Abstract
The present study reports the role of auxin and its transport inhibitor during the establishment of an efficient and optimized protocol for the somatic embryogenesis in Digitalis trojana Ivan. Hypocotyl segments (5 mm long) were placed vertically in the Murashige and Skoog medium supplemented with three sets [indole-3-acetic acid (IAA) alone or 2,3,5-triiodobenzoic acid (TIBA) alone or IAA–TIBA combination] of formulations of plant growth regulators, to assess their differential influence on induction and proliferation of somatic embryos (SEs). IAA alone was found to be the most effective, at a concentration of 0.5 mg/l, inducing ~ 10 SEs per explant with 52% induction frequency. On the other hand, the combination of 0.5 mg/l of IAA and 1 mg/l of TIBA produced significantly fewer (~ 3.6 SEs) and abnormal (enlarged, oblong, jar and cup-shaped) SEs per explant with 24% induction frequency in comparison to that in the IAA alone. The explants treated with IAA–TIBA exhibited a delayed response along with the formation of abnormal SEs. Our study revealed that IAA induces high-frequency SE formation when used singly, but the frequency gradually declines when IAA was coupled with increasing levels of TIBA. Eventually, our findings bring new insights into the roles of auxin and its polar transport in somatic embryogenesis of D. trojana.
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Abbreviations
- DMRT:
-
Duncan multiple range test
- IAA:
-
Indole-3-acetic acid
- MS:
-
Murashige and Skoog (1962)
- PGR:
-
Plant growth regulator
- SE:
-
Somatic embryo
- TIBA:
-
2,3,5-triiodobenzoic acid
References
Calderón-Montaño JM, Burgos-Morón E, Orta ML, Maldonado-Navas D, García-Domínguez I, López-Lázaro M (2014) Evaluating the cancer therapeutic potential of cardiac glycosides. Biomed Res Int 2014:794930. https://doi.org/10.1155/2014/794930
Chée RP, Cantliffe DJ (1989) Inhibition of somatic embryogenesis in response to 2,3,5-triiodobenzoic acid and 2,4-dichlorophenoxyacetic acid in Ipomoea batatas (L.) Lam. cultured in vitro. J Plant Physiol 135(4):398–403. https://doi.org/10.1016/S0176-1617(89)80094-X
Choi Y, Kim H, Soh W, Yang D (1997) Developmental and structural aspects of somatic embryos formed on medium containing 2,3,5-triiodobenzoic acid. Plant Cell Rep 16(11):738–744. https://doi.org/10.1007/s002990050312
Compton ME (1994) Statistical methods suitable for the analysis of plant tissue culture data. Plant Cell Tissue Organ Cult 37:217–242. https://doi.org/10.1007/BF00042336
Cooke TJ, Racusen RH, Cohen JD (1993) The role of auxin in plant embryogenesis. Plant Cell 5(11):1494–1495. https://doi.org/10.1105/tpc.5.11.1494
Davis PH (1978) Digitalis L. In: Davis PH (ed) Flora of Turkey and the East Aegean Islands, vol 6. Edinburg University Press, Edinburg, pp 680–687
De Klerk GJ, Guan H, Huisman P, Marinova S (2011) Effects of phenolic compounds on adventitious root formation and oxidative decarboxylation of applied indoleacetic acid in Malus ‘Jork 9’. Plant Growth Regul 63(2):175–185. https://doi.org/10.1007/s10725-010-9555-9
Dodeman VL, Ducreux G, Kreis M (1997) Zygotic embryogenesis versus somatic embryogenesis. J Exp Bot 48(8):1493–1509. https://doi.org/10.1093/jxb/48.8.1493
Duncan DB (1955) Multiple range and multiple F test. Biometrics 11:1–42. https://doi.org/10.2307/3001478
Friml J (2003) Auxin transport—shaping the plant. Curr Opin Plant Biol 6(1):7–12. https://doi.org/10.1016/S1369526602000031
Friml J, Vieten A, Sauer M, Weijers D (2003) Efflux-dependent auxin gradients establish the apical–basal axis of Arabidopsis. Nature 426(6963):147–153. https://doi.org/10.1038/nature02085
Gantait S, Kundu S (2017) Neoteric trends in tissue culture-mediated biotechnology of Indian ipecac [Tylophora indica (Burm. f.) Merrill]. 3 Biotech 7(3):231. https://doi.org/10.1007/s13205-017-0865-8
Halperin W, Wetherell D (1964) Adventive embryony in tissue cultures of the wild carrot, Daucus carota. Am J Bot 51(3):274–283. http://www.jstor.org/stable/2440296
Hamasaki RM, Purgatto E, Mercier H (2005) Glutamine enhances competence for organogenesis in pineapple leaves cultivated in vitro. Braz J Plant Physiol 17(4):383–389. https://doi.org/10.1590/S1677-04202005000400006
Kreis W (2017) The foxglove (Digitalis) revisited. Planta Med 83(12–13):962–976. https://doi.org/10.1055/s-0043-111240
Kuberski C, Scheibner H, Steup C, Diettrich B, Luckner M (1984) Embryogenesis and cardenolide formation in tissue cultures of Digitalis lanata. Phytochemistry 23(7):1407–1412. https://doi.org/10.1016/S0031-9422(00)80475-6
Kumar A, Shekhawat N (2009) Plant tissue culture and molecular markers: their role in improving crop productivity, 1st edn. IK International Publishing House, New Delhi
Liu C, Xu Z, Chua NH (1993) Auxin polar transport is essential for the establishment of bilateral symmetry during early plant embryogenesis. Plant Cell 5(6):621–630. https://doi.org/10.1105/tpc.5.6.621
Liu Y, Dong Q, Kita D, Huang JB, Liu G, Wu X, Zhu X, Cheung AY, Wu HM, Tao LZ (2017) RopGEF1 plays a critical role in polar auxin transport in early development. Plant Physiol 175(1):157–171. https://doi.org/10.1104/pp.17.00697
Michalczuk L, Ribnicky DM, Cooke TJ, Cohen JD (1992) Regulation of indole-3-acetic acid biosynthetic pathways in carrot cell cultures. Plant Physiol 100(3):1346–1353. https://doi.org/10.1104/pp.100.3.1346
Morris DA (2000) Transmembrane auxin carrier systems–dynamic regulators of polar auxin transport. Plant Growth Regul 32(2):161–172. https://doi.org/10.1023/A:1010701527848
Munkert J, Franco MS, Nolte E, Silva IT, Castilho RO, Ottoni FM, Schneider NF, Oliveira MC, Taubert H, Bauer W, Andrade SF, Alves RJ, Simões CMO, Braga FC, Kreis W, de Pádua RM (2017) Production of the cytotoxic cardenolide glucoevatromonoside by semisynthesis and biotransformation of evatromonoside by a Digitalis lanata cell culture. Planta Med 83(12–13):1035–1043. https://doi.org/10.1055/s-0043-109557
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15(3):473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
Mutschler E, Geisslinger G, Kroemer HK, Ruth P, Schäfer-Korting M (2008) Arzneimittelwirkungen-Lehrbuch der Pharmakologie und Toxikologie. Endo-Praxis 24(03):29 (in German)
Pasternak TP, Prinsen E, Ayaydin F, Miskolczi P, Potters G, Asard H, Van Onckelen HA, Dudits D, Fehér A (2002) The role of auxin, pH, and stress in the activation of embryogenic cell division in leaf protoplast-derived cells of alfalfa. Plant Physiol 129(4):1807–1819. https://doi.org/10.1104/pp.000810
Sahrawat AK, Chand S (2001) Continuous somatic embryogenesis and plant regeneration from hypocotyl segments of Psoralea corylifolia Linn., an endangered and medicinally important Fabaceae plant. Curr Sci 81(10):1328–1331
Schiavone FM, Cooke TJ (1987) Unusual patterns of somatic embryogenesis in the domesticated carrot: developmental effects of exogenous auxins and auxin transport inhibitors. Cell Differ 21(1):53–62. https://doi.org/10.1016/0045-6039(87)90448-9
Sharp WR, Sondahl MR, Caldas LS, Maraffa SB (1980) The physiology of in vitro asexual embryogenesis. In: Janick J (ed) Horticultural reviews, vol 2. Wiley, Hoboken, pp 268–310. https://doi.org/10.1002/9781118060759.ch6
Soriano M, Li H, Boutilier K (2013) Microspore embryogenesis: establishment of embryo identity and pattern in culture. Plant Reprod 26(3):181–196. https://doi.org/10.1007/s00497-013-0226-7
Thorpe TA (1994) Morphogenesis and regeneration. In: Vasil IK, Thorpe TA (eds) Plant cell and tissue culture. Springer, Dordrecht, pp 17–36. https://doi.org/10.1007/978-94-017-2681-8_2
Vanneste S, Friml J (2009) Auxin: a trigger for change in plant development. Cell 136(6):1005–1016. https://doi.org/10.1016/j.cell.2009.03.001
Venkatesh K, Rani AR, Baburao N, Padmaja G (2009) Effect of auxins and auxin polar transport inhibitor (TIBA) on somatic embryogenesis in groundnut (Arachis hypogaea L.). Afr J Plant Sci 3(12):288–293. http://www.academicjournals.org/journal/AJPS/article-full-text-pdf/0F99E4811235
Verma SK, Sahin G, Yucesan B, Eker I, Sahbaz N, Gurel S, Ekrem G (2012) Direct somatic embryogenesis from hypocotyl segments of Digiatlis trojana Ivan and subsequent plant regeneration. Ind Crops Prod 40:76–82. https://doi.org/10.1016/j.indcrop.2012.02.034
Verma SK, Das AK, Cingoz GS, Gurel E (2016) In vitro culture of Digitalis L. (Foxglove) and the production of cardenolides: an up-to-date review. Ind Crops Prod 94:20–51. https://doi.org/10.1016/j.indcrop.2016.08.031
Acknowledgements
The authors are grateful to the Scientific and Technological Research Council of Turkey (TUBITAK) for financial support. A research grant under the visiting scientist programme (Code 2221-TUBITAK) was awarded to Sandeep Kumar Verma.
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Verma, S.K., Das, A.K., Gantait, S. et al. Influence of auxin and its polar transport inhibitor on the development of somatic embryos in Digitalis trojana. 3 Biotech 8, 99 (2018). https://doi.org/10.1007/s13205-018-1119-0
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DOI: https://doi.org/10.1007/s13205-018-1119-0