PRODUCTION Of NUTRIENT SOURCES FOR RHIZOBIUM

More documents

Recommendations

Info

diluted to provide 0.5% (v/v) glycerol as same as the standard rhizobia culture medium. As depicted at Table 16. G5 medium used as the standard glycerol medium in this experiment. The medium containg 5% sucrose fermented by yeast-isolate PIY2 and PUY4 were adjusted. First steps, diluted to 0.5% (v/v) as same as G5. The amounts of glycerol in adjusted cultured medium was tested by using High Performance Liquid Chromatography. Second steps, added the other substances as same as G5 were 0.1% mannitol, 0.02% MgSO4, 7H2O, 0.05% K2HPO4, 0.01% NaCl, 0.1% Glucose, 0.05% Arabinose, 200 µM Fe-EDTA, and pH 6.8 at room temperature. Except glycerol and yeast extract which much more for support the bacterial cell in the cultured medium. After heat sterilization, the adjusted medium was inoculated with two strains: Bradyrhizobium japonicum USDA 110 and Bradyrhizobium spp. THA 5. Table 16. The comparison of media containing glycerol for Bradyrhizobium cultivation. 5%Sucrose G5 PIY2 medium PUY4 medium Mannitol - 1.0 g 1.0 g 1.0 g Glycerol - 4.0 ml 6.0 g 6.0 g Yeast extract 10.0 g 1.0 g - - MgSO4.7H2O - 0.2 g 0.2 g a 0.2 g a K2HPO4 - 0.5 g 0.5 g a 0.5 g a KH2PO4 - - - - NaCl - 0.1 g 0.1 g a 0.1 g a Glucose - 1.0 g 1.0 g a 1.0 g a Arabinose - 0.5 g 0.5 g a 0.5 g a Tryptone 20.0 g - - - Sucrose 50.0 g - - - PVP-40 - 20.0 g - b - b Fe- EDTA - 200 µM (0.073 g/l) 200 µM (0.073 g/l) 200 µM (0.073 g/l) Distilled water 1000 ml 1000 ml 1000 ml 1000 ml pH 5.5 6.8 6.8 6.8 Note that a same as standard medium b no add ��
Table 17. Growth of Bradyrhizobium on media containing glycerol. Bradyrhizobium japonicum USDA 110 Bradyrhizobium spp.THA5 Media 0 day 2 day 4 day 6 day 0 day 2 day 4 day 6 day Control 1.0x10 7 1.1x10 7 9.0x10 6 6.4x10 6 1.0x10 7 1.1x10 7 8.7x10 6 8.0x10 6 G5 (100 ml) 1.0x10 7 7.5x10 8 1.9x10 9 3.4x10 9 1.0x10 7 5.3x10 8 2.2x10 9 5.7x10 9 G5 (500 ml)) 1.0x10 7 8.5x10 8 1.4x10 9 4.0x10 9 1.0x10 7 5.9x10 8 1.4x10 9 6.2x10 9 YEPD containing 5% sucrose prefermented by isolate PIY2 YEPD containing 5% sucrose prefermented by isolate PUY4 �� 1.0x10 7 7.0x10 5 1.0x10 6 6.1x10 6 1.0x10 7 2.5x10 8 2.1x10 9 1.0x10 10 1.0x10 7 4.4x10 7 2.2x10 8 3.7x10 8 1.0x10 7 6.5x10 8 1.7x10 9 5.0x10 9 From the Table 17, B. japonicum USDA 110 could not grow well in PIY2 and PUY4 cultured medium when compared with G5 medium. In PIY2 cultured medium, rhizobial cells decreased in the second days and slowly increased during the day of cultivation which gave the amounts of cell at 10 6 cell/ml at the sixth day of cultivation. PUY4 cultured medium gave the higher cell at 10 8 cells/ml at the sixth day of cultivation. In contrast, the amounts of cells from G5 medium were 10 9 cells/ml at the forth day and the sixth day of cultivation. The quantitation of G5 medium in the flask was not effect to the bacterial cell growth when compared. For the cultivation of Bradyrhizobium spp. THA5. Interestingly, clearly showed that the PIY2 and PUY4 adjusted cultured medium could support the growth of Bradyrhizobium spp. THA5. The best results was obtained with the final number of Bradyrhizobium cells being almost the same as that found with G5 medium. Especially, PIY2 cultured medium gave high cell density at 10 10 cells/ml at the sixth days of cultivation. PUY4 cultured medium gave the amounts of cell number similar to G5 medium. The results presented here clearly demonstrated that sucrose, preferment by yeast, can be used for the cultivation of Bradyrhizobium, in particulary, Bradyrhizobium spp. THA5. However, PIY2 cultured medium could not enhance the growth of Bradyrhizobium japunium USDA 11O, it
Page 1 and 2:
PRODUCTION OF NUTRIENT SOURCES FOR
Page 3 and 4:
PRODUCTION OF NUTRIENT SOURCES FOR
Page 5 and 6:
CHARDCHAI BUROM : PRODUCTION OF NUT
Page 7 and 8:
CONTENTS Page ABSTRACT (THAI)……
Page 9 and 10:
3.6.1� Determination of the suita
Page 11 and 12:
�� 17� Glycerol concent
Page 13 and 14:
40� Time courses of mannitol prod
Page 15 and 16:
59� Effects of various heat shock
Page 17 and 18:
LIST OF TABLES Table Page ��
Page 19 and 20:
LIST OF ABBREVIATIONS Aw water avai
Page 21 and 22:
CHAPTER I INTRODUCTION For mass cul
Page 23 and 24:
available (Burton, 1965). The molec
Page 25 and 26:
1.1.1� Carbon metabolism in Rhizo
Page 27 and 28:
strains unable to use the carbon so
Page 29 and 30:
Figure 2. The fructose bisphosphate
Page 31 and 32:
1.2�Classification of yeasts Mank
Page 33 and 34:
Table 3. (continued). Class Order F
Page 35 and 36:
Ustilaginales (Oberwinkler, 1987).
Page 37 and 38:
f Cystofilobasidium has thick-walle
Page 39 and 40:
Table 5. (continued). Sugar alcohol
Page 41 and 42:
specific cases, but Brown (1978) pr
Page 43 and 44:
KY 6166 was also studied using larg
Page 45 and 46:
Figure 5. Important pathways of gly
Page 47 and 48:
may be present in different ratios.
Page 49 and 50:
amylase systems of the yeast specie
Page 51 and 52:
Table 7. (continued). EC Recommende
Page 53 and 54:
Table 8. (continued). Species a D.p
Page 55 and 56:
Table 9. (continued). Cell product
Page 57 and 58:
2.1.3 Equipment and other materials
Page 59 and 60:
Spotted either the supernatant of c
Page 61 and 62:
amylase action, the actual reaction
Page 63 and 64:
A) Determination of the suitable an
Page 65 and 66:
B) Salt-stress conditions Fifty mil
Page 67 and 68:
2.2.8 Cultivation of Bradyrhizobium
Page 69 and 70:
The cultured medium of yeast. Quant
Page 71 and 72:
intervals for accumulation of gas i
Page 73 and 74:
CHAPTER III RESULTS AND DISCUSSION
Page 75 and 76:
Glycerol Xylitol Glucose Mannitol S
Page 77 and 78:
Glycerol Xylitol Glucose Mannitol S
Page 79 and 80:
Table 10. Primary screening of glyc
Page 81 and 82:
Table 10. (continued). No. Isolate
Page 83 and 84:
Glycerol in cell lysate (g/l) A Gly
Page 85 and 86:
average were 4-10 g of glycerol per
Page 87 and 88:
were isolated from fresh fruits, Ma
Page 89 and 90:
�third day of incubation (Figure
Page 91 and 92: Reducing sugars (g/l) Reducing suga
Page 93 and 94: These results indicated that the co
Page 95 and 96: 3.6.1 Determination of the suitable
Page 97 and 98: g/l 1.8 1.6 1.4 1.2 1.0 0.8 0.6 0.4
Page 99 and 100: Reducing sugars (g/l) 8 6 4 2 0 0 1
Page 101 and 102: Reducing sugars (g/l) 4.0 3.5 3.0 2
Page 103 and 104: g/l 1.4 1.2 1.0 0.8 0.6 0.4 0.2 0.0
Page 105 and 106: g/l g/l g/l 1.5 1.0 0.5 0.0 1.2 1.0
Page 107 and 108: Starch fermentation was conducted i
Page 109 and 110: g/l g/l 7 6 5 4 3 2 1 0 8 6 4 2 0 0
Page 111 and 112: g/l g/l 8 6 4 2 0 8 6 4 2 0 1 2 3 4
Page 113 and 114: g/l g/l 8 6 4 2 0 8 6 4 2 0 0 1 1 2
Page 115 and 116: g/l g/l 1.6 1.4 1.2 1.0 0.8 0.6 0.4
Page 117 and 118: g/l g/l 1.6 1.4 1.2 1.0 0.8 0.6 0.4
Page 119 and 120: The results showed that amounts var
Page 121 and 122: g/l g/l 4.0 3.5 3.0 2.5 2.0 1.5 1.0
Page 123 and 124: �� fermentation, but the inc
Page 125 and 126: g/l g/l 2.0 1.6 1.2 0.8 0.4 0.0 3.2
Page 127 and 128: g/l g/l 1.2 1.0 0.8 0.6 0.4 0.2 0.0
Page 129 and 130: �� intracellularly a decreas
Page 131 and 132: g/l g/l 5.0 4.0 3.0 2.0 1.0 0.0 2.5
Page 133 and 134: 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: Table 15. The comparison of media f
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 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 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 199 and 200:
Page 201 and 202:
Page 203 and 204:
Page 205 and 206:
Page 207 and 208:
Page 209 and 210:
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
show all

PRODUCTION Of NUTRIENT SOURCES FOR RHIZOBIUM

You also want an ePaper? Increase the reach of your titles

Delete template?

Save as template?