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Detoxification of Dissolved SO2 (Bisulfite) by Terricolous Mosses

Group 1 De Juan, Michelle Ligaya E. Gamboa, Domina Flor L. Manalaysay, Jessica Alba G. Matundan, Celine Marie C.

*Terricolous: Land-dwelling *Sulfur dioxide exists in these forms: SO2, SO3-2 and HSO3- (bisulfite)

INTRODUCTION

Pleurozium schreberi - calcifuge moss - moderately tolerant of SO2 Rhytidiadelphus triquetrus - calcareous or calcicole moss - strongly affected by SO2 pollution in the 20th century

Pleurozium schreberi

Source: http://commons.wikimedia.org/wiki/File:Rhytidiadelphus_triquetrus.JPG

Rhytidiadelphus triquetrus

Source: http://commons.wikimedia.org/wiki/File:Pleurozium.schreberi.jpg

Tolerance of SO2 is seen


on the metabolic detoxification of dissolved bisulfite by these mosses

Bryophytes (as well as Lichens) are sensitive to atmospheric pollution, particularly SO2 - Limited cuticle - High surface area - Low metabolic activities - Modest innate growth rates

* Detoxification Resistant bryophytes: relatively high growth rates: which means that they have the ability to detoxify
* In higher vascular plants: tolerance on SO2 is by detoxification mechanisms (excluding tolerance by cuticle and stomata) oxidized to sulfate ion (SO4-) OR reduced to sulfide

*SO2 Phytotoxicity
Intracellular O2production as cause of SO2 phytotoxicity

*Photo-oxidation: oxidation in the presence of radiant energy (light)

*SO2 Phytotoxicity
Intracellular production as cause of SO2 phytotoxicity O2Superoxide dismutase -Active in SO2 tolerant plants - Catalyses decomposition of O2- Inhibits photo-oxidation of SO2

*Photo-oxidation: oxidation in the presence of radiant energy (light)

*SO2 Phytotoxicity
Intracellular production as cause of SO2 phytotoxicity O2Superoxide dismutase -Active in SO2 tolerant plants - Catalyses decomposition of O2- Inhibits photo-oxidation of SO2

Diethyl dithiocarbamate (DETC) - Controls activity of superoxide dismutase

*Photo-oxidation: oxidation in the presence of radiant energy (light)

* Detoxification like higher vascular plants in bryophytes: Sphagnum - Higher tolerance for plants in more polluted areas - Oxidation of bisulfite: brought about by metal cations (Fe3+, Mn2+ and Cu2+)

*Focus of the study Pleurozium schreberi Rhytidiadelphus triquetrus


In dilute bisulfite solutions: Photosynthesis in these mosses was strongly inhibited by short (2-hour) incubations with bisulfite Longer incubations: no effect Shoots have a high capacity to detoxify dissolved SO2

*Hypotheses (a) tolerance of bisulfite by P. schreberi and R. triquetrus depends primarily on detoxification (oxidation) of the pollutant (b) bisulfite detoxification involves metabolic energy (c) Ca2+ and Fe3+ stimulate the detoxification process (d) SOD is involved in bisulfite detoxification

MATERIALS AND METHODS

(1) Pleurozium schreberi: collected from an acid, sandy loam soil under grassland and scrub (2) Rhytidiaelphus triquetrus: was collected from chalk grassland on a rendzina soil

(1) Incubation treatments Short-term incubation experiment Long-term incubation experiment * Reagent used: NaHSO3 (2) Bisulfite disappearance in relation to initial concentration (3) DCMU experiment (3-( 30,40dichlorophenyl)-1,1-dimethylurea) - Inhibits photosynthetic electron transport and oxygen evolution

(4) DETC experiment (diethyldithiocarbamate) - Inhibitor of superoxide dismutase - To see if the enzyme plays a role in bisulfite oxidation (5) Bisulfite oxidation: influence of Ca2+ and Fe3+ (6) Bisulfite and sulfate determinations - Spectrophotometric methods (7) Statistical analyses - One-way ANOVA - Duncans multiple range test

RESULTS

* Disappearance of bisulfite during short-term incubations with moss shoots


- Decay of bisulfite greater in the presence of light for both species - Rhytidiadelphus triquetrus: has a greater capacity in catalyzing disappearance of bisulfite

*Disappearance of bisulfite during long-term incubations


- Absence of mosses: 28% remaining at the end of the fifth day of incubation - Presence of mosses: 95% of bisulfite had disappeared after a three day incubation

*Disappearance of bisulfite in relation to initial concentration


- Disappearance of bisulfite: dependent on initial concentration and presence of light (1) Pleurozium schreberi: initial concentration of bisulfite is roughly proportional to the final; no significant effect of light (2) Rhytidiadelphus triquetrus: total bisulfite lost increased in the presence of light

*Disappearance of bisulfite in relation to acidity


- pH 3 to 5: small reduction in final volume of bisulfite for both species * Much greater reduction at pH 6

*Effects of DCMU on disappearance of bisulfite


- Presence of DCMU in both light and dark: great bisulfite persistence - Twice as much bisulfite remained in the incubation solutions of Pleurozium schreberi as in those of Rhytidiadelphus triquetrus

*Effects of DETC on disappearance of bisulfite


- inhibited bisulfite loss from the incubation medium in both species - Pleurozium schreberi: doubling of the concentration of bisulfite remaining - Rhytidiadelphus triquetrus: even greater increase in concentration of bisulfite remaining

*Evidence for extracellular oxidation of bisulfite to sulfate: influence of Ca and Fe


- Pleurozium schreberi Fe: no difference Ca: more sulfate was detected EDTA: no change or small increase in sulfate - Rhytidiadelphus triquetrus Fe: small but significant increase in sulfate Ca: reduction in sulfate production EDTA: reduction in sulfate

DISCUSSION

- Addition of bisulfite Rapid cessation of photosynthesis Increasing incubation periods: photosynthesis restored Due to oxidation of bisulfite - Presence of the two mosses Greatly accelerates decrease in amount of bisulfite in incubation solutions

- Rate of bisulfite loss depends on: (1) Presence or absence of light (2) Application of metabolic inhibitors (3) Acidity (4) Nature and concentrations of adsorbed metal cations (5) Species of moss

- Light significantly stimulated bisulfite loss from the external solution - Differences in the degree of photoprotection between the two mosses might also explain their different abilities to detoxify bisulfite solutions

- DCMU inhibits photosynthetic electron transport and oxygen evolution. It caused a substantially reduced rate of bisulfite loss from the incubation solution, especially in R. triquetrus

- Experiment with DETC, an inhibitor of SOD, led to a very significant reduction in the rate of bisulfite loss from the incubation solution with both mosses

- Fe(III) catalysed extracellular oxidation of bisulfite as the pretreatment was accompanied by an increase in the sulfate concentration of the external solution.
- The reduced extracellular sulfate observed could indicate that Ca(II) enhances cellular uptake of bisulfite (indirect effect).
- Ca(II) functions for stabilizing cell membranes or embedded portein channels against loss of permeability control

- EDTA pretreatement did not cause lowered bisulfite disappearance - Possibly, EDTA was relatively ineffective in chelating metals such as Fe(III) from the moss shoots. - Alternatively, the EDTA may have disrupted normal membrane function so that bisulfite uptake or retention rates were modified

CONCLUSION

*Loss of bisulfite (1) External oxidation of bisulfite using metabolic (including photooxidative) energy
(2) passive external oxidation of bisulfite catalysed by adsorbed Fe3+ ions (3) cellular uptake and metabolic detoxification of bisulfite

Authors Bhagawan Bharali Jeffrey Bates


Department of Crop Physiology, Assam Agricultural University, Jorhat-785013, Assam, India and 2Division of Biology, Imperial College London, Silwood Park Campus, Ascot, Berkshire SL5 7PY, UK