一株拮抗致病疫霉的黏细菌菌株的分离、鉴定与发酵条件优化

doi:10.3969/j.issn.1004-1524.20241106

浙江农业学报 ›› 2025, Vol. 37 ›› Issue (12): 2554-2562.DOI: 10.3969/j.issn.1004-1524.20241106

一株拮抗致病疫霉的黏细菌菌株的分离、鉴定与发酵条件优化

张玉(), 丁一秀, 李俊达, 尚少杰, 赵晓静, 候可心, 高艳东, 刘惠荣^*()

内蒙古农业大学生命科学学院,内蒙古呼和浩特 010018

收稿日期:2024-12-20 出版日期:2025-12-25 发布日期:2026-01-09
作者简介:张玉(1998—),男,河南南阳人,硕士研究生,研究方向为黏细菌抗马铃薯晚疫病活性分析。E-mail:1602531312@qq.com
通讯作者: *刘惠荣,E-mail:huirong_liu@imau.edu.cn
基金资助:
国家自然科学基金项目(32260696);国家自然科学基金项目(31370058);内蒙古自治区科技计划项目(2021GG0079)

Isolation and identification of a myxobacterial strain against Phytophthora infestans and optimization of its fermentation conditions

ZHANG Yu(), DING Yixiu, LI Junda, SHANG Shaojie, ZHAO Xiaojing, HOU Kexin, GAO Yandong, LIU Huirong^*()

College of Life Sciences, Inner Mongolia Agricultural University, Hohhot 010018, China

Received:2024-12-20 Online:2025-12-25 Published:2026-01-09

摘要/Abstract

摘要：

黏细菌(myxobacteria)是一类重要的功能微生物,其代谢产物种类丰富多样。致病疫霉(Phytophthora infestans)是引起马铃薯晚疫病的病原菌,对我国马铃薯产业造成严重经济损失。因此,从黏细菌代谢产物中筛选对致病疫霉具有拮抗活性的物质具有重要研究价值。本研究从鄂尔多斯地区分离获得一株对致病疫霉具有显著拮抗活性的黏细菌菌株E12,该菌株的子实体呈黄色珊瑚状突起,经鉴定为珊瑚状珊瑚球菌(Corallococcus coralloides)。在MD1培养基中,于32 ℃、无NaCl条件下发酵11 d,并在发酵过程中添加树脂进行吸附浓缩,所得发酵浓缩液的抑菌圈直径最大,达38 mm。研究表明,大孔树脂可有效吸附菌株发酵液中的活性代谢产物并提高其产量。本研究为开发基于黏细菌的抗马铃薯晚疫病生物农药提供了基础数据。

关键词: 黏细菌, 致病疫霉, 珊瑚状珊瑚球菌, 代谢产物, 发酵

Abstract:

Myxobacteria are an important group of functional microorganisms known for their diverse and abundant metabolites. Phytophthora infestans is the pathogen of potato late blight, which causes great losses to the potato industry in China every year. Therefore, screening for active compounds derived from myxobacterial metabolites that exhibit antagonistic activity against P. infestans holds significant research value. In this study, a myxobacterial strain E12 with remarkable antagonistic activity against P. infestans was isolated from the Ordos region. This strain produced yellow, coral-like fruiting bodies and was identified as Corallococcus coralloides. When cultured in MD1 medium at 32 ℃ without NaCl for 11 days, the addition of resin during fermentation yielded in the largest inhibition zone diameter of 38 mm. The study demonstrated that macroporous resin could effectively adsorb active metabolites from the fermentation solution and enhance their yield. These results provide fundamental data for developing myxobacteria-based biopesticides to control potato late blight.

Key words: myxobacteria, Phytophthora infestans, Corallococcus coralloides, metabolite, fermentation

中图分类号:

S436.32

张玉, 丁一秀, 李俊达, 尚少杰, 赵晓静, 候可心, 高艳东, 刘惠荣. 一株拮抗致病疫霉的黏细菌菌株的分离、鉴定与发酵条件优化[J]. 浙江农业学报, 2025, 37(12): 2554-2562.

ZHANG Yu, DING Yixiu, LI Junda, SHANG Shaojie, ZHAO Xiaojing, HOU Kexin, GAO Yandong, LIU Huirong. Isolation and identification of a myxobacterial strain against Phytophthora infestans and optimization of its fermentation conditions[J]. Acta Agriculturae Zhejiangensis, 2025, 37(12): 2554-2562.

图/表 9

表1 菌株E12发酵条件的正交试验设计因素表

Table 1 Factors for the orthogonal experimental design of strain E12 fermentation conditions

水平 Level	因素Factor
水平 Level	(A)温度 Temperature/℃	(B)时间 Time/d	(C)NaCl质量分数 NaCl mass fraction/%
1	30	9	0
2	32	11	0.5
3	34	13	1.0

图1 菌株E12对致病疫霉的拮抗平板和子实体 A,菌株E12拮抗致病疫霉菌的平板;a,E12菌饼;b,空白VY/2培养基对照;B,菌株E12的子实体形态。

Fig.1 Antagonistic plate of the strain E12 against P. infestans and the fruiting bodies of strain E12 A, Plate assay showing the antagonistic activity of strain E12 against Phytophthora infestans; a, Mycelial plug of E12; b, Blank control (VY/2 medium); B, Morphology of the fruiting bodies of strain E12.

图2 菌株E12的系统发育树

Fig.2 Phylogenetic tree of strain E12

图3 菌株E12发酵上清液在不同培养基上的抑菌圈直径柱上无相同小写字母表示差异显著(p<0.05)。下同。

Fig.3 Inhibition zone diameter of the fermentation supernatant of strain E12 on different culture media Bars without the same lowercase letter indicate significant (p<0.05) differences. The same as below.

图4 菌株E12发酵上清液在不同温度、NaCl质量分数和发酵时间下的抑菌圈直径

Fig.4 Inhibition zone diameter of fermentation supernatants of strain E12 at different temperatures, NaCl mass fraction and fermentation time

表2 正交试验优化结果

Table 2 Results of orthogonal experiment optimization

实验编号 No.	因素 Factor				抑菌圈直径 Inhibition zone diameter/mm
实验编号 No.	A	B	C	空白Blank	抑菌圈直径 Inhibition zone diameter/mm
1	3	2	3	1	0
2	3	3	1	2	20.00±3.06
3	2	1	3	2	0
4	2	3	2	1	21.30±0.67
5	2	2	1	3	26.00±1.15
6	1	3	3	3	0
7	1	1	1	1	12.00±2.00
8	3	1	2	3	16.00±2.31
9	1	2	2	2	18.00±3.06

表3 极差分析表

Table 3 Results of range analysis

因素	k₁	k₂	k₃	R
A	10.00	16.00	12.00	6.00
B	9.30	14.60	14.00	5.30
C	19.30	18.70	0	19.30
空白Blank	11.30	12.70	14.00	2.70

图5 不同发酵条件下发酵上清液对致病疫霉的抑菌圈直径 1,单因素试验优化的发酵条件;2,正交试验优化的发酵条件。

Fig.5 Inhibition zone diameter of fermentation supernatants against Phytophthora infestans under different fermentation conditions 1, Fermentation condition optimized through the single-factor experiment; 2, Fermentation conditions optimized through the orthogonal experiment.

图6 E12菌株发酵上清浓缩液对致病疫霉菌的抑菌圈直径 A、B、C分别为未加树脂的发酵上清浓缩液、树脂过滤后的上清浓缩液和树脂洗脱物浓缩液。

Fig.6 Inhibition zone diameter of the concentrated fermentation supernatant of strain E12 against Phytophthora infestans A, B, and C represent the concentrated fermentation supernatant without resin addition, the concentrated supernatant after resin filtration, and the concentrated resin eluate, respectively.

参考文献 31

[1]	杜欣然, 王晶晶, 冉柒, 等. 黏细菌资源及其系统分类[J]. 微生物学通报, 2023, 50(7): 3104-3121.
	DU X R, WANG J J, RAN Q, et al. Resources and taxonomy of myxobacteria: a review[J]. Microbiology China, 2023, 50(7): 3104-3121. (in Chinese with English abstract)
[2]	WANG C Y, HU J Q, WANG D G, et al. Recent advances in discovery and biosynthesis of natural products from myxobacteria: an overview from 2017 to 2023[J]. Natural Product Reports, 2024, 41(6): 905-934.
[3]	王春玲, 吕颖颖, 姚青, 等. 粘细菌资源挖掘与多相分类研究进展[J]. 微生物学通报, 2021(8): 2870-2880.
	WANG C L, LYUY Y, YAO Q, et al. Research progress in resources mining and polyphase classification of myxobacteria[J]. Microbiology China, 2021(8): 2870-2880. (in Chinese with English abstract)
[4]	WANG T H, LV J L, XU J P, et al. The catalase-peroxidase PiCP1 plays a critical role in abiotic stress resistance, pathogenicity and asexual structure development in Phytophthora infestans[J]. Environmental Microbiology, 2023, 25(2): 532-547.
[5]	BERINDEAN I V, TAOUTAOU A, RIDA S, et al. Modern breeding strategies and tools for durable late blight resistance in potato[J]. Plants, 2024, 13(12): 1711.
[6]	祝菊澧, 梁静思, 王伟伟, 等. 马铃薯致病疫霉研究进展[J]. 微生物学通报, 2020, 47(3): 952-966.
	ZHU J L, LIANG J S, WANG W W, et al. Research progress in Phytophthora infestans, pathogen of potato late blight[J]. Microbiology China, 2020, 47(3): 952-966. (in Chinese with English abstract)
[7]	丁一秀. 鄂尔多斯高原地区粘细菌的分离鉴定及其拮抗致病疫霉活性的初步分析[D]. 呼和浩特: 内蒙古农业大学, 2017.
	DING Y X. Isolation and identification of myxobacteria from Ordos Plateau area and preliminary analysis of their antibiotic activities against Phytophthora infestans[D]. Hohhot: Inner Mongolia Agricultural University, 2017. (in Chinese with English abstract)
[8]	武志华, 郭维维, 董晔, 等. 抗马铃薯晚疫病菌的粘细菌菌株X6-Ⅱ-1的分离鉴定、拮抗活性及发酵条件的优化[J]. 农业生物技术学报, 2018, 26(9): 1467-1479.
	WU Z H, GUO W W, DONG Y, et al. Isolation and identification of myxobacterial strain X6-Ⅱ-1 resistant to Phytophthora infestans and its antibiotic activity and optimal fermentation condition[J]. Journal of Agricultural Biotechnology, 2018, 26(9): 1467-1479. (in Chinese with English abstract)
[9]	余培斌, 杜晶, 陈建新. 高温好氧堆肥过程中芽孢杆菌的筛选、鉴定及应用[J]. 食品与发酵工业, 2020, 46(12): 199-205.
	YU P B, DU J, CHEN J X. Study on screening and identification of Bacillus in the process of high-temperature aerobic composting and its application[J]. Food and Fermentation Industries, 2020, 46(12): 199-205. (in Chinese with English abstract)
[10]	DWORKIN M. Recent advances in the social and developmental biology of the myxobacteria[J]. Microbiological Reviews, 1996, 60(1): 70-102.
[11]	GARRITY G M, BELL J A, LILBURN T G. Bergey’s manual of systematic bacteriology[M]. New York: Springer-Verlag, 2004.
[12]	任兴波, 武志华, 崔海辰, 等. 致病疫霉拮抗菌株YR-7的分离鉴定及其活性物质[J]. 微生物学通报, 2016, 43(7): 1513-1523.
	REN X B, WU Z H, CUI H C, et al. Isolation and identification of the strain YR-7 against Phytophthora infestans and study on its antibiotic substances[J]. Microbiology China, 2016, 43(7): 1513-1523. (in Chinese with English abstract)
[13]	肖春芳. 西南地区马铃薯对晚疫病的抗性评价及抗病相关机制的解析[D]. 武汉: 华中农业大学, 2024.
	XIAO C F. Evaluation of potato resistance to late blight in southwest China and dissection of disease resistance mechanisms[D]. Wuhan: Huazhong Agricultural University, 2024. (in Chinese with English abstract)
[14]	张欣杰, 宋文睿, 陈汉, 等. 马铃薯晚疫病化学防控现状与展望[J]. 中国植保导刊, 2021, 41(6): 33-39.
	ZHANG X J, SONG W R, CHEN H, et al. Status and prospects of chemical prevention and control of potato late blight[J]. China Plant Protection, 2021, 41(6): 33-39. (in Chinese with English abstract)
[15]	WANG Y X, LI M S, YING J H, et al. High-efficiency green management of potato late blight by a self-assembled multicomponent nano-bioprotectant[J]. Nature Communications, 2023, 14: 5622.
[16]	CAULIER S, GILLIS A, COLAU G, et al. Versatile antagonistic activities of soil-borne Bacillus spp. and Pseudomonas spp. against Phytophthora infestans and other potato pathogens[J]. Frontiers in Microbiology, 2018, 9: 143.
[17]	张玉. 粘细菌B25-Ⅰ-3菌株制剂对马铃薯晚疫病的田间防治效果[D]. 呼和浩特: 内蒙古农业大学, 2023.
	ZHANG Y. Field control effect of preparation of myxobacterium strain B25-Ⅰ-3 on potato late blight[D]. Hohhot: Inner Mongolia Agricultural University, 2023. (in Chinese with English abstract)
[18]	武志华, 赵璞钰, 丁一秀, 等. 致病疫霉拮抗菌株B25-Ⅰ-3的鉴定及其次级代谢产物[J]. 微生物学通报, 2020, 47(11): 3586-3599.
	WU Z H, ZHAO P Y, DING Y X, et al. Identification and secondary metabolites of strain B25-Ⅰ-3 against Phytophthora infestans[J]. Microbiology China, 2020, 47(11): 3586-3599. (in Chinese with English abstract)
[19]	BADER C D, PANTER F, MÜLLER R. In depth natural product discovery: myxobacterial strains that provided multiple secondary metabolites[J]. Biotechnology Advances, 2020, 39: 107480.
[20]	封成玲, 刘洋, 贾紫伟, 等. 产酶溶杆菌L-43合成抗菌肽的发酵工艺优化[J]. 食品与发酵工业, 2022, 48(19): 196-203.
	FENG C L, LIU Y, JIA Z W, et al. Optimization of fermentation process for production of antimicrobial peptides with Lysobacter enzymogenes L-43[J]. Food and Fermentation Industries, 2022, 48(19): 196-203. (in Chinese with English abstract)
[21]	宋亚迪, 郭婷, 修志君, 等. 马铃薯疮痂病生防链霉菌菌株PBSH9发酵条件优化[J]. 作物杂志, 2024(5): 255-262.
	SONG Y D, GUO T, XIU Z J, et al. Optimization of fermentation conditions for the biocontrol Streptomyces strain PBSH9 against potato scab disease[J]. Crops, 2024(5): 255-262. (in Chinese with English abstract)
[22]	刘晓霞, 伏漪, 王宗成, 等. 立枯丝核菌拮抗菌枯草芽孢杆菌ATCC 6051发酵条件优化及抑菌物质分析[J]. 核农学报, 2024, 38(7): 1326-1334.
	LIU X X, FU Y, WANG Z C, et al. Optimizing fermentation conditions and inhibitory substance analysis of Bacillus subtilis ATCC 6051 antagonistic to Thanatephorus[J]. Journal of Nuclear Agricultural Sciences, 2024, 38(7): 1326-1334. (in Chinese with English abstract)
[23]	白雪莹, 韩剑, 孙博源, 等. 梨火疫病和梨腐烂病生防潜力粘细菌的筛选鉴定及室内防效评价[J]. 中国生物防治学报, 2023, 39(6): 1384-1397.
	BAI X Y, HAN J, SUN B Y, et al. Screening and identification of biocontrol potential myxobacteria strains against fire blight and pear canker diseases and evaluation of indoor control efficacy[J]. Chinese Journal of Biological Control, 2023, 39(6): 1384-1397. (in Chinese with English abstract)
[24]	尚少杰. 菌株CF-9-D抗肿瘤活性物质的分离鉴定及其代谢组与基因组分析[D]. 呼和浩特: 内蒙古农业大学, 2023.
	SHANG S J. Isolation and identification of antitumor active substances from the strain CF-9-D and analysis of its metabolome and genome[D]. Hohhot: Inner Mongolia Agricultural University, 2023. (in Chinese with English abstract)
[25]	SHARMA A, KUMAR A, BABU V, et al. Myxobacteria from animal dung pellets collected from northwestern Himalayas: a new source of di-isobutyl phthalate[J]. Journal of Basic Microbiology, 2022, 62(2): 162-173.
[26]	MOELLER M, NORRIS M D, PLANKE T, et al. Scalable syntheses of methoxyaspartate and preparation of the antibiotic cystobactamid 861-2 and highly potent derivatives[J]. Organic Letters, 2019, 21(20): 8369-8372.
[27]	WANG Y Y, BEGLEY T P. Mechanistic studies on CysS: a vitamin B₁₂-dependent radical SAM methyltransferase involved in the biosynthesis of the tert-butyl group of cystobactamid[J]. Journal of the American Chemical Society, 2020, 142(22): 9944-9954.
[28]	任兴波, 张子良, 赵璞钰, 等. 马铃薯晚疫病菌拮抗粘细菌YR-35的分离鉴定及其代谢产物稳定性[J]. 中国生物防治学报, 2016, 32(3): 379-387.
	REN X B, ZHANG Z L, ZHAO P Y, et al. Isolation and identification of the strain YR-35 resistant to Phytophthora infestans and its metabolites[J]. Chinese Journal of Biological Control, 2016, 32(3): 379-387. (in Chinese with English abstract)
[29]	HOFFMANN T, KRUG D, BOZKURT N, et al. Correlating chemical diversity with taxonomic distance for discovery of natural products in myxobacteria[J]. Nature Communications, 2018, 9: 803.
[30]	BERLEMAN J E, KIRBY J R. Deciphering the hunting strategy of a bacterial wolfpack[J]. FEMS Microbiology Reviews, 2009, 33(5): 942-957.
[31]	YE X F, LI Z K, LUO X, et al. A predatory myxobacterium controls cucumber fusarium wilt by regulating the soil microbial community[J]. Microbiome, 2020, 8(1): 49.

一株拮抗致病疫霉的黏细菌菌株的分离、鉴定与发酵条件优化

Isolation and identification of a myxobacterial strain against Phytophthora infestans and optimization of its fermentation conditions

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