Pigment extraction from lichens with dimethyl sulfoxide (DMSO) and estimation of chlorophyll degradation

The role of carbonic anhydrase in the carbon-concentrating-mechanism of bryophytes of the class Anthocerotae was investigated by comparing the gas-exchange characteristics of material which had been incubated in the membrane-permeable Carbonic Anhydrase inhibitor ethoxyzolamide, with those of untreated material and material which had been incubated in buffer solution. In Phaeoceros laevis (Anthocerotae), incubation in ethoxyzolamide caused a depression in the rate of gross assimilation and a decrease in CO # affinity beyond that which could be attributed to increased diffusion limitation. A range of liverworts and mosses, in which a carbonconcentrating-mechanism is absent, were also investigated. These showed no depression of rates of gross assimilation after incubation in ethoxyzolamide relative to those of untreated material. The CO # compensation point and CO # uptake characteristics of Phaeoceros laevis were significantly affected by incubation in ethoxyzolamide. Values of CO # compensation point for Phaeoceros laevis rose from 2.5 Pa, after incubation in buffer, to 20 Pa after incubation in ethoxyzolamide. The CO # compensation point for the liverworts Pellia epiphylla and Marchantia polymorpha was not significantly affected by incubation in ethoxyzolamide. Measurements of the release of CO # at the end of a short (15 min) period of illumination revealed that, after suppression of carbonic anhydrase activity, the rapid release of a CO # pool occurred in Phaeoceros laevis but not in the liverworts. There were also significant differences between values for fractionation measured in units per mil (=), measured instantaneously, for Phaeoceros laevis incubated in ethoxyzolamide, compared with fractionation values for this species after incubation in buffer. Incubation in ethoxyzolamide caused fractionation values to rise from 12.4-22.7=, indicating that the carbon-concentrating-mechanism of this species had been inactivated. Incubation in ethoxyzolamide had no effect on fractionation values for the liverworts. The convexity of the light saturation curves of liverworts and Phaeoceros laevis was also investigated, but there were no differences between groups before or after the two treatments. The data indicate an important role for carbonic anhydrase in the functioning of the carbon-concentrating-mechanism in the Anthocerotae.

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“…Chlorophyll analysis was carried out on five replicate samples from each species of bryophyte (Ronen & Galun, 1984).…”

Section: Chlorophyll Analysismentioning

confidence: 99%

The role of carbonic anhydrase in photosynthesis and the activity of the carbon‐concentrating‐mechanism in bryophytes of the class Anthocerotae

Smith

Griffiths

2000

New Phytologist

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“…Rates of COj exchange were related to thallus dry weight (4 days at 70 °C), projected thallus area, thallus carbon concentration (elemental analyser; CHNORapid, Foss Heraeus, Hanau, FRG; also for thallus nitrogen determination), and total (a + è) chlorophyll concentration (Ronen & Galun 1984).…”

Section: Experimental Methodsmentioning

confidence: 99%

Lichen carbon gain under tropical conditions: water relations and CO₂ exchange of Lobariaceae species of a lower montane rainforest in Panama

et al. 2004

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Diel (24-h) time courses of microclimate, water relations, and COj exchange were measured under quasi-natural conditions at a forest edge in a lower montane, tropical rainforest in Panama for six Lobariaceae {Lobaria crenulata, L. dissecta^ Pseudocyphellaria aurata^ P. intñcata, Sticta sublimbata, S. weigelii). Responses to experimentally controlled water content (WC), photosynthetic photon fluence rate (PPFR), and temperature were studied in most detail with P. aurata.Photosynthesis was well adapted to high temperatures, and all species exhibited 'shade plant' characteristics with low light compensation points and low light saturation. Lobaria and Pseudocyphellaria species suffered from a strong depression of net photosynthesis (NP) at suprasaturating WC; suprasaturation depression was less in cyphellate Sticta species.Photosynthetic capacity correlated with thallus nitrogen concentration, and maximal NP rates of the cyanobacterial Sticta species was 4 to 5 times higher than that of the green algal Lobaria species. However, high rates of NP were uncommon and brief events under natural conditions; the different environmental factors were rarely optimal at the same time. Similar to earlier observations with other rainforest lichens, NP ceased during the period of highest irradiation on most days due to desiccation. During moist periods low light often limited carbon fixation, and high thallus hydration was often detrimental to NP. In spite of these limitations the maximal daily integrated net photosynthetic carbon income (SNP) was quite high especially for the Sticta species [17-3 and 24-1 mgC (gC) ^ ' day ^ ' for 5. sublimbata and 5. weigelii, respectively]. High nocturnal carbon loss, due to high night temperatures and continuous hydration, resulted in frequent negative diel carbon balances (EC) in all species. The average nocturnal carbon loss amounted to 83 and 70% SNP for P. aurata and P. intricata, respectively and to 64 and 59% of SNP for 5. sublimbata and S. weigelii, respectively. Their average diel SC was as high as 3-7 and 5-3 mgC (gC) ^ ' day ^ '. In contrast, SC were much lower for the other species, it amounted to only 0-18 mgC (gC) ^ ' day^ ' for L. crenulata. Thus, the Sticta species stood out amongst the species studied for their most successful adaptation of their photosynthetic productivity to the habitat conditions in the lower montane rainforest.

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“…Chlorophyll extraction with DMSO as solvent was conducted based on the method described by Ronen and Galun (Hiscox and Israelstam, 1978;Ronen and Galun, 1984) and on methodological adaptions made in the lab of O.L. Lange (pers.…”

Section: Chlorophyll Extraction With Dimethyl Sulfoxide (Dmso)mentioning

confidence: 99%

“…Our overall analytical technique is based on the photometric method established by Ronen and Galun (Ronen and Galun, 1984;Hiscox and Israelstam, 1978) and on methodological adaptions made in the lab of O.L. Lange (pers.…”

mentioning

confidence: 99%

Revisiting chlorophyll extraction methods in biological soil crusts – methodology for determination of chlorophyll a and chlorophyll a + b as compared to previous methods

Caesar¹,

Tamm²,

Ruckteschler³

et al. 2017

Preprint

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Abstract. Chlorophyll concentrations of biological soil crust (biocrust) samples are commonly determined to quantify the 10 relevance of photosynthetically active organisms within these surface soil communities. Whereas chlorophyll extraction methods for freshwater algae and leaf tissues of vascular plants are well established, there is still some uncertainty regarding the optimal extraction method for biocrusts, where organism composition is highly variable and samples comprise major amounts of soil. In this study we analyzed the efficiency of two different chlorophyll extraction solvents, the effect of grinding the soil samples prior to the extraction procedure and the impact of shaking as an intermediate step during 15 extraction. The analyses were conducted on four different types of biocrusts. Our results show, that for all biocrust types chlorophyll contents obtained with ethanol were significantly lower than those obtained with dimethyl sulfoxide (DMSO) as solvent. Grinding of biocrust samples prior to analysis caused a highly significant decrease in chlorophyll content for green algal lichen-and cyanolichen-dominated biocrusts, and a tendency towards lower values for moss-and algae-dominated biocrusts. Shaking of the samples after each extraction step had a significant positive effect on the chlorophyll content of 20 green algal lichen-and cyanolichen-dominated biocrusts. Based on our results we confirm a DMSO-based chlorophyll extraction method without grinding pretreatment and suggest to insert an intermediate shaking step for complete chlorophyll extraction (see supplement S6 for detailed manual). Determination of a universal chlorophyll extraction method for biocrusts is essential for the inter-comparability of studies conducted across all continents.

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Pigment extraction from lichens with dimethyl sulfoxide (DMSO) and estimation of chlorophyll degradation

Cited by 313 publications

References 21 publications

The role of carbonic anhydrase in photosynthesis and the activity of the carbon‐concentrating‐mechanism in bryophytes of the class Anthocerotae

The role of carbonic anhydrase in photosynthesis and the activity of the carbon‐concentrating‐mechanism in bryophytes of the class Anthocerotae

Lichen carbon gain under tropical conditions: water relations and CO₂ exchange of Lobariaceae species of a lower montane rainforest in Panama

Revisiting chlorophyll extraction methods in biological soil crusts – methodology for determination of chlorophyll a and chlorophyll a + b as compared to previous methods

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Pigment extraction from lichens with dimethyl sulfoxide (DMSO) and estimation of chlorophyll degradation

Cited by 313 publications

References 21 publications

The role of carbonic anhydrase in photosynthesis and the activity of the carbon‐concentrating‐mechanism in bryophytes of the class Anthocerotae

The role of carbonic anhydrase in photosynthesis and the activity of the carbon‐concentrating‐mechanism in bryophytes of the class Anthocerotae

Lichen carbon gain under tropical conditions: water relations and CO2 exchange of Lobariaceae species of a lower montane rainforest in Panama

Revisiting chlorophyll extraction methods in biological soil crusts – methodology for determination of chlorophyll a and chlorophyll a + b as compared to previous methods

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Lichen carbon gain under tropical conditions: water relations and CO₂ exchange of Lobariaceae species of a lower montane rainforest in Panama