International Journal of Biotechnology and Molecular Biology Research
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Article Number - D23B6BA56446


Vol.6(7), pp. 48-57 , December 2015
DOI: 10.5897/IJBMBR2014.0207
ISSN: 2141-2154



Full Length Research Paper

Characterization of gamma-irradiated seeds of a wild Namibian marama bean (Tylosema esculentum) with microsatellite markers



Takundwa, M. M.
  • Takundwa, M. M.
  • Department of Biological Sciences, University of Namibia, Windhoek, Namibia.
  • Google Scholar
Ruzvidzo, O.
  • Ruzvidzo, O.
  • Department of Biological Sciences, North-West University, Private Bag X2046, Mmabatho, 2735, South Africa.
  • Google Scholar
Nambahu, F.
  • Nambahu, F.
  • Department of Biological Sciences, University of Namibia, Windhoek, Namibia.
  • Google Scholar
Kawadza, D. T.
  • Kawadza, D. T.
  • Department of Biological Sciences, North-West University, Private Bag X2046, Mmabatho, 2735, South Africa.
  • Google Scholar
Chatukuta, P.
  • Chatukuta, P.
  • Department of Biological Sciences, North-West University, Private Bag X2046, Mmabatho, 2735, South Africa.
  • Google Scholar
Chimwamurombe, P. M.*
  • Chimwamurombe, P. M.*
  • Department of Biological Sciences, University of Namibia, Windhoek, Namibia.
  • Google Scholar







 Received: 31 August 2014  Accepted: 25 November 2015  Published: 31 December 2015

Copyright © 2015 Author(s) retain the copyright of this article.
This article is published under the terms of the Creative Commons Attribution License 4.0


Legumes have been used as food by indigenous people of Africa yet their full potential as food sources has not been completely realized. With modern technologies such as marker-assisted selection and mutational breeding, there is scope for rapid improvement of germplasm in these underutilized species. One approach is to induce genetic mutations through irradiation of plant tissues with neutrons or gamma rays generating molecular changes. The irradiation leaves residual radiation in the exposed tissues creating offspring with new features. Genetic mutations in plants can improve their performance, germination, quality, size and disease resistance. The effects of radiation applied to marama bean (Tylosema esculentum) were explored. The aim of this study was to screen individuals of T. esculentum derived from seeds irradiated with gammarays using microsatellite markers. There were two irradiation treatments -50 and 100 gamma ray irradiations that were compared to non-irradiated seeds as controls. For each treatment, seeds were imbibed and germinated. Seed emergence rates and percentages were ascertained. Each set of seeds was genotyped using a SSR marker - MARA039, located in a sub-unit of a polygalacturonase – an enzyme that mainly regulates seed germination. In regard to emergence, significant differences between the irradiated seeds and non-irradiated counterparts were noted. Irradiated seeds germinated faster and in adosage-dependent manner compared to non-irradiated counterparts (p>0.05). Genotyping revealed that the total number of AGA repeats in the microsatellite region shifted from its original five in the controls to between four and seven in the irradiated seeds. Furthermore, there were several point mutations in the irradiated seeds as compared to the controls. The study findings, suggest that irradiation may induce beneficial mutational changes such as faster germination rate of the marama bean, considering that the mutated microsatellite repeat region is located in gene encoding the enzyme regulating seed germination in plants.

Key words: Marama bean, Tylosema esculentum, mutational breeding, irradiation, seed cycle, SSR.

Azam-Ali SN (1996). Bambara groundnut: How did we get here? Proceedings oftheInternational Bambara Groundnut Symposium, University of Nottingham, UK.

 

Baskin CC, Baskin JM (1998). Seeds: Ecology, biogeography, and evolution of dormancyand germination. California: Academic Press.

 

Batista R, Saibo N, Lourenco T, Oliveira MM (2008). Microarray analysis reveals that plant mutagenesis may induce more transcriptomic changes than transgene insertion. Proc. Nat. Acad. Sci. USA 105:3640-3645.
crossref

 

Bonghi C, Rascio N, Ramina A, Casadoro G (1992). Cellulase and polygalacturonase involvement in the abscission of leaf and fruit explants of peach. Plant Mol. Biol. 20:839-848.
crossref

 

Castro S, Silveira P, Pereira-Coutinho A, Figueiredo E (2005). Systematic studies in Tylosema (Leguminosae). Bot. J. Linn. Soc. 147:99-115.
crossref

 

Chimwamurombe PM (2008). ABS and creation of an enabling environment for innovation, is it an issue for SADC countries? Marama bean domestication: an ABS case. Build. Bridges Poverty Reduct. Sustain. Dev. 3:5-7.

 

Chimwamurombe PM (2010). Domestication of Tylosema esculentum (marama bean) as a crop for Southern Africa-Genetic Diversity of the Omitara marama sub-population of Namibia. Aspects Appl. Biol. 96:37-43.

 

Clarke AE, Gleeson PA (1981). Molecular aspects of recognition and response in pollenstigma interaction. In the Phytochemistry of Cell recognition and Cell Surface Interactions (Loewus, F.A. and Ryan, C.A., eds. New York: Plenum Press. pp. 161-211.
crossref

 

Coetzer LA, Ross JH (1977). Tylosema. Trees in South Africa 28:77-80.

 

Datta SK, Datta K, Chatterjee J (2009). Cyto-palynoloical, Biochemical and Molecular Characterization of original and induced mutants of Garden Chrysanthemum. Rome: Food and Agricultural Organization of the United Nations.

 

Gepts P (2004). Crop domestication as a long term experiment. Plant Breed. Rev. 24:1-44.

 

Ghiani A, Onelli E, Aina R, Cocucci M, Citterio S (2011). A comparative study of melting and non-melting flesh peach cultivars reveals that during fruit ripening endo-polygalacturonase (endo-PG) is mainly involved in pericarp textural changes, not in firmness reduction. J. Exp. Bot. 62(11):4043-4054.
crossref

 

Girija M, Dhanavel D (2009). Mutagenic effectiveness and efficiency of gamma rays, ethyl methane sulphonnate and their combined treatments in cowpea (Vigna unguiculata L. Walp). J. Mol. Sci. 4(2):68-75.

 

Halloran GM, Monaghan BG (1996). RAPD variation within and between naturalpopulations of morama [Tylosema esculentum (Burchell) Schreiber] in Southern Africa. South Afr. J. Bot. 62(6):287-297.
crossref

 

Jenkins E, Paul W, Coupe S, Bell S, Davies E, Roberts J (1996). Characterization of anmRNA encoding a polygalacturonase expressed during pod development in oilseed rape (Brassica napus L.). J. Exp. Bot. 47:1111-1115.
crossref

 

Kanai M, Nishimura M, Ayhashi M (2010). A peroxisomal ABC transporter promotes seedgermination by inducing pectin degradation under the control of ABI5. Plant J. 62(6):936-947.

 

Karssen CM, Zagorski S, Kepczynski J, Groot SPC (1989). Key role for endogenousgibberellins in the control of seed germination. Ann. Bot. 63:71-80.

 

Lawlor D (2004). Improvement of marama bean (Tylosema esculentum) - an under–utilizedgrain and tuber producing legume for southern Africa. ICA4-CT-200- 30010, Final Report. Rothamsted: European Union.

 

Mu JH, Lee HS, Kao TH (1994). Characterization of a pollen-expressed receptor-likekinase gene of petunia. Plant Cell 6:709-721.
crossref

 

Naomab E (2004). Assessment of genetic variation in natural populations ofMarama Bean(Tylosema esculentum) using molecular markers. A thesis submitted to the University of Namibia for the degree of Master of Science. Retrieved from the University of NamibiaLibrary, Special collections.

 

Nepolo E, Chimwamurombe PM, Cullis CA, Kandawa-Schulz MA (2010). Determining genetic diversity based on ribosomal intergenic spacer length variation in Marama Bean (Tylosema esculentum) from the Omipanda area, Eastern Namibia. Afr. J. Plant Sci. 4(9):368-373.

 

Netto AT, Ando A, Figueira A, Latado RR, dos Santos PC, Correa LS, Peres LEP, Hauagge R, Pulcinelli CE, Ishiy T, Ferreira-Filho AWP, Camargo CEO (2011). Genetic Improvement of Crops by Mutation Techniques in Brazil. Plant Mutat. Rep. 2(3):24-37.

 

Parry MAJ, Modgwick PJ, Bayon C, Tearall K, Hernandez-Lopez A, Baudo M, Rakszegi M, Hamada W, Al-Jassin A, Ouabbou H, Labhilili M, Phillips AL (2009). Mutation discovery for improvement. J. Exp. Biol. 60(10):2817-2825.

 

Peretto R, Favaron F, Bettini V, Delorenzo G, Marini S, Alghisi P, Cervone F, Bonfante P (1992). Expression and localization of polygalacturonase during the outgrowth oflateral roots in Allium porrum L. Planta 188:164-172.
crossref

 

Popelka JC, Terryn N, Higgins TJV (2004). Gene technology of grain legumes: can itcontribute to the food challenge in developing countries? Plant Sci. 167:195-206.
crossref

 

Ruane J, Sonnino A (2006). The role of biotechnology in exploring and protectingagricultural genetic resources. Rome: Food and Agriculture Organization of the United Nations.

 

Reynolds M, Foulkes MJ, Slafer GA, Berry P, Parry MAJ, Snape JW, Angus WJ (2009). Raising yield potential in wheat. J. Exp. Bot. 60:1899-1918.
crossref

 

Seetohul S, Puchooa D, Ranghoo-Sanmukhiya VM (2009). Improvement of Taro (Colocasia Esculenta Var Esculenta) through in-vitro mutagenesis. Univ. Mauritius Res. J. 13A:79-89.

 

Sprent JI, Odee DW, Dakora FD (2010). African legumes: a vital but under-utilized resource. J. Exp. Bot. 61(5):1257-1265.
crossref

 

Swain S, Kay P, Ogawa M (2011). Preventing unwanted breakups: usingpolygalacturonasesto regulate cell separation. Plant Signal. Behav. 6(1):93-97.
crossref

 

Taylor JE, Tucker GA, Lasslett Y, Smith CJS, Arnold CM, Watson CF, Schuch W, Grierson D, Roberts JA (1990) Polygalacturonase expression during abscission of normaland transgenic tomato plants. Planta 183:133-138.

 

Takundwa M, Nepolo E, Chimwamurombe PM, Cullis CA, Kandawa-Schulz M, Kunert K (2010). Development and use of microsatellite markers for genetic variation analysisin the Namibian germplasm, both within and between populations of marama bean (Tylosema esculentum). J. Plant Breed. Crop Sci. 2(8):233-242.

 

Van Harten AM (2007). Mutation breeding: Theory and Practical applications. Cambridge: Cambridge University Press.

 

Takundwa et al. 57

 

Wilson KJ, Nessler CL, Mahlberg PG (1976). Pectinase in Asclepias latex and its possible role in laticifer growth and development. Am. J. Bot. 63:1140-1144.
crossref

 


APA Takundwa, M. M., Ruzvidzo, O., Nambahu, F., Kawadza, D. T., Chatukuta, P., & Chimwamurombe, P. M. (2015). Characterization of gamma-irradiated seeds of a wild Namibian marama bean (Tylosema esculentum) with microsatellite markers. International Journal of Biotechnology and Molecular Biology Research, 6(7), 48-57.
Chicago Takundwa, M. M., Ruzvidzo, O., Nambahu, F., Kawadza, D. T., Chatukuta, P. and Chimwamurombe, P. M.. "Characterization of gamma-irradiated seeds of a wild Namibian marama bean (Tylosema esculentum) with microsatellite markers." International Journal of Biotechnology and Molecular Biology Research 6, no. 7 (2015): 48-57.
MLA Takundwa, et al. "Characterization of gamma-irradiated seeds of a wild Namibian marama bean (Tylosema esculentum) with microsatellite markers." International Journal of Biotechnology and Molecular Biology Research 6.7 (2015): 48-57.
   
DOI 10.5897/IJBMBR2014.0207
URL http://academicjournals.org/journal/IJBMBR/article-abstract/D23B6BA56446

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