Sie sind auf Seite 1von 9

2013

University of Toronto Peilu Gan - 997543163

[RADIOACTIVITY AND FUNGI, THE ROLE OF MELANIN]

Introduction
Melanin has long been identified as a key pigment molecule in almost all organisms. It has a many different biological functions, from its use as a defense mechanism against predators, to aiding the ability of cells to protect against harmful UV radiation. As a molecule, it exists in two forms, primarily Eumelanin, a dark brown pigment made of 5,6-dihydroxyindole (DHI) and 5,6-dihydroxyindole-2-carboxylic acid monomers, with relatively low sulfur content (<1%), and Pheomelanin, a reddish-brown pigment made of Benzothiazine/Benzothiazole subunits, with relatively high sulfur content (10-12%) (Ito S, 1985). The slightly different structure of these two melanin molecules conveys slightly different chemical properties, but is typically used in the same way in organisms theyre found in. As well, the widespread use of Melanin can also be attributed to the ease at which its synthesized (Ito S., 1985). Many fungi actively synthesize Melanin, and its two known roles so far are its uses as a virulence factor (Wheeler & Bell A., 1988), and as a method to protect the cells in harsh environments. From the extreme cold (Robinson, 2001) to the high levels of radioactivity of the Chernobyl reactors and coolant water, (Zhdanova et al. 2001), melanin has been identified in almost all fungal organisms living in these harsh environments. Studies looking into Fungal fossils have identified that almost all fungi that survived through the period in the past dubbed Magnetic Zero, where the Earth lost its ability to protect against cosmic radiation, were melanized (Hulot et al, 2003). In addition, lab results suggest melanin has a role on radioprotectivity (Mirchink et al. 1972). However, further analysis showed that a fungicidal dose for non-melanized radiation is 1.7104 Gray, which are orders of magnitude higher than levels found in Chernobyl (Saleh et al, 1988).

This suggested a new role of melanin in Fungi under ionizing radiation. Since pigments like Chlorophyll have long been known to be able to harvest light energy at very specific wavelengths, scientists have brought forth a new theory: that ionizing radiation changes the properties of melanin, possibly allowing fungus to use it as a method of harvesting energy (Dadachova et al., 2007).

Radiotrophic Fungi Isolated from Chernobyl Reactors


Radiotrophic fungi are fungi that are believed to be able to use external radiation as a source of energy. The origins of this were first discovered in 1991, when Zhdanova et al. discovered and extracted 5 samples of fungi, including Cladosporium cladosporioides and Penicillium roseo-purpureum, from soils surrounding the Chernobyl reactor (Zhdanova et al., 1991). When grown in the presence of radioactive particles (from the Chernobyl Reactor), it was seen fungi actively grew towards and into these particles, completely breaking them down over 50-100 days (Zhdanova et al, 1994). In a more recent study conducted by Zhdanova et al, where they used external beams of beta and gamma radiation (in the forms of 32P and 109Cd respectively) to control for confounding factors, researchers showed two-thirds of fungi actively grew towards radioactive sources (Zhdoanova et al, 2004). In total, they showed that two-thirds of all fungi tested showed significant directional growth towards the particles giving off gamma radiation. While not significant, there was still directional growth towards the sources of beta-radiation. From these studies, the authors concluded that fungi actively grew towards ionizing radiation, and proposed that Melanin might act in a similar manner to Chlorophyll, except with the ability to absorb a much wider wavelength.

Radiotrophic Fungi on the Russian Orbital Station, Mir


One unlikely source of Radiotrophic fungi was found on both the former Russian space station Mir, as well as the current International Space Station (Novikova et al, 2006). While it did exist as a microgravity research station, the fungal species isolated were not from the scientific experiments, implicating contamination by flight materials, the crew, or possibly other experiments. It was shown that, as long as some level of humidity was maintained, even in the presence of little or no other biological material, these fungi were able to survive, and even thrive ( Novikova et al). All fungi found on the station were intrinsically melanized, implicating melanin in aiding the survival of the fungi. Another interesting phenomena, regarding the role of fungi in space, was discovered in the Biorisk experiment (Novikova et al., 2007), conducted to identify if any fungal or bacterial organisms would undergo morphological changes that would become dangerous in space. This experiment showed a slew of morphological changes in the fungi tested (Penicillium expansum and Aspergillus versicolor), including increased melanin and increased mitochondrial production (Novikova et al., 2007) under nutrient-poor conditions. Considering that the level of background radiation, 4 centi-Gray per year, is nowhere near fungicidal, this also implies that the role of melanin is not acting as a shield against radiation.

Fungal Growth and Radiation


Another source of Fungi that appeared to thrive off some form of radiation was found at the Evolution Canyon of Israel. This canyon is characterized by the two slopes, with the southern-facing slope receiving up to 8-times more sunlight than the northern facing slope. As such, it was found that melanin levels differed between the northern-facing slopes compared to

the southern facing slope, even when comparing the exact same species. Specifically, the melanized fungi Aspergillus niger found on the northern-facing slopes were found to have melanin levels 3-fold higher compared to their southern counterparts (Singaravelan et al., 2008). In addition, their study found that the northern facing slopes survived much better under the presence of UVA radiation, and researchers proposed that melanin was induced in A. niger as an adaptive trait. However, when 11 fungi from the northern facing slope and similar counterparts were isolated from the southern facing slope, it was found that, up to 400 Gray of Gamma radiation, the growth rate of Fungi isolated from southern slope showed significantly higher growth compared to that found on the northern slope (explained by higher melanin levels), but when radiation was increased to 4,000 gray, the rate of growth of the fungi found on the Northern slope matched up to that found on the Southern slope. Upon 10,000 Gray of radiation, growth rate for both fungi slowed to a stop at about the same time. This suggests that the effect of radiation on growth is affected by the amount of melanin present in the fungi itself. Another study, conducted by Dadachova et al. (2007), looked at the effects of radiation on the growth of 3 separate fungal strains (Cryptococcus neoformans, Cladosporium sphaerospermum, and Wangiella dermatitidis. When they measured the fungis ability to grow under ionizing radiation, they showed that mealanized fungi they studied showed increased growth under radiation (2.4 fold increase in C. neoformans, 1.6 fold increase in C. sphaerospermum, and 1.3 fold increase in W. dermatitidis). However, they while they showed that melanized cells grew better in radiation, they also showed that non-irradiated non-melanized cells grew better than their melanized counterparts in

C. neoformans. As well, while there was a large increase in colony-forming units of melanized fungal cells under radiation, these colonies were a lot more immature compared to their nonmelanized counterparts, and had a lot smaller radii (8-26 fold less mass per colony). When the dry weight was measured for each of the 3 fungi studied, it showed only a 7-9% increase in dry weight under ionizing radiation. This suggested that the process of making melanin had negative effects on fitness for fungi that didnt intrinsically make melanin, likely due to toxic intermediates in the process of making melanin.

Mechanisms for the Metabolism of Radiation


Knowing that fungi showed increased growth under the presence of radiation, several hypothesis were proposed about the precise mechanisms of this action. One such study looked at the ability of radiation to affect the ability of Melanin to act as an electron donor (Gan et al., 1976). Irradiated melanin showed a 4-fold increase in ability to reduce NADH under the presence of melanin(Dadochova et al, 2007). However, what was curious was that all radiation, regardless including thermal, light, and ultraviolet, caused this increase in rate of reduction, showing that this rate was not affected by the level of electron energies, but rather the presence of the energy. Furthermore, they found that exposure to radiation caused an increased ability to metabolize, as measured an XTT (2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide) and MTT ( 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, which is typically used to measure cell viability (Mossman et al., 1983). Because of the inability of XTT to cross the cell membrane, this assay was also used to identify the localization of the increased metabolic activity, which showed that increased metabolism was localized to where melanin was located (Dadochova et al., 2007). However, melanized cells also showed a slight increase in

metabolic activity, which was theorized to be caused by the melanins ability to shrink cellular pores, likely causing the cell to increase cellular metabolism in order to survive. A more recent study looked at the effect of ionizing radiation on the ATP levels of a fungal cell. This experiment showed that, in melanized cells under all types of radiation (Gamma, Ultraviolet, and Sunlight), exposure actually decreased ATP levels (Dadochova et al, 2011). They proposed numerous mechanisms for this, one of which being that Fungus utilize the energy from the EM radiation in a way similar to photosynthesis, which causes ATP consumption but increases the amount of NADH in cells. This is supported by previous studies (Dadochova et al, 2007) (Zhdanova et al, 2008) which all show that melanized cells undergo better growth when exposed to ionizing radiation under limited nutrient conditions.

Conclusion
Overall, only one thing could be said about the role of radioactivity in melanin: that more study is required. Melanin polymers are still poorly understood, and their role in increasing cell size can only be speculated upon as of now, but we can say that the one property of melanin that makes it an amazing pigment is its capability to absorb all types of electromagnetic radiation. This confers the known ability to shield, but a still relatively unknown process in energy transduction. Extreme environments push the boundaries of evolution, some of which we are still discovering now, such as radiotrophic fungi: organisms that use the extremely dangerous radiation to promote their own growth.

Works Cited
Ito S, F. K. (1985). Microanalysis of eumelanin and pheomelanin in hair and melanomas by chemical degradation and liquid chromatography. Analytical Biochem(144), 527-536. Wheeler, M. H., & Bell A. A. (1988). Melanin and their importance in Pathogenic Fungi. Current Topics in Medicine, 238-387. Robinson CH (2001) Cold adaptation in Arctic and Antarctic fungi. New phytologist 151: 341 353 Wember VV, Zhdanova NN (2001) Peculiarities of linear growth of the melanin-containing fungi Cladosporium sphaerospermum Penz. and Alternaria alternata (Fr.) Keissler. Mikrobiol. Z. 63: 312. Mirchink TG, Kashkina GB, Abaturov ID (1972) Resistance of fungi with different pigments to radiation. Mikrobiologiia 41: 8386. Saleh YG, Mayo MS, Ahearn DG (1988) Resistance of some common fungi to gamma irradiation. Appl. Environm. Microbiol. Dadachova, E., Bryan, R. a, Huang, X., Moadel, T., Schweitzer, A. D., Aisen, P., Nosanchuk, J. D., et al. (2007). Ionizing radiation changes the electronic properties of melanin and enhances the growth of melanized fungi. PloS one, 2(5), e457. doi:10.1371/journal.pone.0000457 R. A. Bryan, Z. Jiang, M. Friedman, andE. DadachovaThe effects of gamma radiation, UV and visible light on ATP levels in yeast cells depend on cellular melanization. Fungal Biol. 115(10): 945-949 (2011). Hulot G, Gallet Y. Do superchrons occur without any palaeomagnetic warning? Earth Planetary Sci Lett. 2003;210:191201. Zhdanova, N., Lashkom T., Vasiliveskaya, A., Bosisyuk, L., Sinyaveskaya O., Gavrilyuk, V., and Muzale, P. Interaction of soil micromycetes with hot particles in a model system. Microbiologichny Zhurnal, 1991. Zhdanova NN, Redchitz TI, Krendyasova VG, Lashko TN, Gavrilyuk VI, Muzalev PI, Shcherbachenko AM. Tropism of soil micromycetes under the influence of ionizing radiation. Mycologiya i Fitopatologiya Singaravelan N, Grishkan I, Beharav A, Wakamatsu K, Ito S, Nevo E. Adaptive melanin response of the soil fungus Aspergillus niger to UV radiation stress at Evolution Canyon, Mount Carmel, Israel.PLoS ONE Gan EV, Haberman HF, Menon IA (1976) Electron transfer properties of melanin. Arch. Biochem. Biophys. 173: 666672.

Mosmann, Tim (December 1983). "Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays". Journal of Immunological Methods 65 (1 2): 5563. doi:10.1016/0022-1759(83)90303-4. ISSN 0022-1759. PMID 6606682 Novikova N, De Boever P, Poddubko S, Deshevaya E, Polikarpov N, Rakova N, Coninx I, Mergeay M. Survey of environmental biocontamination on board the International Space Station. Res Microbiol. 2006;157:512 Bryan, R., Jiang, Z., Friedman, M., & Dadachova, E. (2011). The effects of gamma radiation, UV and visible light on ATP levels in yeast cells depend on cellular melanization. Fungal biology, 115(10), 9459. doi:10.1016/j.funbio.2011.04.003 \