Microspore-derived Embryo Formation and Morphological Changes during the Isolated Microspore Culture of Radish (Raphanus sativus L.)

다국어 초록 (Multilingual Abstract)

Raphanus sativus L. cv. Taebaek, a efficiently microspore-derived embryo (MDE)-forming cultivar, and ‘Chungwoon’,a non-MDE-forming cultivar were selected as donor plants for isolated microspore culture. Radish flower bud of 2.0 (small,S), 4.0 (medium, M), and 6.0 (large, L) ± 0.5 mm in length were isolated to determine the temporal relationship between flowerbud size and MED yield. Anatomical observations revealed no difference in the structure of the flower buds between the twocultivars. In both cultivars, the stigmas were much longer than the floral leaf in M-sized flower buds. The MDE yields for ‘Taebaek’per petri dish were 6.6 and 1.3 for M- and L-sized of flower buds, respectively, but MDE formation was not induced in theS flower buds. On the other hand, ‘Chungwoon’ failed to form MDEs in all flower buds. The microspore density of ‘Taebaek’was 1.3 times more than that of ‘Chungwoon’ for M sized flower buds. Of the M-sized buds from ‘Taebaek’ and ‘Chungwoon’,92.1 and 81.6%, respectively, were in the late uninucleate microspore stage, which is characterized by the highest frequencyof MDE formation. Anatomical observations of MDE formation revealed that the microspores were able to divide to form aprimordium from which cell division took place continuously in the ‘Teabeak’ cultivar. However, the microspores of ‘Chungwoon’failed to progress beyond the primodium stage, resulting in lack of MDE formation. By contrast, after the formation of the primordium,various developmental stages of embyos from microspore were observed in the ‘Taebaek’ cultivar. These results can be usedto determine MDE forming potentials of radish cultivars.

참고문헌 (Reference)

1 Forster, B. P., "The resurgence of haploids in higher plants" 12 : 368-375, 2007

2 Touraev, A., "The microspore: A haploid multipurpose cell" 35 : 53-109, 2001

3 Biddington, N. L., "The influence of ethylene in plant tissue culture" 11 : 173-187, 1992

4 Žárský, V., "The expression of a small heat shock gene is activated during induction of tobacco pollen embryogenesis by starvation" 18 : 139-147, 1995

5 Gallie, D.R., "The ethylene biosynthetic and perception machinery is differentially expressed during endosperm and embryo development in maize" 271 : 267-281,

6 Xie, J.H., "The effect of cool-pretreatment on the isolated microspore culture and the free amino acid change of anthers in Japonica rice (Oryza sativa L)" 151 : 79-82, 1997

7 EdwardC.Yeung, "The Canola Microspore-Derived Embryo as a Model System to Study Developmental Processes in Plants" 한국식물학회 45 (45): 119-133, 2002

8 Cloutier, C., "Study of microspore-culture responsiveness in oilseed rape (Brassica napus L.) by comparative mapping of a F2 population and two microspore-derived populations" 91 : 841-847, 1995

9 Touraev, A., "Stress-induced microspore embryogenesis in tobacco: An optimized system for molecular studies" 15 : 561-565, 1996

10 Touraev, A., "Stress as the major signal controlling the developmental fate of tobacco microspores: towards a unified model of induction of microspore/pollen embryogenesis" 200 : 144-152, 1996

11 Smykal, P., "Pollen embryogenesis-The stress mediated switch from gametophytic to sporophytic development. Current status and future prospects" 43 : 481-489, 2000

12 Cho, M.S., "Plant regeneration from isolated microspore of Indica rice" 31 : 881-885, 1990

13 Gamborg, O. L., "Nutrient requirements of suspension culture of soybean root cells" 50 : 151-158, 1968

14 전창후, "Microspore-derived Embryo Formation in Response to Cold Pretreatment, Washing Medium, and Medium Composition of Radish (Raphanus sativus L.)" 한국원예학회 29 (29): 494-499, 2011

15 전창후, "Microspore-Derived Embryo Formation in Radish (Raphanus sativus L.) According to Nutritional and Environmental Conditions" 한국원예학회 52 (52): 530-535, 2011

16 Hansen, M., "Microspore culture of swede (Brassica napus ssp. rapifera) and the effects of fresh and conditioned media" 12 : 496-500, 1993

17 Takahata, Y., "Microspore culture of radish (Raphanus sativus L.): Influence of genotype and culture conditions on embryogenesis" 16 : 163-166, 1996

18 Polsoni, L., "Large-scale microspore culture technique for mutation-selection studies in Brassica napus" 66 : 1681-1685, 1988

19 Heberle-Bors, E, "Isolated pollen culture in tobacco: Plant reproductive development in a nutshell" 2 : 1-10, 1989

20 Babbar, S.B., "Isolated microspore culture of Brassica: An experimental tool for developmental" 3 : 185-202, 2004

21 Lichter, R., "Induction of haploid plants from isolated pollen of Brassica napus Z" 105 : 427-434, 1982

22 Baillie, A.M.R., "In vitro culture of isolated microspores and regeneration of plants in Brassica campestris" 11 : 234-237,

23 Takahashi, Y., "Improvement of microspore culture method for multiple samples in Brassica" 61 : 96-98, 2011

24 Xie, J., "Improved isolated microspore culture efficiency in medium with maltose and optimized growth regulator ombination in Japonica rice (Oryza sativa)" 42 : 245-250, 1995

25 Weber, S., "Improved doubled haploid production protocol for Brassica napus using microsproe colchicine treatment in vitro and ploidy determination by flow cytometry" 124 : 511-513, 2005

26 Seguí-Simarro, J.M., "How microspores transform into haploid embryos: Changes associated with embryogenesis induction and microspore-derived embryogenesis" 134 : 1-12, 2008

27 Raina, S.K., "High-frequency embryogenesis and plantlet regeneration from isolated microspores of Indica rice" 17 : 957-962, 1998

28 Curtis, I. S., "Genetic engineering of radish: Current achievements and future goals" 30 : 733-744, 2011

29 Burnett, L., "Embryogenesis and plant regeneration from isolated microspores of Brassica rapa L. ssp. oleifera" 11 : 215-218, 1992

30 Touraev, A., "Efficient microspore embryogenesis in wheat (Triticum aestivum L.) induced by starvation at high temperature" 9 : 209-215, 1996

31 Chuong, P.V., "Effects of donor genotype and bud sampling on microspore culture of Brassica napus" 66 : 1653-1657, 1988

32 Wu, L. M., "Effect of silver nitrate on the tissue culture of immature wheat embryos" 53 : 530-534, 2006

33 Takahata, Y., "Effect of donor plant age and inflorescence age on microspore culture of Brassica napus L" 58 : 51-55, 1991

34 da Silva Dias, J.C., "Effect of activated charcoal on Brassica oleracea microspore culture embryogenesis" 108 : 65-69, 1999

35 Telmer, C. A., "Determination of developmental stage to obtain high frequencies of embryogenic microspores in Brassica napus" 84 : 417-423, 1992

36 Senratna, T., "Desiccation of microspore derived embryos of oilseed rape (Brassica napus L.)" 10 : 342-344, 1991

37 Žárský, V., "Derepression of the cell cycle by starvation is involved in the induction of tobacco pollen embryogenesis" 5 : 189-194, 1992

38 Tarnowski, B. I., "DAPI as a useful stain for nuclear quantitation" 66 : 297-302, 1991

39 Yeung, E. C., "Comparative development of zygotic and microspore-derived embryos in Brassica napus L. cv. Topas. I. histodifferentiation" 157 : 27-39, 1996

40 Gu, H. H., "Cold pretreatment enhances microspore embryogenesis in oilseed rape (Brassica napus L.)" 42 : 137-143, 2004

41 Curtis, I. S., "Biotechnology in agriculture and forestry" Springer 381-389, 2007

42 Binarova, P., "A short severe heat shock is required to induce embryogenesis in late bicellular pollen of Brassica napus L" 10 : 200-208, 1997