Optimization of regeneration of anise (Pimpinella anisum L) in in vitro condition

Document Type : Research Paper

Authors

1 M.Sc. graduated, Dept. Production Engineering and Plant Genetics, Shirvan Agriculture Faculty, University of Bojnord, Bojnord, I.R. Iran.

2 Assistant professor, Dept. Production Engineering and Plant Genetics, Shirvan Agriculture Faculty, University of Bojnord, Bojnord, Iran

3 Associate Professor, Dept. of Ornamental plants, Plant Science Research Institute, Ferdowsi University of Mashhad. I.R. Iran.

10.22092/ijrfpbgr.2023.362211.1436

Abstract

Extended Abstract
 
Background and objectives: 
Anise Pimpinella anisum L. is a valuable medicinal plant that 2-3% of its seed weight is essential oils, and about 80-90% of its essential oil is trans-antol. This research was carried out to optimize the indirect organogenesis of P. anisum under in vitro conditions.
 
Methodology:
In the first experiment, different methods of seed germinating and disinfection treatments for decontamination of samples were investigated. In the second experiment, the effect of different combinations of BA, Kin and NAA on the direct regeneration of hypocotyl bud explants was investigated. In the third experiment, callus production was investigated from explants of root, hypocotyl, hypocotyl bud, cotyledon leaf and normal leaf, using different concentrations of 2,4-D. In the fourth experiment, the effect of a narrower range of 2,4-D concentration on the callus formation of petiole and root explants was evaluated. In the fifth experiment, the produced calli was exposed to different concentrations of BA and NAA for regeneration. All experiments were conducted in a completely randomized design with three replications. Data analysis was done using Minitab software, and the mean comparison was made using the Tukey method (p<0.05).
 
Results:
The best germination was obtained at a value of 70% in 24 hours of seed soaking in water (hydro priming) and disinfection with 70% alcohol and 1.5% sodium hypochlorite in 1/2 MS culture medium. In direct regeneration, stem-less leaves were produced. The best direct regeneration response in terms of petiole length was related to treatments of 9 (0.5 mg/L BA plus 0.5 mg/L Kin plus 0.5 mg/L NAA), 14 (0.5 mg per liter Kin plus 1mg/L NAA), 17 (0.25 mg/L BA plus 0.25 mg/L Kin plus 1mg/L NAA) and 19 (1mg/L BA plus 1mg/L Kin plus 1mg/L NAA) with about 4 cm. The highest leaf number (47 leaves) was obtained using the treatments of 14 and 17. In the third experiment, petiole and root explants had the best callus formation. In the fourth experiment, using the treatment of (0.5 mg/L BA +0.75 mg/L of 2, 4-D), the highest rate of 97.22% callus formation was observed in petiole explants. In the fifth experiment, the callus obtained from the previous stage induced the highest shoot with a value of 40% in the control treatment without hormones. Some branches were rooted, and the resulting seedlings were transferred to the greenhouse after adaptation.
 
Conclusion:
 It was concluded that for the regeneration and breeding of anise plant, the indirect regeneration of the callus is more suitable than the direct regeneration method. Also, the petiole was a suitable organ as the initial explant. The 1.5% sodium hypochlorite was the best disinfectant for seed decontamination. Hormonal treatment of 0.75 mg/L of 2, 4-D +0.5 mg/L of BA induced the highest rate (97.22%) of callus in a petiole explant. Therefore, it could be used as a successful hormonal compound for callus formation in anise. The production of branches and roots from callus in the control treatment indicated that it was possible to obtain a whole plant without using any plant growth regulators. The results of this research can be used in micropropagation studies, genetic engineering, cell suspension culture and production of secondary metabolites and other projects related to anise improvement and molecular studies.

Keywords

Main Subjects

References

Ahmadian, A., 2012. Important Medicinal Plants of Iran (Vol. 1- Shrub Plants). Publications of Torbat Heydarieh University, Iran 408p. (In Persian).
Amer A. and Omar H. 2019. In vitro propagation of the multipurpose Egyptian medicinal plant Pimpinella anisum. Egyptian Pharmaceutical Journal, 18(3):254-262.
Antony, T., Anees, P. V. M., Kumar, V., Sangamithra, D., Philip, T. and Santhoshkumar, A. V. 2015. Application of mercuric chloride and charcoal in micro-propagation of teak (Tectona grandis). Indian Journal of Tropical Biodiversity, 23(2): 157-166.
Arafa, N. M. and Aly, U. I. 2022. Induced biosynthesis of acephenanthrylene in callus culture of Pimpinella anisum L., by yeast and phenylalanine application. Egyptian Journal of Chemistry, 65(12): 327 – 335
Bansal, S., Sharma, M. K., Joshi, P., Malhotra, E. V., and Malik, S. K. 2023. Meta-topolin mediated enhanced in vitro propagation and genetic integrity assessment in sweet potato (Ipomoea batatas L. Lam). South African Journal of Botany, 157: 27-36.
Bhat, M. S., Rather, Z. A., Nazki, I. T., Banday, N., Wani, T., Rafiq, S., and Darwish, H. 2022. Standardization of in vitro micropropagation of Winter Jasmine (Jasminum nudiflorum) using nodal explants. Saudi Journal of Biological Sciences, 29(5): 3425-3431.
Davazdah emami, S. and Majnoun Hosseini, N., 2018. Cultivation and production of some medicinal and spice plants. Tehran University Publication, Tehran, Iran. 320 pages. (In Persian).
Darroudi H, Akbari Nia M., Safar Nejad A., Hosseini S.M. And Hajian Shahri M. 2017. Acclimatization of tissue cultured plantlets Ribes khorasanicum as an endemic endanger species in different substrates. Plant Research Journal (Iranian Biology Journal). 30(2), 346-356. (In Persian).
De Carlo, A., Tarraf, W., Lambardi, M., and Benelli, C. 2021).Temporary immersion system for production of biomass and bioactive compounds from medicinal plants. Agronomy, 11(12), 2414.
Desai, P., Desai, S., Rafaliya, R., and Patil, G. 2022. Plant tissue culture: Somatic embryogenesis and organogenesis. In Advances in Plant Tissue Culture (pp. 109-130). Academic Press.
Fathi, Sh. and Hasanpour, M., 2018. The effect of osmotic pretreatment on germination indicators of anise medicinal plant under salt stress. Seed Research, 9(32), 45-55. (In Persian).
Fazeli-Nasab, B. and Fooladvand Z., 2017. The effects of plant growth regulators and explants on callus
induction in Ajowan. The journal of Molecular and cellular researches. Iranian Biology Journal, 32(1): 63-75. (In Persian).
Ghazi, A., Ghani, F. H., and Mohsin, N. M. 2019. In Vitro Regeneration of Pimpinella Anisum L. Using Different Plant Growth Regulators. Iraqi Journal of Science, 60(4), 715-723.
Hasnain, A., Naqvi, S. A. H., Ayesha, S. I., Khalid, F., Ellahi, M., Iqbal, S. and Abdelhamid, M. 2022. Plants in vitro propagation with its applications in food, pharmaceuticals and cosmetic industries; current scenario and future approaches. Frontiers in Plant Science, 13, 1009395.
Khatibzadeh, R., Azizi, M., Arouiee, H. and Farsi, M., 2013. Effect of explants types and levels of plant growth substances on callogenesis in Levisticum officinale Koch. on a modified MS medium. Iranian Journal of Medicinal and Aromatic Plants, 29(2): 387-397. (In Persian).
Long, Y., Yang, Y., Pan, G., and Shen, Y. 2022. New insights into tissue culture plant-regeneration mechanisms. Frontiers in plant science, 13, 926752.
Mahdavi, B., 2016. Effects of Priming Treatments on Germination and Seedling Growth of Anise (Pimpinella anisum L.). Agriculture Science Developments, 5(3): 28-32.
Mir Haider, H., 2018. Herbal education, the use of plants in the prevention and treatment of diseases. Islamic culture publishing office, Tehran. Iran (Vol.2). 536p. (In Persian).
Momeni, A., Zaker Tavallaie, F., Shokoohifar, F. and Kheirkhah, M., 2018. Optimization of regeneration in Mountain Ziziphora (Ziziphora clinopodioides Lam). 151 Iranian Journal of Rangelands and Forests Plant Breeding and Genetic Research, 27(1): 143-151. (In Persian).
Mohammadian, A., Barzin, G., Atai Samagh, H. and Safarzadeh, A., 2017. Effect of priming with nitric oxide on germination indicators of medicinal plant anise (Pimpinella anisum L.) under salinity stress conditions, 7th National Conference on Sustainable Agriculture and Natural Resources, Tehran, Institute of Education Excellent Mehr Arvand، Tehran, Iran (In Persian).
Mukherjee, S., and Chakraborty, S. 2021. Plant tissue culture: a review on recent advances and future prospects. Journal of Applied Biology and Biotechnology, 9(02): 1-10.
Narimani, R., Samiei, L., and Moghaddam, M. 2022. Micro propagation optimization of two endangered and endemic of Iran species of Ferulago (Ferulago subvelutina Rech. f. and Ferulago angulata (Schlecht) Boiss). Iranian Journal of Rangelands and Forests Plant Breeding and Genetic Research, 29(2). (In Persian).
Nazari, R., Abedy, B., Balandari, A., Samiei, L., and Tehranifar, A. 2020. Use of response surface methodology for optimizing the media of establishment and proliferation phases of Iranian seedless barberry. Plant Cell, Tissue and Organ Culture, 141, 87-101.
Romdhane, M. and Tizaoui, C. 2005. The kinetic modelling of a steam distillation unit for the extraction of
aniseed (Pimpinella anisum) essential oil. Journal of chemical technology and Biotechnology, 80: 759-766.
Samiei, L., Davoudi Pahnehkolayi, M., Tehranifar, A., and Karimian, Z. 2021. Organic and inorganic elicitors enhance in vitro regeneration of Rosa canina. Journal of Genetic Engineering and Biotechnology, 19, 1-7.
Sadatnoori, S.A., Mortazavian, S.M.M., Ghamari Zare, A., Omidi M. and Hoori M. 2013. Evaluation of Callus Induction and Somatic Embryogenesis in Black Cumin (Bunium persicum Boiss). Journal of Agricultural Biotechnology, 5(2): 69-86. (In Persian).
Sobhanverdi, T. , Chopani, A. and Fatovat, R., 2013, investigating the effect of different hormonal compounds and the type of explant on callus formation and regeneration of the medicinal plant anise (Pimpinella anisum L.), the second national conference of medicinal plants and sustainable agriculture, Hamadan. (In Persian).
Srilakshmi, A. and Gayatri, M.C., 2014. Micropropagation of Pimpinella candolleana wight & ARN. An endemic medicinal herb from South India. International Journal of Pharma and Biosciences, 5(4): 253-259.
Yildiz, M., Fatih Ozcan, S., T Kahramanogullari, C., and Tuna, E. 2012. The effect of sodium hypochlorite solutions on the viability and in vitro regeneration capacity of the Tissue. The Natural Products Journal, 2(4), 328-331.
Yousefzadeh, S., 2018. The response of anise medicinal plant to the effect of salinity and drought stress in the germination stage, the third national biology conference of Payame Noor University, Sari. (In Persian).
Zaker Tavallaie, F., Ebrahimi, S., Zare Mehrjerdi, M. and Ghorbanzadeh Neghab, M., 2019. Investigating the effect of explant type and hormonal composition on Astragalus verus callogenesis, Iranian Journal of Rangelands and Forests Plant Breeding and Genetic Research, 28(1): 102-114. (In Persian)

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