PRODUCTION Of NUTRIENT SOURCES FOR RHIZOBIUM

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APPENDICES A. Reagents 1. Lactophenol-cotton blue Lactic acid 20.0 ml Phenol crystal 20.0 g Glycerol 40.0 ml Distilled Water 20.0 ml Keep in brown bottle and add 0.05 g of cotton blue or methylene blue. 2. Malachite green (spore straining) Malachite green 5.0 g Distilled water 100.0 ml 3. Carbol fuchsin (Ziehl’s) Basic fuchsin 0.3 g Ethanol 95% 10.0 ml Added with Phenol 5% aqueous solution 100.0 ml. 4. Silver nitrate-Sodium hydroxide reagent Preparation of reagent Dipping solution I Make 1 ml of a saturated aqueous silver nitrate solution up to 200 ml with acetone. Redissolve the resulting precipitate by adding 5 ml of water and shaking. Dipping solution II Dissolve 2 g of sodium hydroxide pellets in 2 ml of water with heating, and make up to 100 ml with methanol. Storage Dipping solution I should be prepared freshly for using each time, dipping solution II may be stored for several days. Substances Silver nitrate Sodium hydroxide pellets
Acetone Methanol Solvent for mobile phase 1-Butanol-Acetone-Water (5:4:1) Chromatographic plates Commercial plates of 20x20 cm Silica gel 60 F 254 (MERCK Art. 5554.) were used. The plates were impregnated by soaking in buffered methanol solution and allowing them to dry at 110°C for 30 min. Buffered methanol solution Chloroform-Methanol (50:50) Reaction The ionic silver in the reagent is reduced to metallic silver by reducing carbohydrates. Method The chromatograms are freed from mobile phase in a stream of warm air, immersed in solution I for 1 second or sprayed evenly with it, then dried in a stream of cold air, then immersed in solution II for 1 second or sprayed with it, and finally heated to 100°C for 1-2 min. water. 5. Iodine solution Potassium iodide 6.6 g Iodine 0.66 g Distilled water 165.0 ml Dissolved 6.6 g of potassium iodide and 0.66 g of iodine in 165 ml of distilled 6. Dinitrosalicylic acid reagent Distilled water 1414.0 ml 3,5-Dinitrosalicylic acid (DNS) 10.0 g Sodium hydroxide (NaOH) 19.0 g Potassium sodium tartate 306.0 g Phenol (dissolved at 50°C) 7.6 g Solution I: Dissolved 10 g of DNS and 19 g of NaOH in 1,416 ml of distilled water with warming and vigorously stirring. Solution II: Dissolved 306 g of sodium-potassium tartate in 500 ml of distilled water
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PRODUCTION OF NUTRIENT SOURCES FOR
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PRODUCTION OF NUTRIENT SOURCES FOR
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CHARDCHAI BUROM : PRODUCTION OF NUT
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CONTENTS Page ABSTRACT (THAI)……
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3.6.1� Determination of the suita
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�� 17� Glycerol concent
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40� Time courses of mannitol prod
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59� Effects of various heat shock
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LIST OF TABLES Table Page ��
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LIST OF ABBREVIATIONS Aw water avai
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CHAPTER I INTRODUCTION For mass cul
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available (Burton, 1965). The molec
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1.1.1� Carbon metabolism in Rhizo
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strains unable to use the carbon so
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Figure 2. The fructose bisphosphate
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1.2�Classification of yeasts Mank
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Table 3. (continued). Class Order F
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Ustilaginales (Oberwinkler, 1987).
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f Cystofilobasidium has thick-walle
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Table 5. (continued). Sugar alcohol
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specific cases, but Brown (1978) pr
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KY 6166 was also studied using larg
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Figure 5. Important pathways of gly
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may be present in different ratios.
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amylase systems of the yeast specie
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Table 7. (continued). EC Recommende
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Table 8. (continued). Species a D.p
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Table 9. (continued). Cell product
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2.1.3 Equipment and other materials
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Spotted either the supernatant of c
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amylase action, the actual reaction
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A) Determination of the suitable an
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B) Salt-stress conditions Fifty mil
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2.2.8 Cultivation of Bradyrhizobium
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The cultured medium of yeast. Quant
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intervals for accumulation of gas i
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CHAPTER III RESULTS AND DISCUSSION
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Glycerol Xylitol Glucose Mannitol S
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Glycerol Xylitol Glucose Mannitol S
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Table 10. Primary screening of glyc
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Table 10. (continued). No. Isolate
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Glycerol in cell lysate (g/l) A Gly
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average were 4-10 g of glycerol per
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were isolated from fresh fruits, Ma
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�third day of incubation (Figure
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Reducing sugars (g/l) Reducing suga
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These results indicated that the co
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3.6.1 Determination of the suitable
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g/l 1.8 1.6 1.4 1.2 1.0 0.8 0.6 0.4
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Reducing sugars (g/l) 8 6 4 2 0 0 1
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Reducing sugars (g/l) 4.0 3.5 3.0 2
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g/l 1.4 1.2 1.0 0.8 0.6 0.4 0.2 0.0
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g/l g/l g/l 1.5 1.0 0.5 0.0 1.2 1.0
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Starch fermentation was conducted i
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g/l g/l 7 6 5 4 3 2 1 0 8 6 4 2 0 0
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g/l g/l 8 6 4 2 0 8 6 4 2 0 1 2 3 4
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g/l g/l 8 6 4 2 0 8 6 4 2 0 0 1 1 2
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g/l g/l 1.6 1.4 1.2 1.0 0.8 0.6 0.4
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g/l g/l 1.6 1.4 1.2 1.0 0.8 0.6 0.4
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The results showed that amounts var
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g/l g/l 4.0 3.5 3.0 2.5 2.0 1.5 1.0
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�� fermentation, but the inc
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g/l g/l 2.0 1.6 1.2 0.8 0.4 0.0 3.2
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g/l g/l 1.2 1.0 0.8 0.6 0.4 0.2 0.0
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�� intracellularly a decreas
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g/l g/l 5.0 4.0 3.0 2.0 1.0 0.0 2.5
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g/l g/l 2 1.8 1.6 1.4 1.2 1 0.8 0.6
Page 135 and 136: g/L 7 6 5 4 3 2 1 0 1 2 3 4 5 6 7 0
Page 137 and 138: g/l 8 7 6 5 4 3 2 1 0 1 2 3 4 5 6 7
Page 139 and 140: �� effected by heat shock po
Page 141 and 142: Table 15. The comparison of media f
Page 143 and 144: Table 17. Growth of Bradyrhizobium
Page 145 and 146: �� Rhizobium (Stowers, 1985;
Page 147 and 148: �� 3.9 Identification of yea
Page 149 and 150: KAY1 PIY2 PUY4 Y69 LIY2 COY1 FAY2 C
Page 151 and 152: �� assimilations were observ
Page 153 and 154: Table 20. (continued). Yeast strain
Page 155 and 156: Table 20. (continued). 15. Pichia k
Page 157 and 158: �� 3.9.5 Ascospore formation
Page 159 and 160: (a) (b) (c ) (d) (e) �� Figu
Page 161 and 162: (a) (b) (c) (d) (e) (f) �� F
Page 163 and 164: (c) (a) �� Figure 72. Cell m
Page 165 and 166: �� Metshnikowia (Pitt and Mi
Page 167 and 168: CHAPTER IV CONCLUSION This study ex
Page 169 and 170: only a low capital investment. Ther
Page 171 and 172: �� Barnett, J. A., Payne, R.
Page 173 and 174: �� De Mot, R., and Verachter
Page 175 and 176: �� Jennings, D. H. (1995). T
Page 177 and 178: �� Lie, T. A., Muilenbury M.
Page 179 and 180: �� Onishi, H., and Suzuki, T
Page 181 and 182: �� Sinclair, N. A., and Stok
Page 183 and 184: �� Van Rossum, D., Schuurman
Page 185: �� Young, J. P. W., and Hauk
Page 189 and 190: � 300 µg/ml, 200 µg/ml, and 100
Page 191 and 192: Soluble Starch 20.0 g Distilled wat
Page 193 and 194: 13.�Nitrate broth KNO 3 1.0 g Glu
Page 195 and 196: If the sample had been diluted befo
Page 197 and 198: Table 1D. (continued). Source of ye
Page 212 and 213: E. Additional figures Abs 540 nm (a
Page 214 and 215: Abs 580 nm 1.4 1.2 1 0.8 0.6 0.4 0.
Page 216 and 217: Figure 7E. Calibration curves for H
Page 218 and 219: Figure 9E. Example of HPLC chromato
Page 220: Cell/mL 1.0E+09 8.0E+08 6.0E+08 4.0
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PRODUCTION Of NUTRIENT SOURCES FOR RHIZOBIUM

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