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uffer. DNA pattern and concentration was detected on a 1% agarose gel and by UV spectrophotometer. RAPD The protocol for RAPD analysis was adapted from that of Williams et al. (1990). The volume of the final reaction (25 µl) consisted of 1 X buffer (10 mM Tris-HCl, pH 8.0 and 50 mM KCl), 3.0 mM MgCl2, 1.25 U Taq DNA polymerase, 200 µM dNTP, 10 mM random primer (Operon), 25-100 ng of genomic DNA. Amplifications were made in a Perkin Elmer 9600 thermocycler with an initial denaturing step of 1 min at 94 °C, followed by 45 cycles of 1 min at 94°C, 1 min at 36°C, 2 min at 72°C and a final extension of 5 min at 72°C. PCR products were subjected to 1% agarose gel electrophoresis run at 90 V and DNA bands were visualized by ethidium bromide staining. RFLP One gram of genomic DNA was cut with the restriction enzymes DraI, BamHI, EcoRI, HindIII or stuI and then subjected to 1% agarose gel electrophoresis. The gels were blotted onto a positively charged nylon membrane (Boehringer Mannheim) by vacuum blotter. The probes were amplified by PCR using 18S rRNA, 5S rRNA and 5S rRNA repeat unit primer and PCR products were labeled with digoxigenin according to the protocol of Dig High prime DNA Labeling and Detection Starter Kit II (Roche). Hybridization was detected by enhanced chemiluminescence on Kodax X-ray film with 0.5-2 h exposure time. Data analysis DNA fragments were scored as presence (1) or absence (0) for each primer or restriction enzyme used. These scores were used to calculate their genetic similarity according to Nei and Li (1979), using NTSYS-pc1.80 from which the phenograms were constructed using a UPGMA. Kasetsart J. (Nat. Sci.) 40(1) 109 RESULTS AND DISCUSSION Primer selection and levels of polymorphism Twenty primers were screened for the genomic DNA amplification of nineteen Cycas species. Only 5 primers (Table 1) were found to give polymorphic DNA patterns. The total numbers of 87 bands with the fragments ranging from 0.35- 2.5 kb are shown in Figures 1 and 2. Amplification with primers OPB-1, OPB-8 and OPB-17 revealed unique bands of 0.8 kb, 0.6 kb and 0.7 kb, respectively. These bands could be used as genetic markers to identify the relationships among Cycas since they were specific to certain groups of Cycas (Nicolosi et al., 2000). RAPD analysis RAPD data were subjected to UPGMA and NTSYS-pc (Version 1.8). The similarity index showed that the relationships among all nineteen species fell in the range of 0.816-0.516 (Table 2). Maximum similarity was between C. edentata and C. litoralis (83.9%), while the least similarity were found between C. wadei and C. pranburiensis, C. tansachana and C. bougainvilleana, C. wadei and C. bougainvilleana (50.6 %). The distribution of species within the clusters showed apparent relation with geographical origin. The dendrogram (Figure 3) classified the 19 species into two major clusters, A and B. Cluster A contained all Cycas of Thailand geographical origin while cluster B contained those from other countries. Table 1 Nucleotide sequences of random oligonucleotide primers which showed polymorphism. Primer no. Sequence OPB-1 5′ GTTTCGCTCC 3′ OPB-8 5′ GTCCACACGG 3′ OPB-14 5′ TCCGCTCTGG 3′ OPB-15 5′ GGAGGCTGTT 3′ OPB-17 5′ AGGAACGAAG 3′
110 Cluster A was further separated into two groups, A I and A II. A I was subdivided into two groups, I and II. Group I consisted of C. chamaoensis, C. macrocarpa, C. pectinata, C. clivicola and C. siamensis. It is interesting to find that all Cycas of group I are of Thailand origin especially C. chamaoensis, C. macrocarpa, C. clivicola and C. siamensis were found only in Thailand but C. pectinata found also in China, Thailand and Vietnam was somewhat Kasetsart J. (Nat. Sci.) 40(1) separated as indicated by isozyme (Yang and Meerow, 1996). Group II contained C. micholitzii, C. simplicipinna, C. edentata, C. litoralis and C. pranburiensis. The similarity index suggested that C. micholitzii wass closely related to C. simplicipinna which also belonged to the same Stangerioides section based on their morphology (Pu and Chiu, 1990; Stevenson, 1992) (Table 5). The similarity index showed the close relationship of C. edentata and C. litoralis and both were also Figure 1 RAPD amplification of 19 Cycas species using the primers (A) OPB-1 showing a common band of 0.8 kb, (B) OPB-8 showing a common band of 0.6 kb, (C) OPB-14 having no common band , M: 100 bp DNA size marker, lane 1 C. chamaoensis, lane 2 C. macrocarpa, lane 3 C. clivicola, lane 4 C. micholitzii, lane 5 C. simplicipinna, lane 6 C. pectinata, lane 7 C. pranburiensis, lane 8 C. edentata, lane 9 C. litoralis, lane 10 C. tansachana, lane 11 C. siamensis, lane 12 C. nongnoochiae, lane 13 C. wadei, lane 14 C. seemannii, lane 15 C. bougainvilleana, lane 16 C. parvulus, lane 17 C. chevalieri, lane 18 C. nathorstii, lane 19 C. diannanensis. (B) and (C) indicated the similar DNA pattern found between C. edentata (lane 8) and C. litoralis (lane 9) as well as those of C. parvulus (lane 16) and C. diannanensis. (lane 19).
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January - March 2006 Volume 40 Numb
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KASETSART JOURNAL NATURAL SCIENCE T
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In sacco Degradation Characteristic
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2 and management practices a sorghu
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4 the greater plant height was obta
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6 at Wolenchity (Figure 3) that led
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8 Stover and aboveground biomass Co
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10 Radford, B.J., A.J. Dry, L.N. Ro
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12 At present, new cultivars with h
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14 2000), grapevine (Lavee, 2000),
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16 repeatability of FW, FLT, FLW, S
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18 (Table 5) indicating that select
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Kasetsart J. (Nat. Sci.) 40 : 20 -
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22 measured to define fruit shape i
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24 for higher fruit number and yiel
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28 (Table 2). The numbers of flower
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30 to the color of fruit and seed c
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32 1999. Carrots and related vegeta
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34 MATERIALS AND METHODS Laboratory
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36 DISCUSSION Extensive studies hav
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38 from Gossypium hirsutum. J. Inse
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40 The natural resistance of plants
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42 4. Data collection and statistic
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44 was not significantly different
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46 showed the highest yield compare
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48 Kessmann, H., T. Staub, C. Hofma
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50 There are approximately 60 comme
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52 in PBST containing 2% ovalbumin
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54 Kasetsart J. (Nat. Sci.) 40(1) T
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56 Virus-free orchid plants show fa
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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
Page 107 and 108: 102 The amount of Na + (%) 6 5 4 3
Page 109 and 110: 104 NGE, 7-16 folds). Finally, all
Page 111 and 112: 106 Anal. Biochem. 162: 156-159. Cr
Page 113: 108 grown for academic purpose at N
Page 117 and 118: 112 Table 2 Similarity index of 19
Page 119 and 120: 114 subjected to UPGMA and resultin
Page 121 and 122: 116 Tabel 3 The similarity index of
Page 123 and 124: 118 formation as seen from phylogen
Page 125 and 126: 120 Genet. 101: 860-864. Lerceteau,
Page 127 and 128: 122 INTRODUCTION Methylotrophic yea
Page 129 and 130: 124 RESULTS AND DISCUSSION Screenin
Page 131 and 132: 126 optimum temperature, insignific
Page 133 and 134: 128 medium of C. boidinii (Volfova
Page 135 and 136: 130 was observed soon after 24h cul
Page 137 and 138: 132 Kasetsart J. (Nat. Sci.) 40(1)
Page 139 and 140: 134 ACKNOWLEDGEMENTS This work was
Page 143 and 144: 138 Determination of enzyme activit
Page 145 and 146: 140 Table 1 Optimum and stability o
Page 147 and 148: 142 for example recombinant E. coli
Page 149 and 150: 144 were studied for β-1,3-1,4-glu
Page 151 and 152: 146 ACKNOWLEDGEMENT This work was s
Page 155 and 156: 150 vulcanization times were calcul
Page 157 and 158: 152 RESULTS AND DISCUSSION Mechanic
Page 159 and 160: 154 Kasetsart J. (Nat. Sci.) 40(1)
Page 161 and 162: 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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230 Kasetsart J. (Nat. Sci.) 40(1)
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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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