Volatile Organic Compounds Produced by Cacao Endophytic Bacteria and Their Inhibitory Activity on Moniliophthora roreri

Abstract

Several studies have reported that bacteria produce anti-fungal volatiles. We identified the organic volatile compounds produced by six cacao endophytic bacteria (CEB) strains (Bacillus amyloliquefaciens CFFSUR-B35, Bacillus megaterium CFFSUR-B32, Bacillus muralis CFFSUR-B39, Bacillus pumilus CFFSUR-B34, Bacillus subtilis CFFSUR-B31, and Novosphingobium lindaniclasticum CFFSUR-B36). We evaluated their inhibitory effect on mycelium growth and spore germination of the phytopathogenic fungus Moniliophtora roreri. The volatiles produced by these six CEB, were collected and identified by SPME and GC-MS. Moreover, the inhibitory effect of five synthetic volatile organic compounds, individually and in mixtures (dimethyl disulfide, 2-5 dimethyl pyrazine, α-pinene, 2-heptanone and 2-ethyl hexanol) on M. roreri mycelium growth and spore germination was evaluated. All strains examined produced volatiles in different amounts; 13 to 10 compounds were identified, including sulfide, alcohol benzene derivate, pyrazine, ketone, nitrogen and terpene compounds. The B. subtilis CFFSUR-B31 strain produced the largest number of volatiles, while B. pumilus CFFSUR-B34 produced the fewest and the lowest amounts. The volatile organic compounds produced by B. pumilus CFFSUR-B34, B. muralis CFFSUR-B39 and N. lindaniclasticum CFFSUR-B36 inhibited M. roreri mycelium growth by more than 35%, sporulation by more than 81% and spore germination by more than 74%. However, when synthetic compounds were evaluated individually and in mixtures, 2-ethyl hexanol at 100,000 ppm (20 mg/filter paper disc) inhibited M. roreri mycelium growth by 100%, followed by organic volatile compound mixtures C (dimethyl disulfide, 2,5-dimethyl pyrazine, α-pinene, 2-ethyl-hexanol, 2-Heptanone) and D (only the top four) at 100,000 ppm (4 and 5 mg/filter paper disc) which inhibited spore germination by 97 and 89%, respectively.