Variation in hypothallus width and the growth of the lichen Rhizocarpon geographicum (L.) DC.

Variations in hypothallus width were studied in relation to radial growth in the lichen Rhizocarpon geographicum (L.) DC. in South Gwynedd, Wales, UK. Variations were present both within and between thalli and in successive three-month growth periods, but there was no significant variation associated with thallus size. In individual thalli, there were increases and reductions in hypothallus width in successive three-month growth periods attributable to hypothallus growth and changes at the margin of the areolae. Total radial growth over 18 months was positively correlated with initial hypothallus width. These results suggest: 1) individual thalli of similar size vary considerably in hypothallus width, 2) fluctuations in the location of the margin of the areolae in successive three month periods is an important factor determining this variability, 3) hypothallus width predicts subsequent radial growth over 18 months, and 4) variation in hypothallus; width is a factor determining between thallus variability in radial growth rates in yellow-green species of Rhizocarpon.

Soredial dispersal from individual soralia in the lichen Hypogymnia physodes (L.) Nyl.

Soredial dispersal from individual soralia of Hypogymnia physodes (L.) Nyl. was studied in the field under natural conditions and by exposing the soralia to an electric fan. Individual soralia were placed on the adhesive surface of dust particle collectors which were pinned to vertical boards in the field. The majority of soredia that were deposited on the adhesive strips during the experiments were found within 1 cm of the source soralium. Deposition was studied over 6 successive days under natural conditions. Significantly fewer soredia were deposited from soralia after removal of mature accumulations and from soralia taken from moist thalli compared with soralia from air dry thalli. In addition, there was a decline in soredial deposition over the 6 days. The influence of wind speed and initial thallus moisture content on soredial deposition over short intervals of time was studied using an electric fan. More soredia and larger soredial clusters were deposited from air dry than moist soralia at all wind speeds. Variation in wind speed between 4 and 9 m/sec had little effect on soredial deposition. Deposition of soredia was also studied using the fan over successive 5-min intervals. Large numbers of soredia were deposited during the first 5-min period. Deposition then declined but recovered after about four 5-min periods. In all experiments there were differences between individual soralia in total numbers of soredia deposited and in the pattern of deposition over time. These results suggest (1) soredia accumulate on soralia and these deposits may be gradually or rapidly depleted in the field, (2) that after the release of soredial accumulations some newly exposed soredia may be rapidly dispersed, (3) a high initial thallus moisture content inhibits soredial release and (4) variation in wind speed is less important than moisture in influencing soredial deposition. The results may help to explain the intermittent pattern of soredial deposition and the poor correlations between deposition and climatic factors observed previously in the field. © 1992.

Experimental studies of lobe growth in the lichen Parmelia conspersa (Ehrh. Ex Ach.) Ach.

The radial growth of individual lobes of the foliose lichen, Parmelia conspersa (Ehrh. Ex Ach.) Ach. was studied to determine whether (1) adjacent lobes exchange carbohydrate and (2) marginal competition between lobes influences radial growth. In a survey of thalli of different size, the number of marginal lobes was linearly related to thallus circumference. However, the relationship between mean lobe width and thallus circumference was fitted by a second order polynomial. Hence, mean lobe width may reach a maximum in thalli approx. 3 cm in diameter. The interactions between marginal lobes were studied by either painting single lobes with acrylic paint or by removing lobes from the thallus. Painting the whole lobe virtually stopped its radial growth while partially painted lobes grew less than control lobes. The radial growth of a lobe was unaffected by either completely painting or removing its neighbour. Removal of both neighbouring lobes did not influence the radial growth of a lobe but severing the lobe from the thallus reduced its radial growth. In addition, lobe width increased significantly when both neighbouring lobes were removed. These results suggest that adjacent lobes have a considerable degree of independence and that there is little exchange of carbohydrate between them. In addition, marginal competition between adjacent lobes may restrict the lateral extension of the lobe and this may maintain a more constant mean lobe width in larger thalli. It is possible that the intensity of marginal competition between adjacent lobes may vary with thallus size and this could be a factor determining the growth curve of a foliose lichen throughout its life.

Lobe growth in foliose lichen thalli

The margins of foliose lichen thalli comprise individual lobes which grow radially and divide. This results in a complex marginal structure in which lobes differing in morphology, state of division, and growth pattern are crowded together. Various aspects of the biology of these lobes are reviewed including their carbohydrate supply, morphology, pattern of division and branching, the effect of lobe overcrowding and interactions between neighbouring lobes. As the thalus grows, lobes become increasingly crowded together and this restricts further lateral growth. Restriction of lobe width may be reponsible for the changes in radial growth rate (RGR) with size observed in foliose species. Various aspects of lobe biology may be responsible for lobe growth variation including the physical independence of lobes from their neighbours, the genetic origin of the lobes, and the pattern of lobe branching. Overall symmetry of a thallus is maintained by a fluctuating pattern of growth of individual lobes in successive months together with competition for space at the margin

Competition between three lichen species using a factorial experimental design

Competition between three foliose lichens common on slate rock in south Gwynedd, Wales was studied in the field using a factorial experimental design. The lichens were grown as fragments glued to pieces of slate in two- and three-species mixtures. In the two-species mixtures, Parmelia conspersa (Ehrh. Ex Ach.) Ach. outcompeted Parmelia glabratula ssp. fuliginosa (Fr. ex Duby) Laund. strongly and Physcia orbicularis (Neck.) Poetsch less strongly, while P. orbicularis outcompeted P. glabratula weakly. Significant two-factor interactions indicated that the results from the three-species mixture could not be predicted from the two-species mixtures. Parmelia glabratula and P. orbicularis grew better in the presence of two competitors than one. This result suggests that the three species may co-occur on well-lit rock surfaces at the site.

Growth and regeneration of lichen thalli with the central portions artificially removed

The 12-month radial growth of Parmelia conspersa thalli with isidia or with apothecia and isidia was not influenced by removal of the thalli centres. When large thalli had their centres removed and the thallus perimeter was divided into fragments of about 1.0cm in diameter, growth of the fragments was less than the controls, but recovered to near control values after four or five months growth. These results suggest first, that fixed carbon for radial growth may be made in a narrow annulus at the perimeter and second, that there may be little transfer of fixed carbon between the annulus and the centre of the thallus ar around the annulus. Fragments of the centre and the perimeter regenerated growing points, suggesting that fragmentation may be an important method of vegetative reproduction in some lichens.

Development and growth of the lichen Rhizocarpon geographicum

The development of new areolae on the marginal hypothallus of the lichen Rhizocarpon geographicum (L.) DC was studied after complete or partial removal of the central areolae. New areolae developed slowly on the isolated hypothalli over two years. Development was similar when the areolae were completely removed and when the central areolae were separated from the marginal hypothallus by ‘moats’ 2 to 5 mm in width. However, in intact thalli, the marginal areolae developed rapidly during Jan. – June 1986 but showed periods of retreat from the margin during Oct. - Dec. 1985 and July – Sept. 1986. These results suggested that primary areolae may develop from free-living algal cells trapped by the hypothallus while secondary areolae may develop from zoospores produced by the thallus. Complete removal of the areolae resulted in no measurable radial growth of the marginal hypothallus over 18 months. Removal of the central areolae to within 1 and 2 mm of the hypothallus significantly reduced growth. These results suggest that the areolae may supply the hypothallus with carbon for growth. When the marginal hypothallus was experimentally removed a new hypothallus developed within one year. Regeneration occurred initially by retreat of the marginal areolae and later by new hyphal growth. The concentration of ribitol, arabitol and mannitol was measured in the areolae and marginal hypothallus on four occasions in 1985/6 in a population growing on a steep south facing rock surface. The three carbohydrates were present in significantly higher concentration in the areolae than in the hypothallus. Hence, the slow growth of this species may result from inhibited transport of carbohydrate from areolae to hypothallus.

The biology of the crustose lichen Rhizocarpon geographicum

The crustose lichen Rhizocarpon geographicum (L.) DC. comprises yellow-green lichenized areolae which develop and grow on the surface of a non-lichenized fungal hypothallus, the latter extending beyond the edge of the areolae to form a marginal ring. The hypothallus advances very slowly and the considerable longevity of R. geographicum, especially in Arctic and Alpine environments, has been exploited by geologists in dating the exposure age of rock surfaces (lichenometry). This review explores various aspects of the biology of R. geographicum including: (1) structure and symbionts, (2) lichenization, (3) development of areolae, (4) radial growth rates (RaGR), (5) growth physiology, (6) changes in RaGR with thallus size (growth ratesize curve), (7) maturity and senescence, and (8) aspects of ecology. Lichenization occurs when fungal hyphae become associated with a compatible species of the alga Trebouxia, commonly found free-living on the substratum. Similarly, 'primary' areolae develop from free-living algal cells trapped by the advancing hypothallus. The shape of the growth rate-size curve of R. geographicum is controversial but may exhibit a phase of decreasing growth in larger thalli. Low rates of translocation of carbohydrate to the hypothallus together with allocation for stress resistance results in very slow RaGR, a low demand for nutrients, hence, the ability of R. geographicum to colonize more extreme environments. Several aspects of the biology of R. geographicum have implications for lichenometry including early development, mortality rates, the shape of the growth-rate size curve, and competition. © The Author(s) 2012.

Formulation and characterisation of lyophilised rapid disintegrating tablets using amino acids as matrix forming agents

Despite recent advances in the formulation of lyophilised rapid disintegrating tablets (RDTs), the inclusion of matrix supporting/disintegration enhancing agents has been limited to the use of saccharides and polyols. In this study, the feasibility of using amino acids as matrix forming agents in lyophilised RDTs was investigated. Twelve amino acids were chosen (alanine, arginine, threonine, glycine, cysteine, serine, histidine, lysine, valine, asparagine, glutamine and proline), and the suitability for freeze drying, mechanical properties and disintegration time after inclusion of the amino acids at varied concentration were studied. In addition, the porosity of the RDTs and wettability profile of the amino acids were investigated to understand the mechanisms of disintegration. The results suggest the suitability of these amino acids for the lyophilisation regime, as they displayed satisfactory safety margin between the glass transition and shelf temperature (-40 degrees C), except proline-based formulations. Moreover, the crystallisation behavior of alanine, glycine, cysteine and serine at high concentration increased the stability of the formulation. The characterisation of the RDTs suggests that high concentration of the amino acids is required to enhance the mechanical properties, whereas only optimum concentrations promote the disintegration. Moreover, wetting time of the amino acid and porosity of the tablet are the two factors that control the disintegration of RDTs.

Dispersal in a population of the lichen Hypogymnia physodes

Dispersal of a Hypogymnia physodes (L.) Nyl. population was studied on an isolated Prunus blireiana L. tree at a site in North Seattle, U.S.A. Lichen propagules were trapped on adhesive strips pinned to four sites on the tree for 7 successive days. Soredia of H. physodes were frequently deposited on the strips but thallus fragments were rare. More soredia were deposited on the upper and lower branches than on the trunk, few soredia were deposited on the underside of the branches. The total daily deposition of soredia on the tree was positively correlated with average daily wind speed. Dispersal downwind from the tree was studied with squares of adhesive contact paper pinned to boards and located at intervals up to 25 m from the tree. Soredia and a few thallus fragments were recorded 25 m and 10 m, respectively, downwind on a day when average wind speed was 10.3 m/sec. The dispersal of soredia by wind from four individual thalli was studied over 10 successive days. Soredia were deposited from each thallus on each day mostly within 2 cm of the source. Higher wind speeds were necessary to dispersae soredia on days when the relative humidity was high. Soredia and thallus fragments were also dispersed by splash dispersal. More soredia were splashed furthest at a splash height of 90 cm. These results suggest that initial colonization of the tree by H. physodes may have occurred by wind-dispersed soredia. Subsequent spread probably occurred from established thalli mainly by the dispersal of soredia by wind and rain splash.

Factors associated with lobe division in the lichen Parmelia conspersa (Ehrh. ex. Ach.)Ach.

The factors associated with lobe division were studied in thalli of the lichen Parmelia conspersa (Ehrh. ex Ach.)Ach. Lobe division was studied in sequences of adjacent lobes using spatial pattern analysis. In five large thalli, lobe division within the thallus margin was randomly distributed. Correlations between the degree of lobe division, the radial growth of the lobe and lobe morphology were studied in six thalli. Lobe division was positively correlated with either lobe width or area in four thalli. Correlations were observed with radial growth or morphology of the adjacent lobes in two thalli. Dividing and non-dividing lobes were removed from large thalli and glued to pieces of slate with their tips either at the same level or in front of neighbouring lobes. Dividing lobes divided more rapidly when their tips were glued in front of their neighbours. The levels of ribitol, arabitol and mannitol were measured within a 2 mm region of the tip in dividing and non-dividing lobes on four occasions in 1994. Carbohydrate levels were significantly increased in dividing compared with non-dividing lobes. In addition, the mean size of the algal cells was greater in non-dividing compared with dividing lobes especially at the lobe base. However, the percentage of zoosporangia and aplanosporangia did not vary significantly in dividing and non-dividing lobes. These results suggest that: 1) the pattern of lobe division within the thallus margin may be random, 2) lobe division may be determined by lobe size and the location of the lobe tip relative to the neighbouring lobes and 3) there may be an increase in the productivity of lobes associated with lobe division.

Does radial growth of the lichen Parmelia conspersa depend exclusively on growth processes at the lobe tip?

The objective of this study was to test the hypothesis that the radial growth of lobes of the lichen Parmelia conspersa depends largely on growth processes which occur at the lobe tip. First, individual lobes were removed from thalli and portions of the lobe removed to within various distances from the tip. Radial growth of the lobe was unaffected until less than 2 mm of the lobe tip remained. Second, the surfaces of individual lobes were painted with acrylic paint leaving different portions of the lobe exposed. Painting lobes to within 0.5 mm and 1 mm of the tip substantially reduced radial growth. Third, the levels of ribitol, arabitol and mannitol were measured in different regions behind the lobe tip on four occasions during 1994. The concentration of the three carbohydrates was greatest at the lobe tip and the levels declined linearly with distance from the tip. Fourth, painting one vertical half of the lobe tip did not affect radial growth but artificially bisecting the lobe tip with a scalpel reduced radial growth. Although transport of carbohydrate from other regions of the lobe cannot be ruled out, the results support the hypothesis that radial growth in P. conspersa depends largely on processes within a region approximately 2 mm behind the lobe tip.

Factors determining the growth curve of the foliose lichen Parmelia conspersa

Growth curves of the foliose lichen Parmelia conspersa (Ehrh. Ex Ach.)Ach. Were obtained by plotting radial growth (RGR, mm yr-1) of the fastest measured lobe, the slowest measured lobe, a randomly selected lobe, and by averaging a sample of lobes from each thallus against thallus diameter. Growth curves derived from the fastest-growing lobe and by averaging lobes were asymptotic and could be fitted by the growth model of Aplin and Hill. Mean lobe width increased with thallus size, reaching a maximum at approx. 4.5 cm thallus diameter. In four out of six thalli, radial growth of lobes over four months was positively correlated with initial lobe width or area. The RGR of isolated lobes was unaffected until the base of the lobe was removed to within 1-2 mm of the tip. The concentration (micrograms mg-1 biomass) of ribitol, arabitol and mannitol was greater in the marginal lobes of large than in small thalli. The results suggested that the growth curve of P. conspersa is determined by processes that occur within individual marginal lobes and can be explained by the Aplin and Hill model. Changes in lobe width and in the productive capacity of individual lobes with thallus size are likely to be more important factors than the degree of translocation within the lobe in determining the growth curve.

Experimental studies of hypothallus growth in the lichen Rhizocarpon geographicum

Removal of the areolae of the crustose lichen Rhizocarpon geographicum (L.)DC. resulted in either low or no measurable radial growth of the marginal hypothallus. Radial growth of the hypothallus was also significantly reduced compared with intact thalli when (1) areolae were removed to within 1 and 2 mm of the hypothallus and (2) a 5 mm wide ‘moat’ was created between the areolae and the hypothallus. Adding ribitol (0.01 M) to isolated hypothalli at 3-month intervals over 15 months results in total radial growth c. 60% that of intact thalli. Adding an arabitol/mannitol mixture (0.05 M arabitol, 0.03 M mannitol) increased radial growth compared with deionized water and ribitol treatments. Adding ribitol (0.7 M), arabitol (0.2 M) and mannitol (0.08 M) to the areolae of intact thalli had no significant effects on radial growth of the hypothallus. On a south-facing rock surface, isolated hypothalli grew at a similar rate to intact thalli for 2 months. Growth then declined and the hypothalli disappeared from the rock surface within 6 months. The effects of addition of carbohydrate suggest that the marginal hypothallus has the capacity to utilize exogeneous materials. However, in intact thalli in the field, the radial growth of the hypothallus is likely to be a result of transfer of materials from the areolae through hyphal connections.

The levels of ribitol, arabitol and mannitol in individual lobes of the lichen Parmelia conspersa (Ehrh. ex Ach.) ACH.

The levels of the soluble carbohydrates ribitol, arabitol and mannitol were measured in individual lobes of the lichen Parmelia conspersa (Ehrh. ex Ach.) Ach. Lobes were collected from a north and a south facing slate rock surface in South Gwynedd, Wales, U.K. on 4 days during 1990-1991. On each day sampled, the most significant variation in soluble carbohydrate levels was between the individual lobes of a thallus. In addition, carbohydrate levels were significantly greater on the south facing rock surface on 2 of the 4 days sampled. Factorial analyses of variance suggested that the levels of individual carbohydrates varied significantly between days but not between north and south facing rock surfaces. Mannitol levels varied less between days than arabitol levels. Levels of ribitol, arabitol and mannitol were positively correlated in individual lobes. A stepwise multiple regression suggested that on the north facing rock surface, arabitol and mannitol levels could be explained by variations in the level of ribitol. By contrast, on the south facing rock surface, the levels of fungal carbohydrates were less dependent on the level of ribitol and there was evidence of a relationship between arabitol and mannitol. Variations in carbohydrate production, allocation and metabolism could help to explain lobe growth variation in foliose lichens and the radial growth of lobes over a longer period of time. Greater carbohydrate production rather than differences in allocation and metabolism may explain the increased growth and frequency of P. conspersa on south facing rock surfaces in South Gwynedd. © 1994.