In vitro propagation of Tunisian local garlic (Allium sativum L.) from shoot tip culture

Ayed, Chadha; Bayoudh, Chokri; Rhimi, Awatef; Mezghani, Najla; Haouala, Faouzi; AL Mohandes Dridi, Bouthaina

doi:10.22077/jhpr.2018.1457.1016

Document Type : Original Article

Authors

¹ High Agronomic Institute, 4042 Chatt-Mariem, IRESA-University of Sousse, Tunisia

² Regional Research Centre on Horticulture and Organic Agriculture (CRRHAB)

³ National Gene Bank of Tunisia

⁴ Al Imam Mohammad Ibn Saud Islamic University (IMSIU)

https://doi.org/10.22077/jhpr.2018.1457.1016

Abstract

Purpose: Tunisian garlic is widely threatened by the attack of several viruses genus. For this purpose, a reliable protocol was established for rapid in vitro propagation of local garlic (Allium sativum L.) cultivars for large-scale production of virus-free plants and high quality bulblets. Research method: Well disinfected shoot-tips of 1 mm were used as explants and cultivated on MS basal solid media enriched with various growth regulators: 6-Benzylaminopurine, α-Naphthaleneacetic acid, Kinetin, Indole-3-butyric acid and 2-isopentenyladenine for assessment of shoot formation, shoot proliferation and bulb formation. Findings: Among the different phytohormone concentrations and combinations, MS basal medium without any growth regulators (M₀) was found optimal for shoot-tip initiation (96% explants development) and plantlets elongation (56.26 mm). For shoot proliferation, the M₁culture medium containing 1 mg L^-1 BAP and 0.25 mg L^-1 NAA was the best, giving a multiplication rate of 1.7 plantlets/explant. Shoots on M₀culture medium formed bulblets earlier. Multiple bulblets per explants were obtained on medium M₂₂containing 2 mg L^-1 Kin and 0.1 mg L^-1 NAA. Separated bulblets were transferred individually on bulbification media. Non-dividable bulblet was developed in various sizes. Research limitations: Bulblet acclimatization step needs to be well studied for high quality cloves production. Originality/value: This efficient optimized in vitro protocol will be successfully applied for large multiplication of virus-free garlic cultivars.

Keywords

Main Subjects

Biotechnology and Tissue culture

References

Asha, Devi A., Khar, A., & Lawande, K. E. (2007). Genotypic response of short day garlic (Allium sativum L.) accessions to shoot multiplication. Journal of Spices and Aromatic Crops, 16, 15-21.

Ayabe, M., & Sumi, S. (1998). Establishment of a novel tissue culture method, stem-disc culture, and its practical application to micropropagation of garlic (Allium sativum L.).Plant Cell Reports, 17, 773-779.

Bekheet, S. A. (2006). A synthetic seed method through encapsulation of in vitro proliferated bulblets of garlic (Allium sativum L.).Arab Journal of Biotechnology, 9, 415-26.

Dixit, V., & Chaudhary, B.R. (2013). Allium sativum: four step approach to efficient micropropagation. International Journal of Innovative Biological Research, 2, 6-14.

Gad El-Hak, S.E.N.H., Ahmed, K. Z., Moustafa, Y. M., & Ezzat A. S. (2011). Growth and cytogenetical properties of micro-propagated and successfully acclimatized garlic (Allium sativum L.) clones with a modified shoot tip culture protocol.Journal of Horticultural Science and Ornamental Plants, 3, 115–129.

Ghaemizadeh, F., Dashti, F., Khodakaramian, G., & Sarikhani, H. (2014). Combination of stem-disc dome culture and thermotherapy to eliminate Allexiviruses and Onion yellow dwarf virus from garlic (Allium sativum cv. Hamedan). Archives of Phytopathology and Plant Protection, 47, 499-507.

Gimenez, M.D., Yanez-Santos A.M., Paz, R.C., Quiroga, M.P., Marfil, C.F., Conci, V.C., & Garcia-Lampasona, S.C. (2016). Assessment of genetic and epigenetic changes in virus-free garlic (Allium sativum L.) plants obtained by meristem culture followed by in vitro propagation. Plant Cell Reports, 35, 129-141.

Gull, I., Noreen, A., Aslam, M.S., & Athar, M.A. (2014). Comparative effect of different pyhtohormones on the micropropagation of Allium sativum. Pakistan Journal of Biochemistry and Molecular Biology,47, 121-124.

Haider, S., Hossain, M., Rahman, S., Sultana, S., Quddus, T., Chakraborti, M., Hoque, A., Shahriar, M., & Haque M. (2015). In vitro plantlet regeneration of four local garlic (Allium sativum) accessions of Bangladesh. British Biotechnology Journal, 8, 1-12.

Hammami, I., & El May, M. V. (2012). Impact of garlic feeding (Allium sativum) on male fertility. Andrologia, 45, 217-224.

Haque, M.S., Wada, T., & Hattori, K. (2003). Shoot regeneration and bulblet formation from shoot and root meristem of garlic cv. Bangladesh local. Asian Journal of Plant Sciences,2, 23-27.

Hassan, M.N., Haque, M.S., Hassan, M.M., & Haque, M.S. (2014). An efficient protocol for somatic embryogenesis of garlic (Allium sativum L.) using root tip as explant.Journal of the Bangladesh Agricultural University, 12, 1-6.

Ikeuchi, M., Sugimoto, K., & Iwase, A. (2013). Plant callus: mechanisms of induction and repression. The Plant Cell, 25, 3159-3173.

Izquierdo-Oviedo, H., Disotuar, R., Gonzalez, M. C., & Gonzalez, S.J. (2016).Micropropagation of garlic (Allium sativum L.) and determination of the genetic stability of the plantlets obtained by AFLP markers. Biotecnologia Aplicada, 33, 4211-4218.

Jabbes, N., Geoffriau, E., Le Clerc, V., Dridi, B., & Hannechi, C. (2011). Intersimple sequence repeat fingerprints for assess genetic diversity of Tunisian garlic populations. Journal of Agriculture Science, 3, 77-85.

Khoshtinat, K., Barzegar, M., Sahari, M.A., & Hamidi, Z. (2017). Encapsulation of Iranian garlic oil with β-cyclodextrin: Optimization and its characterization. Journal of Agricultural Science and Technology, 19, 97-111.

Koch, M., & Salomon, R. (1994). Improvement of garlic via somaclonal variation and virus elimination. International Symposium on Alliums for the Tropics. Bangkok, Thailand.

Luciani, F.G., Mary, A.K., Pellegrini, C., & Curvetto, N.R. (2006). Effects of explants and growth regulators in garlic callus formation and plant regeneration. Plant Cell, Tissue and Organ Culture, 87, 139-143.

Mahajan, R. (2016). In vitro and cryopreservation techniques for conservation of snow mountain garlic.In Protocols for in vitro cultures and secondary metabolite analysis of aromatic and medicinal plants, (Jain SM ed.). Humana Press, New York, 335-346.

Manjunathagowda, D.C., Gopal, J., Archana, R., & Asiya, K.R. (2017). Virus–free seed production of garlic (Allium sativum L.): Status and prospects. International Journal of Current Microbiology and Applied Sciences, 6, 2446-2456.

McDonald, M.R., De los Angeles, J.M., & Hovius, M.H. (2004). Management of diseases of onions and garlic. In Diseases of fruits and vegetables: Volume II (Naqvi S.A.M.H ed.). Springer, Dordrecht, 149-200.

Mehta, J., Sharma, A., Sharma, N., Megwal, S., Sharma, G., Gehlot, P., & Naruka. R. (2013). An improved method for callus culture and in vitro propagation of garlic (Allium sativum L.). International Journal of Pure & Applied Bioscience, 1, 1-6.

Metwally, E.I., El-Denary, M.E., Omar, A.M.K., Naidoo, Y., & Dewir.Y.H. (2012). Bulb and vegetative characteristics of garlic (Allium sativum L.) from in vitro culture through acclimatization and field production. African Journal of Agricultural Research, 7, 5792–5795.

Metwally, E. I., El-Denary, E., Dewi, Y. H., & Naidoo Y. (2014). In vitro propagation of garlic (Allium sativum L.) through adventitious shoot organogenesis. African Journal of Biotechnology, 13, 3892-3900.

Mishra, R. K., Jaiswal, R. K., Kumar, D., Saabale, P. R., & Singh A. (2014). Management of major diseases and insect pests of onion and garlic: A comprehensive review. Journal of Plant Breeding and Crop Science, 6, 160-170.

Mukhopadhyay, M.J., Sengupta, P., Mukhopadhyay, S., & Sen, S. (2005). In vitro stable regeneration of onion and garlic from suspension culture and chromosomal instability in solid callus culture. Scientia Horticulturae, 104, 1-9.

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

Parrano, L., Afunian, M., Pagliaccia, D., Douhan, G., & Vidalakis, G. (2012). Characterization of viruses associated with garlic plants propagated from different reproductive tissues from Italy and other geographic regionspra. Phytopathologia Mediterranea, 51, 549-565.

Pramesh, D., & Baranwal, V.K. (2015). Production of virus-free garlic (Allium sativum L.) through meristem tip culture after solar or hot air treatment of cloves. The Journal of Horticultural Science and Biotechnology, 90, 180-186.

Perotto, M.C., Cafrune, E.E., & Conci, V.C. (2010). The effect of additional viral infections on garlic plants initially infected with Allexiviruses. European Journal of Plant Pathology, 126, 489-495.

Roksana, R., Alam, M.F., Islam, R., & Hossain, M.M. (2002). In vitro bulblet formation from shoot apex in garlic (Allium sativum L.). Plant Tissue Culture, 12, 11-17.

Robledo-Paz, A., & Tovar-Soto, M.H. (2012). Biotechnological tools for garlic propagation and improvement. Innovations in Biotechnology, 1-27.

Sarasan, V., Cripps, R., Ramsay, M.M., Atherton, C., McMichen, M., Prendergast, G., & Rowntree J.K. (2006). Conservation in vitro of threatened plants - Progress in the past decade. In Vitro Cellular and Developmental Biology – Plant, 42, 206-214.

Shyab, S. (2017).Response of garlic callus to salt level. Academia Journal of Biotechnology, 5, 006-011.

Taskin, H., Baktemur, G., Kurul, M., & Buyukalaca, S. (2013). Use of tissue culture techniques for producing virus-free plant in garlic and their identification through Real-Time PCR. Scientific World Journal. Volume 2013, Article ID 781282, 5 pages.

Vieira, R.L., da Silva, A.L., Zaffari, G.R., Steinmacher, D.A., de Freitas Fraga, H.P., & Guerra, M.P. (2015). Efficient elimination of virus complex from garlic (Allium sativum L.) by cryotherapy of shoot tips. Acta Physiologiae Plantarum, 37, 1733.

Zheng, S.J., Henken, B., Krens, F.A., & Kik, C. (2003). The development of an efficient cultivar independent plant regeneration system from callus derived from both apical and non-apical root segments of garlic (Allium sativum L.). In Vitro Cellular and Developmental Biology – Plant, 39, 288-292.

Journal of Horticulture and Postharvest Research

In vitro propagation of Tunisian local garlic (Allium sativum L.) from shoot tip culture

References

References

Volume 1, Issue 2 - Serial Number 2
September 2018
September 2018
Pages 75-86

In vitro propagation of Tunisian local garlic (Allium sativum L.) from shoot tip culture

References

References

Volume 1, Issue 2 - Serial Number 2September 2018September 2018Pages 75-86

Volume 1, Issue 2 - Serial Number 2
September 2018
September 2018
Pages 75-86