Abscisic acid and salinity stress induced somaclonal variation and increased histone deacetylase (HDAC) activity in Ananas comosus var. MD2
Somaclonal and phenotypic variation caused by genetic and/or epigenetic modifications, are a valuable source of genetic variation to improve desirable polygenetic traits in crops. In this study, we induced somaclonal variation in vitro pineapple (Ananas comosus var. MD2) through hormonal induction, NaCl, and abscisic acid (ABA) supplementation. Our results showed that supplementation of high concentration of 6-benzylaminopurine (4.0 mg/L BAP) alone or combined with indole-butyric acid (IBA) produced the highest percentage of dwarf variants (100%). Murashige and Skoog (MS) media containing 4.0 mg/L BAP plus 2.0 mg/L IBA produced the shortest plantlets (1.9 ± 0.1 cm). In comparison, MS media containing 1.0% NaCl induced formation of dwarf plantlets with a mean plantlet height of 1.4 ± 0.3 cm, whereas 1.0 mg/L ABA generated plantlets with a mean plantlet height of 1.7 ± 0.1 cm. We then analyzed the histone deacetylase (HDAC) enzyme activity for dwarf and non-dwarf plantlets. In general, dwarf plantlets exhibited higher HDAC activity than non-dwarf plantlets. The highest HDAC activity (109, 333.33 ± 4.40 ng/min/mg) was recorded for dwarf plantlets grown on media supplemented with 1.0 mg/L ABA. The dwarf variants also underwent phenotypic recovery to normal phenotype within 8 months after transferred to MS basal media. No ploidy alteration was detected in these dwarf plantlets after analyzed by flow cytometry. Taken together, although the generated dwarf plantlets showed higher HDAC activity compared to non-dwarf plantlets, their capability of reverting to non-dwarf phenotype suggested that it might be due to epigenetic modulation.
KeywordsFlow cytometry Histone deacetylation Phenotype recovery Pineapple Somaclonal variants
The authors thank the Ministry of Higher Education, Malaysia for the financial support (FRGS Grant No. FP041-2014A) and University of Malaya, Malaysia, for providing the experimental facilities and PPP grant PG117-2015A.
JSY and NK conceived and designed the research. NAAH and MRM conducted the experiments. JSY, NK and MM contributed reagents and analytical tools. NAAH, BCT and JSY analyzed the data. NAAH, BCT and JSY wrote the manuscript. All authors read and approved the manuscript.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- Alwee SS, Van der Linden CG, Van der Schoot J, de Folter S, Angenent GC, Cheah S-C, Smulders MJM (2006) Characterization of oil palm MADS box genes in relation to the mantled flower abnormality. Plant Cell Tissue Organ Cult 85(3):331–344. https://doi.org/10.1007/s11240-006-9084-4 CrossRefGoogle Scholar
- Cuperlovic-Culf M, Culf AS (2014) Role of histone deacetylases in fungal phytopathogenesis: a review. Int J Mod Bot 4(2):48–60Google Scholar
- de Ancos B, Sánchez-Moreno C, Adolfo G (2017) Pineapple composition and nutrition. Handbook of pineapple technology: postharvest science, processing and nutrition. Wiley, ChichesterGoogle Scholar
- Eeuwens CJ, Lord S, Donough CR, Rao V, 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(3):311–323. https://doi.org/10.1023/a:1016543921508 CrossRefGoogle Scholar
- Ephritikhine G, Fellner M, Vannini C, Lapous D, Barbier-Brygoo H (1999) The sax1 dwarf mutant of Arabidopsis thaliana shows altered sensitivity of growth responses to abscisic acid, auxin, gibberellins and ethylene and is partially rescued by exogenous brassinosteroid. Plant J 18(3):303–314. https://doi.org/10.1046/j.1365-313X.1999.00454.x CrossRefPubMedGoogle Scholar
- Fraga HP, Vieira LN, Heringer AS, Puttkammer CC, Silveira V, Guerra MP (2016) DNA methylation and proteome profiles of Araucaria angustifolia (Bertol.) Kuntze embryogenic cultures as affected by plant growth regulators supplementation. Plant Cell Tissue Organ Cult 125(2):353–374CrossRefGoogle Scholar
- Grieve CM, Grattan SR, Maas EV, Wallender W, Tanji K (2012) Plant salt tolerance. Agricultural salinity assessment and management. 2nd edn, vol 71. ASCE manual reports on engineering practice. ASCE, Reston, pp 405–459Google Scholar
- Jain S, Brar D, Ahloowalia B (2013) Somaclonal variation and induced mutations in crop improvement, vol 32. Springer, DordrechtGoogle Scholar
- Latham T, Mackay L, Sproul D, Karim M, Culley J, Harrison DJ, Hayward L, Langridge-Smith P, Gilbert N, Ramsahoye BH (2012) Lactate, a product of glycolytic metabolism, inhibits histone deacetylase activity and promotes changes in gene expression. Nucleic Acids Res 40(11):4794–4803CrossRefPubMedPubMedCentralGoogle Scholar
- Law RD, Suttle JC (2005) Chromatin remodeling in plant cell culture: patterns of DNA methylation and histone H3 and H4 acetylation vary during growth of asynchronous potato cell suspensions. Plant Physiol Biochem 43(6):527–534 doi. https://doi.org/10.1016/j.plaphy.2005.03.014 CrossRefPubMedGoogle Scholar
- Liu X, Chen C-Y, Wang K-C, Luo M, Tai R, Yuan L, Zhao M, Yang S, Tian G, Cui Y (2013) PHYTOCHROME INTERACTING FACTOR3 associates with the histone deacetylase HDA15 in repression of chlorophyll biosynthesis and photosynthesis in etiolated Arabidopsis seedlings. Plant Cell 25(4):1258–1273CrossRefPubMedPubMedCentralGoogle Scholar
- Rastogi J, Siddhant PB, Sharma BL (2015) Somaclonal variation: a new dimension for sugarcane improvement. GERF Bull Biosci 6(1):5–10Google Scholar
- Semal J (2013) Somaclonal variations and crop improvement, vol 20. Springer, BerlinGoogle Scholar
- Tsaftaris AS, Polidoros AN, Koumproglou R, Tani E, Kovacevic N, Abatzidou E (2005) Epigenetic mechanisms in plants and their implications in plant breeding. In: Tuberosa R, Philips R, Gale M (eds) The wake of the double helix: from the green revolution to the gene revolution. Avenue Media, Bologna, pp 157–171Google Scholar
- Van Wyk BE (2005) Food plants of the world: identification, culinary uses and nutritional value. Briza Publications, PretoriaGoogle Scholar
- Wardy W, Saalia FK, Steiner-Asiedu M, Budu AS, Sefa-Dedeh S (2009) A comparison of some physical, chemical and sensory attributes of three pineapple (Ananas comosus) varieties grown in Ghana. Afr J Food Sci 3(4):094–099Google Scholar