PRODUCTION Of NUTRIENT SOURCES FOR RHIZOBIUM

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Distilled water 1000 ml The compositions were suspended in 1 liter of distilled or deionized water and heated to boil until complete dissolving. Final pH 6.8 at room temperature and sterilized by autoclaving for 15 min at 15 lbs pressure (121°C). C. Instruments 1. A typical counting chamber (haemocytometer) (Singleton, 1992) (Figure 1C) The instrument, seen from one side at (a) consists of rectangular glass block in which the central plateau lies precisely 0.1 mm below the level of the shoulders on either side. The central plateau is separated from each shoulder by a trough, and is itself divided into two parts by a shallow trough (seen at (b)). On the surface of each part of the central plateau is an etched grid (c) consisting of a square which is divided into 400 small squares, each 1/400 mm 2 . A glass cover-slip is positioned as shown at (b) and is pressed firmly onto the shoulders of the chamber; to achieve proper contact it is necessary, while pressing, to move the cover-slip (slightly) against the surface of the shoulders. Proper (close) contact is indicated by the appearance of a pattern of color lines (Newton′s rings), shown in black and white at (b). 1.1 Using the chamber. A small volume of a bacterial suspension is picked up in a Pasteur pipette by capillary attraction; the thread of liquid in the pipette should not be more than 10 mm. The is then placed as shown in (b), i. e. with the opening of the pipette in contact with the central plateau, and the side of the pipette against the cover-slip. With the pipette in this position, liquid is automatically drawn by capillary attraction into the space bounded by the cover-slip and part of the central plateau; the liquid should not overflow into the trough. (It is sometimes necessary to tap the end of the pipette, slightly, against the central plateau to encourage the liquid to enter the chamber). A second sample can be examined, if required, in the other half of the counting chamber. The chamber is left for 30 minutes to allow the cells to settle, and counting is the carried out under a high power of the microscope-which is focused on the grid of the chamber. Since the volume between grid and cover-slip is accurately known, the count of cells per unit volume can be calculated. 1.2 A work example. Each small square in the grid is 1/400 mm 2 . As the distance between grid and cover-slip is 1/10 mm, the volume of liquid over each small square is 1/4000 3 mm-i. e. �,000,000 ml. Suppose, for example, that on scanning all 400 small squares, 500 cells were counted; this would give an average of 500/400 (=1.25) cells per small square, i. e. 1.25 cells per �,000,000 ml. The sample therefore contains 1.25*4,000,000 cells/ml, i. e. 5*10 6 cells/ml.
If the sample had been diluted before examination (because it was too concentrated), the count obtained must be multiplied by the dilution factor; for example, if diluted 1-in-10, the count should be multiplied by 10. N. B. The chamber described above is the Thomas chamber; in a Helber chamber the distance between central plateau and cover-slip is 0.02 mm. Figure 1C. A typical counting chamber (haemocytometer). Source: Singleton (1992).
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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
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g/L 7 6 5 4 3 2 1 0 1 2 3 4 5 6 7 0
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g/l 8 7 6 5 4 3 2 1 0 1 2 3 4 5 6 7
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�� effected by heat shock po
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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
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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 and 186: �� Young, J. P. W., and Hauk
Page 187 and 188: Acetone Methanol Solvent for mobile
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: 13.�Nitrate broth KNO 3 1.0 g Glu
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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