Extract of Bacillus velezensis SX-45: The Antifungal Mechanism Against Fusarium oxysporum of Ginseng

doi:10.11924/j.issn.1000-6850.casb2021-0906

Abstract

Abstract:

Ginseng root rot is one of the serious diseases in the production of ginseng. The objective of this study is to clarify the antifungal mechanism of the extract of Bacillus velezensis SX-45 against Fusarium oxysporum of ginseng. In this study, Bacillus velezensis SX-45 and Fusarium oxysporum of ginseng were used as materials, the antifungal mechanism data were determined by thiobarbituric acid, coomassie bright blue G-250 methods etc. The results showed that after being treated with the extract of Bacillus velezensis SX-45 at the concentration of 15 μg/mL, the mycelia of Fusarium oxysporum of ginseng had malformation, the hypha surface protrusion and the hypha tip expansion were observed. The electrical conductivity and malondialdehyde content in the fungal suspension increased significantly, which were respectively 4.24 and 42.0 times those of the control group. The OD₂₆₀value of the fungal suspension also increased from 3.48 to 3.83. With the increase of extract concentration, the soluble protein content, reducing sugar content, NAD-MDH activity and SDH activity decreased significantly by 97.7%, 56.6%, 97.5% and 48.4%, respectively. This study indicates that the extract of Bacillus velezensis SX-45 has a good inhibition effect on Fusarium oxysporum, and can lay a foundation for the research and development of new biocontrol agent against ginseng diseases.

Key words: Fusarium oxysporum of ginseng, Bacillus velezensis, extract, antifungal activity, antifungal mechanism

CLC Number:

S432.1

ZHANG Tingting, MA Guilong, XIE Xiaobao, GAO Xinxin, CAI Qi. Extract of Bacillus velezensis SX-45: The Antifungal Mechanism Against Fusarium oxysporum of Ginseng[J]. Chinese Agricultural Science Bulletin, 2022, 38(33): 124-131.

Figures/Tables 12

References 29

[1]	CHOI S Y, CHO C W, LEE Y, et al. Comparison of ginsenoside and phenolic ingredient contents in hydroponically-cultivated ginseng leaves, fruits, and roots[J]. Journal of ginseng research, 2012, 36(4):57-69.
[2]	KWON S W, HAN S B, PARK I H, et al. Liquid chromatographic determination of less polar ginsenosides in processed ginseng[J]. Journal of chromatography a, 2001, 921(2):335-339. pmid: 11471818
[3]	王铁生. 中国人参[M]. 沈阳: 辽宁科学技术出版社, 2001:28-29.
[4]	CHEN J B, CHEN L X, LI M J, et al. Effects of processing method on the pharmacokinetics and tissue distribution of orally administered ginseng[J]. Journal of ginseng research, 2018, 42(1):27-34. doi: 10.1016/j.jgr.2016.12.008 pmid: 29348719
[5]	YAHARA S, KAJI K, TANAKA O. Further study on dammaran-type saponins of roots, leaves, flower-buds, and fruits of Panax ginseng[J]. Chemical & pharmaceutical bulletin, 1979, 27(1):88-92.
[6]	METORI K, FURUTSU M, TAKAHASHI S, et al. The preventive effect of ginseng with du-zhong leaf on protein metabolism in aging[J]. Biological & pharmaceutical bulletin, 1997, 20(20):237-242.
[7]	张达正, 刘强, 张倩. 人参出苗前病害的预防[J]. 人参研究, 2006(1):39-40.
[8]	刘亚南, 赵东岳, 刘敏, 等. 人参病虫害发生及农药施用现状调查[J]. 中国农学通报, 2014(10):294-298.
[9]	傅俊范. 人参病害防治(上)[J]. 新农业, 2003, 11:44-45.
[10]	吴连举, 关一鸣, 逄世峰, 等. 利用生防微生物防治人参、西洋参土传病害研究进展[J]. 安徽农业科学, 2010, 38(28):15630-15631.
[11]	DILANTHA FERNANDO W G, NAKKEERAN S, PORITSANOS N, et al. Multiple mechanisms of biocontrol by Pseudomonas hlororaphis PA23 affect stem rot of canola caused by Sclerotinia sclerotiorum [A]. 第十二届国际油菜大会论文集[C], 2007.
[12]	ZHANG Y, WANG X Y, LING S B, et al. Fermentation optimization, fungistatic effects and tomato growth promotion of four biocontrol bacterial strains[J]. Agriculture, 2021, 11(7):686-686. doi: 10.3390/agriculture11070686 URL
[13]	LI J Y, ZHAO Q Q, WU R Y H H D, et al. Biocontrol bacteria strains Y4 and Y8 alleviate tobacco bacterial wilt disease by altering their rhizosphere soil bacteria community[J]. Rhizosphere, 2021,19 100390.
[14]	XU S J, WANG Y X, HU J Q, et al. Isolation and characterization of Bacillus amyloliquefaciens MQ01,a bifunctional biocontal bacterium with antagonistic activity against Fusarium graminearum and biodegradation capacity of zearalenone[J]. Food control, 2021, 130:108259. doi: 10.1016/j.foodcont.2021.108259 URL
[15]	杜鹏程, 刘海洋. 棉花苗期根腐类病害生防细菌的筛选与鉴定[J]. 新疆农业科学, 2020(4):686-693. doi: 10.6048/j.issn.1001-4330.2020.04.014
[16]	松正祥, 龙欣钰, 孟祥佳, 等. 枯草芽孢杆菌YZU-S149的分离鉴定及对西瓜枯萎病的生防作用[J]. 长江大学学报:自然科学版, 2021, 18(4):114-120.
[17]	肖敏, 肖彤斌, 严婉荣, 等. 生防细菌在植物病害防治中的应用[J]. 基因组学与应用生物学, 2013, 32(4):533-539.
[18]	张玉姣, 叶强, 郑英杰, 人参土传病害生物防治研究进展[J]. 中国林副特产, 2018(3):84-87
[19]	吴长宝, 回云静, 徐小明, 等. 枯草芽孢杆菌生物菌剂对人参病原菌室内抑菌实验研究[J]. 人参研究, 2010, 22(4):11-13.
[20]	张晓云, 丁万隆, 王蓉, 等. 人参病害生防细菌分离鉴定及其抑菌活性测定[J]. 中国现代中药, 2018, 20(11):1387-1391.
[21]	JIANG Y, RAN C, CHEN C Q, et al. Evaluation of isolation and field effect of ginseng disease resistance and growth-promoting bacteria[J]. China journal of Chinese materia medica, 2018, 43(11):2230-2235.
[22]	杨胜清, 张帆, 马贵龙, 等. 贝莱斯芽孢杆菌S6拮抗物质分离纯化及抑菌机理[J]. 农药, 2017, 56(9):645-648,660.
[23]	刘琳, 王强, 袁仁文, 等. 芥菜乙醇提取物对西瓜枯萎病菌的抑菌机理[J]. 植物保护学报, 2019, 46(4):816-823.
[24]	蒋利荣, 覃玥, 诸葛臣林, 等. 羊踟蹰提取物对金黄色葡萄糖球菌的抑菌机制研究[J]. 广州化工, 2020, 48(12):71-72.
[25]	陈志杰, 谢江辉, 陈宇丰, 等. 一株植物病原拮抗细菌的分离筛选及拮抗物质[J]. 生态学杂志, 2018, 37(5):1595-1604.
[26]	许乐, 王子强, 张爽, 等. 丹参根腐病拮抗细菌筛选、鉴定及生防机理研究[J]. 中国生物防治学报, 2021, 4(11):12.
[27]	王璐. 萎缩芽孢杆菌CP 297和解淀粉芽孢杆菌CP 2014对毛桃致腐霉菌的抑菌机理及保鲜应用[D]. 太谷: 山西农业大学, 2019:39-40.
[28]	赵德庆. 芒果采后炭疽病拮抗菌的筛选、作用机制及抑菌代谢产物分析[D]. 海口: 海南大学, 2019:29-31.
[29]	黄建凤, 张发宝, 逄玉万. 两株香蕉枯萎病拮抗细菌的筛选及抑菌机理[J]. 微生物学通报, 2017, 44(4):835-844.

仪器	生产厂家	仪器型号
组合式全温振动培养箱	金坛市国旺实验仪器厂	HSY-C
智能人工气候培养箱	浙江托普仪器有限公司	RTOP系列
紫外分光光度计	北京普析通用仪器有限公司	T6新世纪
多功能酶标仪	美谷分子仪器（上海）有限公司	SpectraMax i3x
蔡司显微镜	ZEISS	Imager.A2
电导率仪	上海雷磁新泾仪器有限公司	DDS-11A
电热恒温水浴锅	上海一恒科学仪器有限公司	HWS-24型

仪器	生产厂家	仪器型号
组合式全温振动培养箱	金坛市国旺实验仪器厂	HSY-C
智能人工气候培养箱	浙江托普仪器有限公司	RTOP系列
紫外分光光度计	北京普析通用仪器有限公司	T6新世纪
多功能酶标仪	美谷分子仪器（上海）有限公司	SpectraMax i3x
蔡司显微镜	ZEISS	Imager.A2
电导率仪	上海雷磁新泾仪器有限公司	DDS-11A
电热恒温水浴锅	上海一恒科学仪器有限公司	HWS-24型

提取物浓度/(μg/mL)	电导率/(μs/cm)
提取物浓度/(μg/mL)	1 h	2 h	3 h	4 h	5 h
0	15.20±0.15d	15.80±0.21d	17.70±1.12d	21.10±0.23d	23.70±0.38d
5	28.10±0.12c	29.20±0.45c	29.60±0.40c	31.90±0.15c	34.50±0.66c
10	47.30±0.45b	49.10±1.77b	52.70±0.59b	54.20±1.61b	56.40±2.22b
15	54.10±0.06a	55.30±0.57a	57.70±1.10a	60.60±0.50a	64.50±2.46a

提取物浓度/(μg/mL)	电导率/(μs/cm)
提取物浓度/(μg/mL)	1 h	2 h	3 h	4 h	5 h
0	15.20±0.15d	15.80±0.21d	17.70±1.12d	21.10±0.23d	23.70±0.38d
5	28.10±0.12c	29.20±0.45c	29.60±0.40c	31.90±0.15c	34.50±0.66c
10	47.30±0.45b	49.10±1.77b	52.70±0.59b	54.20±1.61b	56.40±2.22b
15	54.10±0.06a	55.30±0.57a	57.70±1.10a	60.60±0.50a	64.50±2.46a

处理浓度/(μg/mL)	人参根腐病菌丙二醛含量/(μmol/g)
0	0.015±0.003d
5	0.212±0.002c
10	0.400±0.002b
15	0.630±0.001a