Callus Induction on True Shallot Seed Explant Using a Combination of BA and 2,4-D

Krisna Dharmayanti(1), Endang Sulistyaningsih(2*), Rani Agustina Wulandari(3)

(1) Department of Agronomy, Faculty of Agriculture, Universitas Gadjah Mada Jln. Flora no. 1, Bulaksumur, Sleman, Yogyakarta 55281
(2) Department of Agronomy, Faculty of Agriculture, Universitas Gadjah Mada Jln. Flora no. 1, Bulaksumur, Sleman, Yogyakarta 55281
(3) Department of Agronomy, Faculty of Agriculture, Universitas Gadjah Mada Jln. Flora no. 1, Bulaksumur, Sleman, Yogyakarta 55281
(*) Corresponding Author


BA and 2,4-D combination were commonly used for in vitro culture of Shallot (Allium cepa L. var agregatum 2n = 2x = 16) to induce callus, but there was no information for callus induction on shallot seed (TSS) explant. Callus could be utilized for in vitro selection and generating of genetic variation. The aims of the research was to identify the response of TSS (Trisula and Tuk Tuk) as explant  and to obtain the optimum combination of BA and 2,4-D (mg.L-1): (0–0, 2–1, 2–2, 2–3, and 2–4) in callus induction. The research had been carried out in the Tissue Culture Laboratory, Faculty of Agriculture, Universitas Gadjah Mada during the year 2015-2016. Factorial treatments of variety and growth regulators were arranged in Completely Randomized Design with four replications. Data of percentage of germination, shoot height, root length, percentage of callus formation, callus weight, and chromosomes number of callus were recorded. The results showed that combination of 2 mg.L-1 BA + (1– 4) mg.L-1 2,4-D induced callus formation on TSS but inhibits shoots and roots growth. The best callus proliferation was at a concentration of 1 mg.L-1 2,4-D. Tetraploid callus chromosomes (2n = 4x = 32) was detected in Trisula grown in the 2 mg.L-1 BA + 4 mg.L-1 2,4-D, but in the Tuk Tuk callus did not detected the changing of chromosomes number.


Chromosome number; plant growth regulator; tetraploid

Full Text:



Bhojwani, S.S. and P.K. Dantu. 2013 Plant Tissue Culture: An Introductory Text. Springer, London. [online] Available at: http: //www. prof/images/Uploaded_files/()Sant Saran Bhojwani, Prem Kumar Dantu (auth.)-Plant Tissue Culture_ An Introductory Text Springer India (2013)[3811896].PDF. [Accessed 15 May 2017].

Chaudhury, A. and R. Qu, 2000. Somatic embryogenesis and plant regeneration of turf-type bermudagrass: effect of 6-benzyladenine in callus induction medium. Plant Cell, Tissue and Organ Culture. 60, pp. 113–120.

Chauhan, M.and  S.L. Kothari. 2004. Optimization ofnutrient levels in the medium increase the efficiency of callus induction and plant regeneration in recalcitrant Indian barley (Hordeum vulgareL.) in vitro. In VitroCellular and Developmental Biology-Plant. 40, pp. 520–527.

Duval Y., T.D. Gosselin, K. Konan and  C. Pannetier. 1988. In vitro vegetative micropropagation of oil palm (Elaeisguineensis Jacq.). Strategy and Results. Proceedings of the 1987 Internation Oil Palm/Palm Oil Conference (Agriculture), Bangi. Malaysia.

Flick, C.E., D.A. Evans, and W.R. Sharp. 1993. Organogenesis in D.A. Evans, W.R. Sharp, P.V. Amirato, and T. Yamada (eds.). Hand book of Plant Cell Culture. London: Collier Macmillan Publisher.

Hailekidan, B., M. Andargie,  and K. Assefa. 2013. In vitro plantlet regeneration from the bulbs of shallot (Allium cepa var. group Aggregatum). Research in Plant Science. 1, pp. 45–52.

Jouanneau, J.P. 1971 Côntrole par les cytokinines de la synchronisation des mitoses dans les cellules de tabac. Exp. Cell Res., 67, pp. 329–337.

Kaeppler,  S.M., H.F.  Kaeppler, and Y. Rhee . 2000. Epigenetic aspects of somaclonal variation in plants. Plant Mol. Biol., 43, pp. 179–188.

Kikuchi A., N. Sanuki , K. Higashi, T. Koshiba, and H. Kamada . 2006. Abscisic acid and stresstreatment are essential for the acquisition ofembryogenic competence  by carrot  somatic cells. Planta 223: 637-645.

Larkin, P.J. and W.R. Scowcroft. 1981. Somaclonal variation a novel source of variability from cell cultures for plant improvement. Theor. Appl. Genet, 60, pp. 197–214.

Leva, A.R., R. Petruccelli & L.M.R. Rinaldi. 2012. Somaclonal Variation in Tissue Culture: A Case Study With Olive. Recent Advance in Plant In Vitro Culture. 10–17.

Niizeki, M. and L. Zhongen. 2003. Somaclonal variation as a tool for plant breeding and genetics. Bull. Fac. Agric. And Life Sci. Hirosaki Univ., 6, pp. 1–7.

Salisbury, F.B and C.W. Ross. 1995. Fisiologi tumbuhan.3 th ed. Bandung: ITB.

Shah, M.I., M. Jabeen,  and I Ilahi . 2003.In vitro callus induction, its proliferation and regeneration inseed explants of Wheat (Triticum aestivum L.) Var.Lu-26s. Pakistan Journal of Botany,35, pp. 209–217.

Sharma, V.K., R. Hansch, R.R. Mendel, and J. Schulze. 2005. Mature embryo axis based high-frequency somatic embryogenesis and plant regeneration from multiple cultivars of barley ( Hordeum vulgare L.). J. Exp. Bot., 56, pp. 1913–1922.

Skirvin, R.M., Norton, M. and Mcpheeter, K.D. (1993). Somaclonal variation: Has it proved useful for plant improvement. Acta Hort., 336, pp. 333−340.

Sukmadjaja, D. and Ade, M. 2011. Pertunasan dan pertumbuhan beberapa varietas tebu (Sacharum officinarum L.) secara in vitro. Bogor: Balai Besar Penelitian dan Pengembangan Bioteknologi Sumberdaya Genetik Pertanian.

Sulistyaningsih, E., Aoyagi, Y.and Tashiro, Y. 2006. Flower bud culture of shallot (Allium cepa L. Aggregatum group) with cytogenetic analysis of resulting gynogenic plants and somaclones. Plant Cell Tiss Organ Cult., 86, pp. 248–25.

Triharyanto, E. and Purnomo, D. 2014. Study of viability and seed structure of shallot. Journal of Agricultural Science and Technology, 4, pp. 121–125.


Article Metrics

Abstract views : 979 | views : 919


  • There are currently no refbacks.

   Ilmu Pertanian (Agricultural Science) ISSN 0126-4214 (print), ISSN 2527-7162 (online) is published by Faculty of Agriculture Universitas Gadjah Mada collaboration with Perhimpunan Sarjana Pertanian Indonesia (PISPI) and licensed under a  Creative Commons Attribution-ShareAlike 4.0 International License.

analytics View My Stats