Screening and identification of Bacillus altitudinis strain CY1 and its control effects against tobacco black shank

doi:10.3969/j.issn.1004-1524.20240188

Acta Agriculturae Zhejiangensis ›› 2025, Vol. 37 ›› Issue (2): 405-416.DOI: 10.3969/j.issn.1004-1524.20240188

• Plant Protection • Previous Articles Next Articles

Screening and identification of Bacillus altitudinis strain CY1 and its control effects against tobacco black shank

GAO Qiang¹(), WANG Lili¹, ZHANG Jianlong¹, YANG Bo¹, LI Feng¹, ZHU Xianzhi¹, LIU Aixin², HAN Chao²^,^*(), TIAN Lei¹^,^*()

1. Linyi Tobacco Company, Linyi 276003, Shandong, China
2. College of Plant Protection, Shandong Agricultural University, Tai’an 271018, Shandong, China

Received:2024-03-01 Online:2025-02-25 Published:2025-03-20
Contact: HAN Chao,TIAN Lei

Abstract

Abstract:

To obtain efficient antagonist strains against tobacco black shank, the rhizosphere soil samples of healthy tobacco plants were collected from tobacco field with tobacco black shank, and the bacterial strain against Phytophthora parasitica was screened out by confrontation culture method. This strain was identified based on morphological, physiological and molecular biology characteristics. The fermentation condition was optimized by single factor experiment, and the available application concentration and antimicrobial mechanism of fermentation broth were explored. Additionally, the control effect of CY1 on tobacco black shank was determined by both indoor pot and field trials. The results showed that a total of 135 soil microbial strains were isolated from the collected rhizosphere soil samples of tobacco, of which a bacterial strain CY1 displayed excellent antagonistic effect against Phytophthora parasitica with an inhibitory rate of 68.6%, and it had a broad spectrum of antifungal function. Based on morphological, physiological and 16S rRNA and gyrB gene sequences analysis, the strain CY1 was identified as Bacillus altitudinis. Its optimal fermentation conditions were as follows: YSP cultural medium with an initial pH value of 7, rotation speed of 220 r·min^-1, at 30 ℃ for 48 h. The fermentation broth with a concentration range from 1×10⁴ CFU·mL^-1 to 1×10⁸ CFU·mL^-1 inhibited mycelial growth, caused mycelial terminal deformity and swell, and induced granular contents in the mycelial tube of Phytophthora. Parasitica. The indoor pot and field control efficacies of the fermentation broth of CY1 with 1×10⁶ CFU·mL^-1 concentration were 71.7% and 74.7%, respectively, which was comparable to those of mancozed, but significantly (P<0.05) higher than those of propamocarb and lower than those of oxathiapiprolin. To sum up, the Bacillus altitudinis strain CY1 has promising biocontrol potential and application prospect, which provided biological control resources of tobacco black shank.

Key words: Bacillus altitudinis, tobacco black shank, screening and identification, fermentation condition, control effect

CLC Number:

S435.72

GAO Qiang, WANG Lili, ZHANG Jianlong, YANG Bo, LI Feng, ZHU Xianzhi, LIU Aixin, HAN Chao, TIAN Lei. Screening and identification of Bacillus altitudinis strain CY1 and its control effects against tobacco black shank[J]. Acta Agriculturae Zhejiangensis, 2025, 37(2): 405-416.

Figures/Tables 8

References 41

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病原菌 Phytopathogen	抑菌率 Inhibition rate/%
烟草疫霉菌Phytophthora parasitica	68.6±1.3a
尖孢镰刀菌Fusarium oxysporum	65.1±2.1ab
茄病镰刀菌Fusarium solani	64.8±1.6ab
立枯丝核菌Rhizoctonia solani	58.9±1.7b
链格孢菌Alternaria alternate	56.3±2.3c
群结腐霉Pythium myriotylum	54.1±1.4c
烟草炭疽菌Collectotrichum nicotianae	52.7±0.9d

病原菌 Phytopathogen	抑菌率 Inhibition rate/%
烟草疫霉菌Phytophthora parasitica	68.6±1.3a
尖孢镰刀菌Fusarium oxysporum	65.1±2.1ab
茄病镰刀菌Fusarium solani	64.8±1.6ab
立枯丝核菌Rhizoctonia solani	58.9±1.7b
链格孢菌Alternaria alternate	56.3±2.3c
群结腐霉Pythium myriotylum	54.1±1.4c
烟草炭疽菌Collectotrichum nicotianae	52.7±0.9d

指标 Items	CY1菌株 CY1 strain	Bacillus altitudinis	指标 Items	CY1菌株 CY1 strain	Bacillus altitudinis
葡萄糖Glucose	+	+	革兰氏染色Gram stain	+	+
葡聚糖Dextrin	-	-	V-P试验V-P test	+	+
半乳糖Galactose	-	-	ONPG试验ONPG test	+	+
蔗糖Saccharose	+	+	接触酶反应Contactase reaction	+	+
甘露醇Mannitol	+	+	甲基红反应Methyl red reaction	+	+
丙三醇Glycerine	+	+	明胶液化Gelatin liquefaction	+	+

指标 Items	CY1菌株 CY1 strain	Bacillus altitudinis	指标 Items	CY1菌株 CY1 strain	Bacillus altitudinis
葡萄糖Glucose	+	+	革兰氏染色Gram stain	+	+
葡聚糖Dextrin	-	-	V-P试验V-P test	+	+
半乳糖Galactose	-	-	ONPG试验ONPG test	+	+
蔗糖Saccharose	+	+	接触酶反应Contactase reaction	+	+
甘露醇Mannitol	+	+	甲基红反应Methyl red reaction	+	+
丙三醇Glycerine	+	+	明胶液化Gelatin liquefaction	+	+

处理 Treatment	盆栽试验结果Pot test result			大田试验Field test result
处理 Treatment	发病率 Incidence of disease/%	病情指数 Disease index	防效 Control efficacy/%	发病率 Incidence of disease/%	病情指数 Disease index	防效 Control efficacy/%
CY1发酵液	32.0±6.0 bc	23.8±3.8 bc	71.7±5.2 b	26.0±2.0 d	18.4±1.8 d	74.7±3.9 b
Fermentation broth of CY1
10%氟噻唑吡乙酮OD	28.0±5.0 c	16.2±2.7 d	80.7±4.5 a	24.0±4.0 d	13.8±2.1 d	81.0±4.2 a
oxathiapiprolin 10% OD
80%代森锰锌WP	37.0±6.0 bc	26.8±2.1 bc	68.2±4.2 b	31.0±4.0 c	20.8±2.3 c	71.4±4.3 b
mancozeb 80% WP
72.2%霜霉威WG	43.0±4.0 b	31.8±3.9 b	61.6±3.4 c	38.0±4.0 b	27.2±3.1 b	62.6±2.8 c
propamocarb 72.2% WG
对照Control	93.0±2.0 a	81.4±3.2 a	—	81.0±5.0 a	72.8±4.6 a	—

Screening and identification of Bacillus altitudinis strain CY1 and its control effects against tobacco black shank

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