Application of a High-throughput Polyacrylamide Electrophoresis Gel System with Ethidium Bromide Staining in Genetic Diversity Estimation of Sunflower Genotypes Using TRAP Markers

Document Type : research

Authors

Assistant Professor, Crop and Horticultural Science Research Department, Ardabil Agricultural and Natural Resources Research and Education Center, AREEO, Ardabil (Moghan), Iran

Abstract

In this research, the application of new electrophoresis method of polyacrylamide gel using ethidium bromide staining for genetic variation of sunflower genotypes using TRAP novel marker has been proposed which has advantages such as low-cost, higher speed of analysis, analysis of a large number of samples per unit time (196×2) per experiment, as well as the reuse of gels in comparison with conventional methods of silver staining and fluorescent primer labeling. Six fixed and six arbitrary primers produced 19 TRAP markers so that from 116 bands, 109 of them were polymorphic. Mean number of polymorphic bands were 5.79. DNA fragments size were between 25-920 bp. Polymorphic Information Content (PIC) of this marker were between 0.03 and 0.50 (with the average of 0.33). Primer combination (MAX3BR and Sa12) with PIC value of 0.49 was the highest polymorphic locus. The results showed that this method could be very useful not only in detecting products of the microsatellite markers, but also in other markers, such as TRAP, which produces many polymorphic bands.

Keywords


Anonymous. 2017. The Compositae Genome Project (CGP) Web Site: http://cgpdb.ucdavis.edu/.
Barata, C., and Carena, M. 2006. Classification of North Dakota maize inbred lines into heterotic groups based on molecular and testcross data. Euphytica, 151(3): 339-349.
Dowling, C.D., Burson, B.L., Foster, J.L., Tarpley, L. and Jessup, R.W. 2013. Confirmation of pearl millet-napiergrass hybrids using EST-derived simple sequence repeat (SSR) markers. American Journal of Plant Sciences, 4(5): 1004.
Fiust, A., Rapacz, M., Wójcik-Jagła, M. and Tyrka, M. 2015. Development of DArT-based PCR markers for selecting drought-tolerant spring barley. Journal of applied genetics, 56(3): 299-309.
Fu, Y.-B., Peterson, G.W., Richards, K.W., Tarn, T.R. and Percy, J.E. 2009. Genetic diversity of Canadian and exotic potato germplasm revealed by simple sequence repeat markers. American Journal of Potato Research, 86(1): 38-48.
Gelin, J., Forster, S., Grafton, K., Mcclean, P. and Rojas-Cifuentes, G. 2007. Analysis of seed zinc and other minerals in a recombinant inbred population of navy bean (Phaseolus vulgaris L.). Crop Science 47(4): 1361-1366.
Genovesi, A.D., Jessup, R.W., Engelke, M.C. and Burson, B.L. 2009. Interploid St. Augustinegrass [Stenotaphrum secundatum (Walt.) Kuntze] hybrids recovered by embryo rescue. In Vitro Cellular Developmental Biology-Plant, 45(6): 659-666.
Henareh, M., Dursun, A., Abdollahi-Mandoulakani, B. and Haliloğlu, K. 2016. Assessment of genetic diversity in tomato landraces using ISSR markers. Genetika, 48(1): 25-35.
Hu, J. and Vick, B.A. 2003. Target region amplification polymorphism: a novel marker technique for plant genotyping. Plant Molecular Biology Reporter, 21(3): 289-294.
Kadaru, S.B., Yadav, A.S., Fjellstrom, R.G. and Oard, J.H. 2006. Alternative ecotilling protocol for rapid, cost-effective single-nucleotide polymorphism discovery and genotyping in rice (Oryza sativa L.). Plant Molecular Biology Reporter, 24(1): 3-22.
Mir, R., Kumar, N., Jaiswal, V., Girdharwal, N., Prasad, M., Balyan, H., and Gupta, P. 2012. Genetic dissection of grain weight in bread wheat through quantitative trait locus interval and association mapping. Molecular Breeding, 29(4): 963-972.
Naghavi, M., Ghareyazie, B. and Hosseini Salekdeh, Gh. 2005. Molecular Markers. Tehran. University Press. p 320.
Quezada, M., Pastina, M.M., Ravest, G., Silva, P., Vignale, B., Cabrera, D., Hinrichsen, P., Garcia, A.A.F. and Pritsch, C. 2014. A first genetic map of Acca sellowiana based on ISSR, AFLP and SSR markers. Scientia Horticulturae, 169: 138-146.
Wang, D., Shi, J. Carlson, S., Cregan, P., Ward, R. and Diers, B. 2003. A low-cost, high-throughput polyacrylamide gel electrophoresis system for genotyping with microsatellite DNA markers. Crop Science, 43(5): 1828-1832.
Weiguo, Z., Zhihua, Z., Xuexia, M., Yong, Z., Sibao, W., Jianhua, H., Hui, X., Yile, P. and Yongping, H. 2007. A comparison of genetic variation among wild and cultivated Morus species (Moraceae: Morus) as revealed by ISSR and SSR markers.  Biodiversity and Conservation, 16(2): 275-290.
Yoshida, T. 2004. Genetic diversity among Japanese cultivated sorghum assessed with simple sequence repeats markers.  Plant Production Science, 7(2): 217-223.
Zeinalzadeh-Tabrizi, H., Haliloglu, K. and Razban Haghighi, A. 2015a. Genetic diversity of sunflower genotypes (Helianthus annuus L.) using TRAP markers. Journal of Crop Biotechnology, 4(12): 39-53.
ZeinalzadehTabrizi, H., Hosseinpour, A., Aydin, M. and Haliloglu, K. 2015b. A modified genomic DNA extraction method from leaves of sunflower for PCR based analyzes. Journal of Biodiversity and Environmental Sciences, 7(6): 222-225.
ZeinalzadehTabrizi, H., Sahin, E. and Haliloglu, K. 2011. Principal components analysis of some F1 sunflower hybrids at germination and early seedling growth stage. Journal of the Faculty of Agriculture (Atatürk Üniversitesi Ziraat Fakültesi Dergisi), 42(2): 103-109.