The effects of substrate water availability on the isolation and growth of fungi

The work reported in this thesis was carried out to contribute to the knowledge of the effects of substrate water availability or water activity (a ) on fungal growth parameters and its implications in the preparationw of materials susceptible to biodeterioration. Fungi were isolated from soils of different ecological sites at a range of substrate aw levels controlled by sodium chloride (NaCl). Three groups of fungi were isolated : firstly, those isolated only at high a (aw about 0.997).secondly, those isolated at high and decreasing aw (aw 0.997 to 0.85) and finally, those isolated at only decreased aw (aw O.95 to 0.80). From these isolations, test fungi were selected to study the effects of pH, temperature, exo-enzyme production and biocide efficacy at decreased aw levels, with glycerol and NaCl as a controlling solutes. The linear extension rates of the fungi increased at all test pH values near optimum a of growth. Test fungi of the Aspergillus glaucus group were found to be most resistant to low aw. Growth and survival of vegetative and fruiting bodies at elevated temperatures were enhanced with the addition of a controlling solutes. A. flavus, A. fumigatus displayed high heat resistance and A. amstelodami, A. versicolor and Penicillium citrinum displayed low heat resistance at high aw levels and vice versa at low aw levels. Amylase, lipase and protease activities were studied at lowered aw , using modifications of the test tube method of Raute11a and Cowling. Amylase and protease production in most xerophilic fungi ceased around 0.80 aw , but lipase production in some xerophilic fungi, including A. glatlcus fungi, was up to and including 0.70 aw with g1ycero1.

Effects of environmental temperature on members of the mucorales

The following investigation characterises the interaction between temperature and growth in psychrophilic, mesophilic and thermophilic fungi in order to gain further insight into the physiological mechanisms underlying fungal growth at extreme temperatures. In the first part of the investigation, the effect of environmental temperature on the growth of vegetative mycelium and sporangiospore production and germination was considered in order to determine the cardinal temperatures of these activities in different thermal groups. Subsequent investigations of plasma membrane permeability suggested that plasma membrane structure and function may be significant in establishing both the upper and lower growth temperature limits characteristic of psychrophiles, mesophiles and thermophiles. Analysis of the plasma membrane fractions revealed significant differences in membrane phospholipid composition between these thermal groups and it is suggested that the differing cardinal growth temperatures characteristic of psychrophilic, mesophilic and thermophilic fungi reflect the temperature ranges over which these organisms exhibit levels of plasma membrane fluidity sufficient to maintain membrane-associated growth processes. In contrast, the membrane protein components appear uniform in both character and thermostability and are therefore unlikely to contribute to this phenomenon.

The breakdown of plant cell biomass by fungi

Three species of fungi Sporotrichum thermophile, Botrytis cinerea and Trichoderma viride were assessed for their ability to utilize a variety of plant cell substrates (methanol extracted), Catharanthus roseus, Daucus carota, re-autoclaved C. roseus, re-autoclaved D. carota) which preliminary studies had indicated contained the necessary nutrients for fungal growth. Incubated in a suitable manner all three fungal species were able to grow on C. roseus and D. carota plant cell biomass in addition to material which had undergone methanol extraction or a re-autoclaving process to remove soluble components. Fungal biomass yields were markedly influenced by substrate, with each fungal species demonstrating a preference for particular plant cell material. Incubation conditions i.e. static or shaken and temperature also proved important. Release of glucose (i.e. values higher than Day 0) promoted by fungal breakdown of plant cell biomass was only noted with methanol extracted, re-autoclaved C. roseus and re-autoclaved D. carota material. A re-autoclaved substrate was also generally associated with high fungal C1, Cx, B-glucosidase and endo-polygalacturonase activity. In addition for each enzyme highest values were usually obtained from a particular fungal species. Buffering cultures at pH 3 or 5 further influenced enzyme activity, however in a majority of cases when flasks were unbuffered and the pH rose naturally to alkaline values higher enzyme activity was recorded. Likewise Tween 80 addition had only a limited beneficial effect. Finally filtrates containing glucose produced both from the re-autoclaving process and through fungal activity on plant cell biomass were utilized for Fusarium oxysporum, Saccharomyces cerevisiae and C. roseus plant cell culture. Although reasonable fungal biomass was obtained the use of such filtrates proved unsuitable for plant cell growth.

Lobe growth variation and the maintenance of symmetry in foliose lichen thalli

The aim of this study was to determine how thallus symmetry could be maintained in foliose lichens when variation in the growth of individual lobes may be high. Hence, the radial growth of a sample of lobes was studied monthly, over 22 months, in 7 thalli of Parmelia conspersa (Ehrh. Ex Ach.) Ach. And 5 thalli of P. glabratula ssp fuliginosa (fr. ex Duby) Laund. The degree of variation in the total radial growth of different lobes within a thallus over 22 months varied between thalli. Individual lobes showed a fluctuating pattern of radial growth from month to month with alternating periods of fast and slow growth. Monthly variations in radial growth of different lobes were synchronized in some but not in all thalli. Few significant correlations were found between the radial growth of individual lobes and total monthly rainfall or shortwave radiation. The levels of ribitol, arabitol and mannitol were measured in individual lobes. All three polyols varied significantly between lobes within a thallus suggesting that variations in algal phostosynthesis and in the partitioning of fungal polyols may contribute to lobe growth variation. The effect on thallus symmetry of lobes which grew radially either consistently faster or slower than average was studied. Slow growing lobes were overgrown, and gaps in the perimeter were eliminated by the growth of neighbouring lobes, in approximately 7 to 9 months. However, a rapidly growing lobe, with its neighbours removed on either side, continued to grow radially at the same rate as rapidly growing control lobes. The results suggested that lobe growth variation results from a combination of factors which may include the origin of the lobes, lobe morphology and the patterns of algal cell division and hyphal elongation in different lobes. No convincing evidence was found to suggest that exchange of carbohydrate occurred between lobes which would tend to equalize their radial growth. Hence, the fluctuating pattern of lobe growth observed may be sufficient to maintain a degree of symmetry in most thalli. In addition, slow growing lobes would tend to be overgrown by faster growing neighbours thus preventing the formation of indentations in the thallus perimeter.

Growth of crustose lichens: a review

Crustose species are the slowest growing of all lichens. Their slow growth and longevity, especially of the yellow-green Rhizocarpon group, has made them important for surface-exposure dating (‘lichenometry’). This review considers various aspects of the growth of crustose lichens revealed by direct measurement including: 1) early growth and development, 2) radial growth rates (RGR, mm yr-1), 3) the growth rate-size curve, and 4) the influence of environmental factors. Many crustose species comprise discrete areolae that contain the algal partner growing on the surface of a non-lichenised fungal hypothallus. Recent data suggest that ‘primary’ areolae may develop from free-living algal cells on the substratum while ‘secondary’ areolae develop from zoospores produced within the thallus. In more extreme environments, the RGR of crustose species may be exceptionally slow but considerably faster rates of growth have been recorded under more favourable conditions. The growth curves of crustose lichens with a marginal hypothallus may differ from the ‘asymptotic’ type of curve recorded in foliose and placodioid species and the latter are characterized by a phase of increasing RGR to a maximum and may be followed by a phase of decreasing growth. The decline in RGR in larger thalli may be attributable to a reduction in the efficiency of translocation of carbohydrate to the thallus margin or to an increased allocation of carbon to support mature ‘reproductive’ areolae. Crustose species have a low RGR accompanied by a low demand for nutrients and an increased allocation of carbon for stress resistance; therefore enabling colonization of more extreme environments.

The biology of the lichen Rhizocarpon geographicum: symbionts, growth, ecology, and lichenometry

Rhizocarpon geographicum (L.) DC. is one of the most widely distributed species of crustose lichens. This unusual organism comprises yellow-green ‘areolae’ that contain the algal symbiont which develop and grow on the surface of a non-lichenized, fungal ‘hypothallus’ that extends beyond the margin of the areolae to form a marginal ring. This species grows exceptionally slowly with annual radial growth rates (RGR) as low as 0.07 mm yr-1 and its considerable longevity has been exploited by geologists in the development of methods of dating the age of exposure of rock surfaces and glacial moraines (‘lichenometry’). Recent research has established some aspects of the basic biology of this important and interesting organism. This chapter describes the general structure of R. geographicum, how the areolae and hypothallus develop, why the lichen grows so slowly, the growth rate-size curve, and some aspects of the ecology of R. geographicum including whether the lichen can inhibit the growth of its neighbours by chemical means (‘allelopathy’). Finally, the importance of R. geographicum in direct and indirect lichenometry is reviewed.