Biotechnology Bulletin ›› 2022, Vol. 38 ›› Issue (6): 136-146.doi: 10.13560/j.cnki.biotech.bull.1985.2021-1124

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Isolation and Identification of Bacillus subtilis K-268 and Its Biological Control Effect on Rice Blast

ZU Xue1,2,3(), ZHOU Hu1,2,3, ZHU Hua-jun1,2,3, REN Zuo-hua1,2,3, LIU Er-ming1,2,3()   

  1. 1. College of Plant Protection,Hunan Agricultural University,Changsha 410128
    2. Hunan Key Laboratory of Plant Pest Biology and Control,Changsha 410128
    3. Southern Grain and Oil Crop Collaborative Innovation Center,Changsha 410128
  • Received:2021-09-01 Online:2022-06-26 Published:2022-07-11
  • Contact: LIU Er-ming E-mail:1762823097@qq.com;ermingliu@163.com

Abstract:

The use of biocontrol bacteria is an effective measure for green prevention and control of rice blast. In order to discover biocontrol bacteria with high efficiency against Magnaporthe oryzae,an antagonistic bacterium K-268 with the optimal inhibitory effect on M. oryzae from the healthy rice leaves of the susceptible variety K020268 was obtained by using the plate confrontation method and its antimicrobial spectrum was determined. Using morphological observation,physiological and biochemical identification 16S rRNA and gyrA sequence analysis,the identity of the strain was determined. In addition,the physical characteristics of the strain and its control effect against rice leaf blast were preliminarily studied. Results showed that strain K-268 had inhibitory rate of(86.30 ± 0.70)% on the mycelial growth of M. oryzae,and it demonstrated the inhibitory effects on 14 plant pathogenic fungi,such as Rhizoctonia solan,Exserohilum turcicum and Diaporthe citri,etc. Strain K-268 was identified to be Bacillus subtilis,its logarithmic growth period was 14-32 h,the optimal;growth temperature was 30℃,the optimal pH was 6.0-7.0,and had good salt tolerance. The results from in vitro inoculation experiments showed that the disease rate in the prevention group and the treatment group was14.81% and 23.46%,respectively,when the bacterial solution was diluted by 10 times to 6×108 CFU;and the effect of which was not significantly different from that of the 750 times diluted 75% tricyclazole diluent(WP). Results of in vivo spray inoculation showed that the control effects on the rice leaf blast were 59.00% and 60.23%,respectively,when the 10 times diluted fermentation liquid(6×108CFU/mL)were sprayed 24 h before and after inoculation with Magnaporthe oryzae conidia suspension(1×106 /mL),respectively. These effects were close to the 40% rice blast spirit(EC)with 500 times dilution. Therefore,K-268 was confirmed to have certain development and application value in the biological control of rice blast.

Key words: rice blast, Bacillus subtilis, isolation and identification, biological control