Genetic Stability of Banana Plant Regenerated from Floral Axis Organogenesis Assessed by Newly Developed SSR Markers

Puji Lestari, I. Roostika, Kristianto Nugroho, Edison HS, Habib Rijzaani, Mastur Mastur


Molecular marker is robust to precisely monitor the genetic stability of in vitro-banana plants. This study examined the genetic stability of 8 monthold banana plants of Soka variety derived from floral axis organogenesis using newly developed SSR markers. The results showed that the same qualitative and similar quantitative morphological characters of pseudostem, leaf and fruit were identified between mother plants and culture plants from floral axis regeneration. Both plants types were quite similar in number of tillers, brix percentage, fruit peel/mesocarp thickness and fruit length. Eleven out of 211 good quality of SSR loci showing high homology with important genes were selected for suitable PCR primers and produced unambiguous bands.The number of total bands was 323 for total SSR primers, in range of 20-60 per primer for total individual plants. Most culture plants showed identical with their mother plants, with very minor variation as reflected by genetic similarity coefficient range of 0.9-1.0. A high similar pattern on SSR to support morphological characters of mother plants and culture plants indicated a successful micropropagation using floral axis to encounter off-type clones.The floral axis organogenesis in this study is able to provide sufficient genetic materials of Soka for varietal registration and other applications.


Banana; Floral axis; Genetic stability; In vitro; SSR

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Ateş Sönmezoǧlu, Ö., Bozmaz, B., Yildirim, A., Kandemir, N., & Aydin, N. (2012). Genetic characterization of Turkish bread wheat landraces based on microsatellite markers and morphological characters. Turkish Journal of Biology, 36, 589–597. crossref

Butiuc-keul, A., Farkas, A., & Cristea, V. (2016). Genetic stability assessment of in vitro plants by molecular markers. Studia Universitatis Babeş-Bolyai Biologia, LXI(1), 107–114. Retrieved from crossref

Condello, E., Palombi, M. A., & Tonelli, M. G. (2008). Genetic stability of wild pear (Pyrus pyraster, Burgsd) after cryopreservation by encapsulation dehydration. Agricultural and Food Science, 18(2), 136–143. crossref

Creste, S., Benatti, T. R., Orsi, M. R., Risterucci, A.-M., & Figueira, A. (2006). Isolation and characterization of microsatellite loci from a commercial cultivar of Musa acuminate. Molecular Ecology Notes, 6(2), 303–306. crossref

Cristea, V., Crăciunaș, C., Marcu, D., Palada, M., & Butiuc-Keul, A. (2014). Genetic stability during in vitro propagation of Dianthus spiculifolius Schur. Propagation of Ornamental Plants, 14(1), 26–31. Retrieved from crossref

Culley, T. M., Stamper, T. I., Stokes, R. L., Brzyski, J. R., Hardiman, N. A., Klooster, M. R., & Merritt, B. J. (2013). An efficient technique for primer development and application that integrates fluorescent labeling and multiplex PCR. Applications in Plant Sciences, 1(10), 1300027. crossref

D’Hont, A., Denoeud, F., Aury, J.-M., Baurens, F.-C., Carreel, F., Garsmeur, O.,...Wincker, P. (2012). The banana (Musa acuminate) genome and the evolution of monocotyledonous plants. Nature, 488, 213–217. crossref

Doyle, J. J., & Doyle, J. L. (1990). Isolation of plant DNA from fresh tissue. Focus, 12, 13–15. Retrieved from website

Elhory, S. M. A., Aziz, M. A., Rashid, A. A., & Yunus, A. G. (2009). Prolific plant regeneration through organogenesis from scalps of Musa sp cv. Tanduk. African Journal of Biotechnology, 8(22), 6208–6213. crossref

FAO. (2003). Chapter 1. Overview of world banana production and trade. Retrieved from website

François-Xavier, C., Sandoval, J. A., Marie, P., & Auboiron, E. (1993). Variations in micropropagated bananas and plantains: literature survey. Fruits, 48(1), 15-23. Retrieved from website

Ge, C., Cui, Y.-N., Jing, P.-Y., & Hong, X.-Y. (2014). An alternative suite of universal primers for genotyping in multiplex PCR. PLoS ONE, 9(3), e92826. crossref

Govindaraju, S., Saravanan, J., Jayanthi, B., Nancy, D., & Indra Arulselv, P. (2012). In vitro propagation of Banana (Musa sp - Rasthali variety) from sword suckers for its commercial production. Research in Plant Biology, 2(5), 01–06. Retrieved from website

Hapsari, L., & Lestari, D. A. (2016). Fruit characteristic and nutrient values of four Indonesian banana cultivars (Musa spp.) at different genomic groups. AGRIVITA Journal of Agricultural Science, 38(3), 303–311. crossref

Hapsari, L., Kennedy, J., Lestari, D. A., Masrum, A., & Lestarini, W. (2017). Ethnobotanical survey of bananas (Musaceae) in six districts of East Java, Indonesia. Biodiversitas, 18(1), 160–174. crossref

Hippolyte, I., Bakry, F., Seguin, M., Gardes, L., Rivallan, R., Risterucci, A.-M., … Glaszmann, J.-C. (2010). A saturated SSR/DArT linkage map of Musa acuminate addressing genome rearrangements among bananas. BMC Plant Biology, 10, 65. crossref

IPGRI. (1996). Descriptors for banana (Musa spp.). Retrieved from pdf

Izquierdo, H., González, M. C., & de la C Núñez, M. (2014). Genetic stability of micropropagated banana plants (Musa spp.) with non-traditional growth regulators. Biotecnología Aplicada, 31(1), 23–27. Retrieved from website

James, E. A., Brown, G. K., Citroen, R., Rossetto, M., & Porter, C. (2011). Development of microsatellite loci in Triglochin procera (Juncaginaceae), a polyploid wetland plant. Conservation Genetics Resources, 3(1), 103–105. crossref

Jarda, L., Butiuc-Keul, A., Höhn, M., Pedryc, A., & Cristea, V. (2014). Ex situ conservation of Dianthus giganteus d’Urv. subsp. banaticus (Heuff.) Tutin by in vitro culture and assessment of somaclonal variability by molecular markers. Turkish Journal of Biology, 38, 21–30. crossref

Kariyana, K., & Nisyawati. (2013). Effect of ascorbic acid, activated charcoal and light duration on shoot regeneration of banana cultivar barangan (Musa acuminate L.) in vitro culture. International Journal of Recent Research and Applied Studies, 16(1), 118–123. Retrieved from pdf

Khan, S., Saeed, B., & Kauser, N. (2011). Establishment of genetic fidelity of in-vitro raised banana plantlets. Pakistan Journal of Botany, 43(1), 233–242. Retrieved from pdf

Kour, B., Kour, G., Kaul, S., & Dhar, M. K. (2014). In vitro mass multiplication and assessment of genetic stability of in vitro raised Artemisia absinthium L. plants using ISSR and SSAP molecular markers. Advances in Botany, 2014(727020), 1–7. crossref

Krikorian, A. D., Irizarry, H., Cronauer-Mitra, S. S., & Rivera, E. (1993). Clonal fidelity and variation in plantain (Musa AAB) regenerated from vegetative stem and floral axis tips in vitro. Annals of Botany, 71(6), 519–535. crossref

Lakshmanan, V., Venkataramareddy, S. R., & Neelwarne, B. (2007). Molecular analysis of genetic stability in long-term micropropagated shoots of banana using RAPD and ISSR markers. Electronic Journal of Biotechnology, 10(1), 106-113. crossref

Lisnandar, D. S., Fajarudin, A., Effendi, D., & Tambunan, I. R. (2015). Organogenesis bunga aksis pisang bergenom AAB dan ABB. Jurnal Hortikultura, 25(1), 1–8. crossref

Martin, G., Baurens, F.-C., Cardi, C., Aury, J.-M., & D’Hont, A. (2013). The complete chloroplast genome of banana (Musa acuminate, Zingiberales): Insight into plastid monocotyledon evolution. PLOS ONE, 8(6), e67350. crossref

Martin, K. P. (2005). Cost effective in vitro propagation of Musa ornate Roxb. through floral tip axis segment culture. Propagation of Ornamental Plants, 5(2), 84–88. Retrieved from pdf

Missiaggia, A., & Grattapaglia, D. (2006). Plant microsatellite genotyping with 4-color fluorescent detection using multiple-tailed primers. Genetics and Molecular Research, 5(1), 72–78. Retrieved from website

Muhammad, A., Rashid, H., Hussain, I., & Naqvi, S. M. S. (2007). Proliferation-rate effects of BAP and kinetin on banana (Musa spp. AAA group) “Basrai.” HortScience, 42(5), 1253–1255. crossref

Nei, M. (1973). Analysis of gene diversity in subdivided populations. Proceedings of the National Academy of Sciences of the United States of America, 70(12 Pt 1-2), 3321–3323. crossref

Panis, B. (2009). Cryopreservation of Musa germplasm. (F. Engelmann & E. Benson, Eds.), Technical Guidelines No. 9 (2nd ed.). Montpellier, France: Bioversity International. Retrieved from pdf

Poerba, Y. S., Imelda, M., & Martanti, D. (2012). Analisa kestabilan genetik pisang kepok “unti sayang” hasil mikroprogasi dengan marka RAPD dan ISSR. Berita Biologi, 11(2), 275–282. Retrieved from website

Procházková, D., Boušová, I., & Wilhelmová, N. (2011). Antioxidant and prooxidant properties of flavonoids. Fitoterapia, 82(4), 513–523. crossref

Ravishankar, K. V, Vidhya, L., Cyriac, A., Rekha, A., Goel, R., Singh, N. K., & Sharma, T. R. (2012). Development of SSR markers based on a survey of genomic sequences and their molecular analysis in banana (Musa spp.). The Journal of Horticultural Science and Biotechnology, 87(1), 84–88. crossref

Ravishankar, K. V., Raghavendra, K. P., Athani, V., Rekha, A., Sudeepa, K., Bhavya, D., … Ananad, L. (2013). Development and characterisation of microsatellite markers for wild banana (Musa balbisiana). The Journal of Horticultural Science and Biotechnology, 88(5), 605–609. crossref

Ray, T., Dutta, I., Saha, P., Das, S., & Roy, S. C. (2006). Genetic stability of three economically important micropropagated banana (Musa spp.) cultivars of lower Indo-Gangetic plains, as assessed by RAPD and ISSR markers. Plant Cell, Tissue and Organ Culture, 85(1), 11–21. crossref

Resmi, L., & Nair, A. S. (2011). Differential effect of cytokinins in the micropropagation of diploid and triploid Musa cultivars. International Journal of Integrative Biology, 11(1), 35–38. Retrieved from website

Rohlf, F. J. (2000). NTSYSpc: Numerical taxonomy and multivariate analysis system, Version 2.1 (Guide user). Setauket, NY, USA: Applied Biostatistics Inc. Retrieved from pdf

Rotchanapreeda, T., Wongniam, S., Swangpol, S. C., Chareonsap, P. P., Sukkaewmanee, N., & Somana, J. (2016). Development of SSR markers from Musa balbisiana for genetic diversity analysis among Thai bananas. Plant Systematics and Evolution, 302(7), 739–761. crossref

Roy, O. S., Bantawa, P., Ghosh, S. K., da Silva, J. A. T., DebGhosh, P., & Mondal, T. K. (2010). Micropropagation and field performance of ‘Malbhog’ (Musa paradisiaca, AAB group): A popular banana cultivar with high keeping quality of North East India. Tree and Forestry Science and Biotechnology, 4(Special Issue 1), 52–58. Retrieved from pdf

Rozen, S., & Skaletsky, H. (2000). Primer3 on the WWW for general users and for biologist programmers. In S. Misener & S. A. Krawetz (Eds.), Bioinformatics Methods and Protocols. Methods in Molecular BiologyTM, vol 132 (pp. 365–386). Totowa, NJ: Humana Press. crossref

Sato, Y. (2009). Ethnobotanical study of local practices maintaining landrace diversity of bananas (Musa spp.) and enset (Ensete ventricosum) in East African Highland (G-COE Series No. 59). Kyoto. Retrieved from website

Schuelke, M. (2000). An economic method for the fluorescent labeling of PCR fragments. Nature Biotechnology, 18, 233–234. crossref

Shankar, C. S., Balaji, P., & Sekar, D. S. (2014). Mass propagation of banana (Musa sp.) cv. grand naine through direct organogenesis by benzyl adenine purine and kinetin. Journal of Academia and Industrial Research, 3(2), 92–97. Retrieved from pdf

Silvarolla, M. B., Mazzafera, P., & de Lima, M. M. A. (2000). Caffeine content of Ethiopian coffea arabica beans. Genetics and Molecular Biology, 23(1), 213–215. crossref

Sultan, M. T., Khan, M. H., Hakim, L., Mamun, A., Morshed, A., Islam, R. M., & Islam, R. M. (2011). In vitro plant regeneration from male flowers of banana. International Journal of Biosciences (IJB), 1(1), 1–11. Retrieved from website

Tiwari, J. K., Chandel, P., Gupta, S., Gopal, J., Singh, B. P., & Bhardwaj, V. (2013). Analysis of genetic stability of in vitro propagated potato microtubers using DNA markers. Physiology and Molecular Biology of Plants, 19(4), 587–595. crossref

Venkataramana, R. K., Sampangi-Ramaiah, M. H., Ajitha, R., Khadke, G. N., & Chellam, V. (2015). Insights into Musa balbisiana and Musa acuminate species divergence and development of genic microsatellites by transcriptomics approach. Plant Gene, 4, 78–82. crossref

Vincent, L., & Anushma, P. L. (2018). Micropropagation in banana using inflorescence: A review. Journal of Cell and Tissue Research, 18(3), 6573–6582. Retrieved from pdf

Wang, J. Y., Zheng, L. S., Huang, B. Z., Liu, W. L., & Wu, Y. T. (2010). Development, characterization, and variability analysis of microsatellites from a commercial cultivar of Musa acuminate. Genetic Resources and Crop Evolution, 57(4), 553–563. crossref

Yu, J., Holland, J. B., McMullen, M. D., & Buckler, E. S. (2008). Genetic design and statistical power of nested association mapping in maize. Genetics, 178(1), 539–551. crossref

Zerihun, D., Vashist, U., & Boora, K. S. (2009). Molecular characterization of citrus cultivars using DNA markers. International Journal of Biotechnology and Biochemistry, 5(3), 271–280. Retrieved from website



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