Isolation and Identification of Bacillus velezensis ZHX-7 and Its Antibacterial and Growth-promoting Effects

doi:10.13560/j.cnki.biotech.bull.1985.2023-0152

Abstract

Abstract:

In order to obtain highly effective biocontrol microorganism for controlling peanut diseases, using Aspergillus niger as the indicator, a highly effective biocontrol strain ZHX-7 was screened from soil samples by agar disk dilution and dual-culture assay. The soil samples were collected from the rhizosphere of diseased and healthy peanut plants in continuous cropping field in Laixi, Shandong province. The strain ZHX-7 was identified as Bacillus velezensis based on the morphological characteristics, 16S rDNA and gyrB sequence analysis. The bacterial suspension, sterile fermentation broth and volatile gas of ZHX-7 significantly inhibited the growth of Sclerotium rolfsii, Fusarium neocosmosporiellum, Phoma arachidicola and Alternaria alternata. The antagonistic experiment results showed that the inhibition rates of ZHX-7 suspension against the growth of S. rolfsii, F. neocosmosporiellum, P. arachidicola and A. alternata were 64.40%, 70.37%, 77.43% and 65.32%, respectively. In the sterile fermentation broth of ZHX-7 diluted 10 times with PDA medium, the inhibition rates of ZHX-7 against the growth of S. rolfsii, F. neocosmosporiellum, P. arachidicola and A. alternata were 67.81%, 52.16%, 23.52% and 69.93%, respectively. The double dish test showed that the inhibition rates of ZHX-7 volatile gas against the growth of S. rolfsii, F. neocosmosporiellum, P. arachidicola and A. alternata were 86.82%, 35.40%, 81.70% and 44.19%, respectively. ZHX-7 produced secondary metabolites such as protease, cellulose and amylase. In addition, under the condition of pot culture in greenhouse, ZHX-7 fermentation broth significantly promoted the growth of peanut. Compared with the control, the fresh mass and dry mass increased by 31.24% and 26.47%, respectively. ZHX-7 fermentation broth improved the resistance of peanut to S. rolfsii, and the control effect reached 51.76%. In conclusion, a strain of B. velezensis ZHX-7 screened in this study may inhibit a variety of peanut pathogenic fungi and promote peanut growth at the same time. It is a multifunctional strain with broad antagonistic spectrum against peanut diseases, and high potential for biocontrol application.

Key words: Bacillus velezensis, peanut diseases, isolation and identification, antibacterial rate, biological control, growth-promoting effect, Sclerotium rolfsii

LI Ying, SONG Xin-ying, HE Kang, GUO Zhi-qing, YU Jing, ZHANG Xia. Isolation and Identification of Bacillus velezensis ZHX-7 and Its Antibacterial and Growth-promoting Effects[J]. Biotechnology Bulletin, 2023, 39(12): 229-236.

Figures/Tables 7

Fig. 1 Antagonistic effect of ZHX-7 against A. niger（A）and its colony morphology（B）

Fig. 2 Phylogenetic analysis of ZXH-7 based on 16S rDNA and gyrB sequences

Fig. 3 Determination of antagonistic activity of ZHX-7 bacterial suspension against four pathogens A: Antagonistic effect of ZHX-7 on four peanut pathogens. B: Statistics of colony growth of pathogens. C: Statistics of inhibition rate of ZHX-7 against four peanut pathogens. ** indicates extremely significant differences at 0.01 level. The same below

Fig. 4 Effects of ZHX-7 fermentation broth on the growth of four pathogens

Fig. 5 Inhibitory effects of ZHX-7 volatiles against four pathogens

Fig. 6 Detection of ZHX-7 secondary metabolite extracellular enzyme activity

Table1 Control effect of ZHX-7 on peanut growth and peanut Sclerotium blight

处理Treatment	鲜质量Fresh mass/g	干质量Dry mass/g	死亡率Mortality/%	病情指数Disease index	防效Efficacy/%
CK	10.47±0.74 A	1.48±0.10 A	56.67±5.77 A	85.42±3.15 A	/
ZHX-7	13.74±0.96 B	1.88±0.11 B	18.33±2.89 B	41.25±3.31 B	51.76±2.13

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