Iranian Journal of Microbiology 2011. 3(4):216-221.

Impact of gamma rays on the Phaffia rhodozyma genome revealed by RAPD-PCR
N Najafi, R Hosseini, AR Ahmadi

Abstract


Background and Objectives: Phaffia rhodozyma is a red yeast which produces astaxanthin as the major carotenoid pigment. Astaxanthin is thought to reduce the incidence of cancer and degenerative diseases in man. It also enhances the immune response and acts as a free-radical quencher, a precursor of vitamin A, or a pigment involved in the visual attraction of animals as mating partners. The impact of gamma irradiation was studied on the Phaffia rhodozyma genome.
Materials and Methods: Ten mutant strains, designated Gam1-Gam10, were obtained using gamma irradiation. Ten decamer random amplified polymorphic DNA (RAPD) primers were employed to assess genetic changes.
Results: Nine primers revealed scorable polymorphisms and a total of 95 band positions were scored; amongst which 38 bands (37.5%) were polymorphic. Primer F with 3 bands and primer J20 with 13 bands produced the lowest and the highest number of bands, respectively. Primer A16 produced the highest number of polymorphic bands (70% polymorphism) and primer F showed the lowest number of polymorphic bands (0% polymorphism). Genetic distances were calculated using Jaccard's coefficient and the UPGMA method. A dendrogram was created using SPSS (version 11.5) and the strains were clustered into four groups.
Conclusion: RAPD markers could distinguish between the parental and the mutant strains of P. rhodozyma. RAPD technique showed that some changes had occurred in the genome of the mutated strains. This technique demonstrated the capability to differentiate between the parental and the mutant strains.


Keywords


astaxanthin; Phaffia rhodozyma; RAPD; UPGMA

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References


Medwid RD. Phaffia rhodozyma is polyploid. J Ind Microbiol Biotechnol 1998; 21: 228-232.

Weber RWS, Davoli P. Teaching techniques for mycology: 20. astaxanthin, a carotenoid of biotechnological importance from yeast and salmonid fish. Mycologist 2003; 17: 30-34.

Sun N, Seunghee L, Gilhwan A, Misun W, Kyung BS.Low-dose gamma irradiation as means of isolating carotenoid-hyperproducing yeast mutant. J Microbiol Biotechnol 2002; 12: 1010-1012.

Sun N, Lee S, Song KB. Characterization of a carotenoid-hyperproducing yeast mutant isolated by low-dose gamma irradiation. Int J Food Microbiol 2004;94: 263- 267.

El-Sherbeny S, Hanafy M, Hassan A, Amin M. RAPD analysis of DNA polymorphism in the yeast Hanssenula anomala and Rhodotorula rubra irradiated by gamma and fast neutrons. Int J Agri Biol 2005; 7: 942-946.

Dhakshanamorthy D, Selvaraj R, Chidambaram ALA.Induced mutagenesis in Jatropha curas L. using gamma rays and detection of DNA polymorphism through RAPD marker. Competes Rendus Biologies 2011; 334:24-30.

Ishii K, Yamada Y, Hase Y, Shikazono N, Tanaka A. RAPD analysis of mutants obtained by ion beam irradiation to hinoki cypress shoot primordia. Nuc Inst Meth Phys Res 2003; 206: 570-573.

Awan MS, Tabassam N, Ayub N, Babar M E, Rahman M, Rana, SM, et al. Gamma irradiation induced mutagenesis in Aspergillus niger to enhance its microbial fermentation activity for industrial enzyme production. Mol Biol Rep 2011; 38: 1367-1374.

Kaberi D, Naresh V. Exposure to low dose of gamma radiation enhances the excision repair in Saccharomyces cerivisiae. J Gen App Microb 1998; 44: 243-249.

Peak M, Peak G, Blazej E. Symposium report radiation induced DNA damage and repair. Int J Biol 1988; 54:513-519.

Fox J, McNally N. Cell survival and DNA double–strand break repair following x-ray or neutron irradiation of V79 cells. Int J Radiat Biol 1999; 54: 1021-1030.

Puchala M, Schessler H. Oxygen effect in the radiolysis of proteins. Int J Radiat Biol 1993; 64: 149-156.

Nagy A, Palagy Z, Ferenczy L, Vagvolgy C. Radiation induced chromosomal rearrangement as an aid to analysis of the genetic constitution of Phaffia rhodozyma FEMS Microb Lett 1997; 152: 249-254.

Hernandez P, Martin A, Dorado G. Development of SCARS of RAPD products. Mol Breed 1999; 3: 245-253.

Manabe M, Ino T, Kasaya M, Takum I, Nakamura O.Segregation distortion through female gametophytes in inter-specific hybrids of wheat as revealed by RAPD A analysis. Herediats 1999; 131: 47-53.

Ahmadi AR, Golkhoo S, Amin P, Aziz-Soltani M, Baran-Talab F, Delaram M. Isolation of astaxanthin overproducing mutant of Phaffia rhodozyma JH-82. Modern Genet J 2007; 1: 41-46.

Chung N (1996). DNA preparation from 10 ml yeast culture. uke.edu/web/ceramide/protocols/.

Atienzar FA, Venier P, Jha AN, Depledge MH.Evaluation of the random amplified polymorphic DNA (RAPD) assay for the detection of DNA damage and mutations. Mut Res 2002; 521: 151-163.

Atienzar F, Jha AN. The amplified polymorphic DNA (RAPD) assay and related techniques applied to genotoxicity and carcinogenesis studies: A critical review. Mut Res 2006; 613: 76-102.

Palágyi Z, Papp T, Takó M, Nagy Á, Pesti M, Vágvölgy C. Genetic variability of astaxanthin-roducing yeasts: random amplified polymorphic DNA (RAPD) analysis of Phaffia rhodozyma and Xanthopyllomyces dendrorhous. Acta Biologica Szegediensis 2004; 48:35-38.

Ginchner T, Zindar I, Szakova J. Evaluation of DNA damage and mutagenecity induced by lead in tobacco plants. Mut Res 2008; 652: 186-190.

Hadrys H, Balick M, Schierwater B. Applications of random amplified polymorphic DNA (RAPD) in molecular ecology. Mol Ecol 1992; 1: 55-63.


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