Document Type : Research Paper - Horticulture


1 Assistant Professor, Department of Horticultural Plant Biotechnology, Iranian Academic Center for Education, Culture and Research, Mashhad, Iran

2 Instructor, Department of Horticultural Plant Biotechnology, Iranian Academic Center for Education, Culture and Research, Mashhad, Iran

3 Ph.D. Student of Horticultural Science, Department of Horticultural science and landscape, Faculty of Agriculture, Ferdowsi University of Mashhad, Iran

4 Assistant Professor, Department of Horticultural Plant Biotechnology, Iranian Academic Center for Education, Culture and Research, Khorasan Razavi, Iran


Haworthia cooperi is a type of ornamental succulent that is propagated by separating the offsets and leaf cuttings. Due to the small number of produced offsets by the mother plant, the rate of propagation in this way is low, so micropropagation can be considered as an effective and efficient way to propagate this ornamental plant.
Materials and Methods
For this purpose, in the present study, the growth response of Haworthia cooperi plants to in vitro culture conditions in 1398 in the research laboratory of the Department of Ornamental Plants Biotechnology was investigated. This research was conducted in the form of three separate experiments based on a completely randomized design with three replications. In the first experiment, the effects of cytokinin type (BA and KIN) and concentration of cytokinin (0, 1, 2 and 3 mg/l) on leaf explant regeneration were evaluated. In the second experiment, the effects of culture medium type (MS and B5) and concentration of culture medium (0.5 and 1) on plantlet propagation were investigated. In the third experiment, plantlet rooting rate was evaluated under the influence of auxin type (NAA, IBA and IAA) and concentration (0, 0.5, 1 and 1.5 mg/l) of auxin.
Results and Discussion
The results showed that application of BA and KIN hormones increased the number of regenerated plantlets. The highest number of plantlets (5.50) was recorded in the MS medium containing 3 mg/l BA with 0.5 mg/l NAA. Plantlets cultured in the hormone-free culture medium had no regenerated symptoms. The number of plantlets increased with increasing the concentration of both hormones, but BA hormone in the culture medium had a greater effect on this factor. Lack of cytokinin hormone application in culture medium led to root production; however, the use of BA and KIN hormones in the culture medium reduced the number of produced roots by plantlets. The highest number of roots was recorded in hormone-free culture medium and the lowest number was recorded in the high concentration of BA hormone. The reduction in the number of roots in the culture medium containing BA was greater than the culture medium containing KIN. The of the second experiment revealed that use of MS culture medium showed better performance compared to B5 culture medium. Thus, the highest dry weight of plantlet and roots was obtained at full concentration of MS culture medium. The results of rooting stage showed that the presence of NAA hormone in the culture medium increased rooting in the plant. However, the produced roots in this culture medium were so thick that these roots are not suitable for plant growth in the acclimation stage. Therefore, considering the number and length of produced roots, the optimal culture medium for plantlet rooting is ½ MS culture medium containing 1 mg/l IAA hormone. Determining the best type of culture medium and optimizing it for regeneration, proliferation and rooting is one of the important factors in success in vitro culture conditions. Based on the results of the current study, application of MS medium containing 3 mg/l BA with 0.5 mg/l NAA was so effective for regeneration phase of this ornamental plant. Also, the efficiency of MS culture medium in the process of propagation of this plant is most likely related to the suitability of the components of this culture medium. In general, due to the presence of plant growth regulators such as NAA, IBA and IAA in the culture medium, rooting is induced in plantlets. In the present study, the use of IAA hormone at a concentration of 1 mg/l was a suitable hormonal combination for rooting in this ornamental plant.
Based on the results of the present study, the application of MS culture medium containing 3 mg/l BA with 0.5 mg/l NAA is recommended for regeneration and proliferation of tissue culture plantlets of Haworthia cooperi. It is also recommended to use ½ MS culture medium containing 1 mg/l of IAA hormone for rooting of tissue culture plantlets.


Main Subjects

Asgharzadeh, R., Moradi, H., Nematzadeh, Gh., & Ghanbari, A. (2016). Evaluation of growth adapter effect on callus induction of Momordica Charantia L. Eco-Phytochemical Journal of Medical Plants, 3(15), 46-56. [In Farsi]
Balapor, Z., Hosseini Moghaddam, H., Zarei, M., & Mollashahi, M. (2020). Micro Propagation of Penta Rootstock (Prunus domestica) in the Two Culture Media (MS and B5). Plant Productions, 42(4), 441-454. [In Farsi]
Balch, P. M., & Figuaroa, C. A. (2002). In vitro propagation of Pelecyphora aselliformis and P. strobiliformis (cactaceae). In Vitro Cellular and Developmental Biology Plant, 38, 73-78.
Cecilia Juarez, M., & Bernardo Passere C. (2002). In vitro propagation of opuntia ellisiana Griff. And a acclimatization ellisiana Griff. and acclimatization to field conditions. Biocell, 26(3), 1-18.
Chien-Ying, K., Al-Abdulkarim, A.M., Al-Jowid, S.M., & Al-Baiz, A. (2009). An effective disinfection protocol for plant regeneration from shoot tip cultures of strawberry. African Journal of Biotechnology, 8(11), 2611-2615.
Darang, M., Hamidoghli, Y., & Ramezani Sayad, A. (2011). Influence of growth regulators on in vitro propagation of cactus (Opuntia ficus-indica). MSc thesis of Horticultural Science, University of Guilan, Rasht.  [In Farsi]
De Oliveira, S.A., Pires, F., Da Silva Machado, M., Jose Prioli, A., & Mangolin C. A. (1995). In vitro propagation of Cereus peruvianus Mill. (Cactaceae), In Vitro Cellular & Developmental Biology-Plant, 31(1), 47-50.
Drew, R.A., & Azimi, M. (2000). Micropropagation of Red Pitaya (Hylocereus undatus). Acta HortiCulture, 575(26), 93-98.
Emara, H. (2008). Factors affecting propagation of strawberry (Fragaria spp.) through tissue cultures. International Journal of Product Development, 13(1), 191-212.
Estrada-Luna, A.A., Martinez-Hernandez, J.J., Estela Torres, M., & Chable-Moreno, F. (2008). In vitro micropropagation of the prickly pear cactus Opuntia lanigera Salm- Dyck and effects of sprayed GA3 after transplantation to ex vitro conditions. Scientia Horticulturae, 117(4), 378-385.
Fathi, Gh., Esmailpour, B., & Jalilvand, P. (2012). Plant growth regulators, Mashhad, JDM press. [In Farsi]
Ghasemi Ghohsare, M., & Kafi, M. (2010). Floriculture Vol 2. Tehran: Tehran University Press. [In Farsi]
Giusti, P., Vitti, D., Fiocchetti, F., Colla, G., Saccardo, F., & Tucci, M. (2002). In Vitro propagation of three endangered cactus species. Scientia Horticulturae, 95(4), 319-332.
Huang, S. H., Xiao, H. Y., & Hong, Y. P. (2014). Tissue culture and rapid propagation of Haworthia cymbiformis. Journal of Fujian Normal University (Natural Science Edition), 30(5), 96-100.
Karimi, N., Naderi, R., Ebrahimi, M., & Mofid, M.R. (2010). Investigation of a Medicinal and ornamental plant propagation: The Cereus Peruvianus Mill. and using tissue culture technique. Journal of Medicinal Plants, 9(2-34), 38-44.  [In Farsi]
Kaul, K., & Sabharwal, P. S. (1992). Morphogenetic studies on Haworthia: Establishment of tissue culture and control of differentiation. American Journal of Botany, 59(4), 377-385.
khalafalla, M. M., Abdellatef, E., Mohamed Ahmed, M. M., & Osman, M. G. (2007). Micropropagation of cactus (Opuntia ficus-indica) as strategic tool to combat desertification in arid and semi-arid regions. International Journal of Sustainable Crop Production, 2(4), 1-8.
Kharrazi, M., Sharifi, A., Keykhah Akhar, F., Bagheri, A., & Moradian, M. (2018). Effect of Hormonal Compositions on Micropropagation of Fifteen Cultivars of Gerbera (Gerbera Jamesonii Bolus ex Hooker f.). Plant Productions, 40(4), 91-102. [In Farsi]
Kim, D. H., Kang, K. W., & Sivanesan, I. (2017). Micropropagation of Haworthia retusa duval. Propagation of Ornamental Plants, 17(3), 77-82.
Koohi, L., Zare, N., Asghari Zakaria, R., & Sheykhzade Mosadegh, P. (2014). The Effect of plant growth regulators and different explants on the response of tissue culture and cell suspension cultures of german chamomile (Matricaria Chamomilla L.). Journal of Crop Ecophysiology (Agriculture Science), 2(30), 203-214. [In Farsi]
Kuykendall, L. D., Stockett, T. M., & Saunders, J. W. (2003). Rhizobium radiobacter conjugation and callus-independent shoot regeneration used to introduce the cercosporin export gene cfp from Cercospora into sugar beet (Beta vulgaris L.). Biotechnology Letter, 25(9), 739-744.
Liu, B., Fang, H., Meng, C., Chen, M., Chai, Q., Zhang, K., & Liu, S. (2017). Establishment of a Rapid and Efficient Micropropagation System for Succulent Plant Haworthia turgida Haw. Hort Science, 52(9), 1278-1282.
Luliana vidican, T., & Cachita-Cosma, D. (2010). Initiation of Opuntia fragilis var. fragilis in vitro cultures. Studia Universitatis Vasile Goldis, Seria stiintele Vietii, 20(3), 35-40.
Mashayekhi, K., & Akbarpour, V. (2008). Mineral and organic minerals used in plant tissue culture (Vol 1). Golestan: Makhtoomgholi Feraghi press (Sarli).  [In Farsi]
Meftahizade, H., Moradkhani, H., Naseri, B., Lotfi, M., & Naseri, A. (2010). Improved in vitro culture and micropropagation of different Melissa officinalis L. genotypes. Journal of Medicinal Plants Research, 4(3), 240-246.
Mycock, D.J., Watt, M.P., Hannweg, K.F., Naicker, K., Makwareka, M., & Berjak, P. (1997). Somatic embryogenesis of two indigenous South African Haworthia spp. (H. limifolia and H. koelmaniorum), South African Journal of Botany, 63(6), 345-350.
Nagarathan, K. C., Prakash, H. S., & Shetty, H. S. (1991). Genotypic effects on the callus formation from different explants of pearl millet B lines. Advances in Plant Sciences, 4(1), 82-86.
Nezamdoost, M., Kalateh Jari, S., & Ebrahimi, A. (2019). Investigation of Haworthia minima organogenesis and rooting under in vitro condition. Presented at the Conference 2nd international & 6th national confrence on organic and conventional agriculture, 25-26 Aug, University of Mohaghegh Ardabili, Ardabil. [In Farsi]
Nourafcan, H., Sefidkon, F., Mousavi, A., Sharifi, M., & Khalighi, A. (2014). Effects of different medium on some morpho-physiological index of lemon verbena under in vitro conditions. Journal of Herbal Drugs, 5(3), 143-150.
Nourafkan, H., & Ansari, F. (2017). The effect of MS and B5 media on growth indices of lemon verbena in in vitro condition. Iranian Journal of Horticultural Science, 48(1), 249-254.  [In Farsi]
Omidi, M., Ydollahi, A., Ahmadi, N., & Beyranvand, F. (2017). Improvement of in vitro micro propagation of Rosa damascene. Iranian Journal of Horticultural Science, 47(4), 773-784.
Pierik, R. L. M. (1997). In Vitro culture of higher plants. Netherlands, Springer science & business media.
Ruvalcaba-Ruiz, D., Rojas-Bravo D., & Valencia-Botin, A. J. (2010). In Vitro propagation of Coryphantha retusa (Britton and Rose) an endemic and endangered cactus. Tropical and Subtropical Agroecosystems, 12(1), 139-143.
Sagastume Tecnoregion, H. (2006). Propagation in vitro cactus cabeza (pilocereus maxonii). Biotecnologia, 23-24.
Sivanesan, I., Song, J. Y., Hwang, S. J., & Jeong, B. R. (2011). Micropropagation of Cotoneaster wilsonii Nakai-a rare endemic ornamental plant. Plant Cell, Tissue and Organ Culture, 105(1), 55-63.
Taji, A. M., Dodd, W. A., & Williams, R. R. (1997). Plant tissue culture practice. Armidale, Australia: University of New England press.
Talebi, M., Etesam, F., Sayed-Tabatabaei, B., & Khaksar, G. (2012). Direct regeneration of Periwinkle (Catharanthus roseus) via node explants culture and different combinations of plant growth regulators. Journal of Science and Technology of Greenhouse Culture Soilless Culture Research Center, 3(2), 55-65.  [In Farsi]
Tripathi, L., & Tripathi, J. N. (2003). Role of biotechnology in medicinal plants. Tropical Journal of Pharmaceutical Research, 2(2), 243-253.
Van Jaarsveld, E. (1999). Indigenous succulent plants for indoors. Veld Flora, 85(2), 82-84.