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Kasetsart J. (Nat. Sci.) 40(1) 51 Figure 1 CymMV-infected Dendrobium Sakura showing mosaic on leaves (A), ORSV-infected Cattleya spp. showing color breaking on flower (arrow)(B), electron micrographs of CymMV particles (C), electron micrographs of ORSV particles (D), symptom on leaves of infected CymMV in vitro plantlets are not smooth, dark green areas raised somewhat above the light green tissue as longitudinal ridges and bumps (E) and CymMV infected plantlets showing mosaic on leaves (arrow)(F).
52 in PBST containing 2% ovalbumin and added to each well, incubated at 37°C for 1 hour and then, repeatedly washed as above. A 100 µl aliquot of freshly prepared substrate (10 mg p-nitrophenyl phosphate; Sigma # N 6260, Sigma Chemical, St. Louis, USA) was dissolved in 10 ml of substrate buffer (9.7% diethanolamine, 0.02% NaN 3, pH 9.6) and added to each well. They were incubated in a moist chamber at 37°C for 30 minute, after that 50 µl of 3 M NaOH was added to all the wells to stop further enzymatic reactions. Absorbance value of each well was measured at 405 nm with an ELISA microplate reader (GDV model DV 990BV4, Italy). The color reactions produced by tested samples were compared with known negative control wells. The mean absorbance of infected samples that exceeded two fold of mean absorbance of the healthy samples was considered as a positive reaction (Satula et al., 1986). 2. Detection of CymMV and ORSV in in vitrocultivated orchids plantlets Forty-four cultivars of in vitro-cultivated orchid plantlets were obtained from commercial tissue culture laboratories in Bangkok, Non Thaburi, Pathum Thani and Nakhon Pathom provinces. One leaf from each of 20 plants of each cultivar was randomly selected and separately assayed for CymMV and ORSV by indirect- ELISA as described previously. 3. Bioassay Bioassay was used to confirm the ELISA results. Eighty samples that gave positive (66) and negative (14) reactions by indirect-ELISA were examined for presence of CymMV. Forty-fiveday-old Cassia occidentalis plants were used as indicator plants for CymMV. The orchid leaves were ground in a 0.01 M phosphate pH 7.0 buffer solutions at ratio 1:2 and used as inoculum. The inoculum was kept cool and used immediately. The C. occidentalis leaves were dusted with 600-mesh carborundum. The inoculum was rubbed gently Kasetsart J. (Nat. Sci.) 40(1) onto the leaf surface with a sterile cotton bud until leaves appears wetted. The inoculated leaves were rinsed with distilled water. Inoculum from infected CymMV orchid plants and plain buffer solution were included as positive and negative controls, respectively. The indicator plants were maintained in a greenhouse and symptoms were observed after incubation for 3-5 days, specifically for small, discrete, brown lesions, which indicated infection. The experiment was triplicate. RESULTS A total of 1,000 tissue-cultured seedlings of Thai native orchid and 880 in vitro cultivated orchid plantlets were assayed for CymMV and ORSV using indirect ELISA. It was found that CymMV and ORSV were not detected in any axenic Thai native orchid seedlings (Table 1). Out of 880 cultivated orchid samples, 243 (27.6%) were positive for CymMV while ORSV was negative. CymMV was detected in 6 out of 10 genera namely Brassolaeliocattleya, Cattleya, Dendrobium, Epicattleya, Oncidium and Mokara. The incidence of CymMV infection was in ranged between 50 % and 100 %. High infection rates were observed in Brassolaeliocattleya Alma Kee, Dendrobium Chanel, Dendrobium Chao Praya, Dendrobium Pravit White, Dendrobium Sakura and Dendrobium Shavin White (Table 2). Leaves of infected CymMV in vitro plantlets are not smooth, dark green areas raised somewhat above the light green tissue as longitudinal ridges and bumps (Figure 1E). CymMV-infected plantlets also showed mosaic on leaves (Figure 1F). All sixty-six positive samples (of eighty) were screened for CymMV using ELISA also positive by bioassay. Local, necrotic lesions appeared on the inoculated leaves of C. occidentalis after 4-6 days of incubation in the greenhouse (Figure 2).
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January - March 2006 Volume 40 Numb
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KASETSART JOURNAL NATURAL SCIENCE T
Page 5 and 6: In sacco Degradation Characteristic
Page 7 and 8: 2 and management practices a sorghu
Page 9 and 10: 4 the greater plant height was obta
Page 11 and 12: 6 at Wolenchity (Figure 3) that led
Page 13 and 14: 8 Stover and aboveground biomass Co
Page 15 and 16: 10 Radford, B.J., A.J. Dry, L.N. Ro
Page 17 and 18: 12 At present, new cultivars with h
Page 19 and 20: 14 2000), grapevine (Lavee, 2000),
Page 21 and 22: 16 repeatability of FW, FLT, FLW, S
Page 23 and 24: 18 (Table 5) indicating that select
Page 25 and 26: Kasetsart J. (Nat. Sci.) 40 : 20 -
Page 27 and 28: 22 measured to define fruit shape i
Page 29 and 30: 24 for higher fruit number and yiel
Page 33 and 34: 28 (Table 2). The numbers of flower
Page 35 and 36: 30 to the color of fruit and seed c
Page 37 and 38: 32 1999. Carrots and related vegeta
Page 39 and 40: 34 MATERIALS AND METHODS Laboratory
Page 41 and 42: 36 DISCUSSION Extensive studies hav
Page 43 and 44: 38 from Gossypium hirsutum. J. Inse
Page 45 and 46: 40 The natural resistance of plants
Page 47 and 48: 42 4. Data collection and statistic
Page 49 and 50: 44 was not significantly different
Page 51 and 52: 46 showed the highest yield compare
Page 53 and 54: 48 Kessmann, H., T. Staub, C. Hofma
Page 55: 50 There are approximately 60 comme
Page 59 and 60: 54 Kasetsart J. (Nat. Sci.) 40(1) T
Page 61 and 62: 56 Virus-free orchid plants show fa
Page 65 and 66: 60 feed source was based on purchas
Page 67 and 68: 62 intake of DM, CP, ether extract
Page 69 and 70: 64 relatively slow and short. The p
Page 71 and 72: 66 week 4. Milk composition was not
Page 73 and 74: 68 Agricultural Experiment Station.
Page 75 and 76: 70 et al., 2002). In contrast to ma
Page 77 and 78: 72 level and showed marked polymorp
Page 81 and 82: 76 loam (Eutric Fluvisol) soil and
Page 83 and 84: 78 Eugenia caryophyllata, Echinops
Page 85 and 86: 80 CONCLUSION In conclusion, out of
Page 87 and 88: 82 of chromic acid titration method
Page 89 and 90: 84 initials at the time of pinning
Page 91 and 92: 86 vessels a point where the number
Page 93 and 94: 88 Kasetsart J. (Nat. Sci.) 40(1) T
Page 95 and 96: 90 CONCLUSIONS In the investigation
Page 97 and 98: 92 1994; Strand et al., 1997). Now
Page 99 and 100: 94 The clones were grown overnight
Page 101 and 102: 96 isozymes. The copy number for nu
Page 103 and 104: 98 Helianthus annuus. Theor. Appl.
Page 105 and 106: 100 MATERIALS AND METHODS Plant mat
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102 The amount of Na + (%) 6 5 4 3
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104 NGE, 7-16 folds). Finally, all
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106 Anal. Biochem. 162: 156-159. Cr
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108 grown for academic purpose at N
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110 Cluster A was further separated
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112 Table 2 Similarity index of 19
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114 subjected to UPGMA and resultin
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116 Tabel 3 The similarity index of
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118 formation as seen from phylogen
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120 Genet. 101: 860-864. Lerceteau,
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122 INTRODUCTION Methylotrophic yea
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124 RESULTS AND DISCUSSION Screenin
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126 optimum temperature, insignific
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128 medium of C. boidinii (Volfova
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130 was observed soon after 24h cul
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132 Kasetsart J. (Nat. Sci.) 40(1)
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134 ACKNOWLEDGEMENTS This work was
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Kasetsart J. (Nat. Sci.) 40 : 136 -
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138 Determination of enzyme activit
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140 Table 1 Optimum and stability o
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142 for example recombinant E. coli
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144 were studied for β-1,3-1,4-glu
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146 ACKNOWLEDGEMENT This work was s
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150 vulcanization times were calcul
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152 RESULTS AND DISCUSSION Mechanic
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156 blend with 0.5 phr Irganox. How
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160 (AA-680 ShiMADZU, Atomic Absorp
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162 warm, relatively shallow and hi
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164 cycle in crustacean (Chen et al
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166 Salaenoi, J. 2004. Changes of e
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168 MATERIALS AND METHODS This rese
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170 DISCUSSION The average total am
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174 repacked and scanned three time
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176 found the most in the shrimp fe
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178 PLS model The comparison betwee
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180 brix value and dry matter of ma
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182 approach to increase immunity t
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184 higher (P
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186 levels were significantly eleva
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190 currents. Zone 4: River outlet
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192 standing waters, thus the occur
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194 and moving to 59%TL in juvenile
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198 thin membrane (Figure 1A, B). T
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200 were clearly seen. The presence
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202 could change to female at 6-12
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206 water until the water became cl
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208 reported by Thu and Preston (19
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210 cavalcade hay. The crude protei
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212 CONCLUSIONS Ruminal disappeared
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214 ∅rskov, E.R. and I. McDonald.
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216 is constrained mainly due to fa
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218 SNF and fat composition in raw
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220 in terms of both manual samplin
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222 Harding, F. 1995. Compositional
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224 proteins described by Barbut an
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226 starch/flour, shaken vigorously
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228 % Stress relaxation 54 49 44 39
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234 categories including puffed sna
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236 Cohesiveness of mass Crispness
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238 Cohesiveness of mass Sweetness
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242 3. Physicochemical properties 3
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246 ACKNOWLEDGEMENTS This research
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248 are now becoming more and more
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250 downstream control. Lam Chae re
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254 Inflow (mcm.) Inflow (mcm.) Inf
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256 were above 0.70. The testing re
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258 1999-2000 could be used for tra
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262 we assume that r ≥ 2 and d r
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266 Figure 1 Prototype domains. num
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268 DISCUSSION Consequences indicat
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272 George, B. K. 1997. The GIS Boo
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discussed; and appropriateness of t
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LITERATURE CITED FORMAT Manuscripts
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PRE-SUBMISSION CHECKLIST (see “In
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Send application materials to Payme
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