PERGAMON

Soil Biology and Biochemistry 31 (1999) 375385

Eects and incidence of volatile organic compound interactions between soil bacterial and fungal isolates
A.E. Mackie, R.E. Wheatley *
Unit of SoilPlant Dynamics, Cellular and Environmental Physiology Department, Scottish Crop Research Institute, Invergowrie, Dundee DD2 5DA, UK Accepted 10 September 1998

Abstract The eects of volatile organic compounds (VOCs) produced by a random selection of soil bacteria on the growth rates and activity of several fungi were assessed using experimental systems that had only atmospheric connections. All of the bacterial isolates either stimulated or inhibited the growth rate of at least one of the fungal species. Growth rates of some of the fungi were inhibited by up to 60% in some cases and stimulated by up to 35% in others (P < 0.05). No single bacterial isolate was eective against all of the fungi. Of the bacterial isolates 54% both inhibited the growth rate of some fungi and stimulated others. Many bacteria, 42% of the total, could only inhibit growth, but none were solely stimulatory (P < 0.05). Growth of some fungi was inhibited within 2 d of exposure to bacterial cultures (P < 0.05) and only resumed when the fungus was placed on fresh PDA plates. Contrastingly, cores taken from the growing margins of cultures did not grow (P < 0.05) when placed onto PDA plates that had previously been exposed to the bacterial cultures. Laccase activity in Phanaerochaete magnoliae ceased completely on exposure to all of the bacterial isolates and was signicantly reduced in Trichoderma viride. Tyrosinase activity in T. viride was not aected by any of the bacterial isolates, but activity in P. magnoliae was increased, inhibited or not aected, depending on the bacterium to which it was exposed. These VOC-mediated eects appeared to be species-specic, with each fungus responding uniquely to the products of each of the bacterial cultures. The interactions varied according to the substrate on which the organisms were cultured, and on the size and age of the bacterial population. Interactions mediated by microbially produced VOCs could be widespread in soils, and the outcome of these interactions will be determined by the microorganisms involved. # 1999 Elsevier Science Ltd. All rights reserved.

1. Introduction Volatile organic compounds (VOCs) occur in soil atmospheres over a range of concentrations (Wheatley et al., 1996) and most are probably microbial in origin (Stotzky and Schenck, 1976). The range, or prole, of compounds produced appears to depend not only on environmental conditions, such as nutrients (Fiddaman and Rossall, 1993; Wheatley et al., 1996, 1997) and temperature (Tronsmo and Dennis, 1978) but also on the organism itself, with individual species producing a reproducible prole of unique VOCs (Zechman and Labows, 1985; Giudici et al., 1990). Exposure to bacterial VOCs can reduce growth (Moore-Landecker and Stotzky, 1972; Wright and Thompson, 1985;
* Corresponding author. Fax: +44-1382-562-426; e-mail: r.wheatley @scri.sari.ac.uk 0038-0717/99/$19.00 # 1999 Elsevier Science Ltd. All rights reserved. PII: S 0 0 3 8 - 0 7 1 7 ( 9 8 ) 0 0 1 4 0 - 0

Fiddaman and Rossall, 1993; Wheatley et al., 1997) and inhibit spore germination (McKee and Robinson, 1988) in a variety of fungal species. Conversely, germination of mycorrhizal spores can be stimulated by VOCs produced by the soil actinomycete Streptomyces orientalis (Tylka et al., 1991). Although some VOCs increase microbial respiration (Owens et al., 1969) others have no eect (Ko and Chow, 1978). These conicting observations may be related to the speciesspecic responses of the soil microbes. In soils, fungal growth and metabolism will be inuenced by surrounding environmental conditions. The eect of VOCs on fungal activity can be quantied by measuring colony diameter or enzyme production. Laccase and tyrosinase are two enzymes that frequently occur in eukaryotic organisms and have been linked to mycelial growth (Lilly and Barnett, 1951; White and Boddy, 1992).

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The majority of these reports on VOC interactions concentrate on a limited number of well documented soil organisms, for example Moore-Landecker and Stotzky (1973) examined the eects of VOCs produced by Agrobacterium spp., and Fiddaman and Rossall (1993) reported on the inhibitory VOCs released by Bacillus subtilis. The frequency of VOC-mediated interactions between soil microbes has not been investigated, so a selection of bacteria, isolated from soil, were screened for the ability to inuence the growth or enzymatic activity of at least one fungal species. This ability was investigated using an experimental system where the only connection between the bacterial and fungal isolates was atmospheric. 2. Materials and methods 2.1. Fungal isolates Five fungal species, representing a range of fungal types; were chosen: Trichoderma viride, a common soil saprophyte, Gaeumannomyces graminis var. tritici, a specic pathogen of wheat, Phytophthora cryptogea, a plant pathogen with a wider host range, Pythium ultimum, a vigorous plant pathogen and Phanaerochaete magnoliae, a nonsoil organism that is a pathogen of beech trees. Stock cultures were maintained on 3.9% potato dextrose agar (PDA) (Oxoid, Unipath) and stored at 48C. For the interaction experiments plugs were removed from the growing edge of a stock plate, placed centrally onto a fresh plate of PDA (0.39%) and incubated at 158C for 96 h. 2.2. Bacterial isolates Bacteria were isolated from an organically cultivated soil of the Craibstone association, previously growing barley. Soil (10 g) was dispersed in 50 ml of sterile distilled water, then diluted 10-fold. Aliquots, 0.2 ml, of the diluted samples were spread onto either 0.3% tryptone soya broth (TSB) or 0.28% nutrient agar (NA), both with added 1.5% technical agar (Oxoid, Unipath), then incubated for 7 d at 158C. All the single colonies were subcultured onto fresh plates of the same medium then, after incubation for 5 d at 158C, the cultures were sealed with paralm and stored at 48C. More than 60 isolates were selected randomly from the collection and screened for VOC activity. 2.3. Bacterialfungal interactions via VOCs Bacteria were inoculated into asks containing 50 ml of tryptone soya broth (0.3%), then incubated at 158C for 72 h. Aliquots, 50 ml, of these bacterial cul-

tures were spread onto plates of tryptone soya broth (0.3%) with added agar (1.5%), then incubated for 24 h at 158C. Then the lid of a plate of 0.39% PDA, inoculated centrally with an agar plug taken from the growing margin of a 96 h fungal culture, was removed and the plate inverted over the top of the 24 h bacterial culture. The plates were sealed together with nescolm so that both organisms grew in a shared atmosphere. The diameters of the fungal colonies, on three replicates, were measured after 5 d incubation at 158C and dierences with the controls assessed by analyses of variance. Control cultures were prepared in the same way except that the TSB plates were not inoculated with bacterial cultures. 2.4. Interaction studies 2.4.1. The eect of bacterial VOCs on fungal growth Three bacterial isolates were selected that showed signicant interactions with the target fungi (P>0.05). The fungal targets and these three selected bacterial cultures were sealed together as described previously, in a series of combinations. Fungi growing on either of two concentrations of PDA, (0.39 or 0.039%) were exposed to one of the three bacteria, growing on either of two concentrations of NA (2.8 and 0.28%). This total of twelve combinations was replicated three times. For the control the bacterial cultures were replaced with uninoculated NA plates. Plates were incubated for 10 d at 158C and fungal radia measured daily. To examine the eects of culture age fungal colonies were either grown to a diameter of 30 mm or the bacteria were incubated on the plate for 96 h, before the plates were sealed together. 2.4.2. Recovery of exposed fungi and VOC adsorption into the fungal medium Cultures of P. magnoliae and the selected bacterial isolates were sealed together with paralm. After fungal growth had stopped the bacterial plates were removed and sterile lids placed over the inhibited fungus. Agar plugs, covered with mycelia, were removed from the edge of these inhibited fungal colonies and placed onto fresh PDA plates of the same strength, at 2, 3, 4 and 5 d after exposure to the bacterial VOCs. Fungal recovery, both on the original exposed culture and the transferred cultures, was monitored daily by measuring colony size. To establish whether VOC adsorbtion into the PDA was preventing fungal recovery, blank PDA plates were sealed with inoculated bacterial plates for 5, 6 and 8 d. Then the bacterial cultures were removed and the PDA plates were inoculated with 7 mm cores taken from the growing margin of an unexposed 96 h culture of P. magnoliae. Control plates were prepared

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by inoculating the fungus onto fresh agar plates. Fungal growth was measured daily on three replicates of these plate combinations.

3. Results Dierences in rates of growth between the control and exposed cultures were calculated for each fungal species and plotted in order of eect (Fig. 1ad). All bacterial cultures inuenced the growth, either by stimulation or inhibition, of at least one fungal species, although none had any eect on the development of P. ultimum which grew to the plate boundaries by d 5. However, no single bacterial isolate was eective against all the fungi. Of the isolates tested 54% were both inhibitory to some fungi and also stimulatory to others (P < 0.05). Some bacteria, 42%, were only inhibitory, but all those that stimulated some fungi also inhibited others, 51% (P < 0.05). The majority of the isolates were inhibitory against P. magnoliae and T. viride (84 and 65%, respectively; P < 0.05). In contrast, fewer of the bacterial isolates had an eect on P. cryptogea or G. graminis, but any eect was predominantly stimulatory. Only 5% of the isolates inhibited P. cryptogea and 10% G. graminis, compared to 42% that stimulated P. cryptogea and 17% G. graminis (P < 0.05). The VOCs produced by the bacterial isolates had a variety of eects on the fungi. For example, bacterial isolate 21, produced VOCs which had no eect on

2.5. Statistical analyses Dierences between the sizes of the fungal colonies in the treatments and controls were assessed by analyses of variance (ANOVA).

2.6. Enzyme activity of the fungal mycelium The eects of bacterial VOCs on the activity of the two enzymes, laccase and tyrosinase, were evaluated on triplicate plates using the method described by White and Boddy (1992). After the fungi and bacteria had been sealed together for 5 d the bacterial cultures were discarded. Laccase activity was assessed by adding 5 ml of 1.44% a-naphthol (w/v) in 96% ethanol and tyrosinase activity by adding 1.08% (w/v) p-cresol in 98% ethanol to the fungal plates. Colour reactions were noted in comparison to the control fungi, which had been exposed to sterile NA plates.

Fig. 1(a). The inuence of VOCs produced by randomly-selected soil bacterial isolates on mean radial growth rate of (a) Trichoderma viride, (b) Phanaerochaete magnoliae, (c) Phytophthora cryptogea and (d) Gaeumannomyces graminis var tritici, expressed as % of the mean growth rate of unexposed cultures. Specic bacterial isolates 2; 21; 24 and 38 are noted; bar is SE, n = 3.

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Fig. 1(b and c). (Continued, caption on previous page.)

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Fig. 1(d). (Continued, caption on page 377.)

either T. viride or G. graminis, inhibited P. magnoliae and stimulated P. cryptogea, whereas bacterial isolate 2 was inhibitory to both T. viride and P. magnoliae, stimulated P. cryptogea and had no eect on G. graminis (P < 0.05). Slightly dierently, bacterial isolate 24 was inhibitory to both T. viride and P. magnoliae, had no eect on P. cryptogea but stimulated G. graminis, and isolate 38 was inhibitory to both T. viride and P. magnoliae, but had no eect on both P. cryptogea and G. graminis (P < 0.05; Fig. 1). 3.1. Inuence of VOCs on the rate of fungal growth Fungal growth rates during exposure to the bacterial VOCs are shown in Fig. 2. Although there was usually a steady increase in colony size, growth rates were signicantly eected (P < 0.05) in many of the combinations. Each fungus reacted dierently to exposure to the bacterial VOCs, and the eects on P. magnoliae were greater than those on T. viride. Similarly, each bacterial culture inuenced fungal growth in a dierent way. The degree of interaction was dependent on the environment, mainly the medium, in which the bacteria and fungi were grown and the age of the bacterial culture. For example, the growth rate of T. viride was not aected by any of the 24 h bacterial cultures but was inhibited by 96 h cultures (P < 0.05) of two of the

three bacteria when the fungus was grown on 0.039% PDA, but only one when grown on 0.39% PDA. The use of larger fungal colonies, i.e. grown to a diameter of approximately 30 mm before use, had no eect on these interactions (Fig. 3). 3.2. Recovery of exposed fungi and VOC adsorption into the fungal medium P. magnoliae growth was inhibited within 2 d of exposure to the bacterial cultures (P < 0.05), and irrespective of whether the bacterial plates were removed after 2, 3, 4 or 5 d, growth never resumed (Fig. 4a). However, when cores were taken from the margins of these inhibited plates and placed onto fresh PDA plates of the same strength, growth resumed immediately (Fig. 4b). Contrastingly, cores from the growing margins of unexposed cultures did not grow (P < 0.05) after being inoculated onto PDA plates (Fig. 4c) that had previously been exposed to the bacterial cultures. 3.3. Enzyme activity of the fungal mycelium Laccase activity in the P. magnoliae cultures ceased completely on exposure to all of the bacterial isolates and was signicantly reduced in T. viride (Table 1).

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Fig. 2. Mean fungal colony diameters, mm, of P. magnoliae () and T. viride (- - -) exposed to the VOCs produced by 24 h (a1d1) and 96 h (a2d2) cultures of the selected interactive bacteria A (r), B (r) and C (w) and sterile NA (q). Bacteria were grown on 2.8% NA (a and c) and 0.28% NA (b and d), and the fungi were grown on 0.39% PDA (a and b) and 0.039% PDA (c and d), bar is SE; n = 3.

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Fig. 3. Mean fungal colony diameters, mm, of T. viride (a) and P. magnoliae (b) grown on 0.39% PDA and exposed to the VOCs produced by bacteria A (r), B (r), and C (w), grown on 2.8% NA and also exposed to sterile NA (q). Fungal and bacterial plates were sealed after the fungal colony reached an approximate diameter of 30 mm, bar is SE; n = 3.

Tyrosinase activity in T. viride was not aected by any of the bacterial isolates, but activity in P. magnoliae was increased on exposure to bacterium A, inhibited by bacterium B and not aected by bacterium C. 4. Discussion As the only connection between the bacterial and fungal isolates was atmospheric any eects reported here can only have been caused by volatile com-

pounds. This work has shown that VOCs produced by bacteria can inuence both fungal mycelial growth rate and enzyme activity. Fungal response to the bacterial VOCs appeared to be species, environment and age specic, both for the fungus and the bacteria. This concept is supported by several reports of diering sensitivity to bacterial volatiles by dierent fungi (Dennis and Webster, 1971; Moore-Landecker and Stotzky, 1972; Rai et al., 1981; Fiddaman and Rossall, 1994). All the bacterial cultures produced VOCs that could inuence the growth of at least one fungal

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Fig. 4. Recovery of P. magnoliae exposed to the VOCs of bacterium A, () and sterile 2.8% NA (- - -), on removal of the bacterial plate (a) after 2 (q), 3 (r), 4 (r) and 5 (w) days and after transferral to a sterile 0.39% PDA plate (b), where q is controls and r bacterium A. Graph c shows the growth of P. magnoliae on 0.39% PDA plates which had previously been exposed to the VOCs of bacterium A () or sterile 2.8% NA (- - -), for 5 (q), 6 (r) and 8 (r) days, bar is SE; n = 3.

species, suggesting that the occurrence of VOC mediated interactive-events could be widespread in soils. Exposure to both larger and older bacterial populations greatly increased both the degree and rate of inhibitory eects on the fungi. This presumably was a result of increased VOC production, suggesting that optimum concentrations are required within a system for these compounds to be eective. Previous research has suggested that, in most cases, the greater the con-

centration of volatile then the greater the inhibition. Robinson et al. (1989) reported that increased amounts of pure trimethylamine increased inhibition of hyphal extension and arthrospore germination in Geotrichum candidum. In our work fungal response was aected by the growth medium. Both fungi were inhibited less when grown on 0.39%, compared to 0.039% PDA. It may be that the increased nutrients supplied allowed the fungi to grow more vigorously, increasing any resist-

A.E. Mackie, R.E. Wheatley / Soil Biology and Biochemistry 31 (1999) 375385 Table 1 The activity of tyrosinase and laccase in T. viride and P. magnoliae cultures that have been exposured to soil bacterial isolates Isolate Bacteria A B C
a b

383

P. magnoliae Laccase 0a 0 0 Tyrosinase + + +d 0 ++

T. viride Laccase +b 0 + Tyrosinase ++c ++ ++

0 means no colour reaction. + a colour reaction less than control. c ++ a colour reaction equal to control. d +++ a colour reaction stronger than control.

ance to volatile compounds. However, established fungal cultures did not show a greater resistance to bacterial VOCs. Growth of established P. magnoliae cultures was still inhibited within 24 h of exposure, suggesting that the size or age of the fungus does not inuence its response. The VOCs that caused inhibition were not fungicidal, as the fungal cultures recovered when removed from the inuence of those VOCs. Persistence of the eects due to VOC adsorption into the agar medium indicated that the active compounds were water soluble. Amounts of enzyme activity in the fungi were also aected by bacterial VOCs. The response of tyrosinase activity in P. magnoliae to each of the bacterial isolates was dierent, being reduced in the presence of bacterium C, increased by bacterium A, but unaected by bacterium B. Laccase activity was reduced by all three bacteria. These changes in fungal enzyme activity might possibly be related to changes in the ambient air in the culture system. Laccase formation in Phlebia spp. has been demonstrated to alter under dierent concentrations of CO2 and O2 with an optimum CO2to-O2 ratio existing (White and Boddy, 1992). As the eects on fungal activity remained the same after the bacterial plates had been removed and aeration increased, it seems probable that absorbed VOCs were responsible for any eects on activity levels, rather than any changes in the CO2-to-O2 ratio. Although there are no reports of VOCs that inhibit these enzymes, there are reports of nonvolatile inhibitors of laccase and tyrosinase, including cucurbitacins that inhibit laccase (Viterbo et al., 1992), and tyrosinase inhibition by cysteine (Kermasha et al., 1993). These phenol-oxidising enzymes play an integral role in fungal morphogenesis and have been correlated to production of aerial mycelium in Phlebia radiata (White and Boddy, 1992) and to production of emergent phases in Hypholoma fasiciculare (Grith et al., 1994). It has been suggested that a complex relationship exists between mycelial morphogenesis and activity of phenol-oxidising enzymes (Grith et al.,

1994) and a link between fungal development and enzyme production may help to explain the decreased fungal growth in the exposed fungal cultures. Patterns of interactions may reect the niche held by fungi in the environment. P. magnoliae was inhibited by VOCs produced by almost all bacterial cultures. This extreme susceptibility may be due to the fact that P. magnoliae was isolated from decaying beech trees and not soil, and so may not be accustomed to the VOCs produced in the soil community. T. viride appeared to be aected in much the same way as P. magnoliae but not to such an extent. T viride is a notoriously resilient saprophyte commonly found in soil and decaying residues. It gains carbon from nonliving organic material, and shares a variety of carbon sources with a wide range of soil biota, so it seems probable that the ability to grow unaected in an atmosphere containing bacterial VOCs would be benecial. P. cryptogea and G. graminis were both stimulated and inhibited by bacterial VOCs. P. cryptogea, a plant pathogen that has a wide host range, was stimulated to the greatest extent and by a large proportion of bacterial cultures, whereas G. graminis, which has a relatively narrow host range was stimulated only by a minority. It may be that VOC interactions between microbial populations enhance nutrient and host location in the rhizosphere. For example, the ability to recognise the presence of a bacterial community associated with a root system, and to respond in a chemotrophic manner to the released VOCs may help the pathogen to locate and infect a potential host. Interactions via VOCs which improve host location have been reported, such as a communication system between the mycorrhizal fungi Gigaspora gigantea and the roots of its host plants (Koske, 1982). Release of VOCs by both members of the symbiosis in the preinfection stage allows directional growth of the fungal hyphae towards roots (Gemma and Koske, 1988). Stimulation of germination and directional growth by host plant VOCs is also seen with the fungal pathogens Sclerotium rolfsii (Tong-Kwee and Keng, 1980), Rhizoctonia solani (Norton and Harman, 1985) and Puccina punctiformis (French et al., 1994). Thus fungal responses to environmental cues, including VOCs, may enhance the ability of the organism to compete and survive within the soil system. Likewise, production of inhibitory VOCs by bacteria may increase their survival rate in soil. As colonisation and subsequent utilisation of a suitable nutrient source is paramount for soil microorganisms, the production of VOCs that are able to inhibit others will help to overcome any potential competitors. Production of VOCs that can inuence the growth of other organisms will directly aect the diversity of a community and as VOCs are metabolic products, the

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A.E. Mackie, R.E. Wheatley / Soil Biology and Biochemistry 31 (1999) 375385 Giudici, P., Romano, P., Zambonelli, C., 1990. A biometric study of higher alcohol production in Saccharomyces cerevisiae. Canadian Journal of Microbiology 36, 6164. Grith, G.S., Rayner, A.D.M., Wildman, H.G., 1994. Interspecic interactions and mycelial morphogenesis of Hypholoma fasciculare (Agaricaceae). Nova Hedwigia 59, 4775. Kermasha, S., Goetghebeur, M., Monfette, A., Metche, M., Rovel, B., 1993. Inhibitory eects of cysteine and aromatic acids on tyrosinase activity. Phytochemistry 34, 349353. Ko, W.H., Chow, F.K., 1978. Soil fungistasis: role of volatile inhibitors in two soils. Journal of General Microbiology 104, 7578. Koske, R.E., 1982. Evidence for a volatile attractant from plant roots aecting germ tubes of a VA mycorrhizal fungus. Transactions of the British Mycological Society 79, 305310. Lilly, V.G., Barnett, H.L., 1951. Enzymes and enzyme action. In: Physiology of the Fungi. McGraw-Hill, London, pp. 4564. McKee, N., Robinson, P.M., 1988. Production of volatile inhibitors of germination and hyphal extension by Geotrichum candidum. Transactions of the British Mycological Society 91, 157190. Moore-Landecker, E., Stotzky, G., 1972. Inhibition of fungal growth and sporulation by volatile metabolites from bacteria. Canadian Journal of Microbiology 18, 957962. Moore-Landecker, E., Stotzky, G., 1973. Morphological abnormalities of fungi induced by volatile microbial metabolites. Mycologia 65, 519530. Norton, J.M., Harman, G.E., 1985. Responses of soil microorganisms to volatile exudates from germinating pea seeds. Canadian Journal of Botany 63, 10401045. Owens, L.D., Gilbert, R.G., Griebel, G.E., Menzies, J.D., 1969. Identication of plant volatiles that stimulate microbial respiration and growth in soil. Phytopathology 59, 14681472. Rai, B., Sirivastava, A.K., Singh, D.B., 1981. Volatile and nonvolatile metabolites of actinomycetes and the growth of some litter decomposing fungi. Soil Biology & Biochemistry 13, 7576. Robinson, P.M., McKee, N.D., Thompson, L.A.A., Harper, D.B., Hamilton, J.T.G., 1989. Autoinhibition of germination and growth in Geotrichum candidum. Mycological Research 93, 214222. Stahl, P.D., Parkin, T.B., 1996. Microbial production of volatile organic compounds in soil microcosms. Proceedings of the Soil Science Society of America 60, 821828. Stotzky, G., Schenck, S., 1976. Volatile organic compounds and microorganisms. CRC Critical Reviews 4, 333381. Tong-Kwee, L., Keng, T.B., 1990. Antagonism in vitro of Trichoderma species against several basidiomycetous soil-borne plant pathogens and Sclerotium rolfsii. Journal of Plant Diseases and Protection 97, 3341. Tronsmo, A., Dennis, C., 1978. Eect of temperature on antagonistic properties of Trichoderma species. Transactions of the British Mycological Society 71, 469474. Tylka, G.L., Hussey, R.S., Roncadori, R.W., 1991. Axenic germination of vesiculararbuscular mycorrhizal fungi: eects of selected Streptomyces species. Phytopathology 81, 754759. Viterbo, A., Bar Nun, N., Mayer, A.M., 1992. The function of laccase from Botrytis cinerea. In: Verhoe, K., Malathrakis, N.E., Williamson, B. (Eds.), Recent Advances in Botrytis Research. Pudoc Scientic Publishers, Wageningen. Wheatley, R.E., Hackett, C., Bruce, A., Kundzewicz, A., 1997. Eect of substrate composition on production and inhibitory activity against wood decay fungi of volatile organic compounds from Trichoderma spp. International Biodeterioration and Biodegradation 39, 199205. Wheatley, R.E., Millar, S.E., Griths, D.W., 1996. The production of volatile organic compounds during nitrogen transformations in soils. Plant and Soil 181, 161168. White, N.A., Boddy, L., 1992. Dierential extracellular enzyme production in colonies of Coriolus versicolor, Phlebia radiata and

type and quantity of a suitable nutrient source within a soil system will dictate the range of VOCs produced by the microbial population. By altering the nutritional status of a soil system, it may be possible to impose a degree of control over the biodiversity and function of a soil community by controlling the quantity and types of VOC produced. Information regarding microbial processes such as nitrication and denitrication can be obtained by collecting the VOCs produced in a soil (Wheatley et al., 1996). Stahl and Parkin (1996) monitored the amounts of geosmin and 2-methylisoborneol produced by active groups of actinomycetes and bacteria within a controlled soil system, and demonstrated that when conditions are constant, key volatiles can be identied that identify microbial activity. Bacterial VOCs aect both fungal mycelial growth and enzyme activity. The VOC eects appear to be species, environment and age specic, as each fungus responded uniquely to the products of the dierent bacteria. Concentrations of nutrients available to bacteria greatly inuenced the ability of the VOCs produced to inhibit the fungi, as did the size of the bacterial population. Our results suggest that interactions mediated by VOCs produced by soil microorganisms may be widespread in soils, and that the outcome of these interactions will dier according to the microorganisms involved. Acknowledgements This work was funded jointly by the BBSRC under the SoilPlantMicrobe Interactions Initiative and the Scottish Oce Agriculture, Environment and Fisheries Department. References
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