Callus induction and indirect organogenesis in Chrysanthemum morifolium Ramat.

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Research Paper 01/09/2016
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Callus induction and indirect organogenesis in Chrysanthemum morifolium Ramat.

Rawnak Jahan Swarna, DilrubaYeasmin, Md. Mostafizur Rahman, Md. Firoz Alam
Int. J. Biosci.9( 3), 139-149, September 2016.
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A procedure for callus induction and indirect organogenesis of Chrysanthemum morifolium Ramat., using leaf segments as explant has been developed. For callus induction, leaf segments were cultured in MS medium (Murashige and Skoog’s, 1962) supplemented with 2,4-D singly (0.1, 0.5, 1.0, 1.5, 2.0 mg/litre) and IAA (0.5 mg/litre) in combination with BAP (0.1, 0.2, 0.5, 1.0, 1.5 mg/litre). Effective callus induction and growth were obtained in the medium of MS supplemented with 2.0 mg/l 2,4-D and 0.5 mg/l IAA + 0.2 mg/l BAP. 90% callus induced in both medium. Induced callus was transferred to the regeneration medium for effective shoot regeneration in 1.5 mg/l NAA + 1.0 mg/l BAP combination. The shoot regeneration frequency reached 90%. In order to induce roots, the shoots were transferred to the root induction media, and 1.0 mg/l IBA was recorded for maximum (100%) number of root induction on calli regenerated shoots. So, the indirect organogenesis of Chrysanthemum morifolium Ramat. was achieved through callus cultures and it may be possi​ble to regenerate a plant from genetically variable cells of the callus tissue.


Awal SMA, Alam MJ, Ali MR, Hasan MNU. 2005. In vitro Propagation of Pointed Gourd (Trichosanthes dioica Roxb.) from Shoot Tips. Biotechnology 4(3), 221-224.

Bajaj YPS, Shidhu MMS, Gill APS. 1992. Micropropagation of Chrysanthemum. In: Biotechnology in Agriculture and Forestry, 20. High-tech and Micropropagation iv Bajaj, Y.P.S. (Ed.). Springer-Verlag, Berlin. Heidelberg, New York. 69-80 P.

Bajaj YPS, Shidhu MMS, Gill APS. 1992. Some factors enhancing micropropagation of Chrysanthemum morifolium Ramat. Plant Tissue Culture 2(1), 41-47.

Ben-Jaacov J, Langhans RW. 1972. Rapid multiplication of chrysanthemum plant by stem tip proliferation. Horticultural Science 7, 289-290.

Bhattacharya P, Dey S, Das N, Bhattacharya BC. 1990. Rapid mass propagation of Chrysanthemum morifolium by callus derived from stem and leaf explants. Plant Cell Reports 9(8), 439-442.

Bhojwani SS, Rajdan MK. 1983.  Plant tissue  culture: Theory and Practice. Elsevier Sci. Pub. Amsterdam, Oxford, New York, Tokeyo. 313-327 P.

Chrysanthemum Flowers: Health Benefits, Medical Uses, & Nutritional Properties. 2015.

Das B, Haque MM, Islam AS, Rahman MH, Hoque MI. 1986. In vitro plantlet development in Jute. Prop. Nayudamma Mem. Sym.On Agricultural Application on Biotechnology. Madras, India. 106-114 P.

Din MSU, Nasir UJK, Zaman S, Reza MA. 2005. Regeneration of Multiple Shoots from Different Explants viz. Shoot Tip, Nodal Segment and Cotyledonary Node of in vitro Grown Seedlings of Peltophorumm pterocarpum (DC.) Backer ex K. Heyne. Biotechnology 4(1), 35-38.

Earle ED, Langhans RW. 1973. Propagation of Chrysanthemum in vitro. 1. Multiple plantlets from shoot tips and the establishment of tissue culture. Journal of Horticultural Science 99, 128-132.

Gautheret RJ. 1939. Sur Ia possibility de realiserIa culture indefinie des tissue de tubercules de carrotle. C. R. Hebd, Seanc. Acad. Sci. Paris 208, 118-121.

Haberlandt G. 1902. Kulturversuche mit isolierten pflanzenzellen. Sitzungsber. Akad. Wiss. Wien. Math-Naturwiss. Kl. Abt. J. 111, 69-92.

Hitmi A, Barthomeuf C, Sallanon H. 1999. Rapid mass propagation of Chrysanthemum cinerariaefolium Vis. by callus culture and ability to synthesisepyrethrins. Plant Cell Report 19, 156–160.

Hoque MI. 1995. In vitro multiple shoot regeneration in Chrysanthemum morifolium Ramat. Plant Tissue Culture 5(2), 153-162.

Hutchinson JF, Kaul U, Maheswaran G,  Moran JR, Craham MW, Richards D. 1982. Genetic improvement of floricultural crops using biotechnology. Australian Journal of Botany 40, 765-787.

Karim MZ, Amin MN, Azad MAK, Begum F, Islam MM, Alam R. 2002. Effect of different Plant Growth regulators on in vitro Shoot Multiplication of Chrysanthemum morifolium. Online Journal of Biological Sciences 3(6), 553-560.

Komalavalli N, Rao MV. 2000. In vitro micro-propagation of Gymnemam slyvestre. A multipurpose medicinal plant. Plant Cell Tissue Organ Culture 61, 97-105.

Lindiro C, Kahia J, Asiimwe T, Mushimiyimana I, Waweru B, Kouassi M, Koffi E, Kone S, Sallah PY. 2013. In vitro regeneration of pyrethrum (Chrysanthemum cinerariaefolium) plantlets from nodal explants of in vitro raised plantlets. International Journal of Application or Innovation in Engineering & Management 2(7), 207-213.

Litz RE, Jaiswal VS. 1990. Micropropagation of tropical and sub-tropical fruits. In: Debergh and Zimmerman RH (eds). Kluwer Academic Publishers, Dordrecht. Micropropagation Technol. Application. 247-266 P.

Munshi MK, Hakim L, Islam MR, Ahmed G. 2004. In vitro clonal propagation of Banyan (Ficus benghalensis L.) through axillary bud culture. International Journal Agriculture Biology 6(2), 321-323.

Murashige T, Skoog F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum 15, 473-497.

Nahid JS, Shyamali S, Kazumi H. 2007. High frequency shoot regeneration from petal explants of Chrysanthemum morifolium In vitro. Pakistan Journal of Biological Sciences 10(19), 3356-3361.

Nhut DT, Don NT, An TTT, Van TPT, Vu NH, Huyen PX, Khiem DV. 2005. Microponic and hydroponic techniques in disease free chrysanthemum (Chrysanthemum sp.) production. Journal of Applied Horticulture 7, 67-71.

Rajani H, Patil SS. 2009. In vitro response of different explants’ types on shoot and root development of Ginger. ISHS Acta Horticulturae 829: VI International Symposium on In Vitro Culture and Horticultural Breeding.

Salinger JP. 1991. Production commercial de  Flores. Zaragoza: Acribia. 371 P.

Sarker RH, Shaheen I. 2001. In vitro Propagation of Chrysanthemum (Chrysanthemum morifolium Ramat.) Through Callus Culture. Plant Tissue Culture 11(1), 85-91.

Sauvaire D, Galgy R. 1978. Multiplication of vegetative dela Cannal a sourse par bounturage in vitro. Comptes rendus de l’Académie des Sciences 287, 446-470.

Shatnawi M, Al-Fauri A, Megdadi H, Kair M, Al-Shatnawi, Saead RS, Abu-Romman, Al-Ghzawi AL. 2010. In Vitro Multiplication of Chrysanthemum morifolium Ramat. and it is Responses to NaCl Induced Salinity. Jordan Journal of Biological Sciences 3(3), 101-110.

Vantu S. 2006. Organogenesis in Chrysanthemum morifolium Ramat. (Cultivar “Romica”) Callus Cultures. Analelestiintifice ale Universitatii “Al. l. Cuza” lasiTomul LII, s. ll a. Biologievegetala. Faculty of Biology.  71 P.

Wankhede KN, Narkhede MN, Shivankar RS, Rathod TH. 2000. Callus induction and micropropagation studies in chrysanthemum. Annals of Plant Physiology 14(2).