Distribution of Nuclei of Different Ploidy Levels during Ovule, Seed and Protocorm Development in Phalaenopsis aphrodite subsp. formosana (Orchidaceae)
Goamg-Tyng Jean, Yu-Lin Kao, Ching-Yan Tang, Wen-Huei Chen
DOI: 10.4236/ajps.2011.23037   PDF    HTML     5,571 Downloads   10,050 Views   Citations


Distribution of nuclei of different ploidy levels was studied at different developmental stages in the embryonic tissue of the ovule, seed and protocorm of Phalaenopsis aphrodite subsp. formosana (Miwa) E.A. Christ. by a combination of flow cytometry and fluorescence microscopy with Apo Tome Slider. Three stages of ploidy patterns were identified in the ovular tissue at different days after pollination (DAP). Firstly, between pollination and fertilization (0 to 50 DAP), 2C nuclei were dominant over 4C nuclei and resulted in low level of cycle value. Secondly, between fertilization and seed maturation (50 to 110 DAP), amount of 4C nuclei increased rapidly, maintained at a high level and then decreased gradually to a low level. Small amount of 8C nuclei was also detected at this stage. Thirdly, at seed maturation (110 to 130 DAP), 2C nuclei became dominant over 4C nuclei again and the cycle value remained at a low but significant level at this stage. After seed sowing, nuclei with ploidy levels of 2C, 4C and 8C were observed in the developing protocorms as early as at 4 DAS (days after sowing). Nuclei with high ploidy levels (8C and 16C) increased gradually until 40 DAS in this study. Significant level of cycle value at this stage of protocorm development indicated the presence of endopolyploidy. 4,6-diamido-2-phenylindol (DAPI) staining showed large and prominent nuclei in the basal portions of the mature seeds before sowing and in the developing protocorms at 20 DAS. These findings clearly demonstrate the occurrence of different distribution patterns of nuclei with different ploidy levels during ovule, seed and protocorm de-velopment in Phalaenopsis aphrodite. These observations will provide fundamental information for further studies in Phalaenopsis orchids.

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Jean, G. , Kao, Y. , Tang, C. and Chen, W. (2011) Distribution of Nuclei of Different Ploidy Levels during Ovule, Seed and Protocorm Development in Phalaenopsis aphrodite subsp. formosana (Orchidaceae). American Journal of Plant Sciences, 2, 325-333. doi: 10.4236/ajps.2011.23037.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] G. Grafi, “Cell Cycle Regulation of DNA Replication: The Endoreduplication Perspective,” Experimental Cell Research, Vol. 244, No. 2, 1998, pp. 372-378. doi:10.1006/excr.1998.4213
[2] S. van den Heuvel, “Cell-Cycle Regulation,” WormBook, The C. elegans Research Community, 2005.
[3] D. Francis, “The Plant Cell Cycle-15 Years on,” New Phytologist, Vol. 174, No. 2, 2007, pp. 261-278. doi:10.1111/j.1469-8137.2007.02038.x
[4] B. A. Larkins, B. P. Dilkes, R. A. Dante, C. M. Coelho, Y. M. Woo and Y. Liu, “Investigating the Hows and Whys of DNA Endoreduplication,” Journal of Experimental Botany, Vol. 52, No. 355, 2001, pp. 183-192. doi:10.1093/jexbot/52.355.183
[5] M. Barow, “Endopolyploidy in Seed Plants,” BioEssays, Vol. 28, No. 3, 2006, pp. 271-281.
[6] D. P. Biradar, A. Lane Rayburn and D. G. Bullock, “Endopolyploidy in Diploid and Tetraploid Maize (Zea mays L.),” Annals of Botany, Vol. 71, No. 5, 1993, pp. 417-421. doi:10.1006/anbo.1993.1053
[7] M. J. M. Smulders, W. Rus-Kortekaas and L. J. W. Gilssen, “Development of Polysomaty during Differentiation in Diploid and Tetraploid Tomato (Lycopersicon esculentum) Plants,” Plant Science, Vol. 97, No. 1, 1994, pp. 53-60. doi:10.1016/0168-9452(94)90107-4
[8] K. A. Pyke, L. L. Marrison and R. M. Leech, “Temporal and Spatial Development of the Cells of the Expanding First Leaf of Arabidopsis thaliana (L.) Heynh.,” Journal of Experimental Botany, Vol. 42, No. 11, 1991, pp. 1407-1416. doi:10.1093/jxb/42.11.1407
[9] P. D. Griffiths, H. J. Ougham and R. N. Jones, “Genotypic and Environmental Effects on Endopolyploidy in the Epidermal Tissues of Lolium perenne L. and Lolium multiflorum Lam,” New Phytologist, Vol. 128, No. 2, 1994, pp. 339-345. doi:10.1111/j.1469-8137.1994.tb04018.x
[10] M. Barow and A. Meister, “Endopolyploidy in Seed Plants is Differently Correlated to Systematics, Organ, Life Strategy and Genome Size,” Plant, Cell & Environment, Vol. 26, No. 4, 2003, pp. 571-584. doi:10.1046/j.1365-3040.2003.00988.x
[11] W. Nagl, “DNA Endoreduplication and Polyteny Understood as Evolutionary Strategies,” Nature, Vol. 261, 1976, pp. 614-615.
[12] F. D’Amato, “Chromosome Endoreduplication in Plant Tissue Development and Function,” In: J. A. Bryant and D. Chiatante, Ed., Plant Cell Proliferation and Its Regulation in Growth and Development, John Wiley & Sons, Chichester, 1998, pp. 153-166.
[13] N. Kudo, Y. Kimura, “Nuclear DNA Endoreduplication during Petal Development in Cabbage: Relationship between Ploidy Levels and Cell Size,” Journal of Experimental Botany, Vol. 53, No. 371, 2002, pp. 1017-1023. doi:10.1093/jexbot/53.371.1017
[14] G. Jovtchev, V. Schubert, A. Meister, M. Barow and I. Schubert, “Nuclear DNA Content and Nuclear and Cell Volume Are Positively Correlated in Angiosperms,” Cytogenetic and Genome Research, Vol. 114, No. 1, 2006, pp. 77-82.
[15] S. Lin, H. C. Lee, W. H. Chen, C. C. Chen, Y. Y. Kao, Y. M. Fu, Y. H. Chen and T. Y. Lin, “Nuclear DNA Contents of Phalaenopsis sp. and Doritis pulcherrima,” Journal of the American Society for Horticultural Science, Vol. 126, No. 2, 2001, pp. 195-199.
[16] W. L. Lim and C. S. Loh, “Endopolyploidy in Vanda Miss Joaquim (Orchidaceae),” New Phytologist, Vol. 159, No. 1, 2003, pp. 279-287. doi:10.1046/j.1469-8137.2003.00797.x
[17] M. Yang and C. S. Loh, “Systemic Endopolyploidy in Spathoglottis plicata (Orchidaceae) Development,” BMC Cell Biology, Vol. 5, 2004, p. 33. doi:10.1186/147-2121-5-33
[18] W. H. Chen, C. Y. Tang and Y. L. Kao, “Ploidy Doubling in Vitro Culture of Excised Protocorms or Protocorm-Like Bodies in Phalaenopsis Species,” Plant Cell, Tissue and Organ Culture, Vol. 98, No. 2, 2009, pp. 229-238. doi:10.1007/s 11240-009-9557-3
[19] H. C. Lee, D. W. Chiou, W. H. Chen, A. H. Markhart, Y. H. Chen and T. Y. Lin, “Dynamics of Cell Growth and Endoreduplication during Orchid Flower Development,” Plant Science, Vol. 166, No. 3, 2004, pp. 659-667. doi:10.1016/j.plantsci.2003.10.034
[20] M. R. Alvarez, “Quantitative Changes in Nuclear DNA Accompanying Postgermination Embryonic Development in Vanda (Orchidaceae),” American Journal of Botany, Vol. 55, No. 9, 1968, pp. 1036-1041. doi:10.2307/2440469
[21] J. A. Nadeau, X. S. Zhang, J. Li and S. D. O’Neill, “Ovule Development: Identification of Stage-Specific and Tissue-Specific cDNAs,” The Plant Cell, Vol. 8, No. 2, 1996, pp. 213- 239.
[22] X. S. Zhang and S. D. O'Niell, “Ovary and Gametophyte Development Are Co-Ordinately Regulated by Auxin and Ethylene Following Pollination,” The Plant Cell, Vol. 5, No. 4, 1993, pp. 403-418.
[23] Y. I. Lee, E. C. Yeung, N. Lee and M. C. Chung, “Embryology of Phalaenopsis amabilis var. formosa: Embryo Development,” Botanical Studies, Vol. 2, 2008, pp. 139-146.
[24] T. Murashige and F. Skoog, “A Revised Medium for Rapid Growth and Bioassays with Tobacco Tissue Culture,” Physiologia Plantarum, Vol. 15, No. 3, 1962, pp. 473-497. doi:10.1111/j.1399-3054.1962.tb08052.x
[25] W. T. Tsai, W. H. Chen, R. M. Hsieh, M. S. Chyou and C. C. Wu, “An Important Factor Affecting the Germination and Growth of Phalaenopsis Seeds,” In: S. Huang, S. Hsieh and T. Liu, Ed., The Impact of Biological Research on Agricultural Productivity, Taichung District Agricultural Improvement Station-Taichung, Taiwan, 1992, pp. 219-228.
[26] N. Kudo and Y. Kimura, “Flow Cytometric Evidence for Endopolyploidy in Seedlings of Some Brassica Species,” Theoretical and Applied Genetics, Vol. 102, No. 1, 2001, pp. 104-110. doi:10.1007/s001220051624
[27] E. C. Yeung and D. W. Meink, “Embryogenesis in Angiosperms: Development of the Suspensor,” The Plant Cell, Vol. 5, No. 10, 1993, pp. 1371-1381. doi:10.2307/3869789
[28] A. Kladnik, P. S. Chourey, D. R. Pring and M. Dermastia, “Development of the Endosperm of Sorghum bicolor during the Endoreduplication-Associated Growth Phase,” Journal of Cereal Science, Vol. 43, No. 2, 2006, pp. 209-215. doi:10.1016/j.jcs.2005.09.004
[29] K. Robinson-Beers, R. E. Pruitt and C. S. Gasser, “Ovule Development in Wild-Type Arabidopsis and Two Female-Sterile Mutants,” The Plant Cell, Vol. 4, No. 10, 1992, pp. 1237-1249.
[30] W.-H. Chen, C.-Y. Tang, T.-Y. Lin, Y.-C. Weng and Y-L. Kao, “Changes in the Endopolyploidy Patterns of Different Tissues in Diploid and Tetraploid Phalaenopsis aphrodite subsp. formosana (Orchidaceae),” Plant Science, Vol. 181, No. 1, 2011, pp. 31-38. doi:10.1016/j.plantsci.2011.03.006

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