Isolation and Identification of Biocontrol Bacteria for Potato Anthracnose in Three Strains and Optimization of Fermentation Parameters for Cultures with High Antibacterial Activity

doi:10.13560/j.cnki.biotech.bull.1985.2025-0209

Abstract

Abstract:

Objective Potato anthracnose, caused by Colletotrichum coccodes, severely impacts both yield and quality. Bacillus spp. are commonly used as biocontrol agents, making the search for efficient and low-toxicity strains to manage this disease highly significant. This study focused on three Bacillus strains with strong antibacterial activity and their composite microbial community. By optimizing the fermentation parameters of the composite Bacillus, the aim is to enhance its inhibitory effect against the pathogen, providing a reference for the development of composite biocontrol agents. Method Endophytic bacteria were isolated from potato tubers by tissue isolation and scribing methods, strains with strong inhibitory effect on potato anthrax were screened by drug-sensitive paper slide method, the taxonomic status of the active strains was determined by combining morphological and molecular biological identification, and the optimal ratio of the three strains was determined by drug-sensitive paper slide method, and the composite group was optimized by one-factor test and response surface design. The fermentation medium and fermentation conditions of HQS6513 were optimized by one-way test and response surface design. Result Three strains QS10-6, QS2-5 and QS2-13 with obvious inhibitory effect on potato coccodes (Colletotrichum coccodes) were screened out from potato tubers and identified as Bacillus atrophaeus, Bacillus atrophaeus and Bacillus velezensis, response surface method analysis was optimized to obtain the optimal fermentation medium for the composite colony HQS6513 as bran 2.5%, yeast extract powder 2.6%, NaCl 1.0%, and the optimal fermentation conditions as incubation temperature of 32 ℃, pH 5, inoculum amount of 2.0%, and shaking table speed of 190 r/min, and the optimized bacterial inhibition rate of the composite colony HQS6513 reached 69.66%. Conclusion Three strains of Bacillus spp. were isolated, and the composite colony HQS6513 composed of these three strains of Bacillus spp. had high inhibitory activity against C. coccodes, and the optimization of fermentation formula and culture conditions for it resulted in the increase of cell production and inhibitory activity of the composite flora HQS6513.

Key words: Colletotrichun coccodes, complex flora, Bacillus atrophicus, Bacilhas velezensis, response surface methodology, fermentation parameters

LUO Ming-kai, ZHANG Hao-jie, SHI Hui-qin, LI Ya-nan, FENG Rui-chao, SHEN Shuo. Isolation and Identification of Biocontrol Bacteria for Potato Anthracnose in Three Strains and Optimization of Fermentation Parameters for Cultures with High Antibacterial Activity[J]. Biotechnology Bulletin, 2025, 41(12): 313-327.

Figures/Tables 16

References 34

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处理 Treatment	QS10-6∶QS2-5∶QS2-13
T1	1∶1∶1
T2	1∶2∶1
T3	1∶3∶3
T4	2∶1∶2
T5	2∶2∶3
T6	2∶3∶1
T7	3∶1∶3
T8	3∶2∶1
T9	3∶3∶2

处理 Treatment	QS10-6∶QS2-5∶QS2-13
T1	1∶1∶1
T2	1∶2∶1
T3	1∶3∶3
T4	2∶1∶2
T5	2∶2∶3
T6	2∶3∶1
T7	3∶1∶3
T8	3∶2∶1
T9	3∶3∶2

菌株 Strain	菌落直径 Colony diameter （mm）	抑菌率 Inhibition rate （%）
QS10-5	29.52±0.27d	50.55±0.46c
QS10-6	34.48±0.14b	42.25±0.24e
QS2-4	31.49±1.54c	47.25±2.58d
QS2-5	25.82±0.16e	56.75±0.27a
QS2-7	26.43±0.45e	55.74±0.77b
QS2-11	29.91±0.37d	49.91±0.06c
QS2-13	23.90±0.33f	59.97±0.55a
QS10-15	30.25±0.12cd	49.35±0.20cd
QS10-16	27.02±0.21e	54.75±0.34b
QS2-15	26.67±0.13e	55.34±0.21b
QS2-17	26.37±0.18e	55.83±0.29b
QS2-19	26.55±0.41e	55.53±0.07b
QS10-18	29.66±0.12d	50.33±0.21c
QS10-19	29.48±0.27d	50.07±0.22c
QS10-20	34.59±0.12b	42.07±0.19e
CK	59.71±0.50a

菌株 Strain	菌落直径 Colony diameter （mm）	抑菌率 Inhibition rate （%）
QS10-5	29.52±0.27d	50.55±0.46c
QS10-6	34.48±0.14b	42.25±0.24e
QS2-4	31.49±1.54c	47.25±2.58d
QS2-5	25.82±0.16e	56.75±0.27a
QS2-7	26.43±0.45e	55.74±0.77b
QS2-11	29.91±0.37d	49.91±0.06c
QS2-13	23.90±0.33f	59.97±0.55a
QS10-15	30.25±0.12cd	49.35±0.20cd
QS10-16	27.02±0.21e	54.75±0.34b
QS2-15	26.67±0.13e	55.34±0.21b
QS2-17	26.37±0.18e	55.83±0.29b
QS2-19	26.55±0.41e	55.53±0.07b
QS10-18	29.66±0.12d	50.33±0.21c
QS10-19	29.48±0.27d	50.07±0.22c
QS10-20	34.59±0.12b	42.07±0.19e
CK	59.71±0.50a

试验 Test	序号 Number	处理 Treatment	菌圈直径 Colony diameter （mm）	抑菌率 Inhibition rate （%）
A：平板试验	1	QS10-6	35.97±0.34b	36.31±0.60e
	2	QS2-5	24.27±0.33e	57.04±0.58b
	3	QS2-13	23.54±0.19e	58.32±0.34ab
	4	HQS65	28.07±0.35c	50.29±0.63b
	5	HQS613	25.41±0.17d	54.99±0.29c
	6	HQS513	26.18±0.32d	53.64±0.57c
	7	HQS6513	23.20±0.23e	58.91±0.42a
	8	CK	56.48±0.71a
B：防治（发酵液）	1	QS10-6	21.41±1.28b	23.11±4.59c
	2	QS2-5	7.99±0.07d	71.32 ±0.26a
	3	QS2-13	7.87±0.04d	71.73±0.14a
	4	HQS65	10.09±0.03c	63.77±0.11b
	5	HQS613	7.87±0.03d	71.73±0.10a
	6	HQS513	8.06±0.03d	71.06±0.12a
	7	HQS6513	7.80±0.03d	71.99±0.09a
	8	百菌清	8.11±0.03d	70.88±0.12a
	9	多菌灵	8.17±0.03d	70.65±0.10a
	10	H₂O	7.91±0.02d
	11	CK	27.85±0.46a
C：防治（菌悬液）	1	QS10-6	21.10±0.26ef	26.61±0.91c
	2	QS2-5	20.77±0.15fj	27.77±0.54c
	3	QS2-13	22.05±0.14cd	23.31±0.49d
	4	HQS65	22.63±0.15c	21.30±0.51e
	5	HQS613	28.45±0.06a	1.04±0.20f
	6	HQS513	21.64±0.10de	24.74±0.36d
	7	HQS6513	20.22±0.29j	29.68±1.01b
	8	百菌清	8.11±0.03h	71.79±0.11a
	9	多菌灵	8.17±0.03h	71.58±0.09b
	10	H₂O	7.91±0.02h
	11	CK	27.85±0.46b
D：预防（发酵液）	1	QS10-6	8.29±0.09b	71.08±0.32f
	2	QS2-5	8.00±0.03b	72.11±0.10c
	3	QS2-13	8.00±0.01 b	72.09±0.04cd
	4	HQS65	8.01±0.01b	72.06±0.04cde
	5	HQS613	8.12±0.02b	71.68±0.06e
	6	HQS513	8.11±0.03b	71.70±0.09de
	7	HQS6513	7.95±0.03b	72.26±0.09bc
	8	百菌清	7.77±0.01b	72.89±0.04a
	9	多菌灵	7.85±0.02b	72.63±0.07ab
	10	H₂O	7.91±0.02b
	11	CK	28.67±0.61a
E：预防（菌悬液）	1	QS10-6	28.20±0.04a	1.65±0.13d
	2	QS2-5	8.31±0.38b	71.01±1.31c
	3	QS2-13	8.31±0.02b	71.00±0.07c
	4	HQS65	28.35±0.08a	1.10±0.26d
	5	HQS613	7.99±0.16bc	72.13±0.56bc
	6	HQS513	8.18±0.13bc	71.48±0.45c
	7	HQS6513	8.18±0.01bc	71.47±0.02c
	8	百菌清	7.64±0.08bc	73.34±0.29ab
	9	多菌灵	7.42±0.17c	74.13±0.58a
	10	H₂O	7.91±0.02bc
	11	CK	28.67±0.61a