Study on the Control and Induced Resistance in Cucumber with Bacillus subtilis B579 against Cucumber Fusarium Wilt

doi:10.13560/j.cnki.biotech.bull.1985.2024-0160

Abstract

Abstract:

【Objective】 To explore the control effect and mechanism of Bacillus subtilis B579 on cucumber Fusarium wilt. 【Method】 Plate confrontation test and greenhouse pot experiment were used to detect the growth and morphological changes of the pathogen, plant growth, activity of defense enzymes in leaves and malondialdehyde content, and expression of defense-related related genes. 【Result】 The B. subtilis B579 had a strong inhibitory effect on cucumber Fusarium wilt, and the mycelium around the inhibition zone showed uneven thickness, swelling and breakage. Pot experiment demonstrated that B579 achieved the disease control efficiency against cucumber wilt, reaching 78.80%. The plant height, stem diameter, fresh weight, and dry weight of cucumber seedlings in B579 treatment increased by 13.87%, 4.17%, 15.15% and 12.77% compared with the control group, respectively. After pretreatment with B579, pathogen inoculation significantly increased the activities of phenylalanine ammonilyase（PAL）, polyphenol oxidase（PPO）, peroxidase（POD）, and catalase（CAT）and reduced the content of malondialdehyde（MDA）in cucumber leaves on day 4 and 7 compared with the control group. The results of real-time fluorescence quantitative PCR showed that the expressions of LOX, PR1, PR3 and CAT genes in cucumber leaves after inoculation with B579 and then challenge with pathogen were significantly higher than those in other treatments at 4 and 7 d. 【Conclusion】 B. subtilis B579 can control cucumber Fusarium wilt by directly inhibiting the growth of pathogen and inducing resistance in cucumber.

Key words: Bacillus subtilis B579, promoting growth, cucumber Fusarium wilt, induced resistance, biological control

FAN Zong-qiang, FENG Jing-han, ZHENG Li-xue, WANG Shuo, PENG Xiang-qian, CHEN Fang. Study on the Control and Induced Resistance in Cucumber with Bacillus subtilis B579 against Cucumber Fusarium Wilt[J]. Biotechnology Bulletin, 2024, 40(7): 226-234.

Figures/Tables 9

Table 1 Primer information for the real-time quantitative PCR reaction

基因Gene	基因序列Sequence（5'-3'）	登录号Accession number	参考文献Reference
ACTIN	F: TGGACTCTGGTGATGGTGTTA	DQ641117	[24]
	R:CAATGAGGGATGGCTGGAAAA	DQ641117	[24]
LOX	F:GAGAGCGTAAGGAATGGGATAGAA	NM_001305708	[8]
	R: CACCGGGTTCGGAAAGG	NM_001305708	[8]
PR1	AACTCTGGCGGACCTTAC	DQ641122	[24]
	GACTTCCTCCACACTACT	DQ641122	[24]
PR3	GCCTTACTCCATAACATCACTCC	DQ641104	[25]
	GATTTCGATATCGAGTCTGGCT	DQ641104	[25]
CAT	AACCCAACCCAAAATCCCA	GU248529	[26]
	TCTAATAGCCTCCTCTTCCAGCA	GU248529	[26]

Fig. 1 Effect of B579 fermentation broth on the growth of pathogen Data are mean ± SE，different small letters on the bar chart indicate significant differences between each treatment group and the control group（P<0.05）；the same below

Fig. 2 Mycelium morphology of pathogens in control group（A）and treatment group（B, C）under optical microscope A: Mycelium folding; B: hyphal expansion; C: hyphal rupture

Fig. 3 Cucumber plants under different treatment

Table 2 Control effect of B579 treatment on cucumber Fusarium wilt

组别 Treatment	病情指数 Disease index	防治效果 Control efficiency/％
CK	0c	-
B579	0c	-
B579+FOC	11.12±2.41b	78.80％
FOC	52.78±4.81a	-

Table 3 Effects of different treatments on cucumber seedling growth

组别Treatment	株高Plant height/cm	茎粗Stem diameter/mm	鲜重Fresh weight/g	干重Dry weight/g
CK	9.95±1.30b	4.80±0.39b	9.11±1.07b	0.47±0.06b
B579	11.33±0.79a	5.00±0.57a	10.49±0.87a	0.53±0.06a
B579+FOC	9.32±0.62b	4.66±0.45b	7.98±0.76c	0.42±0.07b
FOC	7.42±1.10d	3.97±0.58c	6.41±1.19d	0.33±0.07d

Fig. 4 Activities of 4 defensive enzymes in cucumber leaves under different treatments

Fig. 5 MDA content in cucumber leaves under different treatments

Fig. 6 Expressions of four defensive genes in cucumber leaves under different treatments

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