Paternal Contribution to Banana (Musa sapientum L.) & Plantain (Musa paradisiaca L.) Progenies & Progeny Ploidy Composition in a Polycross Mating System Using RAPD & Flow Cytometry

Main Article Content

Victoria Wilson
Abdou Tenkouano
Michael Pillay


Aims: A 4x – 2x polycross mating design of 4 tetraploid female parents was established to determine paternal contributions of 3 diploid male parents to resulting progenies, their ploidy composition and genetic diversity of synthetic hybrids.

Study Design: The polycross mating design comprised 2 blocks having both maternal and paternal selections, with seed parents replicated at 12 plants per clone. Each crossing block had 31 plants of each of the three male parents.

Place and Duration of Study: International Institute of Tropical Agriculture (IITA) High Rainfall Station, Onne (4º51’N, 7º03’E, 10 m above sea level), Rivers State, South-South Nigeria for a period of 24 months.

Methodology: At maturity of maternal parents (TMPx 2796-5; TMPx 1658-4; TMPx 5511-2; and TMPx 7152-2), fruit bunches were harvested, ripened and the seeds extracted. Hard seeds obtained were germinated in vivo in seed trays and emerging seedlings transplanted to perforated nursery bags. At 12 weeks, DNA was extracted from candle leaf for RAPD analysis of 80 progenies and the 3 pollen parents. Ploidy status of progenies was determined using flow cytometry method.

Results: There was significant unequal paternal contribution to Musa polycross progenies with 3 maternal parents; TMPx 2796-5, TMPx 5511-2, and TMPx 1658-4. Two of the 3 paternal parents had progenies with all 4 maternal parents while TMB2x 5105-1 did not have any progeny with TMPx 2796-5. Progenies exhibited 4 ploidy levels with frequency differing with each female parent: TMPx 7152-2 produced 100% 3x progeny; TMPx 5511-2, 63% 3x and 37% 2x; TMPx 2796-5, 91% 3x and 9% 2x and TMPx 1658-4, 82% 3x, 9% 2x, 6% 4x and 3% 5x. The 5x progeny was recorded in the first ratoon crop. The second ratoon crop had only triploids.

Conclusion: The high frequency of 3x progenies from all maternal types in this study, suggests the effectiveness of the polycross mating design in Musa improvement.

Musa spp., tetraploid hybrids, heterosis, ploidy composition, RAPD.

Article Details

How to Cite
Wilson, V., Tenkouano, A., & Pillay, M. (2019). Paternal Contribution to Banana (Musa sapientum L.) & Plantain (Musa paradisiaca L.) Progenies & Progeny Ploidy Composition in a Polycross Mating System Using RAPD & Flow Cytometry. Asian Journal of Biochemistry, Genetics and Molecular Biology, 2(4), 1-14.
Original Research Article


Wilson V, Tenkouano A, Pasberg-Gauhl C, Gauhl F. Heterotic responses of tetraploid and triploid plantain hybrids in Southeast Nigeria. African Crop Science. 1999; 7(2):117-123.

Acquaah, G. Principles of plant genetics and breeding. 2nd ed. Wiley-Blackwell, Oxford; 2012

Kumar LP, Selvarajan R, Iskra-Caruana M, Chabannes M, Hanna R. Biology, etiology, and control of virus diseases of banana and plantain. In: Loebenstein G, Katis NI (eds) Advances in Virus Research. Academic, Burlington. 2015;91:229– 269.

Mok DWS, Peloquin SJ. Breeding value of 2n pollen (diplandroids) in tetraploid x diploid crosses in potato. Theoretical and Applied Genetics. 1975:46:307-314.
DOI: 10.1007

Jane Muthoni, Hussein Shimelis, Rob Melis and Jackson Kabira. Reproductive biology and early generation's selection in conventional potato breeding. AJCS. 2012;6(3):488-497.

Jauhar PP. Synthesis and meiotic studies of triploids of meadow fescue. Agronomy Abstracts, American Society of Agronomy, Fort Collins, Colorado. 1979:64-65.

Burton GW. Meeting human needs through plant breeding: Past progress and prospects for the future. In: Frey, K.J. (Ed.), Plant Breeding. Iowa State University, Ames, Iowa. 1981;433-465.

Nduwumuremyi Athanase. Pangirayi Tongoona, Slyvestre Habimana. Mating Designs: Helpful Tool for Quantitative Plant Breeding Analysis J. Plant Breed. Genet. 2013;117-129.

Varghese Cini, Eldho Varghese, Seema Jaggi, Arpan Bhowmik. Experimental designs for open pollination in polycross trials. Journal of Applied Statistics. 2015; 42(11):2478-2484.

Kumar S, Gerber S, Richardson TE, Gea L. Testing for unequal paternal contributions using nuclear and chloroplast SSR markers in polycross families of radiata pine. Tree Genet Genomes. 2007; 3:207–214.

Amini F, Mirlohi AF, Majidi MM. The possibility of use of AFLP Molecular Markers and phenotypic traits to increase forage yield in Tall Fescue (Festuca arundinacea Schreb.) Breeding. Journal of Agricultural Science and Technology. 2016;18(5):1419-1429.

Mayo O. The theory of plant breeding. Oxford University Press, New York. 1980;293. ISBN:9780231051125

Atanda AS,Olaoye G, Amuda A. Efficacy of modified polycross method in development of sugar cane progenies. Ethiopian Journal of Environmental Studies & Management 2015;8(1):97– 106.

Uzoechi AO, Egesi CN, Ikeogu UN, Abah SP. Polycross as experimental design for open pollination for heterotic cassava development. Paper presented at the World Congress on Root and Tuber Crops Nanning, Guangxi, China; 2016.

Ortiz R, Crouch JH. The efficiency of natural and artificial pollinators in plantain (Musa spp. AAB group) hybridization and seed production. Annals of Botany. 1997;80:693–695.

Ortiz R. Conventional banana and plantain breeding. Acta Hoticulturae. 2013;986:1-20.

Till BJ, Jankowicz-Cieslak J, Sági L, Huynh OA, Utsushi H, Swennen R, Terauchi, R, Mba C. Discovery of nucleotide polymorphisms in the Musa gene pool by Ecotilling. Theoretical and Applied Genetics. 2010;121:1381–1389.
DOI: 10.1007/s00122-010-1395-5

Ortiz R, Swennen R. Review: From crossbreeding to biotechnology-facilitated improvement of banana and plantain. Biotechnological Advances. 2014;32:158-169.

Ortiz R, Vuylsteke D. Recent advances in Musa genetics, breeding and biotechnology. Plant Breeding Abstracts. 1996;66:1355–1363.

Sipen Philip, John K. Chubo, Patricia JH. King, Ong K Huat, Michael R Davey. Genetic Improvement of Banana Using Conventional and In Vitro Technologies, Journal of Crop Improvement. 2011;25:6, 697-727.
DOI: 10.1080/15427528.2011.603406

Tenkouano A. Current issues and future directions for Musa genetic improvement research at the International Institute for Tropical Agriculture. In: Molina AP, Roa VN, Maghuyop MAG (Eds.), Proceedings of the 10th INIBAP-ASPNET Regional Advisory Committee Meeting held at Bangkok, Thailand. 2000;11-23.

Tenkouano A, Crouch JH, Crouch HK, Vuylsteke D, Ortiz R. A comparison of DNA marker and pedigree methods for genetic analysis in plantain and banana (Musa spp.) clones. I. Estimation of genetic relationships. Theoretical and Applied Genetics. 1999a;98:62–68.

Tenkouano A, Crouch JH, Crouch HK, Vuylsteke D, Ortiz R. A comparison of DNA marker and pedigree methods for genetic analysis in plantain and banana (Musa spp.) clones. II. Predicting hybrid performance. Theoretical and Applied Genetics. 1999b;98:69–75.

Brown Allan, Robooni Tumuhimbise, Delphine Amah, Brigitte Uwimana, Moses Nyine, Hassan Mduma, David Talengera, Deborah Karamura, Jerome Kuriba, and Rony Swennen. Bananas and Plantains (Musa spp). In: Genetic Improvement of Tropical Crops, H. Campos, P.D.S. Caligari, Springer International Publishing AG. 2017;219-240.
DOI: 10.1007/978-3-319-59819-2_7

Tenkouano A, Crouch JH, Crouch HK, Vuylsteke D. Ploidy determination in Musa germplasm using pollen and chloroplast characteristics. American Journal of Horticultural Science. 1998a;33(5):889-890.

Tenkouano A, Ortiz R, Vuylsteke D. Combining ability for yield and plant phenology in plantain-derived populations. Euphytica. 1998b;104:151-158.

Fasahat P, Rajabil A, Rad JM, Derera J. Principles and utilization of combining ability in plant breeding. Biom Biostat Int J. 2016;4(1):1-24.

Aguilar Morán JF. Improvement of Cavendish banana cultivars through conventional breeding. Acta Hortic. 2013;986:205–208.

Wei Z, Qu Z, Hou C, Liu Y, Zhang L, Yang C, Wei H. Genetic diversity and paternal analysis of open-pollinated progenies of Larix olgensis seed orchard. Journal of Nature and Science. 2015;1(1):19:1-6.

Moriguchi Y, Ishiduka D, Kaneko T, Itoo S, Taira H, Tsumura Y. The contribution of pollen germination rates to uneven paternity among polycrosses of Cryptomeria japonica. Silvae Genetica. 2009;58(3):139-144.

Bardakci F. Random amplified polymorphic DNA (RAPD) markers. Turkish Journal of Biolog. 2001;25:185-196.

Salem HH, Ali BH, Huang TH, Qin DN, Nang XM Xie QD. Use of random amplified polymorphic DNA analysis for economically important food crops. Journal of Integrative Plant Biology. 2007;49:1670-1680.

Kumar P, Gupta VK, Misra AK, Modi DR, Pandey BK. Potential of molecular markers in plant biotechnology. Plant Omics Journal. 2009;2:141-162. ISSN:1836-3644.

Samantaray S, Geetha KA, Hidyath KP, Maiti S. Identification of RAPD markers linked to sex determination in guggal Commiphora wightii (Armott.) Bhandari. Plant Biotechnology Reports. 2010;4:95-99.

Wheeler N, Payne P, Hipkins V, Saich R, Kenny S, Tuskan G. Polymix breeding with paternity analysis in Populus: a test for differential reproductive success (DRS) among pollen donors. Tree Genetics & Genomes. 2006;2:53–60.
DOI: 10.1007/s11295-005-0033-x

Ortiz R, Austin PD, Vuylsteke D. IITA High Rainfall Station African humid forest. American Journal of Horticultural Science. 1997;32:969-972.

Hullugale NR, Lal R, Gichuru MP. Effect of five years of no-tillage and mulch on soil properties and tuber yield of cassava on an acid Ultisol in South-Eastern Nigeria. Experimental Agriculture. 1990;26:235-240.

Plantain and Banana Improvement Program Annual Report. Crop Improvement Division, International Institute of Tropical Agriculture, Nigeria. 1994;160.

Crouch JH, Vuylsteke D, Ortiz R. Perspectives on the application of biotechnology to assist the genetic enhancement of plantain and banana (Musa spp.). Electronic Journal of Biotechnology. 1998;1:11-22.

Crouch JH, Vuylsteke D, Ortiz R. Perspectives on the application of biotechnology to assist the genetic enhancement of plantain and banana (Musa spp.). Electronic Journal of Biotechnology. 1998;1:11-22.

Mabonga L, M. Pillay, SCAR Marker for the A Genome of Bananas (Musa spp. L.) Supports Lack of Differentiation between the A and B Genomes. Journal of Agricultural Science. 2017;9(6).

Pillay M, Nwakanma DC, Tenkouano A. Identification of RAPD markers linked to A and B genome sequences in Musa L. Genome. 2000;43:763–767.

Dolezel J, Lysak MA, Van den Houwe I, Dolezelova M, Roux N. Use of flow cytometry for rapid ploidy determination in Musa species. INFOMUSA. 1997;6:6-9.

Karamura D, Tumuhimbise R, Muhangi S, Nyine M, Pillay M, Tendo SR, Talengera D, Namanya P, Kubiriba J, Karamura E. African Journal of Biotechnology. 2016; 15(31):1692-1698.
DOI: 10.5897/AJB2016.15442

Basha SD, Sujatha M. Inter and intra-population variability of Jatropha curcas (L.) characterized by RAPD and ISSR markers and development of population-specific SCAR markers. Euphytica. 2007;156:375-386. Available:

Kumar Sambhav Verma, Shamshad ul Haq, Sumita Kachhwaha, S. L. Kothari RAPD and ISSR marker assessment of genetic diversity in Citrullus colocynthis (L.) Schrad: a unique source of germplasm highly adapted to drought and high-temperature stress. 3 Biotech. 2017; 7(288):1-24.
DOI: 10.1007/s13205-017-0918-z

Fernandes L, Rocheta M, Cordeiro J, Pereira S, Gerber S, Oliveira MM, Ribeiro MM. Genetic variation, mating patterns and gene flow in a Pinus pinaster Aiton clonal seed orchard. Ann. For. Sci. 2008; 65:706.

Thomas L, Tew, Yong-Bao Pan. Microsatellite (Simple sequence repeat) marker–based paternity analysis of a seven-parent sugarcane polycross. Crop Sci. 2010;50:1401–1408.
DOI: 10.2135/cropsci2009.10.0579

Hansen OK, Kjaer ED. Paternity analysis with microsatellites in a Danish Abies nordmanniana clonal seed orchard reveals dysfunctions. Can. J. For. Res. 2006;36: 1054–1058.

Wiselogel AE, Van Buijtenen JP. Probability of equal mating in polymix pollinations of loblolly pine. Silvae Genetica. 1988;37:184–187.

Seido K, Yamagishi K, Ida K, Murakami H, Shiraishi S. Investigation of selective fertilization in hinoki (Chamaecyparis obtusa) using a chloroplast DNA marker. Journal of Japanese Forestry Society. 2000;82:380–383. (in Japanese with English summary).

Nikkanen T, Aronen T, Haggman H, Venalainen M. Variation in pollen viability among Picea abies genotypes—Potential for unequal paternal success. Theor. Appl.Genet. 2000;101:511–518.

Moriguchi Y, Tani N, Itoo S, Kanehira F, Tanaka K, Yomogida H, Taira H, Tsumura Y. Gene flow and mating system in fi ve Cryptomeria japonica D. Don seed orchards as revealed by analysis of microsatellite markers. Tree Genet. Genomes. 2005;1:174–183.

Teixeira S, Bernasconi G. High prevalence of multiple paternity within fruits in natural populations of Silene latifolia, as revealed by microsatellite DNA analysis. Mol. Ecol. 2007;16;4370–4379.

Austerlitz F, Gleiser G, Teixeira S, Bernasconi G. The effects of inbreeding, genetic dissimilarity and phenotype on male reproductive success in a dioecious plant. Proceedings of the Royal Society B: Biological Sciences. 2012; 279:91–100.

Varis S, Reiniharju J, Santanen A, Ranta H, Pulkkinen P. Interactions during in vitro germination of Scots pine pollen. Trees. 2010;24:99–104.

Duffy KJ, Johnson SD. Effects of pollen reward removal on fecundity in a self-incompatible hermaphrodite plant. Plant Biology 2011;13:556–560.

Ashman TL, Arceo-Go´mez G. Toward a predictive understanding of the fitness costs of heterospecific pollen receipt and its importance in co-flowering communities. American Journal of Botany. 2013;100: 1061–1070.

Ramesha BT, Yetish MD, Ravikanth G, Ganeshaiah KN, Ghazoul J, Shaanker RU. Stylish lengths: mate choice in flowers. Journal of Biosciences. 2011; 36:229–234.

Rodrigo J, Herrero M, Hormaza JI. Pistil traits and flower fate in apricot (Prunus armeniaca). Annals of Applied Biology. 2009;154:365–375.

Madjidian JA, Andersson S, Lankinen A. Estimation of heritability, evolvability and genetic correlations of two pollen and pistil traits involved in a sexual conflict over timing of stigma receptivity in Collinsia heterophylla (Plantaginaceae). Annals of Botany. 2012a;110:91–99.

Cocucci AA, Marino S, Baranzelli M, Wiemer AP, Sersic A. The buck in the milkweed: evidence of male-male interference among pollinaria on pollinators. New Phytologist. 2014;203: 280–286.

Forrest JRK. Plant size, sexual selection, and the evolution of protandry in dioecious plants. The American Naturalist. 2014;184: 338–351.

Lankinen A, Karlsson Green K. Using theories of sexual selection and sexual conflict to improve our understanding of plant ecology and evolution. AoB PLANTS 2015;7:plv008.