Identification of Bacillus velezensis ZN-S10 and its antification effect on tomato bacterial wilt

doi:10.3969/j.issn.1004-1524.20230864

Acta Agriculturae Zhejiangensis ›› 2023, Vol. 35 ›› Issue (11): 2636-2644.DOI: 10.3969/j.issn.1004-1524.20230864

• Plant Protection • Previous Articles Next Articles

Identification of Bacillus velezensis ZN-S10 and its antification effect on tomato bacterial wilt

WANG Xiaonan¹(), FENG Xiaoxiao², SHI Bin¹, CHEN Enlei¹, CHEN Mengli¹, ZHENG Yongli³^,^*(), WU Huiming¹^,^*()

1. College of Advanced Agricultural Science, Zhejiang A & F University, Hangzhou 311300, China
2. Agricultural Test Station, Zhejiang University, Hangzhou 310058, China
3. Zhejiang Agricultural Products Green Development Center, Hangzhou 310003, China

Received:2023-07-12 Online:2023-11-25 Published:2023-12-04

Abstract

Abstract:

In order to identify an endophytic bacterium from tomato root and its antibacterial effect, the antagonistic activity of tomato endophytic bacteria against different pathogens was tested, providing insights for the biological control of common bacterial diseases. Endophytic bacteria were isolated from tomato roots and identified by morphological, fatty acid, physiological characteristics determination as well as a dual-gene system of gyrA and rpoB phylogenetic trees. The antagonistic effect of endophytic bacteria on five common plant pathogenic bacteria was tested using the plate method, and the efficacy of the bacterium against tomato bacterial wilt disease was evaluated using the pot culture experiment. The strain ZN-S10 was identified as Bacillus velezensis. The antibacterial assay showed that B.velezensis ZN-S10 exhibited different degrees of bacteroistatic activity against Pseudomonas viridifiava, Dickeya dadantii, Erwinia carotovora, Ralstonia solanacearum and Xanthomonas campestris. The greenhouse pot culture experiment demonstrated that the control effect of B.velezensis ZN-S10 on tomato bacterial wilt was 57.14%, which was not significantly different from that of 3% Zhongshengmycin WP, and could effectively reduce the development of tomato bacterial wilt. These research findings provided a theoretical basis for the effective use utilization of endophytic bacteria in the control of common bacterial diseases and highlighted the promising application prospects of B. velezensis ZN-S10 in the management of bacterial diseases in tomatoes.

Key words: Bacillus velezensis, antagonistic activity, Ralstonia solanacearum, control efficiency, biological control

CLC Number:

S436.412

WANG Xiaonan, FENG Xiaoxiao, SHI Bin, CHEN Enlei, CHEN Mengli, ZHENG Yongli, WU Huiming. Identification of Bacillus velezensis ZN-S10 and its antification effect on tomato bacterial wilt[J]. Acta Agriculturae Zhejiangensis, 2023, 35(11): 2636-2644.

Figures/Tables 9

Fig.1 Colony morphology and Gram staining of ZN-S10

Table 1 Physiological and biochemical identification results of endophytic bacterium ZN-S10

生理生化指标测试 Physiological and biochemical index	ZN-S10菌株 ZN-S10 strain
氧化酶 Oxidase	+
卵磷脂酶 Lecithinase	+
过氧化氢酶 Catalase	+
淀粉水解 Amylohydrolysis	+
V-P 测定 V-P test	+
甲基红 Methyl red	-
吲哚 Indole	+
氯霉素 Chloromycetin	-
链霉素 Streptomycin	-
卡那霉素 Kanamycin	-
氨苄青霉素 Ampicillin	-

Fig.2 ZN-S10 phylogenetic tree based on rpoB and gyrA gene sequences The value at the branch is the support rate of each node obtained by repeating the Bootstrap method 1 000 times, with a scale of 0.050 as the evolution distance.

Fig.3 Antibacterial effect of Bacillus velezensis ZN-S10 against different pathogens The filter paper on the left side of the plate in the picture is CK, and the filter paper on the right side is ZN-S10. From left to right are Erwinia carotovora ZJUP0154-1, Pseudomonas viridifiava ZJUP0398-2, Dickeya dadantii ZAFU0898, Ralstonia solanacearum ZAFU0005, and Xanthomonas campestris ZJUP0463-1.

Fig.4 Inhibition rate of Bacillus velezensis ZN-S10 against five pathogens The data is mean ± standard error, and the different lowercase letters on the column indicate significant differences (P<0.05). The same as below.

Table 2 Effect of Bacillus velezensis ZN-S10 and 3% Zhongshengmycin WP on control of Ralstonia solanacearum

处理 Treatment	病情指数 Disease index	相对防治效果 Relative control effect/%
ZN-S10	31.25	57.14
3%中生菌素 WP	31.25	57.14
3% Zhongshengmycin WP
MOCK2	72.92

Fig.5 Control effect of ZN-S10 on Ralstonia solanacearum ZAFU0005 A, Clear water; B, Inoculate Ralstonia solanacearum ZAFU0005; C, Inoculate Ralstonia solanacearum ZAFU0005 after 7 days of root irrigation with 3% mycorrhizal WP; D, After 7 days of root irrigation with ZN-S10, Ralstonia solanacearum ZAFU0005 was inoculated.

Fig.6 MDA content changes of tomato MOCK, Clean water; T1, ZN-S10; T2, ZAFU0005; T3, ZN-S10+ZAFU0005. Under the same treatment time, the bars of different treatments marked with different lowercase letters showed significant difference (P<0.05).The same as below.

Fig.7 GSH content and PAL, SOD, CAT and POD activity changes of tomato

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Identification of Bacillus velezensis ZN-S10 and its antification effect on tomato bacterial wilt

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