Inoculation Concentration of Bacillus subtilis Strain R31 Influenced the Roots Reactive Oxygen Species Generation of Banana (Musa spp. AAA Cavendish cv. Baxi) and Its Biological Control Effect on FusariumWilt

doi:10.16409/j.cnki.2095-039x.2017.02.013

Abstract

Abstract: The purpose of this paper is to study the effects of the inoculation concentrationsof Bacillus subtilis strain R31 on banana roots reactive oxygen generation, its own roots colonization and their biological control on Fusarium wilt. The Mn-SOD activities and H₂O₂ concentrations of banana (Musa spp. AAA Cavendish cv. Baxi) seedlings roots were investigated followed by the inoculation with 1.0×10⁶, 1.0×10⁷or 1.0×10⁸cfu/mL R31, respectively. The influence of different concentrations of H₂O₂ on R31 pellicle formation in MSgg medium, and the colonization of different concentrations R31-gfp on Baxi seedlings roots were also studied. Then, the biological control effects of different concentrations of R31 on Fusarium wilt were tested in fields. Results showed that significantly changed Mn-SOD activities and improved H₂O₂ concentrations of Baxi seedlings roots were observed after inoculated by 1.0×10⁶ or 1.0×10⁷cfu/mL of R31, respectively. R31 inoculation at 1.0×10⁸ cfu/mL could also significantly influence the Mn-SOD activities, which kept at 4-5 U/g at all tested time. However, there is no significant H₂O₂burst of Baxi roots after 1.0×10⁸ cfu/mL R31 inoculation. The concentration of H₂O₂higher than 80 or 120 μmol/mL could significantly inhibit the pellicle formation of R31 in MSgg medium with 1.0×10⁵ or 1.0×10⁶cfu/mL cell density at 24 h, but significantly improve the pellicle formation with the same treatment at 72 h. In the treatments with1.0×10⁷ and 1.0×10⁸cfu/mL R31, 40 to 240 μmol/mL H₂O₂concentrations had no influence on the pellicle formation of R31 at 24 h, but significantly improved the pellicle formation at 72 h. Green fluorescence was observed in the cuticle of Baxi seedlings roots inoculated by R31-gfp at 1.0×10⁶cfu/mL after 5 days, and those inoculated by R31 on 1.0×10⁷or 1.0×10⁸cfu/mL after 3 days. From December 2014 to May 2015, the Fusarium wilt control efficiency were 35.41% and 72.96%, respectively, after the Baxi seedlings were irrigated by 2000 mL 2.0×10⁶ and 2.0×10⁷cfu/mL R31 spores wettable powder suspension broth and water for 4 times in the 3 experiment areas set in Xuwen county of Zhanjiang City. It was showed that low inoculation concentrations of B. subtilis strain R31 could activate the Baxi roots immune system and caused reactive oxygen generation, then delay its own colonization on roots cuticle, which further influenced the biological control efficiency of fusarium wilt.

Key words: Fusarium wilt, biological control, Bacillus subtilis, root, immune system

CLC Number:

S476

CHENG Chuanyan, WANG Yan, YU Guohui, LI Yiping, WEN Shuheng, JI Liang. Inoculation Concentration of Bacillus subtilis Strain R31 Influenced the Roots Reactive Oxygen Species Generation of Banana (Musa spp. AAA Cavendish cv. Baxi) and Its Biological Control Effect on FusariumWilt[J]. journal1, 2017, 33(2/3): 226-233.

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