Inhibitory Effect of Dimethyl Disulfide from Burkholderia pyrrocinia WY6-5 on Aspergillus flavus and Aflatoxins in Peanuts During Storage Period

GONG AnDong;DONG FeiYan;WU NanNan;KONG XianWei;ZHAO Qian;YAN JianLi;Cheelo DIMUNA;College of Life Science, Xinyang Normal University/Henan Key Laboratory of Tea Plant Biology;

Burkholderia pyrrocinia;;Aspergillus flavus;;aflatoxin;;dimethyl disulfide;;antifungal;;storage period

【Objective】The objective of this study is to verify the antifungal effect of Burkholderia pyrrocinia WY6-5, evaluate its control efficacy against Aspergillus flavus and aflatoxins in peanuts during storage period, analyze the inhibitory mechanism, identify antifungal volatiles and detect the minimal inhibitory concentration to A. flavus, so as to provide novel strategies for the prevention and control of fungal diseases and mycotoxin during storage period.【Method】Face-to-face dual cultural test was conducted to analyze the antifungal activity of volatiles from WY6-5. Active charcoal as volatile adsorbent was added into the tests to verify the antifungal activity of volatiles. Dimethyl disulfide(DMDS) emitted form strain WY6-5 was challenged with peanut kernels inoculated with A. flavus conidia in sealed airspace without physical contact. A. flavus cells on peanut coat were collected, fixed in osmic acid, and analyzed through scanning electron microscope(SEM). Transmission electron microscope(TEM) was used to test the inner structure of A. flavus cell affected by volatiles from WY6-5. The commercial DMDS was purchased, serially diluted, and co-cultured with A. flavus conidia and mycelia to detect the minimal inhibitory concentration, respectively.【Result】B. pyrrociniaWY6-5 isolated from rhizosphere soil of tea plants could produce volatile DMDS, prevent the growth of A. flavus mycelia, the inhibition rate was over 95%. Additionally, under the condition of high water activity(aw), WY6-5 could also inhibit the A. flavus infection and aflatoxins production in peanuts during storage period. In peanuts of control treatment, the disease incidence was 100%, and the total concentration of aflatoxins was 399.32 μg·kg~(-1)(aw 0.859) and 3 143.19 μg·kg~(-1)(aw 0.923), respectively. When WY6-5 was added in the treatment, the disease incidence decreased to 2%(aw 0.859) and 21%(aw 0.923), respectively. The concentration of aflatoxins decreased to 4.86 μg·kg~(-1)(aw 0.859) and 121.37 μg·kg~(-1)(aw 0.923), respectively. The inhibition rate of WY6-5 against aflatoxins contamination was 98.78% and 96.14% compared to the control treatment. SEM analysis proved that DMDS from WY6-5 inhibited the germination of A. flavus conidia. TEM analysis further proved that the inner cell structures of A. flavus conidia were not severely damaged by volatiles. Volatile DMDS showed great antifungal activity. The minimal inhibitory concentration against mycelia growth was 100 μL·L~(-1)(compound volume/airspace volume). The minimal inhibitory concentration against conidia germination was 50 μL·L~(-1)(compound volume/airspace volume). 【Conclusion】 B. pyrrocinia WY6-5 can produce valid antifungal volatile DMDS, which can completely inhibit the mycelia growth and conidia germination of A. flavus at low concentration, and greatly prevent the development of A. flavus disease and aflatoxins contamination in peanuts during storage period. WY6-5 and the produced DMDS provide novel bio-active agents for fungal diseases control and mycotoxins during storage period.