微生物源挥发性物质及其生物防治作用研究进展

近年来,人们对微生物源挥发性有机物的研究日益感兴趣,不仅因为释放挥发性物质的微生物种类超出人们现有的认识,而且这些微生物挥发性物质成分复杂、功能多样。本文从现有微生物挥发性物质收集、分析方法以及微生物挥发性物质在植物病害生物防治方面的研究进展进行综述。针对释放挥发性物质真菌和细菌的种类多样性、抗菌挥发性物质成分的多样性以及在生物防治中的应用进行了系统阐述。此外,作者还对产生挥发性物质的微生物新种类,特殊生物活性物质结构、功能、作用机制以及应用前景等研究方面提出了展望。     

细菌挥发性代谢产物的类型、检测技术及应用

细菌挥发性代谢产物是细菌代谢产物的重要组成部分,与细菌生命活动和细菌生长数量密切关联,是细菌与周围各种生物进行交流的重要信息物质。细菌挥发性代谢产物类型和特征表现复杂,其浓度低、种类多,从极性到非极性跨度大,需要依赖特殊检测技术进行分析测定。归纳了常见细菌挥发性代谢产物的类型,介绍了气相色谱-质谱和电子鼻两种检测挥发性有机化合物的方法,例举了细菌挥发性代谢产物检测在医学和食品科学研究领域的应用,并提出了细菌挥发性代谢产物检测研究中的几个关键问题。     

生防细菌产生的拮抗物质及其在生物防治中的作用

利用生防细菌防治植物病害是生物防治的一个主要内容．生防细菌防治植物病害发生发展的一个重要机制是产生拮抗物质．生防细菌的拮抗物质种类多,作用范围广谱．同一种拮抗物质可以由多种细菌菌株产生,而同一细菌也可以产生多种不同结构的拮抗物质．运用现代分子生物学技术和先进的分析测试手段可以加快对产生拮抗物质生防细菌的研究,了解生防细菌在寄主植物根围和叶围的定植效果。明确拮抗物质在生物防治中的作用．拮抗物质的产生除与细菌基因型有关外,一些外在的生物和非生物因素如病原菌存在与否、温度、pH和C、N营养等也影响拮抗物质产生．文中论述了生防细菌应用中存在的问题。指出混合菌剂的研制对防止病原菌抗性产生具有重要作用,应是今后生防菌剂研制中的重点．     

脱皮与不脱皮青稞酿造营养黄酒风味差异的研究

以青稞为原料酿造新型黄酒,为研究原料脱皮与否对新型黄酒风味的影响,利用气相色谱质谱联用(GC-MS)测定脱皮青稞黄酒、不脱皮青稞黄酒及青稞麸皮中挥发性和半挥发性风味物质。结果表明未脱皮青稞黄酒中香气物质总量为71 134.67μg/L;脱皮青稞酿造黄酒香气物质含量较少为39 208.99μg/L。挥发性香气成分在含量上差异主要来源于,3-甲基丁醇、苯乙醇、2-甲基丙醇。通过香气活力值OVA分析发现,对两种青稞黄酒香气贡献的较大的物质种类均为辛酸乙酯、乙酸异戊酯、苯乙醇和苯甲醛,贡献值的大小有差异。对麸皮挥发性风味成分分析发现,青稞黄酒中香气成分均是通过微生物作用产生,麸皮中富含的苯丙氨酸、缬氨酸和亮氨酸通过代谢产生苯乙醇、2-甲基丙醇、3-甲基丁醇,是造成两种青稞黄酒中挥发性风味物质差异的主要原因。研究表明青稞麸皮对黄酒风味的形成有积极意义。     

固相微萃取-气质法测定土壤挥发性抑菌物质*

运用固相微萃取 气相色谱 质谱法 (SPME GC MS) ,测定了参与土壤抑真菌作用的土壤挥发性成分和土壤细菌挥发性代谢物。通过比较土壤来源和土壤细菌来源的挥发性抑菌成分 ,发现在强挥发性抑菌土壤和土壤细菌代谢物中普遍存在着三甲胺、二甲基二硫醚、3 甲基 2 戊酮、甲基吡嗪、 2 ,5 二甲基吡嗪、N ,N 二甲基辛胺、十九烷等化合物。这些化合物很有可能就是参与土壤抑菌作用 ,特别是挥发性物质抑菌作用的主要成分。另外 ,为深入了解土壤中参与抑菌作用的挥发性化合物提供了简便有效的方法。     

基于GC-IMS比较不同干燥方式对柴达木枸杞中挥发性物质的影响CSCD

柴达木枸杞作为青藏高原特有的中药材之一,为探究不同干燥方式对柴达木枸杞品质以及挥发性成分的影响。本文采用自然阴干、热风烘干、真空冷冻干燥以及微波干燥的方式处理柴达木枸杞鲜果,通过气相色谱-离子迁移谱联用(GC-IMS)法分析枸杞鲜果和各类枸杞干果的挥发性成分的变化,构建挥发性成分的差异谱图,并进行主成分分析。从样品中共鉴别出了52种挥发性有机物质以及部分物质的单体和二聚体,包括醛类、烯醛类、酯类、酮类和醇类等。差异图和主成分分析图表明枸杞干燥后挥发性有机物发生了显著的变化,冷冻干燥后的枸杞中挥发性有机物质的种类和含量与枸杞鲜果较为相近,但与其他干燥方式干燥后的枸杞干果差异较大,而热风烘干和微波干燥后的枸杞干果中挥发性有机物种类非常相似。通过建立一种利用GC-IMS检测分析枸杞中挥发性成分的方法,结果表明与其他干燥方式相比,冷冻干燥法能够有效保留枸杞中的挥发性成分,使枸杞保持较高的品质,该研究结果为后续柴达木枸杞的质量评价和加工提供一定的参考依据。     

药用植物内生细菌多样性及其生物学作用

药用植物内生细菌资源丰富,其产生的多种活性成分,在医药业、农业等方面表现出良好的应用前景,是人类生存与发展的重要资源。该文综述了近年来药用植物内生细菌的研究进展情况,特别是对其多样性、产生的活性成分及其生物学作用进行了系统总结,并从药用植物内生细菌研究的特殊性出发,对其前景进行了展望。该文对于药用植物功能内生细菌的研究、开发与利用具有重要的理论指导和现实意义。     

固相微萃取-气质法测定土壤挥发性抑菌物质

运用固相微萃取．气相色谱,质谱法(SPME-GC／MS),测定了参与土壤抑真菌作用的土壤挥发性成分和土壤细菌挥发性代谢物。通过比较土壤来源和土壤细菌来源的挥发性抑菌成分,发现在强挥发性抑菌土壤和土壤细菌代谢物中普遍存在着三甲胺、二甲基二硫醚、3-甲基-2-戊酮、甲基吡嗪、2,5-二甲基吡嗪、N,N-二甲基辛胺、十九烷等化合物。这些化合物很有可能就是参与土壤抑菌作用,特别是挥发性物质抑菌作用的主要成分。另外,为深入了解土壤中参与抑菌作用的挥发性化合物提供了简便有效的方法。     

柠檬和莱檬果皮精油挥发性成分分析

利用气质联用技术(GC-MS),对云南德宏栽培的小莱檬、塔西提2个莱檬品种和费米耐劳、尤力克、艾伦尤力克3个柠檬品种的果皮精油挥发性成分进行研究。结果表明,小莱檬、塔西提、费米耐劳、尤力克、艾伦尤力克柠檬果皮精油中分别检测到17、24、23、20、27种挥发性成分,烯烃类物质是莱檬和柠檬果皮精油挥发性的主要组分,其中特征香气成分D-柠檬烯含量最高,β-蒎烯、γ-萜品烯次之。塔西提莱檬特征香气成分物质种类和含量高于小莱檬,艾伦尤力克柠檬中烃类、醇类、醛类、酯类和酮类物质的种类和含量均高于费米耐劳柠檬和尤力克柠檬,主要特征香气成分含量也最高。     

9 种海洋硅藻挥发性成分的比较分析

采用顶空固相微萃取气相色谱一质谱联用技术,通过SIMCA．P分析软件,对硅藻门下处于生长平台后期的9种海洋硅藻的挥发性成分进行比较分析研究。结果表明：在平台后期硅藻的挥发性成分中,醛类物质的含量和种类占据了很大的优势,壬醛、2,4-辛二烯醛、2-己烯醛、2-壬烯醛、2,4-庚二烯醛等是硅藻挥发性成分的主要构成物质,十四烷、十五烷、1-十五烯、十六烷等烷烃类是硅藻中含量仅次于醛类物质的挥发性成分,3,5-辛烯基-3-酮等短链酮类和1-辛烯基-3-醇是硅藻中含量较高的酮类和醇类物质,9种硅藻中均检测二甲基硫的存在。差异性较大的挥发性物质主要有壬醛、8-十七碳烯、1-戊烯．3．酮、1-己烯-3．醇、2,6-二甲基一苯酚,它们决定了不同的硅藻有着各自不同的风味。研究结果表明这些挥发性物质对硅藻的饵料价值、养殖生物的肉质风味、水环境的异味形成有着重要意义。     

Bacterial volatiles and their action potential

During the past few years, an increasing awareness concerning the emission of an unexpected high number of bacterial volatiles has been registered. Humans sense, intensively and continuously, microbial volatiles that are released during food transformation and fermentation, e.g., the aroma of wine and cheese. Recent investigations have clearly demonstrated that bacteria also employ their volatiles during interactions with other organisms in order to influence populations and communities. This review summarizes the presently known bioactive compounds and lists the wide panoply of effects possessed by organisms such as fungi, plants, animals, and bacteria. Because bacteria often emit highly complex volatile mixtures, the determination of biologically relevant volatiles remains in its infancy. Part of the future goal is to unravel the structure of these volatiles and their biosynthesis. Nevertheless, bacterial volatiles represent a source for new natural compounds that are interesting for man, since they can be used, for example, to improve human health or to increase the productivity of agricultural products.     

Volatile affairs in microbial interactions

Microorganisms are important factors in shaping our environment. One key characteristic that has been neglected for a long time is the ability of microorganisms to release chemically diverse volatile compounds. At present, it is clear that the blend of volatiles released by microorganisms can be very complex and often includes many unknown compounds for which the chemical structures remain to be elucidated. The biggest challenge now is to unravel the biological and ecological functions of these microbial volatiles. There is increasing evidence that microbial volatiles can act as infochemicals in interactions among microbes and between microbes and their eukaryotic hosts. Here, we review and discuss recent advances in understanding the natural roles of volatiles in microbe–microbe interactions. Specific emphasis will be given to the antimicrobial activities of microbial volatiles and their effects on bacterial quorum sensing, motility, gene expression and antibiotic resistance.     

Bacteria associated with truffle‐fruiting bodies contribute to truffle aroma

Truffles, symbiotic fungi renown for the captivating aroma of their fruiting bodies, are colonized by a complex bacterial community of unknown function. We characterized the bacterial community of the white truffle Tuber borchii and tested the involvement of its microbiome in the production of sulphur‐containing volatiles. We found that sulphur‐containing volatiles such as thiophene derivatives, characteristic of T. borchii fruiting bodies, resulted from the biotransformation of non‐volatile precursor(s) into volatile compounds by bacteria. The bacterial community of T. borchii was dominated by α‐ and β‐Proteobacteria. Interestingly, all bacteria phyla/classes tested in this study were able to produce thiophene volatiles from T. borchii fruiting body extract, irrespective of their isolation source (truffle or other sources). This indicates that the ability to produce thiophene volatiles might be widespread among bacteria and possibly linked to primary metabolism. Treatment of fruiting bodies with antibacterial agents fully suppressed the production of thiophene volatiles while fungicides had no inhibitory effect. This suggests that during the sexual stage of truffles, thiophene volatiles are exclusively synthesized by bacteria and not by the truffle. At this stage, the origin of thiophenes precursor in T. borchii remains elusive and the involvement of yeasts or other bacteria cannot be excluded.     

Volatile-mediated antagonism of soil bacterial communities against fungi

Competition is a major type of interaction between fungi and bacteria in soil and is also an important factor in suppression of plant diseases caused by soil-borne fungal pathogens. There is increasing attention for the possible role of volatiles in competitive interactions between bacteria and fungi. However, knowledge on the actual role of bacterial volatiles in interactions with fungi within soil microbial communities is lacking. Here, we examined colonization of sterile agricultural soils by fungi and bacteria from non-sterile soil inoculums during exposure to volatiles emitted by soil-derived bacterial communities. We found that colonization of soil by fungi was negatively affected by exposure to volatiles emitted by bacterial communities whereas that of bacteria was barely changed. Furthermore, there were strong effects of bacterial community volatiles on the assembly of fungal soil colonizers. Identification of volatile composition produced by bacterial communities revealed several compounds with known fungistatic activity. Our results are the first to reveal a collective volatile-mediated antagonism of soil bacteria against fungi. Given the better exploration abilities of filamentous fungi in unsaturated soils, this may be an important strategy for bacteria to defend occupied nutrient patches against invading fungi. Another implication of our research is that bacterial volatiles in soil atmospheres can have a major contribution to soil fungistasis.     

Roles of plant volatiles in defence against microbial pathogens and microbial exploitation of volatiles

Plants emit a large variety of volatile organic compounds during infection by pathogenic microbes, including terpenes, aromatics, nitrogen‐containing compounds, and fatty acid derivatives, as well as the volatile plant hormones, methyl jasmonate, and methyl salicylate. Given the general antimicrobial activity of plant volatiles and the timing of emission following infection, these compounds have often been assumed to function in defence against pathogens without much solid evidence. In this review, we critically evaluate current knowledge on the toxicity of volatiles to fungi, bacteria, and viruses and their role in plant resistance as well as how they act to induce systemic resistance in uninfected parts of the plant and in neighbouring plants. We also discuss how microbes can detoxify plant volatiles and exploit them as nutrients, attractants for insect vectors, and inducers of volatile emissions, which stimulate immune responses that make plants more susceptible to infection. Although much more is known about plant volatile–herbivore interactions, knowledge of volatile–microbe interactions is growing and it may eventually be possible to harness plant volatiles to reduce disease in agriculture and forestry. Future research in this field can be facilitated by making use of the analytical and molecular tools generated by the prolific research on plant–herbivore interactions.     

Pseudomonas Strains Naturally Associated with Potato Plants Produce Volatiles with High Potential for Inhibition of Phytophthora infestans

Bacteria emit volatile organic compounds with a wide range of effects on bacteria, fungi, plants, and animals. The antifungal potential of bacterial volatiles has been investigated with a broad span of phytopathogenic organisms, yet the reaction of oomycetes to these volatile signals is largely unknown. For instance, the response of the late blight-causing agent and most devastating oomycete pathogen worldwide, Phytophthora infestans, to bacterial volatiles has not been assessed so far. In this work, we analyzed this response and compared it to that of selected fungal and bacterial potato pathogens, using newly isolated, potato-associated bacterial strains as volatile emitters. P. infestans was highly susceptible to bacterial volatiles, while fungal and bacterial pathogens were less sensitive. Cyanogenic Pseudomonas strains were the most active, leading to complete growth inhibition, yet noncyanogenic ones also produced antioomycete volatiles. Headspace analysis of the emitted volatiles revealed 1-undecene as a compound produced by strains inducing volatile-mediated P. infestans growth inhibition. Supplying pure 1-undecene to P. infestans significantly reduced mycelial growth, sporangium formation, germination, and zoospore release in a dose-dependent manner. This work demonstrates the high sensitivity of P. infestans to bacterial volatiles and opens new perspectives for sustainable control of this devastating pathogen.     

Volatile Emissions from Mycobacterium avium subsp. paratuberculosis Mirror Bacterial Growth and Enable Distinction of Different Strains

Control of paratuberculosis in livestock is hampered by the low sensitivity of established direct and indirect diagnostic methods. Like other bacteria, Mycobacterium avium subsp. paratuberculosis (MAP) emits volatile organic compounds (VOCs). Differences of VOC patterns in breath and feces of infected and not infected animals were described in first pilot experiments but detailed information on potential marker substances is missing. This study was intended to look for characteristic volatile substances in the headspace of cultures of different MAP strains and to find out how the emission of VOCs was affected by density of bacterial growth. One laboratory adapted and four field strains, three of MAP C-type and one MAP S-type were cultivated on Herrold’s egg yolk medium in dilutions of 10^-0, 10^-2, 10^-4 and 10^-6. Volatile substances were pre-concentrated from the headspace over the MAP cultures by means of Solid Phase Micro Extraction (SPME), thermally desorbed from the SPME fibers and separated and identified by means of GC-MS. Out of the large number of compounds found in the headspace over MAP cultures, 34 volatile marker substances could be identified as potential biomarkers for growth and metabolic activity. All five MAP strains could clearly be distinguished from blank culture media by means of emission patterns based on these 34 substances. In addition, patterns of volatiles emitted by the reference strain were significantly different from the field strains. Headspace concentrations of 2-ethylfuran, 2-methylfuran, 3-methylfuran, 2-pentylfuran, ethyl acetate, 1-methyl-1-H-pyrrole and dimethyldisulfide varied with density of bacterial growth. Analysis of VOCs emitted from mycobacterial cultures can be used to identify bacterial growth and, in addition, to differentiate between different bacterial strains. VOC emission patterns may be used to approximate bacterial growth density. In a perspective volatile marker substances could be used to diagnose MAP infections in animals and to identify different bacterial strains and origins.     

Emission of volatile compounds by Erwinia amylovora: biological activity in vitro and possible exploitation for bacterial identification

Several analytical techniques such as gas chromatography–mass spectrometry, proton transfer reaction–mass spectrometry and laser photoacoustic detection, were used to characterize the volatiles emitted by Erwinia amylovora and other plant-pathogenic bacteria. Diverse volatiles were found to be emitted by the different bacterial species examined. The distinct blend of volatiles produced by bacteria allowed their identification using an electronic nose (e-nose). The present study reports the discrimination of E. amylovora, the fire blight pathogen, from other plant-associated bacteria using an e-nose based on metal oxide semiconductor sensors. Two different approaches were used for bacterial identification. The first one was the direct comparison of the odorous profiles of unknown bacterial isolates with four selected reference species. The second approach was the use of previously developed databases representing the odorous variability among several bacterial species. Using these two strategies, the e-nose successfully identified the isolates in 87.5 and 62.5% of the cases, respectively. Finally, the profiling of the volatiles emitted by E. amylovora lead to identify some metabolic markers with a potential biological activity in vitro.     

Queen-specific volatile in a higher termite Nasutitermes takasagoensis (Isoptera: Termitidae)

In social insect colonies, queen-produced pheromones have important functions in social regulation. These substances influence the behavior and physiology of colony members. A queen-produced volatile that inhibits differentiation of new neotenic reproductives was recently identified in the lower termite Reticulitermes speratus. However, there are no known queen-specific volatiles of this type in any other termite species. Here, we report volatile compounds emitted by live queens of the higher termite Nasutitermes takasagoensis. We used headspace gas chromatography mass spectroscopy (HS GC-MS) to analyze volatiles emitted by live primary queens, workers, soldiers, alates, and eggs collected in a Japanese subtropical forest. Among 14 detected compounds, 7 were soldier-specific, 1 was alate-specific, 1 was egg-specific, and 1 was queen-specific. The queen-specific volatile was phenylethanol, which is different than the compound identified in R. speratus. The identification of this queen-specific volatile is the first step in determining its functions in higher termite social regulation. Comparisons of queen pheromone substances regulating caste differentiation among various termite taxa will contribute to a better understanding of the evolution of social systems in termites.