产香真菌GS-1挥发性物质特征分析及抑菌作用观察

doi:10.16409/j.cnki.2095-039x.2021.02.018

摘要/Abstract

摘要： 产香真菌GS—1为分离自构树枯枝的一株丛赤壳Nectria cinnabarina，该菌株产生浓郁的香味，其挥发性物质对多种重要植物病原菌均有明显的熏蒸抑制作用。为明确菌株GS—1挥发性物质对植物病原菌的熏蒸抑菌机制、挥发性物质化学组分及菌株生长过程中的挥发性物质释放量的动态规律，利用扫描和透射电镜观察GS—1菌株挥发性物质对灰葡萄孢菌丝和细胞结构的影响，通过HS—SPME法萃取产香真菌GS—1挥发性物质，并通过GC—MS进行挥发性物质释放时间规律的测定和组分鉴定。结果表明，菌株GS—1挥发性物质处理后，灰葡萄孢菌丝出现皱缩、畸形、顶端尖细等现象，同时细胞质含量减少且空腔增多。菌株GS—1挥发性物质释放量随培养时间的延长总体呈先升高后略降低逐渐趋于平稳的趋势，在第12 d达到最大峰值。从菌株GS—1挥发性物质中分离到34个化学组分，鉴定了其中18个组分，主要组分为[3S—（3α，3aβ，5α）]—1，2，3，3a，4，5，6，7—八氢—α，α，3，8—四甲基—5—薁甲醇、表荜澄茄油烯醇和α—广藿香烯，分别占挥发性物质总量34.22%、10.70%和5.60%。这些结果为菌株GS—1挥发性物质在植物病害防控中的合理开发和高效利用提供了理论依据。

关键词: 挥发性物质, 产香真菌, 抑菌机制, 气相色谱-质谱联用

Abstract: The aroma-producing fungus GS−1 strain was isolated from the dead branches of Broussonetia papyrifera, and identified as Nectria cinnabarina. The strain produced rich aroma and its volatile organic compounds (VOCs) had obvious fumigation antifungal activity on many important plant pathogens. In order to determine the fumigation antifungal mechanism, chemical components and dynamic release regularity of the VOCs from GS−1 strain, the fumigation antifungal activity on mycelium and cell structure of Botrytis cinerea were observed by scanning and transmission electron microscopy. The VOCs of GS−1 strain were extracted by HS−SPME. The analysis of dynamic release regularity and identification of the VOCs were carry out by GC−MS. The results showed that the hyphae of B. cinerea was shrunk, deformed and top pointed after treated with VOCs of GS−1 strain. Meanwhile, the cytoplasmic content decreased and the cavities increased. With the passage of cultivation time, the VOCs of GS−1 strain increased firstly, then decreased slightly, tended to be stable at last. It reached the maximum value on the 12^th day. Thirty-four chemical components were isolated from the VOCs of GS−1 strain, and 18 of them were identified. The main components were 5−Azulenemethanol,1,2,3,3a,4,5,6,7− octahydro−.alpha.,.alpha.,3,8−tetramethyl−,[3S−(3.alpha.,3a.beta.,5.alpha.)]−, epicubenol and α−patchoulene, respectively accounting for 34.22%, 10.70% and 5.60% in the total VOCs. These results provided a theoretical basis for the rational development and efficient utilization of the VOCs from GS−1 strain in plant disease control.

Key words: VOCs, aroma-producing fungus, antifungal mechanism, GC-MS

中图分类号:

S476

王春生, 智亚楠, 陈利军, 刘铭, 侯梦圆. 产香真菌GS-1挥发性物质特征分析及抑菌作用观察[J]. 中国生物防治学报, 2021, 37(3): 547-554.

WANG Chunsheng, ZHI Yanan, CHEN Lijun, LIU Ming, HOU Mengyuan. Characteristic Analysis and Fumigation Fungistasis Observation of Volatile Organic Compounds from Aroma-producing Fungus GS-1 Strain[J]. Chinese Journal of Biological Control, 2021, 37(3): 547-554.

参考文献

[1] Korpi A, Järnberg J, Pasanen A. Microbial volatile organic compounds[J]. Critical Reviews in Toxicology, 2009, 39(2):139-193.
[2] Morath S U, Hung R, Bennett J W. Fungal volatile organic compounds:A review with emphasis on their biotechnological potential[J]. Fungal Biology Reviews, 2012, 26(2-3):73-83.
[3] Toffano L, Fialho M B, Pascholati S F. Potential of fumigation of orange fruits with volatile organic compounds produced by Saccharomyces cerevisiae to control citrus black spot disease at postharvest[J]. Biological Control, 2017, 108:77-82.
[4] Kai M, Haustein M, Molina F, et al. Bacterial volatiles and their action potential[J]. Applied Microbiology and Biotechnology, 2009, 81(6):1001-1012.
[5] Zhang X Y, Gao Z F, Zhang X X, et al. Control effects of Bacillus siamensis G-3 volatile compounds on raspberry postharvest diseases caused by Botrytis cinerea and Rhizopus stolonifer[J]. Biological Control, 2020, 141:e104135.
[6] Kanchiswamy C N, Malnoy M, Maffei M E. Chemical diversity of microbial volatiles and their potential for plant growth and productivity[J]. Frontiers in Plant Science, 2015, 6:151.
[7] Hung R, Lee S, Bennett J W. Fungal volatile organic compounds and their role in ecosystems[J]. Applied Microbiology and Biotechnology, 2015, 99(8):3395-3405.
[8] 张清华, 黄丽丽, 连鑫坤, 等. 微生物源挥发性物质及其生物防治作用研究进展[J]. 生态学杂志, 2017, 36(7):2036-2044.
[9] 李贞景, 张春慧, 路来风, 等. 白黄链霉菌TD-1菌株挥发性有机物对灰霉孢菌的抑制作用[J]. 食品科学技术学报, 2016, 34(3):46-52.
[10] 陈利军, 郭世保, 田雪亮, 等. 产香真菌GS-1菌株鉴定及其挥发性物质对番茄灰霉病的生防效果[J]. 植物保护学报, 2016, 43(4):608-613.
[11] Schalchli H, Tortella G R, Rubilar O, et al. Fungal volatiles:an environmentally friendly tool to control pathogenic microorganisms in plants[J]. Critical Reviews in Biotechnology, 2013, 36(1):144-152.
[12] Sridharan A P, Sugitha T, Karthikeyan G, et al. Comprehensive profiling of the VOCs of Trichoderma longibrachiatum EF5 while interacting with Sclerotium rolfsii and Macrophomina phaseolina[J]. Microbiological Research, 2020, 236:e126436.
[13] Lemfack M C, Gohlke B, Toguem S M T, et al. mVOC 2.0:a database of microbial volatiles[J]. Nucleic Acids Research, 2018, 46(1):1261-1265.
[14] 傅若农. 固相微萃取(SPME)近几年的发展[J]. 分析试验室, 2015, 34(5):602-620.
[15] 陈奕鹏, 杨扬, 时涛, 等. 内生真菌HND5挥发性物质组分分析及其抑菌作用测定[J]. 热带作物学报, 2017, 38(4):689-694.
[16] Contarino R, Brighina S, Fallico B, et al. Volatile organic compounds (VOCs) produced by biocontrol yeasts[J]. Food Microbiology, 2019, 82:70-74.
[17] 陈利军, 智亚楠, 陈思宇, 等. 留兰香挥发油熏蒸抑菌活性研究[J]. 植物保护, 2019, 45(2):75-80.
[18] 王静, 曹建敏, 陈德鑫, 等. 短小芽孢杆菌AR03挥发性有机物的抑菌活性及其组分分析[J]. 中国农业科学, 2018, 51(10):1908-1919.
[19] 萨仁高娃, 胡文忠, 冯可, 等. 植物精油及其成分对病原微生物抗菌机理的研究进展[J]. 食品科学, 2020, 41(11):285-294.
[20] Inoue Y, Shiraishi A, Hada T, et al. The antibacterial effects of terpene alcohols on Staphylococcus aureus and their mode of action[J]. FEMS Microbiology Letters, 2004, 237(2):325-331.
[21] Trombetta D, Castelli F, Sarpietro M G, et al. Mechanisms of antibacterial action of three monoterpenes[J]. Antimicrob Agents Chemother, 2005, 49(6):2474-2478.
[22] Giorgio A, De Stradis A, Lo Cantore P, et al. Biocide effects of volatile organic compounds produced by potential biocontrol rhizobacteria on Sclerotinia sclerotiorum[J]. Frontiers in Microbiology, 2015, 6:1056.
[23] Medina-Romero Y M, Roque-Flores G, Macías-Rubalcava M L. Volatile organic compounds from endophytic fungi as innovative postharvest control of Fusarium oxysporum in cherry tomato fruits[J]. Applied Microbiology and Biotechnology, 2017, 101(22):8209-8222.
[24] Strobel G A, Dirske E, Sears J, et al. Volatile antimicrobials from Muscodor albus, a novel endophytic fungus[J]. Microbiology, 2001, 147(11):2943-2950.
[25] Strobel G A. Muscodor species-endophytes with biological promise[J]. Phytochemistry Reviews, 2011, 10(2):165-172.
[26] Siddiquee S, Cheong B E, Taslima K, et al. Separation and identification of volatile compounds from liquid cultures of Trichoderma harzianum by GC-MS using three different capillary columns[J]. Journal of Chromatographic Science, 2012, 50(4):358-367.
[27] Jeleń H H. Volatile sesquiterpene hydrocarbons characteristic for Penicillium roqueforti strains producing PR toxin[J]. Journal of Agricultural and Food Chemistry, 2002, 50(22):6569-6574.
[28] Yan D H, Song X Y, Li H C, et al. Antifungal activities of volatile secondary metabolites of four Diaporthe strains isolated from Catharanthus roseus[J]. Journal of Fungi, 2018, 4(2):65.
[29] Zhang Z B, Chen X Y, Chen H B, et al. Anti-inflammatory activity of β-patchoulene isolated from patchouli oil in mice[J]. European Journal of Pharmacology, 2016, 781:229-238.
[30] Usami A, Motooka R, Nakahashi H, et al. Characteristic odorants from bailingu oyster mushroom (Pleurotus eryngii var. tuoliensis) and summer oyster mushroom (Pleurotus cystidiosus)[J]. Journal of Oleo Science, 2014, 63(7):731-739.
[31] Rösecke J, Pietsch M, König W A. Volatile constituents of wood-rotting basidiomycetes[J]. Phytochemistry, 2000, 54(8):747-750.
[32] Takao Y, Kuriyama I, Yamada T, et al. Antifungal properties of Japanese cedar essential oil from waste wood chips made from used sake barrels[J]. Molecular Medicine Reports, 2012, 5(5):1163-1168.