Evaluation of somaclonal variation of Arnebia pulchra (Boraginaceae) calli versus seeds using ISSR markers

Document Type : Research paper

Authors

Department of Agricultural Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.

Abstract

This is the first report on the analysis ISSR markers for assessing the genetic variation in Arnebia pulchra. As the rate of cell division and propagation in A. pulchra is very low, the achievement of biomass in this valuable plant requires techniques and strategies that can guarantee the survival of the callus. The present study was performed to investigate genetic variation in two in vitro culture samples of the medicinal plant A. pulchra including new (one month old) and old (four years old) calli versus seeds by Inter Simple Sequence Repeat (ISSR) marker. In total, 79 bands were produced by 10 ISSR primers. The total expected heterozygosity (He) was calculated 0.028 and the greatest value was contributed to seeds samples (0.054) followed by old and new calli, respectively. The maximum values of different alleles (Na), effective alleles (Ne) and Shannon’s index were also observed in seeds. Cluster analysis was performed in the form of dendogram to indicate the genetic stability of the samples. UPGMA tree discriminated samples in two major groups and principal component analysis (PCoA) confirmed the clustering result. The first major cluster included seeds and the second involved new and old calli. Although it was thought that the morphological changes of A. pulchra could be due to the somaclonal variation caused by successive subcultures, however, the results indicated that the new and old calli were clustered in the same group. Therefore, it can be concluded that the morphological changes of the old and new calli could be due to environmental parameters or epigenetic changes but not directly due to somaclonal variation.

Keywords

References

Azizi P., Hanafi M. M., Sahebi M., Harikrishna J. A, Taheri S., Yassoralipour A., and Nasehi A. (2020). Epigenetic changes and their relationship to somaclonal variation: a need to monitor the micropropagation of plantation crops. Functional Plant Biology, 47(6): 508-23.
Ahmed T. A., Alsamaraee S. A., Zaidan H. Z., and Elmeer K. (2012). Inter-simple sequence repeat (ISSR) analysis of somaclonal variation in date palm plantlets regenerated from callus. In: Second International Conference on Environment and Industrial Innovation, IPCBEE, 126-130.
Aliasl J., Khoshzaban F., Barikbin B., Naseri M., Kamalinejad M., Emadi F., Razzaghi Z., Talei D., Yousefi M., Aliasl F., Barati M., Mohseni-Moghaddam P., Hasheminejad S. A., and Nami H. E. (2014). Comparing the healing effects of Arnebia euchroma ointment with petrolatum on the ulcers caused by fractional CO2 laser: a single-blinded clinical trial. Iranian Red Crescent Medical Journal, 16(10): e16239. DOI: 10.5812/ircmj.16239.
Armijos-González R., Espinosa-Delgado L., and Cueva-Agila A. (2021). Indirect shoot regeneration using 2,4-D induces somaclonal variations in Cinchona officinalis. Floresta e Ambiente, 28(3): e20210017.
Asadi A., and Shooshtari L. (2021). Assessment of somaclonal variation in micropropagation of Damask Rose (Rosa damascena Mill.) using molecular markers. Modern Genetics Journal (MGJ), 15(4): 327-335. (In Persian).
Bairu M. W., Aremu A. O., and Van Staden J. (2011). Somaclonal variation in plants: causes and detection methods. Plant Growth Regulation, 63: 147-173.
Bairu M. W., Fennell C. W., and Van Staden J. (2006). The effect of plant growth regulators on somaclonal variation in Cavendish banana (Musa AAA cv.‘Zelig’). Scientia Horti-culturae, 108: 347-351.
Bennici A., Anzidei M., and Vendramin G. G. (2004). Genetic stability and uniformity of Foeniculum vulgare Mill. regenerated plants through organogenesis and somatic embryogenesis. Plant Science, 166: 221-227.
Côte F. X., Teisson C., and Perrier X. (2001). Somaclonal variation rate evolution in plant tissue culture: Contribution to understanding through a statistical approach. In Vitro Cellular & Developmental Biology-Plant, 37: 539-542.
De la Rosa R., James C. M., and Tobutt K. R. (2002). Isolation and characterization of polymorphic microsatellites in olive (Olea europaea L.) and their transferability to other genera in the Oleaceae. Molecular Ecology Notes, 2(3): 265-267. DOI: https://doi.org/10.1046/j.1471-8286.2002.00217.x.
Devi S. P., Kumaria S., Rao S. R., and Tandon P. (2014). Single primer amplification reaction (SPAR) methods reveal subsequent increase in genetic variations in micropropagated plants of Nepenthes khasiana Hook. f. maintained for three consecutive regenerations. Gene, 538: 23-29. DOI: 10.1016/j.gene.2014.01.028.
Erişen S., Kurt-Gür G., and Servi H. (2020). In vitro propagation of Salvia sclarea L. by meta-Topolin, and assessment of genetic stability and secondary metabolite profiling of micropropagated plants. Industrial Crops and Products, 157: 112892.
Ezati T., Marefatjoo M. J., Haghbeen K., and Ahmadkhaniha R. (2015). Successful indirect regeneration of Arnebia pulchra (Roemer and Schultes) as medicinal plant. Journal of Medicinal Plants and By-products, 4(2): 233-242. DOI: 10.22092/JMPB.2015.108914.
Goodarzi F., Darvishzadeh R., and Hassani A. (2015). Genetic analysis of castor (Ricinus communis L.) using ISSR markers. Journal of Plant Molecular Breeding, 3: 18-34.
Gostimsky S., Kokaeva Z., and Konovalov F. (2005). Studying plant genome variation using molecular markers. Russian Journal of Genetics, 41: 378-388.
Haghbeen K., Mozaffarian V., Ghaffari F., Pourazeezi E., Saraji M., and Joupari M. (2006). Lithospermum officinale callus produces shikalkin. Biologia, 61: 463-467.
Hosseini A., Mirzaee F., Davoodi A., Jouybari H. B., and Azadbakht M. (2018). The traditional medicine aspects, biological activity and phytochemistry of Arnebia spp. Medicinski Glasnik, 15(1): 1-9. Doi: 10.17392/926-18.
Hrahsel L., Basu A., Sahoo L., and Thangjam R. (2014). In vitro propagation and assessment of the genetic fidelity of Musa acuminata (AAA) cv. Vaibalhla derived from immature male flowers. Applied Biochemistry and Biotechnology, 172(3): 1530-1539.
Jin S., Mushke R., Zhu H., Tu L., Lin Z., Zhang Y., and Zhang X. (2008). Detection of somaclonal variation of cotton (Gossypium hirsutum) using cytogenetics, flow cytometry and molecular markers. Plant Cell Reports, 27: 1303-1316.
Krishna H., Alizadeh M., Singh D., Singh U., Chauhan N., Eftekhari M., and Sadh R. K. (2016). Somaclonal variations and their applications in horticultural crops improvement. 3 Biotech, 6(1): 54. Doi: 10.1007/s13205-016-0389-7.
Mohanty S., Panda M., Subudhi E., and Nayak S. (2008). Plant regeneration from callus culture of Curcuma aromatica and in vitro detection of somaclonal variation through cytophoto-metric analysis. Biologia Plantarum, 52: 783-786
Monfared M. A., Samsampour D., Sharifi-Sirchi G. R., and Sadeghi F. (2018). Assessment of genetic diversity in Salvadora persica L. based on inter simple sequence repeat (ISSR) genetic marker. Journal of Genetic Engineering and Biotechnology, 16: 661-667.
Nei M. (1978). Estimation of average heterozygosity and genetic distance from a small number of indivisuals. Genetics, 23: 341-369.
Noormohammadi Z., Kangarloo-Haghighi B., Sheidai M., Farahani F., and Ghasemzadeh-Baraki S. (2014). Genetic stability versus somaclonal variation in tissue culture regenerated olive plants (Olea europea cv. Kroneiki). European Journal of Experimental Biology, 4: 135-142.
Peng X., Zhang T-t., and Zhang J. (2015). Effect of subculture times on genetic fidelity, endogenous hormone level and pharmaceutical potential of Tetrastigma hemsleyanum callus. Plant Cell, Tissue and Organ Culture (PCTOC), 122: 67-77.
Podwyszynska M. (2005). Somaclonal variation in micropropagated tulips based on phenotype observation. Journal of Fruit and Ornamental Plant Research, 13: 109-122.
Rajan R. P., and Singh G. (2021). A review on application of somaclonal variation in important horticulture crops. Plant Cell Biotechnology and Molecular Biology, 10: 161-75.
Razani M., Kayat F., Redwan R. M., and Susanto D. (2020). Detection of abnormal banana plantlets produced by high BAP concentration and number of subcultures using representational difference analysis. International Journal of Agriculture and Biology, 23(3): 541-8.
Shen X., Chen J., Kane M. E., and Henny R. J. (2007). Assessment of somaclonal variation in Dieffenbachia plants regenerated through indirect shoot organogenesis. Plant Cell Tissue and Organ Culture, 91: 21-27.
Smýkal P., Valledor L., Rodriguez R., and Griga M. (2007). Assessment of genetic and epigenetic stability in long-term in vitro shoot culture of pea (Pisum sativum L.). Plant Cell Reports, 26: 1985-1998.
Tikendra L., Potshangbam A. M., Dey A., Devi T. R., Sahoo M. R., and Nongdam P. (2021). RAPD, ISSR, and SCoT markers based genetic stability assessment of micropropagated Dendrobium fimbriatum Lindl. var. oculatum Hk. f.-an important endangered orchid. Physiology and Molecular Biology of Plants, 27(2): 341-357. DOI: https://doi.org/10.1007/s12298-021-00939-x.
Yadav K., Aggarwal A., and Singh N. (2013). Evaluation of genetic fidelity among micropropagated plants of Gloriosa superba L. using DNA-based markers—A potential medicinal plant. Fitoterapia, 89: 265-270.
Zhao F., Liu F., Liu J., Ang P. O., and Duan D. (2008). Genetic structure analysis of natural Sargassum muticum (Fucales, Phaeophyta) populations using RAPD and ISSR markers. Journal of Applied Phycology, 20(2): 191-198. DOI: 10.1007/s10811-007-9207-2.

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History

  • Receive Date: 31 October 2021
  • Revise Date: 25 April 2022
  • Accept Date: 06 June 2022

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