蜡样芽孢杆菌挥发物对苹果轮纹病菌的拮抗活性

doi:10.16420/j.issn.0513-353x.2023-0453

园艺学报 ›› 2024, Vol. 51 ›› Issue (6): 1403-1412.doi: 10.16420/j.issn.0513-353x.2023-0453

蜡样芽孢杆菌挥发物对苹果轮纹病菌的拮抗活性

覃艮红¹^,²^,^*, 袁洪波¹^,³^,^*, 王卓妮¹, 史冰柯¹, 范洋洋¹, 王丽¹, 张猛², 涂洪涛¹^,³, 徐超²^,^**(), 侯珲¹^,³^,^**()

¹ 中国农业科学院郑州果树研究所，郑州 450009
² 河南农业大学植物保护学院，郑州 450009
³ 中国农业科学院中原研究中心，河南新乡 453004

收稿日期:2024-01-30 修回日期:2024-04-23 出版日期:2024-12-18 发布日期:2024-06-22
通讯作者:
** E-mail：houhui@caas.cn，
chaoxu01@163.com
作者简介:
* 共同第一作者
基金资助:
中央级公益性科研院所基本科研业务费专项(1610192023311); 中央级公益性科研院所基本科研业务费专项(ZGS202205); 中国农业科学院科技创新工程项目(CAAS-ASTIP-2016-RIP); 河南省重点研发与推广专项(232102110052)

Antagonistic Activity of Volatile Organic Compounds of Bacillus cereus Strain G3-17 Against Apple Ring Rot

QIN Genhong¹^,², YUAN Hongbo¹^,³, WANG Zhuoni¹, SHI Bingke¹, FAN Yangyang¹, WANG Li¹, ZHANG Meng², TU Hongtao¹^,³, XU Chao²^,^**(), HOU Hui¹^,³^,^**()

¹ Zhengzhou Fruit Research Institute，Chinese Academy of Agricultural Sciences，Zhengzhou 450009，China
² College of Plant Protection，Henan Agricultural University，Zhengzhou 450009，China
³ Zhongyuan Research Center，Chinese Academy of Agricultural Sciences，Xinxiang，Henan 453004，China

Received:2024-01-30 Revised:2024-04-23 Published:2024-12-18 Online:2024-06-22

摘要/Abstract

摘要：

为了探索苹果轮纹病生物防治方法，分析了蜡样芽孢杆菌菌株G3-17挥发性有机化合物对苹果轮纹病菌的拮抗活性。平皿对扣试验显示，在前期研究基础上菌株G3-17不同浓度的挥发物均能显著抑制苹果轮纹病菌的生长。顶空固相微萃取—气相色谱—质谱联用分析显示，菌株G3-17主要分泌6种挥发性有机化合物，其中苯甲醛对苹果轮纹病菌的抑制效果最好，在浓度为125 ~ 625 μL · L^-1范围内抑制率为57.24% ~ 100.00%。苯甲醛处理还能破坏苹果轮纹病菌细胞膜的完整性，并能诱导苹果果实病程相关蛋白基因（MdPR1和MdPR5）的表达。防效试验结果显示，苯甲醛处理能有效防治苹果轮纹病，在浓度为125、250和500 μL · L^-1时的防效分别为39.80%、84.77%和90.67%。研究表明，蜡样芽孢杆菌菌株G3-17挥发物及主要成分苯甲醛对苹果轮纹病具有较好的防治潜力。

关键词: 芽孢杆菌, 苹果轮纹病, 挥发性有机化合物, 苯甲醛, 生物防治

Abstract:

The purpose of this study is to analyze the antagonistic activity of volatile organic compounds（VOCs）of Bacillus cereus strain G3-17 against Botryosphaeria dothidea，and to explore the biological control methods of apple ring rot. The plate-to-plate experiment showed that the volatiles of Bacillus cereus strain G3-17 at different concentrations could significantly inhibit the growth of Botryosphaeria dothidea. Headspace solid-phase microextraction-gas chromatography-mass spectrometry analysis showed that the strain G3-17 mainly secreted six VOCs. Among them，benzaldehyde had the best inhibitory effect on Botryosphaeria dothidea，with an inhibition rate of 57.24%-100.00% in the concentration range of 125-625 μL · L^-1. Further results also showed that benzaldehyde treatment also destroyed the integrity of the cell membrane of Botryosphaeria dothidea. In addition，benzaldehyde treatment could induce the expression of apple fruit pathogenesis-related protein genes（MdPR1 and MdPR5）. Benzaldehyde treatment also could effectively control apple ring rot caused by Botryosphaeria dothidea，and the control effects were 39.80%，84.77% and 90.67% at the concentrations of 125，250 and 500 μL · L^-1，respectively. In summary，the volatiles of Bacillus cereus strain G3-17 and its main VOCs have good control potential against apple ring rot.

Key words: Bacillus cereus, apple ring rot, volatile organic compounds, benzaldehyde, biological control

覃艮红, 袁洪波, 王卓妮, 史冰柯, 范洋洋, 王丽, 张猛, 涂洪涛, 徐超, 侯珲. 蜡样芽孢杆菌挥发物对苹果轮纹病菌的拮抗活性[J]. 园艺学报, 2024, 51(6): 1403-1412.

QIN Genhong, YUAN Hongbo, WANG Zhuoni, SHI Bingke, FAN Yangyang, WANG Li, ZHANG Meng, TU Hongtao, XU Chao, HOU Hui. Antagonistic Activity of Volatile Organic Compounds of Bacillus cereus Strain G3-17 Against Apple Ring Rot[J]. Acta Horticulturae Sinica, 2024, 51(6): 1403-1412.

图/表 8

表1 引物序列

Table 1 Primer sequences

基因 Gene	引物名称 Primer name	引物序列（5′-3′） Primer sequences
MdEF1α	MdEF1α-q_R	CAGTCAGCCTGTGATGTTCC
	MdEF1α-q_F	ATTCAAGTATGCCTGGGTGC
MdPR1	MdPR1-q_R	CCAGTGCTCATGGCAAGGTTTT
	MdPR1-q_F	GCAGCAGTAGGCGTTGGTCCCT
MdPR5	MdPR5-q_R	TTGGATGCTAGTTCGAAGC
	MdPR5-q_F	CTCACCTTGGCCATCCTCTT

图1 蜡样芽孢杆菌菌株G3-17挥发物对苹果轮纹病菌菌株Bd220的抑制作用图中数据为平均数 ± 标准差，不同字母表示经邓肯氏新复极差法检验在P < 0.05水平差异显著。

Fig. 1 Suppression of VOCs of Bacillus cereus strain G3-17 on Botryosphaeria dothidea pathogen strain Bd220 Data are mean ± SD，different letters indicate significant difference at P < 0.05 level by Duncan’s multiple range test.

表2 蜡样芽孢杆菌菌株G3-17挥发性有机化合物的GC-MS检测结果

Table 2 GC-MS detection results of volatile organic compounds produced by strain G3-17 of Bacillus cereus

序号 Order number	保留时间/min Retention time	挥发性有机化合物 VOCs	相对峰面积 Relative hazard peak area	相对含量/% Relative content	CAS编号 CAS No.
1	19.551	苯甲醛 Benzaldehyde	1 603 584	9.64	100-52-7
2	23.206	十六烷 Hexadecane	90 232	0.54	544-76-3
3	35.584	正二十一烷 Heneicosane	116 292	0.70	629-94-7
4	41.109	4-喹啉甲醛 4-Quinolinecarboxaldehyde	403 829	2.43	4363-93-3
5	41.949	二苯甲酮 Benzophenone	77 652	0.47	119-61-9
6	43.171	邻苯二甲酸二异丁酯 Diisobutyl phthalate	95 757	0.58	84-69-5

图2 不同浓度的6种挥发性有机化合物对苹果轮纹病菌菌丝生长的抑制作用

Fig. 2 Inhibitory effects of six volatile organic compounds with different concentration on mycelial growth of Botryosphaeria dothidea

图3 苯甲醛熏蒸对苹果轮纹病的抑制作用

Fig. 3 Suppression of benzaldehyde on apple ring rot

表3 苯甲醛熏蒸对苹果轮纹病的防治效果

Table 3 Control effect of benzaldehyde on apple ring rot

苯甲醛/（μL · L^-1） Benzaldehyde	病斑直径/cm Lesion diameter	防治效果/% Control effect
0（对照Control）	2.78 ± 0.26 a
125	1.65 ± 0.17 b	39.80 ± 0.09 b
250	0.43 ± 0.06 c	84.77 ± 0.07 a
500	0.26 ± 0.11 c	90.67 ± 0.08 a

图4 苯甲醛半最大效应浓度（EC50）和95%有效浓度（EC95）熏蒸处理对苹果轮纹病菌细胞内容物泄漏量的影响同一时间点不同字母表示经邓肯氏新复极差法检验在P < 0.05水平差异显著。

Fig. 4 The effects of fumigation treatment with benzaldehyde half maximum effect concentration（EC50）and 95% effective concentration （EC95）on the leakage of cell contents of Botryosphaeria dothidea. Different letters at the same time point indicated significant difference at the P < 0.05 level by Duncan’s new multiple range test.

图5 苯甲醛熏蒸处理对苹果果皮MdPR1和MdPR5表达的影响同一时间点不同字母表示经邓肯氏新复极差法检验在P < 0.05水平差异显著。

Fig. 5 Effects of benzaldehyde fumigation on the expression of MdPR1 and MdPR5 in apple peel Different letters at the same time point indicated significant difference at the P < 0.05 level by Duncan’s new multiple range test.

参考文献 24

[1]	Alla A, El-Sayed, Ziedan H, El-Mohamedy S. 2008. Control of rhizopus rot disease of apricot fruits(Prunus armeniaca L.) by some plant volatiles aldehydes. Research Journal of Agriculture and Biological Sciences, 4 (5):424-433.
[2]	Chavarria-Carvajal J A, Rodriguez-Kabana R, Kloepper J W, Morgan-Jones G. 2001. Changes in populations of microorganisms associated with organic amendments and benzaldehyde to control plant-parasitic nematodes. Nematropica, 31:165-180.
[3]	Cui Jie,Wang Cui-cui,Liu Yong-guang,Zhao Hong,Zhang De-zhen,Chi Wen-juan,Yang Yuan-yuan. 2022. Identification of bacterial strain SG-11 against Botrytis cinerea and its bacteriostasis. Shandong Agricultural Sciences, 54 (10):104-109. (in Chinese)
	崔杰, 王翠翠, 刘永光, 赵红, 张德珍, 迟文娟, 杨园园. 2022. 番茄果实灰霉病拮抗细菌菌株SG-11的鉴定及其抑菌作用. 山东农业科学, 54 (10):104-109.
[4]	Fernando W G D, Ramarathnam R, Krishnamoorthy A S, Savchuk S C. 2005. Identification and use of potential bacterial organic antifungal volatiles in biocontrol. Soil Biology & Biochemistry, 37 (5):955-964.
[5]	Feng Rong, Liu Li, Chen Hai-nian, Yang Sheng-zhu, Lei Qin-qing, Ding Zhi-feng. 2021. Study on antifungal activity of Bacillus amyloliquefaciens F11. Journal of Agricultural Resources and Environment, 38 (5):849-857. (in Chinese)
	冯蓉, 刘丽, 陈海念, 杨胜竹, 雷钦卿, 丁志峰. 2021. 解淀粉芽孢杆菌F11抗真菌活性研究. 农业资源与环境学报, 38 (5):849-857.
[6]	He Xiao-wen, He Ping, Chang Yuan-sheng, Wang Sen, Wang Hai-bo, Li Lin-guang. 2020. Advances in the research on resistance mechanism of apple to ring rot disease. Acta Horticulturae Sinica, 47 (8):1585-1594. (in Chinese) doi: 10.16420/j.issn.0513-353x.2019-0646
	何晓文, 何平, 常源升, 王森, 王海波, 李林光. 2020. 苹果抗轮纹病机制研究进展. 园艺学报, 47 (8):1585-1594. doi: 10.16420/j.issn.0513-353x.2019-0646
[7]	Jiang Mu-lan, Zhao Rui, Wang Guo-ping. 2006. Identification and purification of effective components of endophytic Bacillus subtilis BY-2 against Sclerotinia sclerotiorum in rape. Chinese Journal of Oil Crop Sciences,(4):453-456. (in Chinese)
[8]	江木兰, 赵瑞, 王国平. 2006. 油菜内生枯草芽孢杆菌BY-2抗油菜菌核病菌有效成分的鉴定和分离提纯. 中国油料作物学报,(4):453-456.
[9]	Ling L, Luo H, Zhao Y, Yang C, Cheng W, Pang M. 2022. Fungal pathogens causing postharvest fruit rot of wolfberry and inhibitory effect of 2,3-butanedione. Frontiers in Microbiology, 13:1068144.
[10]	Li Teng-jie, Liang Sun-yan, Guo Jian-heng, Xu Yue, Zhang Chun-mei, Qin Sheng, Xing Ke. 2022. Inhibitory effects of volatile 2-methylbutyric acid produced by Bacillus tequilensis XK 29 on Ceratocystis fimbriata. Acta Microbiologica Sinica, 62 (12):5018-5028. (in Chinese)
	李腾杰, 梁孙妍, 郭健衡, 徐悦, 张春媚, 秦盛, 邢珂. 2022. 特基拉芽孢杆菌XK29挥发物2-甲基丁酸对甘薯长喙壳菌的抑制作用研究. 微生物学报, 62 (12):5018-5028.
[11]	Liang Yan-qiong, Li Rui, Wu Wei-huai, Tian Ping-fang. 2022. Inhibitory mechanism of volatilized active components from Bacillus subtilis Czk 1 on Ganoderma pseudoferreum. Journal of Agricultural Science and Technology, 24 (2):152-159. (in Chinese)
	梁艳琼, 李锐, 吴伟怀, 田平芳. 2022. 枯草芽孢杆菌Czk1挥发物混合活性组分对橡胶灵芝菌的抑菌机理. 中国农业科技导报, 24 (2):152-159. doi: 10.13304/j.nykjdb.2020.0991
[12]	Liu Jin-rong, Ge Xi-zhen, Tian Guo-ying, Tian Ping-fang. 2019. Research advances in molecular mechanisms of apple response to Botryosphaeria dothidea. China Fruits, 198 (4):11-15. (in Chinese)
	刘金蓉, 葛喜珍, 田国英, 田平芳. 2019. 苹果响应轮纹病菌侵染的分子机制研究进展. 中国果树, 198 (4):11-15.
[13]	Liu Wei-wei. 2009. Determination of antagonistic activity of volatiles from Bacillus spp. against sevsral soilborne pathogens and components analysis[M. D. Dissertation]. Tai’an: Shandong Agricultural University. (in Chinese)
	刘玮玮. 2009. 芽孢杆菌所产挥发物对几种土传病原物的拮抗活性测定与成分分析[硕士论文]. 泰安: 山东农业大学.
[14]	Marzouk T, Chaouachi M, Sharma A, Jallouli S, Mhamdi R, Kaushik N, Djébali N. 2021. Biocontrol of Rhizoctonia solani using volatile organic compounds of Solanaceae seed-borne endophyticbacteria. Postharvest Biology and Technology, 181:111655.
[15]	Muthukumar A, Eswaran A, Sanjeevkumas K. 2011. Exploitation of Trichoderma species on the growth of Pythium aphanidermatumin chilli. Brazilian Journal of Microbiology, 42 (4):1598-1607. doi: 10.1590/S1517-838220110004000047 pmid: 24031794
[16]	Qiao Cai-hong, Zhang Zhong. 2009. Antibacterial activity of essential oil from Angelica sinensis leaf and the mechanism against Escherichia coli. Food and Fermentation Science & Technology, 58 (5):14-18. (in Chinese)
	乔彩红, 张忠. 2022. 当归叶精油的抑菌活性及对大肠杆菌的抑制机理研究. 食品与发酵科技, 58 (5):14-18.
[17]	Wang Jing, Cao Jian-min, Chen De-xin, Qiu Jun, Wang Xiao-qiang, Feng Chao, Wang Wen-jing. 2018. Antimicrobial effect and components analysis of volatile organic compounds from Bacillus pumilus AR03. Scientia Agricultura Sinica, 51 (10):1908-1919. (in Chinese) doi: 10.3864/j.issn.0578-1752.2018.10.010
	王静, 曹建敏, 陈德鑫, 邱军, 王晓强, 冯超, 王文静. 2018. 短小芽孢杆菌AR03挥发性有机物的抑菌活性及其组分分析. 中国农业科学, 51 (10):1908-1919. doi: 10.3864/j.issn.0578-1752.2018.10.010
[18]	Wang Zhuo-ni, Qin Gen-hong, Wang Li, Hou Hui, Yuan Hong-bo, Zhou Hou-cheng, Gong Wen-feng, Tu Hong-tao. 2023. Screening of antagonistic bacteria against strawberry disease and its control effect on strawberry brown leaf spot. China Vegetables,(2):63-71. (in Chinese)
	王卓妮, 覃艮红, 王丽, 侯珲, 袁洪波, 周厚成, 巩文峰, 涂洪涛. 2023. 草莓病害拮抗细菌的筛选及其对草莓褐色叶斑病的防效. 中国蔬菜,(2):63-71.
[19]	Yang T, Xin Y, Liu T, Li Z, Liu X, Wu Y, Wang M, Xiang M. 2022. Bacterial volatile-mediated suppression of root-knot nematode(Meloidogyne incognita). Plant Disease, 106 (5):1358-1365.
[20]	Yuan H B, Shi B K, Wang L, Huang T X, Zhou H C, Hou H, Tu H T. 2021. Isolation and characterization of Bacillus velezensis strain P2- 1 for biocontrol of apple postharvest decay caused by Botryosphaeria dothidea. Frontiers in Microbiology, 12:808938.
[21]	Yang Dong-ya, Qi Rui-xue, Li Zhao-xuan, Lin Wei, Ma Hui, Zhang Xue-yan. 2023. Screening,identification and growth-promoting effect of antagonistic Bacillus spp. against cucumber Fusarium solani. Biotechnology Bulletin, 39 (2):211-220. (in Chinese) doi: 10.13560/j.cnki.biotech.bull.1985.2022-0522
	杨东亚, 祁瑞雪, 李昭轩, 林薇, 马慧, 张雪艳. 2023. 黄瓜茄病镰刀菌拮抗芽孢杆菌的筛选、鉴定及促生效果. 生物技术通报, 39 (2):211-220. doi: 10.13560/j.cnki.biotech.bull.1985.2022-0522
[22]	Zou Man, Chen Yu-shi, Xu Chun-yan, Yang Xiang-zheng, Bao Wen-ting, Wang Chuan-zeng, Zhu Pei-pei, Ma Ya-nan, Zhang Yong, Xin Li,Chen Yi-lun,Zhang Qian. 2023. Research progress of biocontrol Bacillus on postharvest diseases of fruits and vegetables. Technology of Food Industry, 44 (4):488-495. (in Chinese)
	邹曼, 陈雨诗, 许春艳, 杨相政, 包汶婷, 王传增, 朱配配, 马亚男, 张勇, 辛力, 陈义伦, 张倩. 2023. 生防芽孢杆菌对果蔬采后病害防治的研究进展. 食品工业科技, 44 (4):488-495.
[23]	Zhang D, Qiang R, Zhao J, Zhang J, Cheng J, Zhao D, Fan Y, Yang Z, Zhu J. 2022. Mechanism of a volatile organic compound (6-methyl-2-heptanone)emitted from Bacillus subtilis ZD01 against Alternaria solani in potato. Frontiers in Microbiology, 12:808337.
[24]	Zou X R, Wei Y Y, Jiang S, Cao Z, Xu F, Wang H F, Zhan P P, Shao X F. 2021. Volatile organic compounds and rapid proliferation of Candida pseudolambica W 16 are modes of action against gray mold in peach fruit. Postharvest Biology and Technology, 12:111751.

蜡样芽孢杆菌挥发物对苹果轮纹病菌的拮抗活性

Antagonistic Activity of Volatile Organic Compounds of Bacillus cereus Strain G3-17 Against Apple Ring Rot

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

图/表 8

参考文献 24

相关文章 15

编辑推荐

Metrics

本文评价

访问总数:　当日访问总数:　当前在线人数:

[1]	张文静, 徐大勇, 吴倩琳, 杨佛, 信丙越, 曾昕, 李峰. 拮抗番茄灰霉病的贝莱斯芽孢杆菌XDY66基因组分析[J]. 园艺学报, 2024, 51(6): 1413-1425.
[2]	史冰柯, 王卓妮, 覃艮红, 黄天祥, 王丽, 涂洪涛, 袁洪波, 侯珲. 苹果轮纹病拮抗真菌Pa2的分离与鉴定[J]. 园艺学报, 2024, 51(5): 1113-1125.
[3]	韩琴, 高惠惠, 马香, 苏建宇, 徐春燕. 枸杞根际根腐病拮抗细菌的筛选及其防效检测[J]. 园艺学报, 2024, 51(5): 1162-1172.
[4]	强然, 张岱, 杨喆, 陈明玥, 赵景, 朱杰华. 解淀粉芽胞杆菌L19对马铃薯枯萎病菌的抑制及植株的促生作用[J]. 园艺学报, 2024, 51(4): 875-892.
[5]	贾淑鑫, 李金, 闫思远, 郭苗苗, 王若彤, 顾沛雯. 嗜线虫镰刀菌NQ8GⅡ4对枸杞根腐病的防效及其机制研究[J]. 园艺学报, 2024, 51(3): 631-642.
[6]	关思慧, 刘晨旭, 姚祝平, 万红建, 刁明, 程远. 腐植酸处理对樱桃番茄挥发性有机化合物成分和含量的影响[J]. 园艺学报, 2024, 51(2): 346-360.
[7]	吴琼, 张甜甜, 李茂营, 吴慧玲, 郭绍贵, 张洁, 任毅, 张海英, 宫国义. 解淀粉芽孢杆菌5号防控西瓜CGMMV机制初探[J]. 园艺学报, 2024, 51(10): 2427-2438.
[8]	李素平, 李微, 韩冷, 黄建国. 贝莱斯芽孢杆菌HY19挥发物对采后柑橘的防腐效果[J]. 园艺学报, 2024, 51(1): 162-174.
[9]	刘仕达, 邱泽澜, 李晓刚, 钟杰, 朱俊子. 辣椒白绢病拮抗放线菌SD-64的鉴定及作用机制研究[J]. 园艺学报, 2024, 51(1): 175-189.
[10]	邓成凤, 李素平, 张瑞轩, 韩冷, 李勇. 产酶溶杆菌LE16对采后柑橘青霉绿霉病的防治作用[J]. 园艺学报, 2023, 50(7): 1574-1586.
[11]	贺旭, 韩剑, 盛强, 罗明, 包慧芳, 黄伟. 梨火疫病拮抗细菌FX1及其抑菌物质的防病作用[J]. 园艺学报, 2023, 50(5): 1118-1129.
[12]	潘纪源, 董庆龙, 温海彬, 刘亚南, 王晓洁, 张雪梅, 刘文菊, 陆秀君. 巨大芽孢杆菌菌剂对苹果产量、品质及土壤微生物的影响[J]. 园艺学报, 2023, 50(11): 2453-2465.
[13]	高云静, 王天好, 胡旺成, 洪霓, 王国平, 王利平. 梨轮纹病菌弱毒株携带产黄青霉病毒水平传染力测定[J]. 园艺学报, 2023, 50(11): 2477-2488.
[14]	史洪丽, 李腊, 郭翠梅, 余婷婷, 简伟, 杨星勇. 番茄灰霉病生防菌株TL1的分离、鉴定及其生防能力分析[J]. 园艺学报, 2023, 50(1): 79-90.
[15]	杜玉玲, 杨凡, 赵娟, 刘书琪, 龙超安. 新鱼腥草素钠对柑橘指状青霉的抑菌作用[J]. 园艺学报, 2023, 50(1): 145-152.

蜡样芽孢杆菌挥发物对苹果轮纹病菌的拮抗活性

Antagonistic Activity of Volatile Organic Compounds of Bacillus cereus Strain G3-17 Against Apple Ring Rot

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

图/表 8

参考文献 24

相关文章 15

编辑推荐

Metrics

本文评价

访问总数: 当日访问总数: 当前在线人数:

访问总数:　当日访问总数:　当前在线人数: