Nitrogen and carbon isotope variability in the green‐algal lichen Xanthoria parietina and their implications on mycobiont–photobiont interactions

Stable isotope patterns in lichens are known to vary largely, but effects of substrate on carbon and nitrogen stable isotope signatures of lichens were previously not investigated systematically. N and C contents and stable isotope (δ¹⁵N, δ¹³C) patterns have been measured in 92 lichen specimens of Xanthoria parietina from southern Bavaria growing on different substrates (bark and stone). Photobiont and mycobiont were isolated from selected populations and isotopically analyzed. Molecular investigations of the internal transcribed spacer of the nuclear ribosomal DNA (ITS nrDNA) region have been conducted on a subset of the specimens of X. parietina. Phylogenetic analysis showed no correlation between the symbionts X. parietina and Trebouxia decolorans and the substrate, isotope composition, or geographic origin. Instead specimens grown on organic substrate significantly differ in isotope values from those on minerogenic substrate. This study documents that the lichens growing on bark use additional or different N sources than the lichens growing on stone. δ¹⁵N variation of X. parietina apparently is controlled predominantly by the mass fraction of the mycobiont and its nitrogen isotope composition. In contrast with mycobionts, photobionts of X. parietina are much more ¹⁵N‐depleted and show less isotopic variability than mycobionts, probably indicating a mycobiont‐independent nitrogen acquisition by uptake of atmospheric ammonia.     

Differenze Ultrastrutturali Di Alcune Specie Di Trebouxia Poste in Condizioni Di Illuminazione Differenti

Abstract

Ultrastructural changes in some species of « Trebouxia » under different light conditions. — Some species of the phycobiont alga Trebouxia (Tr. decolorans and Tr. albulescens), both isolated and grown on synthetic medium and still in the lichen, were examined in order to study the effect of light on the plastid ultrastructures. The species isolated from Buellia punctata and Xanthoria parietina were very sensitive to light condition and lost their chlorophyll content quickly. Striking ultrastructural changes were found in the algae grown under small light intensities and those which become achloric owing to strong light. In the latter, modifications of the Iamellar System were observed. The disappearance of Chlorophyll pigments was followed by a reduced electron density of the whole Iamellar system, as if were lacking the Iipidic compounds which are usually present and absorb fixators and dyers, thus allowing a good view. On the contrary, normal light conditions did not affect cultures of Trebouxia humicola, a free living alga. In the chloroplasts of the phycobiont species, unlike in the free living alga, grana were very close and sometimes formed very thick masses towards the edge of the chloroplast. It could not be ascertained whether such changes corresponded to a different composition of the lipoproteic compounds of the lamellar system.

Xanthoria parietina could grow in very lighted environments with no damage of the algae present in its thallus. The lichen thalluses, under different light conditions, showed very different colourings: the overlighted ones were rusty-red and the shadowed ones deep green. The chlorophyll content of the lichen thalluses with various shades (table 1) were very similar. The ultrastructural changes induced by strong light intensities in the phycobiont algae, kept in the lichen, were very small in respect of those observed in the same algae isolated and grown on synthetic medium and concerned the Iamellar system and the pyrenoid, above all. The rusty-red lichen showed a great number of stromatic lamellae, often with a parallel trend, so as to simulate a Iamellar system not organized in grana and often presented groups of lamellae concentrically arranged. In the pyrenoid of the algae from rusty-red thalluses, compared with the green ones, a much greater number of electron dense masses was observed, which are very thick and occupy the whole stromatic portion of the pyrenoid. But the Chlorophyll content did not decrease. Unlike the results of PEVELING, we noted that the electron dense masses (cited by the Author as « osmiophilic plastoglobules) were visible even after fixation with permanganate; the different numbers of these globules might depend on environmental factors. The phycobiont alga, when in the lichen thallus, could perhaps support strong light intensities, because pigments or compounds formed with the mycobiont or by it alone prevented the photooxidation of chlorophyll. Hypothetically a relationship might exist between the sensitivity of the phycobiont algae to light intensities and the content in antraquinonic pigments in the lichen thallus. But also using filters with absorption maxima similar to those of these pigments, the « in vitro » cultures of the phycobiont algae became achloric in the same time as the control ones.

Some Authors had found in Trebouxia humicola a different relationship between Chlorophyll pigments and carotinoids from that observed in the phycobiont species and had ascribed to it the greater resistence to strong light of the free living alga. Pigments or other substances present in the mycobiont can have a protective action on the Chlorophyll content and on the ultrastructures. In the phycobiont algae the resistence to strong light might be explained by an exchange of compounds between mycobiont and phycobiont, ending with the structural changes of the pyrenoid.     

Proteins from the liehenXanthoria parietina which bind to phycobiont cell walls Localization in the intact liehen and cultured mycobiont

Summary A protein fraction, previously isolated from the lichenXanthoria parietina and known to bind to the appropriate culturedTrebouxia phycobiont, was visualized in the intact lichen thallus and cultured mycobiont by an indirect immunoperoxidase assay. The protein was localized in both the upper and lower cortices of the lichen thallus; it was also present in the cell walls of the mycobiont culturedin vitro. The possible role of this protein in the recognition, or initial interaction, between separated lichen symbionts is discussed.     

Sterols of the lichen Pseudevernia furfuracea

A mixture of C₂₇, C₂₈ and C₂₉ sterols was isolated from the lichen Pseudevernia furfuracea and characterized by means of GLC and MS. Mono-, di- and tri-unsaturated sterols were identified as well as a small amount of fully saturated sterols (stanols). Only a part of the total sterols present in the lichen tissue was easily extractable with organic solvents. Another portion was only obtained after saponification of the lichen residue remaining after extraction with organic solvents. The composition of these two fractions difrered considerably, the former contained predominantly 5a,8a-epidioxy-5a-ergosta-6,22-dien-3β-ol (ergosterol peroxide) and 24-ethylcholesta-5,22-dien-3β-ol while in the latter 24-ethylcholesta-5,22-dien- 3β-ol and C₂₈ triene sterols were the main components.     

Sterols of Xanthoria parietina: Evidence for two sterol ‘pools’ and the identification of a novel C,8 triene,ergosta-5,8,22-trien-3β-ol

Sterols extracted from Xanthoria parietina with organic solvents and released by saponification of the residual lichen tissue were analysed by GC-MS. The main components of the solvent-extractable sterols were two C₂₈ trienes and those of the more tightly bound sterols were ergost-5-en-3β-ol and two C₂₉ compounds. The structures of the C₂₈ compounds were shown to be ergosta-5,7,22-trien-3β-ol, Ia (ergosterol) and the previously unreported ergosta-5,8,22-trien-3β-ol, IIa, for which the name lichesterol is proposed. The main C₂₉ sterol was identified as (24R)-24-ethylcholesta-5,22-dien-3β-ol (poriferasterol).     

OBSERVATIONS ON LICHENIZED AND FREE-LIVING PHYSOLINUM (CHLOROPHYTA,TRENTEPOHLIACEAE)1

Lichenized Physolinum Printz and free-living Physolinum from a dimly lit cave were studied from fresh collections and cultures, preserved specimens fixed in situ, and cultures that had persisted for 5 years in an environmental chamber. The branched filamentous association consists of a phycobiont and a characteristic ascomycetous mycobiont of one layer that completely ensheathes the algal partner. Epiphytic blue-green algae commonly occur attached to the mycobiont. The phycobiont, Physolinum monilia (De Wildem.) Printz, produces thick-walled, green spiny cells, some of which enlarge and contact the sheathing mycobiont cells; the phycobiont and mycobiont may then develop into new lichenized filaments. The hyaline mycobiont cells extend haustoria bound by the fungus wall deeply into the phycobiont chloroplasts. The epiphytes, Synechocystis-like colonies, are firmly attached to the outer walls of the mycobiont and are associated with several-celled extensions of the fungus beyond the apical phycobiont cells. Free-living Physolinum monilia filaments are branched and moniliform; the search-containing uninucleate cells are spherical to pyriform and have walls of cellulose. Each cell has a single massive chloroplast with plastoglobuli among tightly packed thylakoids. Except for their larger cells, P. monilia filaments appear to be identical to the phycobiont of lichenized Physolinum.     

The major sterols from three species of polyporaceae

The free sterols of the fungi Ganoderma applanatum, Ganoderma lucidum and Polyporus sulfureus were isolated and characterized by means of GC and GC/MS techniques. 24-Methylcholesta-7,22-dien-3β-ol was the main component of the sterol mixtures while 24-methylcholesta-5,7,22-trien-3β-ol ergosterol) and 24-methylcholest-7-en-3β-ol were also present although in lower amounts. P. sulfureus, besides the mentioned sterols, also contained 24 ethylcholestan-3β-ol.     

COMPARATIVE STUDIES ONXANTHORIA PARIETINA,A POLLUTION-RESISTANT LICHEN,ANDRAMALINA DURIAEI,A SENSITIVE SPECIES. II. EVALUATION OF POSSIBLE AIR POLLUTION-PROTECTION MECHANISMS

Surveys of the distribution of the lichensXanthoria parietinaandRamalina duriaeiin Israel showed that environments with air pollution had no damaging effects onX. parietina, whereasR. duriaeihad disappeared from polluted environments: physiological studies supported this relative sensitivity. Investigations of possible defence mechanisms protectingX. parietinafrom the damaging effects of air pollution showed a multitude of possible protective systems. These included constitutive avoidance such as: efficient buffering capacity; a relatively high potassium content; and antioxidation by parietin, and induced tolerance such as: SO₂oxidation to non-toxic sulphate; increased glutathione content; induced proline and arginine synthesis; and increased detoxification of active oxygen forms.     

SURFACEN-ALKANE VARIABILITY IN XANTHORIA PARIETINA

Surface alkanes were extracted from thalli of populations ofXanthoria parietinagrowing in two different Piedmont (Italy) valleys: Susa and Vermenagna. The mainn-alkanes detected in the Susa Valley were C₂₇, C₂₈, C₂₉and C₃₁, whereas C₂₅, C₂₇and C₂₉were the most abundant in Vermenagna Valley. The results indicate that then-alkane qualitative composition ofX. parietinawas affected both by elevation and climatic characteristics typical of the two valleys considered.     

The ultrastructure of the symbionts ofRhizocarpon geographicum,Parmelia conspersa andUmbilicaria pustulata growing under dryness conditions

Summary The dryness-induced ultrastructural changes of both myco- and phycobiont of three lichen species (R. geographicum, P. conspersa, andU. pustulata) have been studied over three months and half, period of time. During this time other ecological factors, such as rock substratum, temperature, light and gas interchange were unaltered compared to the natural conditions. A large number of ultrastructural changes were observed in the mycobiont as well as in the phycobiont (Trebouxia) and often, cells showed a highly disorganized morphology. The most important ultrastructural modifications were: 1. pyrenoglobuli of the algae were peripheral, 2. new and unknown structures were observed in the phycobionts of bothR. geographicum andU. pustulata as well as in the mycobiont of the latter species.     

Dehydrodinosterol,dinosterone and related sterols of a non-photosynthetic dinoflagellate, Crypthecodinium cohnii

The heterotrophic dinoflagellate Crypthecodinium cohnii contained the 4α-methyl sterols, dinosterol, dehydrodinosterol (4α,23,24-trimethylcholesta-5,22-dien-3β-ol) and the tentatively identified 4α,24-dimethyl-cholestan-3β-ol and 4α,24-dimethylcholest-5-en-3β-ol. The major 4-demethyl sterol was cholesta-5,7-dien-3β-ol which was accompanied by a smaller amount of cholesterol and traces of several other C₂₇,C₂₈ and C₂₉ sterols. In addition, a 3-oxo-steroid fraction was isolated and the major component identified as dinosterone (4α,23,24-trimethylcholest-22-en-3-one). The possible biosynthetic relationships of these compounds are discussed.     

Proteins from the lichenXanthoria parietina which bind to phycobiont cell walls. Correlation between binding patterns and cell wall cytochemistry

Summary A protein fraction was isolated from the lichenXanthoria parietina which bound to the appropriate cultured phycobiont, but not to the freshly isolated symbiotic alga. The protein also appeared to discriminate between five other strains of cultured phycobionts from different lichens; phycobionts isolated from lichens in the familyTeloschistaceae bound the protein whereas phycobionts isolated from lichens in other families did not. Using cytochemical techniques, it was shown that protein binding ability was correlated with high levels of acidic polysaccharide in the cell wall, and the presence of a protein coat on the cell wall surface of the phycobiont. The possible role of this protein in recognition between lichen symbionts is briefly discussed.     

24β-ethylsterols,n-alkanes and n-alkanols of Clerodendrum splendens

The sterols of Clerodendrum splendens, an angiosperm belonging to the family Verbenaceae, were found to possess a 24β-ethyl group. No other sterols were detected. The major sterol was 24β-ethylcholesta-5,22E,25(27)-trien-3β-ol [also known as 25(27)-dehydroporiferasterol]. A very small amount of what may have been its 22-dihydroderivative, clerosterol [also known as 25(27)-dehydroclionasterol] was also found. The dominant n-alkane was C₂₉ (n-nonacosane) and the dominant n-alkanol was C₂₈ (n-octacosanol).     

Investigations on the Δ23-, Δ24(28)- and Δ25-Sterols of zea mays

The sterols of Zea mays shoots were isolated and characterized by TLC, HPLC, GC/MS and ¹H NMR techniques. In all, 22 4-demethyl sterols were identified and they included trace amounts of the Δ²³-, Δ²⁴- and Δ²⁵-sterols, 24-methylcholesta-5,E-23-dien-3β-ol, 24-methylcholesta-5,Z-23-dien-3β-ol, 24-methylcholesta-5,25-dien-3β-ol, 24-ethylcholesta-5,25-dien-3β-ol and 24-ethylcholesta-5,24-dien-3β-ol. In the 4,4-dimethyl sterol fraction, cycloartenol and 24-methylenecycloartanol were the major sterol components but small amounts of the Δ²³-compound, cyclosadol, and the Δ²⁵-compound, cyclolaudenol, were recognized. These various Δ²³- and Δ²⁵-sterols may have some importance in alternative biosynthetic routes to the major sterols, particularly the 24β-methylcholest-5-en-3β-ol component of the C₂₈-sterols. Radioactivity from both [2-¹⁴C]MVA and [methyl-¹⁴C]methionine was incorporated by Z. mays shoots into the sterol mixture. Although 24-methylene and 24-ethylidene sterols were relatively highly labelled, the various Δ²³- and Δ²⁵-sterols contained much lower levels of radioactivity, which is possibly indicative of their participation in alternative sterol biosynthetic routes. (24R)-24-Ethylcholest-5-en-3β-ol (sitosterol) had a significantly higher specific activity than the 24-methylcholest-5-en-3β-ol indicating that the former is synthesized at a faster rate.     

Photosynthates stimulate the UV-B induced fungal anthraquinone synthesis in the foliose lichen Xanthoria parietina

Synthesis of the cortical anthraquinone pigment parietin (= physcion) was studied in acetone‐rinsed, parietin‐free Xanthoria parietina thalli. UV‐B induced the synthesis, which increased linearly with UV‐B (log‐transformed) to the highest applied UV‐B level (1.8 W m^?2). At natural UV‐B levels (0.75 W m^?2), parietin resynthesis occurred at a constant pace (106 mg m^?2 d^?1) during a 14‐d period at 220 µmol m^?2 s^?1 PAR. Under these conditions, 56% of the natural parietin content prior to extraction was resumed, accounting for 10% of total net carbon gain. In the presence of UV‐B, the remaining results were consistent with the hypothesis assuming that photosynthates regulate the pace at which parietin is synthesized by the mycobiont. Resynthesis was rapid when photosynthesis was activated by light, or when certain carbohydrates were added exogenously. Additions of ribitol, the carbohydrate delivered from the photobiont, increased the parietin resynthesis substantially. Mannitol, the main fungal polyol, was significantly less effective. Furthermore, parietin resynthesis in X. parietina was depressed at high and low hydration when net photosynthesis is depressed. Therefore, the photobiont regulates the parietin resynthesis pace in its mycobiont partner by the delivery of photosynthates. In conclusion, both lichen bionts play important roles in the synthesis of parietin, which probably acts as a PAR‐ rather than a UV‐B‐screen.     

Stereochemical aspects of the biosynthesis of the side chain of 9beta, 19-cyclopropyl sterols in maize seedlings treated with tridemorph

9β, 19-Cyclopropyl sterols such as 24-methyl pollinastanol accumulate dramatically in maize (Zea mays L. var LG 11) seedlings treated with Tridemorph (2,6-dimethyl-N-tridecyl-morpholine), a systemic fungicide (M. Bladocha, P. Benveniste, Plant Physiol 1983 41: 756-762). In contrast to the situation in control plants where 24-ethyl sterols predominate largely, 24-methyl sterols were more than 98% of total cyclopropyl sterols. In addition, 24-methyl cyclopropyl sterols were a mixture of (24-R)- and (24-S)-24-methyl epimers and are similar in that respect to the 24-methyl cholesterol of control plants. The presence of two epimers at C-24 has been previously explained by the operation of two routes (M. Zakelj, L. J. Goad, Phytochemistry 1983 22: 1931-1936). One may proceed via Δ²⁴⁽²⁸⁾- and Δ²⁴⁽²⁵⁾-sterols to produce the (24-R)-24-methyl epimer. The other route may involve reduction of either a Δ²⁴⁽²⁸⁾-, a Δ²³-, or a Δ²⁵-sterol intermediate to give the (24-S)-24-methyl epimer. Such intermediates have been searched for in excised Zea mays axes grown aseptically in the presence of Tridemorph and either [5-¹⁴C]mevalonic acid, or [Me-¹⁴C]-l-methionine. Whereas Δ²⁴⁽²⁸⁾- and Δ²⁴⁽²⁵⁾-cyclopropyl sterols were found in relatively large amounts, only traces of radioactivity were associated with Δ²⁵-sterols. Gas chromatography/mass spectrometry analysis of the sterols from axes grown in the presence of [Me-²H₃]-l-methionine showed that Δ²⁴⁽²⁸⁾-cyclopropyl sterols contained only two ²H atoms at C-28 as expected and that the 24-methyl pollinastanol fraction contained species with two ²H atoms and no species with three ²H atoms. These results indicate that both (24-R)- and (24-S)-epimers originate from a common Δ²⁴⁽²⁸⁾ precursor. After incubation of the axis with [5-¹⁴C,(4-R)-4-³H₁]mevalonic acid, the 24-methyl pollinastanol had a ³H:¹⁴C atomic ratio of 4:6 which is consistent with the intermediacy of a Δ²⁴⁽²⁵⁾-sterol. All these data are in accordance with a pathway where Δ²⁴⁽²⁸⁾-cyclopropyl sterols are isomerized to give Δ²⁴⁽²⁵⁾-cyclopropyl sterols which in turn would be reduced nonregiospecifically to yield both (24-R)- and (24-S)-24-methyl pollinastanols. A plausible mechanism for the reduction step is discussed.     

STEROLS OF 14 SPECIES OF MARINE DIATOMS (BACILLARIOPHYTA)1

The sterol compositions of 14 species of marine diatoms were determined by gas chromatography and gas chromatography-mass spectrometry. A variety of sterol profiles were found. The sterols 24-methylcholesta-5,22E-dien-3β-ol, cholest-5-en-3β-ol, and 24-methylcholesta-5,24(28)-dien-3β-ol, previously described as the most common sterols found in diatoms, were major sterols in only a few of the species. In light of this and other recent data, it is clear that these three sterols are not typical constituents of many diatom species. Most of the centric species examined had 24-methylcholesta-5,24(28)-dien-3β-ol and 24-methylcholest-5-en-3β-ol as two of their major sterols. The exception was Rhizosolenia setigera, which possessed cholesta-5,24-dien-3β-ol as its single major sterol. In contrast to the centric species, the pennate diatoms examined did not have any particular sterols common to most species. Minor levels ofΔ⁷-sterols, rarely found in large amounts in diatoms, were found in four species. C₂₉sterols were found in many species; seven contained 24-ethylcholest-5-en-3β-ol and three contained 24-ethylcholesta-5,22E-dien-3β-ol, reinforcing previous suggestions that C₂₉ sterols are not restricted to higher plants and macroalgae. 24-Ethylcholesta-5,22E-dien-3β-ol may prove to be useful for taxonomy of the genus Amphora and the order Thalassiophysales. A major sterol of Fragilaria pinnata was the uncommon algal sterol 23,24-dimethylcholesta-5,22E-dien-3β-ol. Cholesta-5,24-dien-3β-ol was the only sterol found in the culture of Nitzschia closterium. This differed from previous reports of 24-methylcholesta-5,22E-dien-3β-ol as the single major sterol in N. closterium. Two C₂₈ sterols possessing an unusual side chain were found in Thalassi-onema nitzschioides, a C_28:2 sterol (16%) and a C_28:1 sterol in lower abundance (2.5%), which may be 23-methylcholesta-5,22E-dien-3β-ol and 23-methyl-5α-cholest-22E-en-3β-ol, respectively. The species Cylindrotheca fusiformis, T. nitzschioides, and Skeletonema sp. may be useful as direct sources of cholesterol in mariculture feeds due to their moderate to high content of this sterol.     

Hydrocarbons,phenols and sterols of the testa and pigment strand in the grain of Hordeum distichon

Material from the testa of decorticated barley grains contained hydrocarbons, esters, triglycerides, free sterols, 5-n-alkylresorcinols, and traces of free alcohols, carbonyl compounds, and various polar, acidic materials. The hydrocarbon fraction was mainly a series of n-alkanes, extending at least from C₁₁ to C₃₆, in which the C₂₉ and C₃₁ components were prominent. Two minor series of alkanes were also present. Sometimes a trace of an unsaturated hydrocarbon was detected. The ester fraction contained sterols and alkanols esterified by fatty acids, which differed in relative amounts from the fatty acids found in the triglycerides. The triglycerides were thought to have leached from within the grain. At least five free sterols were present, including sitosterol and campesterol. The 5-n-alkylresorcinols were at least twelve members of a homologous series, of which four, C₂₅, C₂₇, C₂₉, and C₃₁, made 98% of the total. Members of the series with even numbers of carbon atoms were also present. It is suggested that they are partly responsible for excluding microorganisms from the interior of the grain. The testa membrane, with the associated pigment strand, contained an estolide of fatty acids and various hydroxyacids, a polysaccharide component, and uncharacterized material.     

ULTRASTRUCTURE OF THE LICHEN COENOGONIUM INTERPLEXUM NYL.

Coenogonium interplexum Nyl. is a green to yellow-orange filamentous lichen commonly found on tree bark, rocks, and soil. The mycobiont is the ascomycetous fungus Coenogonium. The ultrastructure of the lichenized phycobiont, Trentepohlia, closely resembles that of the non-lichenized form, a filamentous subaerial green alga. The mycobiont has a typical fungal ultrastructure, and the cell wall sometimes appears thinner at points of contact with the phycobiont wall. Several branched fungal hyphae are usually randomly arranged around a Trentepohlia filament, and may in some cases completely ensheath the alga. Although no haustoria were observed, this relationship may still be termed a lichen since there is some modification of the alga and the lichen is structurally distinct from the two symbionts.     

Carbon-dioxide exchange in lichens: determination of transport and carboxylation characteristics

Measurements were made of net rates of CO₂ assimilation in lichens at various ambient concentrations of CO₂ in air and in helox (79% He, 21% O₂). Because of the faster rate of CO₂ diffusion in the pores of lichen thalli when filled with helox than when filled with air, a given net rate of assimilation was achieved at a lower ambient concentration of CO₂ in helox. The differences were used to estimate resistances to diffusion through the gas-filled pore systems in lichens. The technique was first tested with five lichen species, and then applied in a detailed study with Ramalina maciformis, in which gas-phase resistances were determined in samples at four different states of hydration and with two irradiances. By assuming, on the basis of previous evidence, that the phycobiont in R. maciformis is fully turgid and photosynthetically competent at the smallest hydration imposed (equilibration with vapour at 97% relative humidity), and that, with this state of hydration, diffusion of CO₂ to the phycobiont takes place through continuously gas-filled pores, it was possible also to determine both the dependence of net rate of assimilation in the phycobiont on local concentration of CO₂ in the algal layer, and, with the wetter samples, the extents to which diffusion of CO₂ to the phycobiont was impeded by water films. In equilibrium with air of 97% relative humidity, the thallus water content being 0.5 g per g dry weight, the resistance to CO₂ diffusion through the thallus was about twice as large as the resistance to CO₂ uptake within the phycobiont. Total resistance to diffusion increased rapidly with increase in hydration. At a water content of 2 g per g it was about 50 times as great as the resistance to uptake within the phycobiont and more than two-thirds of it was attributable to impedance of transfer by water. The influences of water content on rate of assimilation at various irradiances are discussed. The analysis shows that the local CO₂ compensation concentration of the phycobiont in R. maciformis is close to zero, indicating that photorespiratory release of CO₂ does not take place in the alga, Trebouxia sp., under the conditions of these experiments.Symbols and Units rate of CO₂ diffusion in air relative to that in carrier gas (unity if the carrier gas is air and 0.43 if is helox) - A₁ net rate of CO₂ uptake by the lichen - A_p gross rate of carboxylation minus photorespiratory decarboxylation in the phycobiont, i.e. net rate of light-activated CO₂ exchange - A_* maximum, CO₂-saturated magnitude of A_p - c concentration of CO₂ - c_a ambient concentration of CO₂ - c_i c_a minus difference in CO₂ concentration across air-filled pore space in the thallus - c₈ CO₂ concentration equivalent to partial pressure of CO₂ at the surface of the phycobiont - ₁ magnitude of c_a at which A₁ = 0 - _* magnitude of c_* at which A_p = 0 - R rate of dark respiration in the lichen (mycobiont and phycobiont) - R rate of dark respiration in region between the surface of the lichen and an arbitrary distance from the surface within the thallus - r resistance to CO₂ transfer from lichen surface to the surface of the phycobiont - r resistance to CO₂ transfer between effective source of dark respiration in the lichen and the surface of the phycobiont - r_g, r _g components of r and r, respectively, attributable to transfer in air-phase - r_w, r _w components of r and r, respectively, attributable to transfer in water-phase - r component of r between surface of lichen and an arbitrary distance from the surface within the thallus - r_* resistance to CO₂ transfer and carboxylation in the phycobiont - RH relative humidity