Abstract
The optimum physical and chemical microenvironment for micropropagation of Limonium sinensis (Girard) Kuntze, var. Golden Diamond was established from immature inflorescence segments as explant. The highest frequency (62 %) of axillary shoot induction was obtained on MS medium (Murashige and Skoog Physiol Plant 15:473–497, 1962) supplemented with 8.88 μM BA, 1.34 μM of NAA and two growth additives cysteine (142.33 μM), and glutamine (684.22 μM). In the subsequent culture maximum average number of shoots (11.13 ± 0.34) were obtained from micro-shoot explant on MS medium supplemented with the same additives and 2.22 μM BA. During subcultures the problem of vitrification was mitigated through increasing agar concentration from 0.8 % to 1.0 % and providing better ventilation. The in vitro developed shoots were rooted on MS medium supplemented with 2.46 μM IBA and 0.88 μM BA. Rooted plants were acclimatized successfully in the greenhouse with 80 % survival rate. RAPD analysis using 15 random decamer primers generated monomorphic banding pattern in micropropagated plants and similar to those of mother plant revealing the genetic integrity of regenerants.
Abbreviations
- BA:
-
N6 -Benzylaminopurine
- IBA:
-
Indole 3-butyric acid
- Kn:
-
Kinetin
- MS:
-
Murashige and Skoog
- NAA:
-
αNaphthalene acetic acid (NAA)
- PGRs:
-
Plant growth regulators
- TDZ:
-
Thidiazuron
- DD:
-
Double distilled
References
Bernier G (1992) Attempts to bridge the molecular genetics and physiology of inflorescence and flower morphogenesis are an exciting experience. Flower Newslett 14:34–40
Bruna S, De Benedetti L, Mercuri A, Schiva T, Burchi G, Pecchiona N, Agrimonti C (2004) Use of RAPD markers for the genetic characterization of Limonium species. ISHS Acta Horticult 651:155–160
Casanova E, Moysett L, Trillas MI (2008) Effects of agar concentration and vessel closure on the organogenesis and hyperhydricity of adventitious carnation shoots. Biol Plant 52:1–8
Dam A, Paul S, Bandyopadhyay TK (2010) Direct somatic embryogenesis and plant regeneration from leaf explants of Limonium sinensis (Girard) Kuntze. Sci Hortic 126:253–260
Deklerk GJ, Hanecakova J, Jasik J (2001) The role of cytokinins in rooting of stem slices cut from apple microcuttings. Plant Biosyst 135:79–84
Doyle JJ, Doyle JL (1990) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull 19:11–15
Huang CL, Hsieh MT, Hsieh WC, Sagare AP, Tsay HS (2000) In vitro propagation of Limonium wrightii (Hance) Ktze. (Plumbaginaceae), an ethnomedicinal plant, from shoot-tip, leaf- and inflorescence-node explants. In Vitro Cell Dev Biol Plant 36:220–224
Larkin PJ, Scowcroft W (1981) Somaclonal variation: a novel source of variability from cell cultures for plant improvement. Theor Appl Genet 60:197–214
Li Y (2008) Effect of salt stress on seed germination and seedling growth of the salinity plants. Pak J Biol Sci 11:1268–1272
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol Plant 15:473–497
Rout GR, Das G (2002) An assessment of genetic integrity of micropropagated plants of Plumbago zeylanica by rapd markers. Biol Plant 45(1):27–32
Samantaray S, Kumar VS, Maiti S (2009) Direct shoot regeneration from immature inflorescence cultures of Chlorophytum arundinaceum and Chlorophytum borivilianum. Biologia 64(2):305–309
Topoonyanont N, Ampawan R, Pierre CD (2000) Reversion of Limonium hybrid ‘misty blue’ inflorescence development and its applicability in micropropagation. Sci Hortic 83:283–299
Trifi M, Rhouma A, Marrakchi M (2000) Phylogenetic relationships in Tunisian date-palm (Phoenix dactylifera L.) germplasm collection using DNA amplification fingerprinting. Agronomie 20:665–671
Ziv M (1991) Vitrification: morphological and physiological disorders of in vitro plants. In: Debergh PC, Zimmerman RH (eds) Micropropagation: technology and application. Kluwer, Dordrecht, pp 45–69
Acknowledgments
This research was financially supported from the “University Research Scholarship”, University of Kalyani, Kalyani, India.
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Dam, A., Paul, S., Bhattacharya, C. et al. Effects of culture conditions on multiple shoot induction from inflorescence and RAPD analysis of cloned plants in Limonium sinensis (Girard) Kuntze, var. Golden Diamond. J. Plant Biochem. Biotechnol. 22, 348–352 (2013). https://doi.org/10.1007/s13562-012-0164-8
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DOI: https://doi.org/10.1007/s13562-012-0164-8