b	1.66±0.12b
XD-1	0.47±0.08a	1.53±0.10bc	2.41±0.09a	1.71±0.06b
XD-2	0.47±0.05a	1.84±0.11a	2.53±0.15a	1.85±0.12ab
XD-3	0.47±0.18a	1.75±0.16a	2.44±0.13a	2.07±0.17a
XD-4	0.46±0.05a	1.44±0.09bc	2.22±0.10b	1.86±0.09ab
XD-5	0.45±0.05a	1.32±0.09c	2.00±0.14c	1.48±0.11c

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