Induction of in vitro polyploidy in ornamental flowers of orchid species (Phalaenopsis amabilis)

Document Type : Research Paper

Authors

1 Assis. Prof. of Biotechnology, Complex of Higher Education of Shirvan, Shirvan, I.R. Iran

2 M.Sc., Azad University of Shirvan, Shirvan, I.R. Iran

Abstract

Phalaenopsis amabilis is one of the best and well-known orchid species that is easy to grow and has good compatibility. This study was done for induction of polyploidy in Phalaenopsis amabilis by in vitro conditions. The study was conducted as factorial based on a completely randomized design with three replicates and each replicate  including three explants of auxiliary buds (approximately 3 mm ) containing two leaves using seven different concentrations of colchicine  (0, 0.01, 0.05, 0.1, 0.15, 0.2 and 0.25 percent) in two time periods (24 and 48 hours). Explants were cultured on MS medium without hormones after treatment with colchicine. After 10 weeks  sevral morphological traits that were under influence of polyploidy, including number and size of leaves, number and size of roots, length and width of stomata guard cells, number of chloroplast, and survival rate of seedlings were evaluated. Cytological observations of new developed leaves were evaluated by flow cytometry to assess the number of chromosomes in plants. The results showed that the highest percentage of polyploidy was obtained using 0.15 and 0.2 percent of colchicine concentrations. Correlation of some morphological traits with polyploidy levels showed that in addition to flow cytometry method, some important morphological traits such as stomata characteristics and its density can be used to recognize polyploidy in orchid plant. Polyploidy was resulted to enhancement in size and reduction in density of guard cells. Colchicine is usable to produce ornamental polyploid orchid.
 

Keywords

Main Subjects


-      Aalishah, A. and Omidi, M., 2008. Cytogenetics Laboratory Methods. Tehran University Press. (In Farsi)
-      Ahmadi Majd, M., Sarikhani, H., Chaiechi, M. and Kashi, A.K., 2012. Investigation of micropropagation and effect of colchicine to induce in vitro polyploidy in Narcissus tazetta. Plant Production, 35: 93-103 (In Farsi).
-      Afshar Mohammadian, M., Omidi, Z., Poorakbari, R. and Asadi Abkenar, A., 2013. Investigation of effect of polyploidy on some anatomical characteristics and antioxidant components of Citrus ourantifolia. Journal of Plant Researches, 26: 238-246. (In Farsi).
-      Alan, A.R., Lim, W., Mutschler, M.A. and Earle, E.D., 2007. Complementary strategies for ploidy manipulations in gynogenic onion (Allium cepa L.). Plant Science, 173: 25-31.
-      Atichart, P., 2007.  Polyploid induction in Dendrobium secundum (Bl.) Lind. by in vitro techniques. Thai Journal of Agricultureal Science, 40: 91-95.
-      Borghai, S.F., Sarikhani, H., Chaiechi, M. and Kashi, A.K., 2010. Induction of in vitro polyploidy in medicinal plant of Melissa officinalis L. Iranian Journal of Medical and Aromatic Plants. 26: 259-283. (In Farsi)
-      Chen, L.L. and Gao, S.H.L., 2007. In vitro tetraploid induction and generation of tetraploids from mixoploid   Astragalus memberanaceus. Scientia Horticulture, 112: 339-344.
-      Chen, W.L. and Yeh, D.M. 2006. Elimination of in vitro contamination shoot multiplication, and ex vitro rooting of Aglaonma. Horticulture Science. 42: 629-632.
-      Escandon, A.S., Alderet, L.M. and Hagwara, J.C., 2007. In vitro polyploidization of Mecardonia tenella, a native plant from South America. Science Horticulture, 115: 56-61.
-      Hesamzadeh Hejazi, S.M. and Ziyaeinasab, M., 2008. Cytogenetic analysis of some of diploid populations of Onobrychis genus in Natural Resources Gene Bank of Iran. Iranian Journal of Rangelands and Forests Plant Breeding and Genetic Research. 16: 158-171. (In Farsi).
-      Kerdsuwan, N. and Te- chato, S., 2012. Effects of colchicine on survival rate, morphological, physiological and cytological characters of Changdaeng orchid (Rhynchostylis gigantean var. rubrum Sagarik) In Vitro. Journal of Agricultural Technology 8: 1451-1460.
-      Madon, M., Clyde, M.M., Hashim, H., Mohd yusuf, Y., Mat, H. and Saratha, S., 2005. Polyploidy induction of oil palm through colchicine and oryzalin treatments. Journal of Oil Palm Research 7: 110-123.
-      Malekzadeh Shafaroodi, S., Gani, A., Habibi, M. and Amiri, A., 2011.  The possibility polyploidy induction in basil (Ocimum basilicum L.) using colchicine. Journal of Horticultural Science, 25: 461-469. (In Farsi).
-      Omidbaigi, R., Mirzaee, M., Hassani, M.E. and Sedghi Moghadam, M., 2010. Induction and identification of polyploidy in basil (Ocimum basilicum L.) medicinal plant by colchicine treatment. International Journal of Plant Production, 4: 87-98.
-      Rafezi, A., Farshadfar, M. and farshadfar, E., 2009.  Determination of intra-specific karyotypic variation in 17 genotypes of Agropyron elongatum L., Iranian Journal of Rangelands and Forests Plant Breeding and genetic Research, 17: 61-69.
-      Rodica, B., Bavaru, A. and Broască, L. 2011. Anatomical aspects of Phalaenopsis amabilis L. Blume. Annals of the Romanian Society for Cell Biology, 16:102-109.
-      Roy, A.T., Leggett, G. and Koutoulis, A., 2001. In vitro tetraploid induction and generation of tetraploids from mixoploids in hop (Humulus lupulus L.). Plant Cell Reports, 20: 489- 495.
-      Shahriari Ahmadi, F., Dehghan, E., Farsi, M. and Azizi, M., 2008. Tetraploid induction of Hyoscyamus muticus L. using colchicines treatment. Pakistan Journal of Biological Sciences, 11: 2653-2659.
-      Thao, N.T.P., Ureshino, K., Miyajima, I., Ozaki, Y. and Okuba, H., 2003. Induction of tetraploids in ornamental Alocasia through colchicines and oryzaline treatments. Plant Cell Tissue and Organ Culture, 72:19-25.