In vitro induction of tetraploidy in Linum album Kotschy ex Boiss. using oryzalin treatment

Javadian, N.; Karimzadeh, Gh.; Sharifi, M.; Moeini, A.; Tahsili, Zh.

doi:10.22092/ijmapr.2016.106568

Document Type : Research Paper

Authors

¹ Ph.D. Student, Plant Breeding and Biotechnology Department, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran

² Plant Breeding and Biotechnology Department, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran

³ Plant Biology Department, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran

https://doi.org/10.22092/ijmapr.2016.106568

Abstract

Linum album Kotschy ex Boiss. is one of the endemic species of Linaceace family in Iran, having podophyllotoxin (PTOX) metabolite. This substance is a pharmaceutically important natural product belonging to the lignans group. It is used as a precursor for the synthesis of important anti-cancer drugs. Industrial production of this component is not economically feasible because of its complex chemical structure. Therefore, various strategies have been used to increase the production of PTOX in plants. In this study, tetraploidy was induced in nodal segments with different concentrations of oryzalin (0, 15, 25, 50 µM) for 24, 48, 72 and 96 h. The results indicated that the viability and the number of produced shoots in each nodal segment were reduced by increasing concentrations and times of oryzalin. Tetraploid plants were identified by flow cytometry, and 2C DNA content of diploids and tetraploids were estimated to be 4.34 and 8.36 pg, respectively. The highest induction efficiency percentage (19%) of tetraploidy was achieved from 25 µM of oryzalin for 48 h. Tetraploid induction caused 1.66 and 1.48 fold-increase in PTOX content in the leaves and stems of tetraploids compared with those of diploids, respectively. The results of this study indicated that the induction of tetraploidy could be an effective technique to increase this valuable metabolite in Linum album.

Keywords

References

Abdoli, M., Moieni, A. and Naghdi Badi, H., 2013. Morphological, physiological, cytological and phytochemical studies in diploid and colchicine-induced tetraploid plants of Echinacea purpurea (L.). Acta Physiologiae Plantarum, 35(7): 2075-2083.

- Ascough, G.D., Van Staden, J. and Erwin, J.E., 2008. Effectiveness of colchicine and oryzalin at inducing polyploidy in Watsonia lepida NE Brown. HortScience, 43(7): 2248-2251.

- Caperta, A.D., Delgado, M., Ressurreição, F., Meister, A., Jones, R.N., Viegas, W. and Houben, A., 2006. Colchicine-induced polyploidization depends on tubulin polymerization in c-metaphase cells. Protoplasma, 227: 147-153.

- Chen, L.L. and Gao, Sh.L., 2007. In vitro tetraploid induction and generation of tetraploids from mixoploids in Astragalus membranaceus. Scientia Horticulturae, 112: 339-344.

- de Carvalho, J.F.R.P., de Carvalho, C.R. and Otoni, W.C., 2005. In vitro induction of polyploidy in annatto (Bixa orellana). Plant Cell Tissue and Organ Culture, 80: 69-75.

- Dehghan, E., Häkkinen, S.T., Oksman-Caldentey, K.M. and Ahmadi, F.Sh., 2012. Production of tropane alkaloids in diploid and tetraploid plants and in vitro hairy root cultures of Egyptian henbane (Hyoscyamus muticus L.). Plant Cell, Tissue and Organ Culture, 110: 35-44.

- De Jesus-Gonzalez, L. and Weathers, P.J., 2003. Tetraploid Artemisia annua hairy roots produce more artemisinin than diploids. Plant Cell Report, 21: 809-813.

- Dhooghe, E., Grunewald, W., Leus, L. and Van Labeke, M.C., 2009. In vitro polyploidisation of Helleborus species. Euphytica, 165: 89-95.

- Dhooghe, E., Van Laere, K., Eeckhaut, T., Leus, L. and Van Huylenbroeck, L., 2011. Mitotic chromosome doubling of plant tissues in vitro. Plant Cell Tissue and Organ Culture, 104: 359-373.

- Doležel, J. and Bartoš, J. 2005. Plant DNA flow cytometry and estimation of nuclear genome size. Annals of Botany, 95: 99-110.

- Doležel, J., Greilhuber, J. and Suda, J., 2007. Estimation of nuclear DNA content in plants using flow cytometry. Nature Protocols, 2: 2233-2244.

- Esmailzadeh, S., 2011. Investigation of fungal elicitors effect on podophyllotoxin content, gene expression and enzymatic activity of phenylalanine ammonia-lyase and cinnamyl alcohol-dehydrogenase in Linum album. Ph.D. Thesis, Department of Plant Biology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.

- Evans, G.M., Rees, H., Snell, C.L. and Sun, S., 1972. The relationship between nuclear DNA amount and the duration of the mitotic cycle. Chromosomes Today, 3: 24-31.

- Gantait, S., Mandal, N., Bhattacharyya, S. and Das, P.K., 2011. Induction and identification of tetraploids using in vitro colchicine treatment of Gerbera jamesonii Bolus cv. Sciella. Plant Cell Tissue and Organ Culture, 106: 485-493.

- Kaensaksiri, T., Soontornchainaksaeng, P., Soonthornchareonnon, N. and Prathanturarug, S., 2011. In vitro induction of polyploidy in Centella asiatica (L.) Urban. Plant Cell Tissue and Organ Culture, 107: 187-194.

- Lavania, U.C., 1988. Enhanced productivity of the essential oil in the artificial autopolyploid of vetiver (Vetiveria zizanioides L. Nash). Euphytica, 38: 271-276.

- Lavania, U.C., 2005. Genomic and ploidy manipulation for enhanced production of phytopharmaceuticals. Plant Genetic Resources, 3: 170-177.

- Lehrer, J.M., Mark, H.B. and Jessica, D., 2008. Lubell induction of tetraploidy in meristematically active seeds of Japanese barberry (Berberis thunbergii var. Atropurpurea) through exposure to colchicine and oryzalin. HortScience, 119: 67-71.

- Levy, M., 1976. Altered glycoflavone expression in induced autotetraploids of Phlox drummondi. Biochemical Systematics and Ecology, 4: 249-259.

- Loureiro, J., Rodriguez, E., Doležel, J. and Santos, C., 2007. Two new nuclear isolation buffers for plant DNA flow cytometry, Annual of Botany, 100: 875-888.

- Mahdavi, S. and Karimzadeh, G., 2010. Karyological and nuclear DNA content variation in some Iranian endemic Thymus species (Lamiaceae). Journal of Agricultural Science and Technology (JAST), 12: 447-458.

- Majdi, M., Karimzadeh, G., Malboobi, M.A., Omidbaigi, R. and Mirzaghaderi, G., 2010. Induction of tetraploidy to feverfew (Tanacetum parthenium Schulz-Bip.): morphological, physiological, cytological, and phytochemical changes. HortScience, 45: 16-21.

- Mohagheghzadeha, A., Hemmatia, Sh. and Alfermann, A.W., 2006. Quantification of aryltetralin lignans in Linum album organs and in vitro cultures. Iranian Journal of Pharmaceutical Sciences, 2: 47-56.

- Murashige, T. and Nakano, R., 1966. Tissue culture as a potential tool in obtaining polyploid plants. Journal of Heredity, 57: 114-118.

- Osborn, T.C., Pires, J.C., Birchler, J.A., Auger, D.L., Chen, Z.J., Lee, H.S., Comai, L., Madlung, A., Doerge, R.W., Colot, V. and Martienssen, R.A., 2003. Understanding mechanisms of novel gene expression in polyploids. Trends in Genetics, 19: 141-147.

- Parisod, C., Holderegger, R. and Brochmann, C., 2010. Evolutionary consequences of autopolyploidy. New Phytologist, 186: 5-17.

- Planchais, S., Glab, N., Inzé, D. and Bergounioux, C., 2000. Chemical inhibitors: a tool for plant cell cycle studies. FEBS Letters, 476: 78-83.

- Rowson, J.M., 1944. Increased Alkaloidal Contents of Induced Polyploids of Datura. Nature, 154: 81-82.

- Shao, J., Chen, C. and Deng, X., 2003. In vitro induction of tetraploid in pomegranate (Punica granatum). Plant Cell Tissue and Organ Culture, 75: 241-246.

- Shariat, A., Karimzadeh, G. and Asareh, M.H., 2013. Karyology of Iranian endemic Satureja (Lamiaceae) species. Cytologia, 78(3): 305-312.

- Sharifnia, F. and Assadi, M., 2001. Flora of Iran (Linaceae). Research Institute of Forests and Rangelands, Tehran, 181p.

- Siljak-Yakovlev, S., Pustahija, F., Solic, E. M., Bogunic, F., Muratovic, E., Basic, N., Catrice, O. and Brown, S. C., 2010. Towards a genome size and chromosome number database of Balkan flora: C-values in 343 taxa with novel values for 242. Advanced Science Letters, 3: 190-213.

- Thao, N., Ureshino, K., Miyajima, I., Ozaki, Y. and Okubo, H., 2003. Induction of tetraploids in ornamental Alocasia through colchicine and oryzalin treatments. Plant Cell Tissue and Organ Culture, 72: 19-25.

- Tosca, A., Pandoifi, R., Citterio, S., Fasoli, A. and Sgorbati, S., 1995. Determination by flow cytometry of the chromosome doubling capacity of colchicine and oryzalin in gynogenetic haploids of Gerbera. Plant Cell Report, 14: 455-458.

- Väinölä, A., 2000. Polyploidization and early screening of Rhododendron hybrids. Euphytica, 112: 239-244.

- Yermanos, D.M. and Gill, K.S., 1967. Induction of polyploidy in Linum species. Crop Science, 7: 423-427.

- Yousefzadi, M., 2010. The influence of Tyrosine, Trypthophan, Light and Salicylic acid on production of Podophyllotoxin and some genes expression and enzymatic activity in biosynthetic pathway of Podophyllotoxin in Linum album. Ph.D. Thesis, Department of Plant Biology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.

- Yousefzadi, M., Sharifi, M., Ahmadian Chashmi, N., Behmanesh, M. and Ghasempour, A., 2010a. Optimization of podophyllotoxin extraction method from Linum album cell cultures. Pharmaceutical Biology, 12: 1421-1425.

- Yousefzadi, M., Sharifi, M., Behmanesh, M., Moyano, E., Bonfill, M., Cusido, R.M. and Palazon, J., 2010b. Podophyllotoxin: Current approaches to its biotechnological production and future challenges. Engineering in Life Sciences, 10: 281-292.

Iranian Journal of Medicinal and Aromatic Plants Research

In vitro induction of tetraploidy in Linum album Kotschy ex Boiss. using oryzalin treatment

References

References

- Rowson, J.M., 1944. Increased Alkaloidal Contents of Induced Polyploids of Datura. Nature, 154: 81-82.

Volume 32, Issue 2 - Serial Number 76
June 2016
Pages 346-359

In vitro induction of tetraploidy in Linum album Kotschy ex Boiss. using oryzalin treatment

References

References

- Rowson, J.M., 1944. Increased Alkaloidal Contents of Induced Polyploids of Datura. Nature, 154: 81-82.

Volume 32, Issue 2 - Serial Number 76June 2016Pages 346-359

Volume 32, Issue 2 - Serial Number 76
June 2016
Pages 346-359