甜瓜镰刀菌果腐病菌拮抗菌筛选及其拮抗性研究

doi:10.16420/j.issn.0513-353x.2022-0741

园艺学报 ›› 2023, Vol. 50 ›› Issue (10): 2257-2270.doi: 10.16420/j.issn.0513-353x.2022-0741

甜瓜镰刀菌果腐病菌拮抗菌筛选及其拮抗性研究

何亚芳¹^,², 包慧芳²^,^*(), 王宁², 詹发强², 张学军³, 史应武², 杨蓉², 侯新强², 龙宣杞²^,^*()

¹ 新疆大学生命科学与技术学院，乌鲁木齐 830046
² 新疆农业科学院微生物应用研究所，新疆特殊环境微生物重点实验室，乌鲁木齐 830091
³ 新疆农业科学院哈密瓜研究中心，乌鲁木齐 830091

收稿日期:2023-04-21 修回日期:2023-09-03 出版日期:2023-10-25 发布日期:2023-10-31
通讯作者: *（E-mail：779556527@qq.com；609160628@qq.com）
基金资助:
新疆自治区科技援疆项目(2020E02094); 农业科技创新稳定支持项目(xjnkywdzc-2022004)

Screening of Antagonistic Bacteria Against Fusarium spp. Causing Melon Fruit Rot and the Antagonistic Properties

HE Yafang¹^,², BAO Huifang²^,^*(), WANG Ning², ZHAN Faqiang², ZHANG Xuejun³, SHI Yingwu², YANG Rong², HOU Xinqiang², LONG Xuanqi²^,^*()

¹ College of Life Sciences and Technology，Xinjiang University，Urumqi 830046，China
² Xinjiang Key Laboratory of Special Environmental Microbilogy，Institute of Microbiology，Xinjiang Academy of Agricultural Sciences，Urumqi 830091，China
³ The Research Center of Hami-melon，Xinjiang Academy of Agricultural Sciences，Urumqi 830091，China

Received:2023-04-21 Revised:2023-09-03 Published:2023-10-25 Online:2023-10-31
Contact: *（E-mail：779556527@qq.com；609160628@qq.com）

摘要/Abstract

摘要：

以新疆喀什地区伽师县盐碱地土壤为材料，通过平板对峙法和琼脂扩散法筛选获得对腐皮镰刀菌（Fusarium solani）BM1具有显著拮抗作用的菌株JS6-1和JS3-4。两菌株发酵上清液对甜瓜果腐病病原菌的抑菌圈直径分别为22.90和21.05 mm，通过形态学观察、生理生化试验和16s rRNA基因序列分析初步鉴定为贝莱斯芽孢杆菌Bacillus velezensis JS6-1和暹罗芽孢杆菌Bacillus siamensis JS3-4。抑菌谱测定结果表明两株菌对尖孢镰刀菌Fusarium oxysporum KW1、香梨黑斑病菌链格孢菌Alternatia alternata HB2、香梨腐烂病菌Valsa mali var. pyri FL1等9种植物病原菌均有较强的抑制作用。酶活及产铁载体测定结果表明，两株菌可产生纤维素酶、蛋白酶、淀粉酶、葡聚糖酶和铁载体等物质。采用酸沉淀法提取两株菌脂肽类物质，并以腐皮镰刀菌Fusarium solani BM1为指示菌进行抑菌试验结果表明，两株菌脂肽类粗提物对病原菌的抑菌圈直径分别为20.55和19.05 mm，对病原菌孢子萌发抑制率分别为100%和93.77%。综上，拮抗菌JS6-1和JS3-4对镰刀菌属病原菌具有较好的拮抗效果，具有进一步开发为生防菌剂的潜力。

关键词: 甜瓜, 镰刀菌, 果腐病, 生防菌, 采后保鲜

Abstract:

In this study，the antagonistic bacteria JS6-1 and JS3-4 against Fusarium solani BM1 were screened from the saline-alkali soil of Jiashi County，Kashgar，Xinjiang by plate confrontation method and the agar diffusion method. The diameters of the inhibition zone of the melon and fruit rot pathogens were 22.90 and 21.05 mm，respectively. JS6-1 and JS3-4 were identified as Bacillus velezensis and Bacillus siamensis by morphological observation，physiological and biochemical tests as well as 16s rRNA gene sequence analysis，respectively. The results of bacteriostatic spectrum showed that the two strains had strong inhibitory effect to nine plant pathogens，such as Fusarium oxysporum KW1，Alternatia alternata HB2，fragrant pear rot pathogen Valsa mali var. pyri FL1 and so on. The results of enzyme activity and iron carrier determination showed that the two strains could produce cellulase，protease，amylase，glucanase and iron carrier. Lipopeptides of two strains were extracted by acid precipitation method，and bacteriostatic experiments were carried out against Fusarium solani BM1. The results showed that the diameters of bacteriostatic zone of the lipopeptides were 20.55 and 19.05 mm，and the inhibition rate against spore germination of pathogenic bacteria were 100% and 93.77%，respectively. In conclusion，JS6-1 and JS3-4 have good antagonistic effect against Fusarium solani and are expected to be used in the biological control of melon fruit rot.

Key words: melon, Fusarium spp., fruit rot, biocontrol bacteria, postharvest preservation

何亚芳, 包慧芳, 王宁, 詹发强, 张学军, 史应武, 杨蓉, 侯新强, 龙宣杞. 甜瓜镰刀菌果腐病菌拮抗菌筛选及其拮抗性研究[J]. 园艺学报, 2023, 50(10): 2257-2270.

HE Yafang, BAO Huifang, WANG Ning, ZHAN Faqiang, ZHANG Xuejun, SHI Yingwu, YANG Rong, HOU Xinqiang, LONG Xuanqi. Screening of Antagonistic Bacteria Against Fusarium spp. Causing Melon Fruit Rot and the Antagonistic Properties[J]. Acta Horticulturae Sinica, 2023, 50(10): 2257-2270.

图/表 13

参考文献 37

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菌株 Strain	抑菌带宽度/mm Inhibition zone width	发酵上清液抑菌圈直径/mm The fermentation supernatant inhibition zone diameter
JS6-1	7.39 ± 0.07 a	22.90 ± 0.74 a
JS3-4	6.67 ± 0.08 b	21.05 ± 0.28 b
JS1-1	5.74 ± 0.17 c	13.29 ± 0.11 c
JS55-2	5.12 ± 0.11 d	9.81 ± 0.47 d
JS15	5.08 ± 0.21 d	9.27 ± 0.30 d

菌株 Strain	抑菌带宽度/mm Inhibition zone width	发酵上清液抑菌圈直径/mm The fermentation supernatant inhibition zone diameter
JS6-1	7.39 ± 0.07 a	22.90 ± 0.74 a
JS3-4	6.67 ± 0.08 b	21.05 ± 0.28 b
JS1-1	5.74 ± 0.17 c	13.29 ± 0.11 c
JS55-2	5.12 ± 0.11 d	9.81 ± 0.47 d
JS15	5.08 ± 0.21 d	9.27 ± 0.30 d

病原菌	抑菌率/% Inhibition rate
Pathogen	JS6-1	JS3-4
香梨腐烂病菌 Valsa mali var. pyri FL1	58.02 ± 1.28 c	59.26 ± 1.44 b
马铃薯黑痣病菌立枯丝核菌 Rhizoctonia solani ZN5	51.85 ± 0.42 de	56.79 ± 1.28 bc
番茄早疫病菌茄链格孢菌 Alternatia solani ZN18	58.02 ± 1.26 c	53.09 ± 1.91 de
核桃腐烂病 Cytcospora sp. ZN19	70.41 ± 2.22 a	75.37 ± 1.18 a
甜瓜枯萎病 Fusarium oxysporum KW1	59.32 ± 0.84 c	54.60 ± 0.74 cd
水稻恶苗串珠镰刀菌 F. moniliforme ZN12	54.26 ± 1.48 d	58.52 ± 1.29 b
马铃薯早疫病菌茄病链格孢菌 A. solani ZN6	62.28 ± 1.18 b	57.72 ± 2.17 bc
西瓜枯萎病尖孢镰刀菌 F. oxysporum ZN10	50.12 ± 0.65 e	50.65 ± 1.01 e
香梨黑斑病菌链格孢菌 A. alternata HB2	58.25 ± 1.26 c	57.98 ± 1.87 bc

病原菌	抑菌率/% Inhibition rate
Pathogen	JS6-1	JS3-4
香梨腐烂病菌 Valsa mali var. pyri FL1	58.02 ± 1.28 c	59.26 ± 1.44 b
马铃薯黑痣病菌立枯丝核菌 Rhizoctonia solani ZN5	51.85 ± 0.42 de	56.79 ± 1.28 bc
番茄早疫病菌茄链格孢菌 Alternatia solani ZN18	58.02 ± 1.26 c	53.09 ± 1.91 de
核桃腐烂病 Cytcospora sp. ZN19	70.41 ± 2.22 a	75.37 ± 1.18 a
甜瓜枯萎病 Fusarium oxysporum KW1	59.32 ± 0.84 c	54.60 ± 0.74 cd
水稻恶苗串珠镰刀菌 F. moniliforme ZN12	54.26 ± 1.48 d	58.52 ± 1.29 b
马铃薯早疫病菌茄病链格孢菌 A. solani ZN6	62.28 ± 1.18 b	57.72 ± 2.17 bc
西瓜枯萎病尖孢镰刀菌 F. oxysporum ZN10	50.12 ± 0.65 e	50.65 ± 1.01 e
香梨黑斑病菌链格孢菌 A. alternata HB2	58.25 ± 1.26 c	57.98 ± 1.87 bc

指标分类 Index classification	测定指标 Identification index	JS6-1	JS3-4
氮源利用 Nitrogen source utilization	硫酸铵Ammonia sulfate	+	+
	草酸铵Sodium oxalate	+	+
	柠檬酸二铵Ammonium citrate	+	+
	L-酪氨酸L-tyrosine	+	+
	磷酸二氢铵Ammonium dihydrogen phosphate	+	+
	谷氨酸Glutamate	+	-
碳源利用 Carbon source utilization	D-葡萄糖D-glucose	+	+
	甘油Glycerinum	+	+
	蔗糖Saccharose	+	+
	D-果糖D-fructose	+	+
	乳糖Lactose	+	-
	淀粉Amylum	+	+
	甘露醇Mannitol	+	+
	鼠李糖Rhamnose	-	-
	壳聚糖Chitosan	-	-
	海藻糖Trehalose	+	+
	麦芽糖Maltose	+	+
其他指标 Other indicators	接触酶反应Contact enzyme test	+	+
	柠檬酸盐利用Citrate utilization	-	-
	甲基红Methy red	-	+
	明胶液化Liquefy glutin	+	+
	淀粉水解Starch hydrolysis	+	+
	V-P试验V-P test	+	+
	吲哚试验Indole test	-	-
	硫化氢Hydrogen sulfide	-	-
生长特性 Growth characteristics	NaCl浓度/% NaCl concentration	2.5 ~ 9.5	2.5 ~ 9.0
	生长温度/℃ Growth temperature	15 ~ 50	4 ~ 50
	pH	5.0 ~ 11.0	5.0 ~ 9.0

甜瓜镰刀菌果腐病菌拮抗菌筛选及其拮抗性研究

Screening of Antagonistic Bacteria Against Fusarium spp. Causing Melon Fruit Rot and the Antagonistic Properties

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菌株编号Strain number	As/Ar	铁载体活性/% Siderophore activity
JS6-1	0.689 ± 0.008	31.10 ± 0.06
JS3-4	0.789 ± 0.015	21.11 ± 0.11

稀释倍数 Dilution multiple	Bacillus velezensis JS6-1		B. siamensis JS3-4
稀释倍数 Dilution multiple	孢子萌发率/% Spore germination rate	萌发抑制率/% Spore germination inhibition rate	孢子萌发率/% Spore germination rate	萌发抑制率/% Spore germination inhibition rate
原液 Stock solution	0	100.00	5.45	93.77
10^-1	8.69	91.64	12.12	86.15
10^-2	25.58	83.28	33.82	61.35
10^-3	34.21	67.79	40.54	53.67
10^-4	58.33	36.91	67.56	22.79
对照Control	87.50	—	87.50	—

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甜瓜镰刀菌果腐病菌拮抗菌筛选及其拮抗性研究

Screening of Antagonistic Bacteria Against Fusarium spp. Causing Melon Fruit Rot and the Antagonistic Properties

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摘要/Abstract

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参考文献 37

相关文章 15

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