Bioluminescence in FungiINTRODUCTIONWhat is Bioluminescence?The current paper main focus is on bioluminescent Fungi but the basic featuresof bioluminescence discussed are common to all bioluminescent organisms.
Bioluminescence is simply light created by living organisms. Probably the mostcommonly known example of bioluminescence by North Americans is the firefly,which lights its abdomen during its mating season to communicate with potentialmates. This bioluminescent ability occurs in 25 different phyla many of whichare totally unrelated and diverse with the phylum Fungi included in this list(an illustration of a bioluminescent fungi is displayed in figure 1).
One of thefeatures of biological light that distinguishes it from other forms of light isthat it is cold light. Unlike the light of a candle, a lightbulb, bioluminescentlight is produced with very little heat radiation. This aspect ofbioluminescence especially interested early scientists who explored it. Thelight is the result of a biochemical reaction in which the oxidation of acompound called “Luciferin” and the reaction was catalyzed by an enzyme called”Luciferase”.
The light generated by this biochemical reaction has beenutilized by scientists as a bioindicator for Tuberculosis as well as heavymetals. On going research involving bioluminescence is currently underway inthe areas of evolution, ecology, histology, physiology, biochemistry, andbiomedical applications.
History of Bioluminescent FungiThe light of luminous wood was first noted in the early writings ofAristotle which occurred in 382 B.C.(Johnson and Yata 1966 and Newton 1952) Thenext mention of luminous wood in the literature occurred in 1667 by RobertBoyle who noticed glowing earth and noted that heat was absent from the light.
Many early scientists such as Conrad Gesner, Francis Bacon, and Thomas Bartolinall observed and made notation of luminous earth(Johnson and Yata 1966 andNewton 1952 ). These early observers thought that the light was due to smallinsects or animal interactions. The first mention that the light of luminouswood was due to fungi occurred from a study of luminous timbers used as supportsin mines by Bishoff in 1823. This opened the way for further study by many otherscientists and by 1855 modern experimental work began by Fabre ( Newton 1952).
Fabre established the basic parameters of bioluminescent fungi, those being:- The light without heat – The light ceased in a vacuum, in hydrogen, andcarbon dioxide – The light was independent of humidity, temperature, light,and did not burn anybrighter in pure oxygenThe work by Herring (1978) found that the luminescent parts of the includedpileus(cap), hymenium(gills) and the mycelial threads in combination orseparately(figure 2) also the individual spores were also seen to be luminescent.
Herring also stated that if the fruiting body (mushroom) was bioluminescentthen the mycelial threads were always luminescent as well but not vice versa.
From the 1850’s to the early part of the 20th century theidentification of the majority of fungal species exhibiting bioluminescenttraits was completed. The research of bioluminescent fungi stagnated from the1920’s till 1950’s (Newton 1952 and Herring 1978 ). After which extensiveresearch began involving the mechanisms of bioluminescence and is still carriedout to the present.
The Process of BioluminescenceBioluminescence results because of a certain Biochemical reaction. This can bedescribed as a chemiluminescent reaction which involves a direct conversion ofchemical energy transformed to light energy( Burr 1985, Patel 1997 andHerring1978). The reaction involves the following elements:- Enzymes (Luciferase) – biological catalysts that accelerate and control therate of chemical reactions in cells. – Photons – packs of light energy. – ATP -adenosine triphosphate, the energy storing molecule of all living organisms. -Substrate (Luciferin) – a specific molecule that undergoes a chemical chargewhen affixed by an enzyme. – Oxygen – as a catalystA simplified formula of the bioluminescent reaction:ATP(energy) + Luciferin (substrate)+ Luciferase(enzyme) + O2(oxidizer) ==== light (protons)The bioluminescent reaction occurs in two basic stages:1) The reaction involves a substrate (D-Luciferin), combining with ATP, andoxygen which is controlled by the enzyme(Luciferase). Luciferins and Luciferasediffer chemically in different organisms but they all require molecular energy(ATP) for the reaction. 2) The chemical energy in stage one excites a specificmolecule (The Luminescent Molecule: the combining of Luciferase and Luciferin).
The excitement is caused by the increased energy level of the luminescentmolecule. The result of this excitement is decay which is manifested in the formof photon emissions, which produces the light. The light given off does notdepend on light or other energy taken in by the organism and is just thebyproduct of the chemical reaction and is therefore cold light. Thebioluminescence in fungi occurs intracellulary and has been noted at thespore level(Burr 1985, Newton 1952 and Herring 1978). This may at times bemistaken for a extracellular source of light but this is due to the diffusionof the light through the cells of the fungus. In examining the photograph infigure 1, it appear that the cap of the fungus is glowing but after study, itwas observed that just the gill structures that emits the light and the cap(which is thin) emits the light of the gills by diffusion(Herring 1978). Theenergy in photons can vary with the frequency (color) of the light. Differenttypes of substrates(Luciferins) in organisms produce different colors. Marineorganisms emit blue light, jellyfish emit green, fireflies emit greenish yellow,railroad worms emit red and fungi emit greeny bluish light (Patel 1997).
Fungal Families Exhibiting BioluminescenceThe phylum Fungi is composed of the following 5 divisions (Newton 1952):- Myxomycetes (slime molds) – Schizomycestes (bacteria) – Phycomycetes (moulds)- Ascomycetes ( yeasts, sac fungi and some molds) – Basidiomycetes (smuts, rusts,and mushrooms)Of the above divisions the majority of bioluminescence occurs in theBasidiomycetes and only one observation has been made involving the Ascomycetes;specifically in the Ascomycete genus Xylaria (Harvey 1952). At present thereare 42 confirmed bioluminescent Basidiomycetes that occur world wide and shareno resemblance to each other visually, other than the ability to bebioluminescent. Of these 42 species that have been confirmed 24 of these havebeen identified just in the past 20 years and as such many more species mayexhibit this trait but are yet to be found. The two main genus that displaybioluminescence are the genus Pleurotus which have at present 12 species whichoccur in continental Europe and Asia. The genus Mycena have 19 speciesidentified to date with a world wide distribution range. In North America only5 species of bioluminescent basiodiomycetes have been reported. These includethe Honey mushroom -Armillaria mellea (illustrated in figure 3), the commonMycena -Mycena galericulata (illustrated in figure 1), the Jack O’Latern -Ophalalotus olearius (pictured in figure 4), Panus styticus and Clitocybeilludens. The question of whether bioluminescent mushrooms were all poisonouswas raised in the discussions between my laboratory partner and myself. Afterexamining the literature and a mushroom field guide book it was evident thatthere was no correlation between the edibility of the mushroom and itsbioluminescence. Some mushrooms such as Armillaria mellea the Honey mushroomwas listed as being excellent to eat. While the Jack O’Latern – Omphalalotusolearius was listed as poisonous and caused sever gastrointestinal cramps. Theedible merits of the common Mycea were unknown and while Panus stypticus waslisted as poisonous it was found to contain a clotting agent and useful instopping bleeding (Lincoff 1981, Newton 1952 and Herring 1978). As it only afield guide to North American mushrooms was available, only the North Americanvarieties were examined. If all 42 species of bioluminescent basidiomyceteswere included in the search, a possible correlation may have been found.
Bioluminescence Research ApplicationsLuminescence has a unique advantages for scientific studies as it is the onlybiochemical process that has a visible indicator than can be measured. Thelight given off in the bioluminescent reaction is now able to be accuratelymeasured with the use of a luminometer. This ability to easily and accuratelydetect small amounts of light has led to the use of the bioluminescent reactionin scientific research involving biological process applications. The followingare just a few applications, some of which have been developed in only the lastfew years (Johnson and Yata 1966, and Patel 1997). The following are twoexamples of which have been recently developed.
The Tuberculosis TestTesting for tuberculosis has long been a problem because of the long time ittakes for the species to grow to a size that is detectable by modern medicine.
Typically growing a culture of Mycobacterium tuberculosis large enough todetermine the strain that a particular patient has can take up to three months.
Of course, this poses a problem because the patient often can not wait for thediagnosis and must be given drugs that his strain may be resistant to. This isfurther complicated because there are 11 drugs used to combat TB, picking theright one before determining the strain has a 1/11 chance of success. Recentlya way of incorporating bioluminescence into the TB tests has been found and cansharply reduce the diagnosis time to as little as 2 days. The techniqueinvolves inserting the gene that codes for luciferase into the genome of the TBbacterial culture taken from the patient. The gene is introduced through a viralvector and once incorporated, the bacteria produces the luciferase. Whenluciferin is added to the culture, light is produced. Since less than 10,000bacteria are needed to code for enough luciferase to produce a detectable amountof light, the culture time is reduced to only 2-3 days. Since the luciferase-luciferin reaction requires ATP, the resistance of the strain in the culture canbe tested by adding a drug and watching for light. This will indicate which ofthe 11 drugs therapy’s will be effective in treating Tuberculosis. By reducingthe time needed to prescribe the correct drugs for treatment, this applicationof bioluminescence will someday be ready to save some of the 3 million killedeach year by tuberculosis (Patel 1997).
BiosensorsBioluminescence has also been used for several years as a biosensor of manysubstances. As seen in the tuberculosis example, bioluminescence can be used asensor for the presence of ATP because ATP is needed in the light producingreaction. Other techniques have been used for detecting ions of mercury andaluminum, among others, by using bacteria with light genes fused to their ion-resistant regulons. For example, if a bacteria that is resistant to Hg is in thepresence of Hg, the genes coding for its Hg resistance will be activated. Theactivation of that gene will also activate the luciferase gene fused to it, sothe bacteria will produce luciferase whenever Hg is present. Adding luciferinand testing for light production with a luminometer reveals the presence of themetal ion in the solution. This technique is especially useful in testing forpollutants in the water supply when concentrations are too low to detect byconventional means(Herring 1978, and Patel 1997). Other areas that are currentlyusing bioluminescence in scientific research include evolution, ecology,histology, physiology, biochemistry, biomedical applications, cytology andtaxonomy. Any area that involves a living organism can utilize bioluminescenttechnology as a biosensor.
ConclusionThe glow light generated by bioluminescent Fungi has for centuriesgenerated interest from philosophers and scientists and has benefited science byproviding problems to solve -How does it work and does it have a practicalapplication? The answers to those basic problems that have been discoveredtoday and have resulted in benefiting mankind, by bettering our livesespecially in regard to it’s biomedical applications. Further research withbioluminescent Fungi is being conducted on a world wide scale and include NorthAmerica, Japan, and Europe. Future research may lead to new discoveries anduses from bioluminescent organisms such as the Fungi group.
ReferencesBurr, G.J. 1985. Chemiluminescence and Bioluminescence. Marcel Dekker, Inc. NewYork, U.S.A.
Johnson, F. H. and Yata, H. 1966. Bioluminescence in progress. Princton, NewJersey, Princeton University Press.
Lincoff,G.H. 1981. The Audubon Society field guide to North American Mushrooms.
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Newton, H.E. 1952. Bioluminescence. Academic Press. New York. U.S.A.
Herring, P.J. 1978. Bioluminescence in Action. Academic Press. New York. U.S.A.
Patel, P.Y. 1997. Bioluminescence in scientific research. Jan 10, 1997.
Http://www. emailprotectedWood, M.F. and Stevens, F. 1997. The Myko web page -Fungi Photos. Jan 10, 1997.
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