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308 Kunasukdakul and Smitamana, 2003). However, all raphids and debris could be successfully removed by centrifugation of protoplasts with 16 % sucrose solution. For the protoplasts culture of C. wendtii, the culture media, culture method and plant growth regulators were important factors affecting plating efficiency and survival rate. The protoplasts could divide in liquid as well as in agarose bead culture. However, microcolonies were formed only in agarose bead culture. The agarose bead culture methods have been found to be an efficient method for cell division and microcolony formation in many crop species including Lavatera thuringiaca (Vazquez-Tello et al., 1995); Rosa hybrida (Marchant et al., 1997) and Cucumis melo ‘Green Delica’ (Sutiojono et al., 1998). The enhanced protoplast division observed in bead culture was due to the dilution of substances having inhibitory effects on protoplast division which are secreted from the cell to the medium (Mizuhiro et al., 2001). Colony formation was observed after culturing protoplasts in MS medium supplemented with 0.2 mg/l 2,4-D, 1 mg/l NAA, 0.5 mg/l Zeatin, 0.3 M mannitol, and 0.15 M sucrose for 30 days. However, it did not form a callus but turned brown and finally died. It has been reported that the protoplasts isolated directly from leaves of monocotyledons, except rice, was very difficult to culture (Kuehnle and Nan, 1990). It was suggested that leaf cells rapidly lose totipotency thus preventing cells from dedifferentiating and reentering the cell cycle (Krautwig and Lörz, 1995). Plant regeneration has been found possible when callus and cell suspension were used as the source of protoplast isolation and culture (Kobayashi et al., 1993; Pauk, et al., 1994). CONCLUSION The procedure for simple and reliable isolation and culture of C. wendtii protoplasts has Kasetsart J. (Nat. Sci.) 41(2) been described for the first time. It might lead to the improvement of the Cryptocoryne through somatic hybridization, somaclonal variation and genetic engineering by using the protoplast technique. Even though the viable protoplasts of C. wendtii could form microcolonies, further research is needed to develop the efficient procedure for the protoplast regeneration. ACKNOWLEDGMENTS This research was financially supported by the Department of Fisheries, Ministry of Agriculture and Co-operatives, Bangkok, Thailand. We thank Dr. Sureeya Tantiwiwat, Department of Botany, Kasetsart University, Dr. Yuphin Khentry and Mr. Adrian Hillman, Graduate School, Kasetsart University for editing this manuscript and their helpful suggestions. LITERATURE CITED Allgayer, R. and J. Teton. 1986. Aquarium Plants. Worlds Lock, Ltd., London. 157 p. Babaoˇglu, M. 2000. Protoplast isolation in Lupin (Lupinus mutabilis Sweet): determination of optimum explant sources and isolation conditions. Turk. J. Bot. 24: 177-185. Balestri, E. and F. Cinelli. 2001. Isolation and cell wall regeneration of protoplasts from Posidonia oceanic and Cymodocea nodosa. Aqua. Bot. 70: 237-242. Cheng, Y.J., W.W. Gou and X.X. Deng. 2003. Molecular characterization of cytoplasmic and nuclear genomes in phenotypically abnormal Valencia orange (Citrus sinensis) plus Meiwa kumquat (Furtunella crassifolia) intergeneric somatic hybrids. Plant Cell Rep. 21: 445-451. Davey, M.R., P. Anthony, J.B. Power and K.C. Lowe. 2005. Plant protoplasts: status and biotechnological perspectives. Biotechnol. Advances 23: 131-171. Eriksson, T.R. 1985. Protoplast isolation and
culture, pp. 1-20. In L.C. Fowke and F. Constabel (eds.). Plant Protoplasts. CRC Press, Florida. Fu, C.H., W.W. Guo, J.H. Liu, X.X. Deng. 2003. Regeneration of Citrus sinensis (+) Clausena lansium intergeneric tripliod and tetraploid somatic hybrids and their identification by molecular markers. In Vitro Cell. Dev. Biol- Plant 39: 360-364. Gleddie, S.C. 1995. Protoplast isolation and culture, pp. 167-180. In O.L. Gamborg and G.C. Phillips (eds.). Plant Cell Tissue and Organ Culture : Fundamental Methods. Springer-Verlag, Berlin Heidelberg. Kane, M.E., G.L. Davis, D.B. McConnell and J.A. Gargiulo. 1999. In vitro propagation of Cryptocoryne wendtii. Aquat. Bot. 63: 197- 202. Kao, K.N. and M.R. Michayluk. 1975. Nutritional requirements for growth of Vicia hajastana cells and protoplasts at a very low population density in liquid media. Planta 126: 105-110. Kobayashi, R., T. Kameya and S. Ichihashi. 1993. Plant regeneration from protoplasts derived from callus of Phalaenopsis spp. Plant Tiss. Cult. Lett. 10: 267-270. Krautwig, B. and H. Lörz. 1995. Review article cereal protoplasts. Plant Sci. 111: 1-10. Kuehnle, A.R. and G.L. Nan. 1990. Factors influencing the isolation and culture of protoplasts from Hawaiian Dendrobium cultivars, pp. 259-262. In J. Kernohan, D.G. Bonhanm, N. Bonhanm and L. Cobb (eds.). Proceedings of the 13 th World Orchid Conference. 13 th World Orchid Conference Proceeding Trust, Auckland. Kunasakdakul, K. and P. Smitamana. 2003. Dendrobium Pratum Red protoplast. Thai J. Agric. Sci. 36: 1-8. Liu, J.H., X. Xu and X.X. Deng. 2005. Intergeneric somatic hybridization and its appplication to crop genetic improvement. Plant Cell Tiss. Org. Cult. 82: 19-44. Kasetsart J. (Nat. Sci.) 41(2) 309 Marchant, R., M.R. Davey and J.B. Power. 1997. Isolation and culture of mesophyll protoplasts from Rosa hybrida. Plant Cell Tiss. Org. Cult. 50: 131-134. Mizuhiro, M., K. Ito and M. Mii. 2001. Production and characterization of interspecific somatic hybrids between Primula malacoides and P. obconica. Plant Sci. 161: 489-496. Mühlberg, H. 1982. The Complete Guide to Water Plants. Sterling Publishing Co., Inc., German Democratic Republic. 392 p. Murashige, T. and F. Skoog. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant 15: 473-497. Nagata, T. and S. Ishii. 1979. A rapid method for isolation of mesophyll protoplast. Can. J. Bot. 57: 1820-1823. Pan, Z.G., C.Z. Liu, S.J. Murch, M. El-Demerdash and P.K. Saxena. 2003. Plant regeneration from mesophyll protoplasts of the Egyptian medicinal plants Artemisia judaica L. and Echinops spinosissimus Turra. Plant Sci. 165: 681-687. Pauk, J., Z. Kertész, B. Jenes, L. Purnhauser, O. Manninen, S. Pulli, Z. Barabás and D. Dudits. 1994. Fertile wheat (Triticum aestivum L.) regenerants from protoplasts of embryogenic suspension culture. Plant Cell Tiss. Org. Cult. 38: 1-10. Phansiri, S., H. Miyake and E. Maeda. 1992. Studies on cell wall regeneration and cell division in soybean protoplasts using fluorescence and scaning electron microscope. Jpn. J. Crop Sci. 61: 487-493. Price, G.C. and E.D. Earle. 1984. Source of orchid protoplasts for fusion experiments. Amer. Orchid Soc. Bull. 53: 1035-1043. Rajaj, K. and T.J. Horeman. 1977. Aquarium Plant: Their Identification, Cultivation and Ecology. T.F.H. Pulb. Inc., West Sylvania. 448 p. Sinha, A., A.C. Wetten and P.D.S. Caligari. 2003.
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April - June 2007 Volume 41 Number
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KASETSART JOURNAL NATURAL SCIENCE T
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Kasetsart J. (Nat. Sci.) 41 : 205 -
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harvesting time which started from
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y Chen and Paull (2000). Figure 1 a
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pineapple fruit allowed the accumul
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Kasetsart J. (Nat. Sci.) 41(2) 215
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Cluster analysis Cluster analysis u
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Table 3 (Continued) 1 2 3 4 5 6 7 8
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CONCLUSION AFLP markers classified
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difference of soil texture used in
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under wet soil condition planting.
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tropics. Following the expansion, w
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sealed in a plastic box with 1 cm w
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moisture content increment after so
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Frequency Frequency 30 20 10 0 20.0
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School, Kasetsart University. The a
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for combining ability of lines (Lam
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selective mass sibbing within indiv
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Although most of S 2-interfamily hy
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composite sets as used in this stud
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days. The numbers of calli producin
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the report of Ching (1982) showing
Page 57 and 58: clearly amplified bands generated b
Page 59 and 60: caused by either the recombination
Page 61 and 62: Kuhlmann, V. and B. Foroughi-Wehr.
Page 63 and 64: Xanthomonas fuscans subsp. aurantif
Page 65 and 66: Kasetsart J. (Nat. Sci.) 41(2) 265
Page 67 and 68: was collected and washed with 70% e
Page 69 and 70: expected size was amplified with 35
Page 71 and 72: eaction technique has been used for
Page 73 and 74: Schaad, N.W., D. Opgenorth and P. G
Page 75 and 76: MATERIALS AND METHODS Model descrip
Page 79 and 80: calculated TF value calculated TF v
Page 81 and 82: sincere gratitude and deep apprecia
Page 83 and 84: 1989). In monogastrics, the inclusi
Page 85 and 86: of all LLS samples in this study we
Page 87 and 88: Melbourne, Parkville, Victoria. Goe
Page 89 and 90: considered responsible for low prod
Page 93 and 94: mineral supplementation suggested b
Page 95 and 96: Body weight chane (kg/head) 32 30 2
Page 97 and 98: CONCLUSION AND RECOMMENDATION With
Page 99 and 100: SAS. (Statistical Analysis System).
Page 101 and 102: genes covering a variety of desirab
Page 103 and 104: mg/l NAA and 0.5 mg/l zeatin) incub
Page 105 and 106: 5. Purification by various sucrose
Page 107: as the culture period increased. So
Page 111 and 112: Kasetsart J. (Nat. Sci.) 41 : 311 -
Page 113 and 114: GC. Analytical methods Total carboh
Page 115 and 116: ash. The crude hot water polysaccha
Page 117 and 118: spirulan (H-SP), and a desulfated c
Page 121 and 122: cells often displayed an increased
Page 123 and 124: LITERATURE CITED Bell, T.A. and D.V
Page 125 and 126: for alternative sources of supply.
Page 127 and 128: BR2.1.2 formed the same clade with
Page 129 and 130: supplement with glutamate (Iida et
Page 131 and 132: optimal for S. limacinum BR2.1.2 fo
Page 133 and 134: fluorescent lamp at 1.5 kLux develo
Page 137 and 138: Wisconsin, USA). Total extracted RN
Page 139 and 140: RESULTS Cloning and sequencing of m
Page 141 and 142: Expression of rmIL-2 and purificati
Page 143 and 144: other strains of mice have differen
Page 145 and 146: dose interleukin-2 therapy promotes
Page 147 and 148: The chitosano-oligosaccharides are
Page 149 and 150: enzyme and cells were maximized by
Page 151 and 152: of crude chitosanases. Fortunately,
Page 153 and 154: Relative activity (%) 100 80 60 40
Page 155 and 156: Uchida, K. Tateishi, O. Shida and K
Page 157 and 158: MATERIALS AND METHODS 1. Materials
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without coating and coated with pec
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in Table 5-7. It was found that the
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the cultures at 10,200 × g for 15
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incubation and reached the final pH
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y lysine- and tyrosine-decarboxylas
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during the stages of ripening as su
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As the 25 companies were divided in
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All the data comes up with informat
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as piece “A” or straight as pie
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et al. (1998); Zeng (2000); and Tal
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p 1, i * Kasetsart J. (Nat. Sci.) 4
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c ≤ c2 jq2 j n q p k p * 2, j , ,
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algorithm would consider RM 5, prio
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the maximum deviation of about 10%.
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Stock Quantities Using Heuristic Op
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which dynamically and automatically
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2. Framework architecture and compo
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Risk (VaR) calculation which was im
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Throughput (job/sec) 7 6 5 4 3 2 1
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Speed up 40 35 30 25 20 15 10 5 0 F
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and discovery, resource selection a
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