AJPS  Vol.4 No.6 A , June 2013
High Frequency Sporophytes Regeneration from the Spore Culture of the Endangered Aquatic Fern Isoetes coreana
Abstract: Using a mixed culture of megaspores and microspores from I. coreana, we established high frequency sporophyte regeneration system. After 20 days of culturing in MS basal medium, microscopic examination showed significant morphological changes and the microspore released numerous small vesicles into the culture medium. Megaspores also showed dramatic morphological changes during its incubation time in culture. The spore wall was cracked by the expansion of the megaspore (about 2 times increase in diameter). Simultaneously, brown spots were observed on the surface of the megaspores. The frequency of female gametophytes developing from immature megaspores cultured in MS basal liquid medium (pH 7) supplemented with 1 mgl-1 GA3 was 46%. However, these female gametophytes derived from megaspore only culture could not differentiate into sporophytes. The mixed culture of microspores and megaspores resulted in successful sporophyte regeneration. The highest frequency (12.3%) of green sporophyte regeneration from mixed spore culture occurred when the cultures were maintained at 25℃ under cool-white fluorescent light (40 μmol·m-2·s-1) with a 16 h photoperiod. Regenerated sporophytes were transferred to a test tube containing vermiculite and a sand mixture and left there until they had three leaves. After root growth and the fifth leaf had emerged, more than 95% of the regenerated sporophytes were successfully transferred to the soil and grown to mature plants. The sporophyte regeneration system established in this study could be successfully used for the restoration of the endangered aquatic species, I. coreana.
Cite this paper: M. Oh, C. Kim, H. Na, H. Shin, J. Liu, H. Choi and S. Kim, "High Frequency Sporophytes Regeneration from the Spore Culture of the Endangered Aquatic Fern Isoetes coreana," American Journal of Plant Sciences, Vol. 4 No. 6, 2013, pp. 14-20. doi: 10.4236/ajps.2013.46A003.

[1]   W. C. Taylor and R. J. Hickey, “Habitat, Evolution and Speciation in Isoetes,” Annals of the Missouri Botanical Garden, Vol. 79, No. 3, 1992, pp. 613-622. doi:10.2307/2399755

[2]   W. C. Taylor, N. T. Luebke, D. M. Britton, R. J. Hickey and D. F. Brunton, “Isoetaceae,” FNA Editorial Committee, Flora of North America, Oxford University Press, New York, 1993, pp. 64-75.

[3]   J. E. Keeley, “CAM Photosynthesis in Submerged Aquatic Plants,” Botanical Review, Vol. 64, No. 2, 1998, pp. 121-175. doi:10.1007/BF02856581

[4]   N. S. Parihar, “The Biology and Morphology of Pteridophytes,” Indian Universities Press, Allahabad, 1996.

[5]   K. R. Stern, “Introductory Plant Biology”, 3rd Edition, W. C. Brown, Publishers, Dubuque, 1985, p. 515.

[6]   COSEWIC, “COSEWIC Assessment and Status Report on the Prototype Quillwort Isoetes prototypus in Canada,” Committee on the Status of Endangered Wildlife in Canada, Ottawa, 2005.

[7]   C. A. Caplen and C. R. Werth, “Isozymes of the Isoetes riparia Complex, I. Genetic Variation and Relatedness of Diploid Species,” Systematic Botany, Vol. 25, No. 2, 2000, pp. 235-259. doi:10.2307/2666641

[8]   S. J. Sam, “A Germination Method for Isoetes,” American Fern Journal, Vol. 72, No. 2, 1982, p. 61. doi:10.2307/1547062

[9]   W. C. Taylor and N. T. Luebke, “Germinating Spores and Growing Sporelings of Aquatic Isoetes,” American Fern Journal, Vol. 76, No. 1, 1986, pp. 21-24. doi:10.2307/1547396

[10]   H. Fernandez and M. A. Revilla, “In Vitro Culture of Ornamental Ferns,” Plant Cell, Tissue and Organ Culture, Vol. 73, No. 1, 2003, pp. 1-13. doi:10.1023/A:1022650701341

[11]   D. Cachita-Cosma, “Metode in Vitro la Plantele de Cultura,” Edit. Ceres, Bucuresti, Agricultural Sciences, University of Florida, 1987.

[12]   B. M. Boom, “Intersectional Hybrids in Isoetes,” American Fern Journal, Vol. 70, No. 1, 1980, pp. 1-4. doi:10.2307/1546200

[13]   L. S. Koot and D. M. Britton, “A Comparative Study of Spore Germination of Some Isoetes Species of Northeastern North America,” Canadian Journal of Botany, Vol. 60, No. 9, 1982, pp. 1679-1687. doi:10.1139/b82-218

[14]   Y. H. Chung and H.-K. Choi, “ Isoetes coreana, a New Species from Korea,” Korean Journal of Plant Taxonomy, Vol. 16, 1986, pp. 1-12 (in Korean, with English Abstract).

[15]   C. Kim, H. R. Na and H.-K. Choi, “Genetic Diversity and Population Structure of Endangered Isoetes coreana in South Korea Based on RAPD Analysis,” Aquatic Botany, Vol. 89, No. 1, 2008, pp. 43-49. doi:10.1016/j.aquabot.2008.02.004

[16]   T. Murashige and F. Skoog, “A Revised Medium for Rapid Growth and Bioassays with Tobacco Tissue Culture,” Plant Physiology, Vol. 15, No. 3, 1962, pp. 473-497. doi:10.1111/j.1399-3054.1962.tb08052.x

[17]   K. P. Martin, S. Sini, C.-L. Zhang, A. Slater and P. V. Madhusoodanan, “Efficient Induction of Apospory and Apogamy in Vitro in Silver Fern (Pityrogramma calomelanos L.),” Plant Cell Reports, Vol. 25, No. 12, 2006, pp.1300-1307. doi:10.1007/s00299-006-0215-5

[18]   S Hedge, “Propagation of Some Ornamental Plants Using Biotechnology,” Dissertation, Mangalore University, Mangalore, 1998.

[19]   E. A. Simabukuro, A. F. Dyer and G. M. Felippe, “The Effect of Sterilization and Storage Conditions on the Viability of Spores of Cyathea delgadii,” American Fern Journal, Vol. 88, No 2, 1998, pp. 124-132. doi:10.2307/1547226

[20]   C. C. Macluf, M. A. Morbelli and G. E. Giudice, “Microspore Morphology of Isoetes Species (Lycophyta) from Southern South America,” Botanical Review, Vol. 72, No. 2, 2006, pp. 121-134. doi:10.1663/0006-8101(2006)72[121:MMOISL]2.0.CO;2