RAPD ANALYSIS TO DETECT SOMACLONAL VARIATION OF PINEAPPLE IN VITRO CULTURES DURING MICROPROPAGATION
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Plant off-type formation has become a concern in pineapple micropropagation, and reliable methods are required to detect and minimize this problem. This study was conducted to confirm the occurrence of somaclonal variation during the micropropagation of pineapple clone Simadu. The effects of culture period (long vs. short duration) and regeneration methods (direct organogenesis, indirect organogenesis, and somatic embryogenesis) were evaluated to determine their contribution to somaclonal variation. RAPD analysis using ten primers was performed to confirm genetic variation. The results showed that RAPD assays could be applied for early detection of somaclonal variation in pineapple, with OPA primers performing better than OPJ primers. Phenotypic variation observed in four-year-old field-grown pineapple plants and plantlets was associated with genetic variation. The findings demonstrated that the long culture period was the main factor contributing to somaclonal variation, while regeneration method and plant growth regulators also played a role in inducing genetic changes. Newly initiated cultures showed a higher level of genetic similarity. Therefore, applying an appropriate micropropagation strategy is necessary to minimize plant off-types. It is recommended to avoid using long-term cultures as mother stock and to apply direct organogenesis rather than indirect organogenesis or somatic embryogenesis for pineapple micropropagation.
Abdellatif KF, Hegazy AE, Aboshama HM, Emara HA, El-Shahed AA. 2012. Morphological and molecular characterization of somaclonal variation in tissue culture-derived banana plants. J Genet Eng Biotechnol 10: 47–53.
Ahmed ABA, Rao AS, Rao MV, Taha M. 2011. Effect of picloram, additives and plant growth regulators on somatic embryogenesis of Phyla nodiflora (L.) Greene. Braz Arch Biol Technol 63: 147–173.
Bairu MW, Fennell CW, van Staden J. 2006. The effect of plant growth regulators on somaclonal variation in Cavendish banana (Musa AAA cv. 'Zelig'). Sci Hortic 108: 347–351.
Bairu MW, Adeyemi O, van Staden J. 2011. Somaclonal variation in plants: causes and detection methods. Plant Growth Regul 63: 147–173.
Chen WH, Chen TM, Fu YM, Hsieh RM, Chen WS. 1998. Studies on somaclonal variation in Phalaenopsis. Plant Cell Rep 18: 7–13.
Coppens d’Eeckenbrugge G, Leal F. 2003. Morphology, anatomy, and taxonomy. In: Bartholomew DP, Paull RE, Rohrbach KG, editors. The Pineapple: Botany, Production and Uses. Wallingford (UK): CABI Publishing. p. 13–32.
Eeuwens CJ, Lord SCR, Donough VR, Vallejo G, Nelson S. 2002. Effects of tissue culture conditions during embryoid multiplication on the incidence of ‘‘mantled’’ flowering in clonally propagated oil palm. Plant Cell Tissue Organ Cult 70: 311–323.
Feuser S, Meler K, Daquinta M, Guerra MP. 2003. Genotype fidelity of micropropagated pineapple (Ananas comosus) plantlets assessed by isozymes and RAPD markers. Plant Cell Tissue Organ Cult 72: 221–227.
Firoozabady E, Moy Y. 2004. Regeneration of pineapple via somatic embryogenesis and organogenesis. In Vitro Cell Dev Biol-Plant 40: 67–74.
International Board for Plant Genetic Resources. 1991. Descriptors for Pineapple. Rome.
Khoddamzadeh AA, Sinniah UR, Kadir MA, Kadzimin SB, Mahmood M, Sreeramanan S. 2010. Detection of somaclonal variation by random amplified polymorphic DNA analysis during micropropagation of Phalaenopsis bellina (Rchb.f.) Christenson. Afr J Biotechnol 9(40): 6632–6639.
Koornneef M. 1991. Variation and mutant selection in plant cell and tissue culture. In: Jones L, editor. Biotechnological Innovation in Crop Improvement. London (UK): Butterworth-Heinemann. p. 100–115.
Leva AR, Petruccelli R, Rinaldi LMR. 2012. Somaclonal variation in tissue culture: A case study with olive. In: Leva AR, editor. Recent Advances in Plant In Vitro Culture. InTech. Available from: (URL removed for formatting). doi: 10.5772/150367.
Masoud S, Hamta A. 2008. Cytogenetic analysis of somaclonal variation in regenerated plants of berseem clover (Trifolium alexandrinum L.). Cytologia 61(4): 392–396.
Miguel C, Marum L. 2011. An epigenetic view of plant cells cultured in vitro: somaclonal variation and beyond. J Exp Bot 62(11): 3713–3725.
Mohamed AE. 2007. Morphological and molecular characterization of some banana micropropagated variants. Int J Agric Biol 9(5): 707–714.
Murashige T, Skoog F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–497.
Noormi R, Murugaiyah V, Subramaniam S. 2012. Optimization of callus induction medium for Hymenocallis littoralis using root and bulb explants. J Med Plants Res 6(12): 2309–2316.
Nwauzoma AB, Jaja ET. 2013. A review of somaclonal variation in plantain (Musa spp.): mechanisms and applications. J Appl Biosci 67: 5252–5260.
Rani V, Raina SN. 2000. Genetic fidelity of organized meristem-derived micropropagated plants: A critical reappraisal. In Vitro Cell Dev Biol-Plant 36: 319–330.
Roostika I, Mariska I, Khumaida N, Wattimena GA. 2012a. Indirect organogenesis and somatic embryogenesis of pineapple induced by dichlorophenoxyacetic acid. J Agrobiogen 8(1): 8–18.
Roostika I, Mariska I, Khumaida N, Wattimena GA. 2012b. Effect of picloram and light on somatic embryogenesis regeneration of pineapple. IJAS 13(2): 43–53.
Roostika I. 2012c. Pemantapan Metode Organogenesis dan Embriogenesis Somatik Nenas (Ananas comosus (L.) Merr.) serta Deteksi dan Reduksi Keragaman Somaklonal. Disertasi. Bogor (ID): Institut Pertanian Bogor.
Rout GR, Senapati SK, Aparajita S, Palai SK. 2009. Studies on genetic identification and genetic fidelity of cultivated banana using ISSR markers. POJ 2(6): 250–258.
Santos MDM, Buso GCS, Torres AC. 2008. Evaluation of genetic variability in micropropagated propagules of ornamental pineapple (Ananas comosus var. bracteatus) using RAPD markers. Genet Mol Res 7(4): 1097–1105.
Shahid MTH, Khan FA, Saeed A, Aslam M, Rasul F. 2014. Development of somaclones in sugarcane genotype BF-162 and assessment of variability by RAPD and SSR markers in selected red-rot resistant somaclones. Afr J Biotechnol 11(15): 3502–3513.
Smith MK, Ko H-L, Hamill SD, Sanewski GM, Graham MW. 2003. Biotechnology. In: Bartholomew DP, Paull RE, Rohrbach KG, editors. The Pineapple: Botany, Production and Uses. Wallingford (UK): CABI Publishing. p. 57–68.
Soneji JR, Rao PS, Mhatre M. 2002. Suitability of RAPD for analyzing spined and spineless variant regenerants of pineapple (Ananas comosus (L.) Merr.). Plant Mol Biol Rep 20: 307a–307i.
Soniya EV, Banerjee NS, Das MR. 2001. Genetic analysis of somaclonal variation among callus-derived plants of tomato. Curr Sci 80(9): 1213–1215.
Suminar E. 2010. Induksi Keragaman Genetik dengan Mutagen Sinar Gamma pada Nenas (Ananas comosus (L.) Merr.) Secara In Vitro. Tesis. Bogor (ID): Institut Pertanian Bogor.
Tawar PN, Sawant RA, Dalvi SG, Nikam AA, Kawar PG, Deviarumath RM. 2008. An assessment of somaclonal variation in micropropagated plants of sugarcane by RAPD markers. Sugar Tech 10(2): 124–127.
Wakasa K. 1979. Variation in the plants differentiated from tissue culture of pineapple. Jpn J Breed 29(1): 13–22.
Williams JGK, Kubelik AR, Livak KJ, Rafalski JA. 1990. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res 18: 6531–6535.
Zucchi MI, Arizono H, Morais VA, Fungaro MHP, Vieira MLC. 2002. Genetic instability of sugarcane plants derived from meristem cultures. Genet Mol Biol 25(1): 91–96.
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