APPENDIX 1 MEDIA PREPARATION

Transcription

1 155 APPENDIX 1 MEDIA PREPARATION 1 Algal culture medium NaNO g K 2 HPO g KgSO g NH 4 Cl CaCl 2 FeCl 3 Distilled water 0.05 g 0.01 g g 1000ml 2 Algal proteose medium Proteose peptone Bristol s solution 1g 1 Lit. Bristol s solution (for 1000 ml) NaNO 3- K 2 HPO 4 KH 2 PO 4 10g 3g 7g

2 156 CaCl 2 MgSO 4. 7H 2 O FeCl 3 solution 1g 3g 0.1ml FeCl 3 solution (for 100 ml) FeCl 3 1g 3 BG-11 Medium Component Stock Solution Quantity Used (g L-1 dh2o) Fe Citrate solution 1 ml Citric acid 6 1 ml Ferric ammonium citrate 6 1 ml NaNO g K 2 HPO 4 3H 2 O 40 1 ml MgSO 4 7H 2 O 75 1 ml CaCl 2 2H 2 O 36 1 ml Na 2 CO ml MgNa 2 EDTA H2O ml Trace metals solution 1 ml H 3 BO g MnCl 2 4H 2 O g ZnSO 4 7H 2 O g CuSO 4 5H 2 O ml Na 2 MoO 4 2H 2 O g Co(NO3) 2 6H 2 O ml

3 Bristol s medium Macro nutrients NaNO 3- K 2 HPO 4 KH 2 PO 4 CaCl 2 NaCl MgSO. 4 7H 2 O 250 mg 75 mg 175 mg 25 mg 25 mg 75 mg Micro nutrients FeCl 3 MnSO. 4 4H 2 O ZnSO. 4 7H 2 O H 3 BO 3 CuSO. 4 5H 2 O 0.3 mg 0.3 mg 0.2 mg 0.2mg 0.06 mg 5 George s medium Peptone KNO g 1.00 g KHPO g K 2 HPO g MgSO 4 7H 2 O 0.02g FeCl 3 EDTA Mg or Ca 0.04 g 0.20 g

4 158 Oligo elements H 3 BO 3 MnCl 2. 2H 2 O ZnSO 4. 7 H 2 O Na 2 MoO 4. 2 H 2 O CuSO 4. 5 H 2 O Co (NO 3 ) 2. 6 H 2 O Distilled water 2.9 mg 1.8 mg 0.2 mg 0.4 mg 0.08 mg 0.05 mg 1000 ml 6 GO algal medium Soil water Peptone KNO ml g 2.05 g KHPO g K 2 HPO g MgSO 4 7H 2 O 0.02g FeCl g Soil water preparation 20 g of loamy soil was air dried and dissolved in 1000ml of distilled water along with a pinch of CaCO 3 was added to soil water prior to sterilization. The above contents were mix well and kept in a steaming condition for 3 hours for two successive days. Collect the extract by filtering through

5 159 Whatman No.1 filter paper and sterilize the extract under 15 psi at 121 C minutes in an autoclave for further use. 7 Potato glucose agar Potato 300.0g Glucose 20.0g Agar 15.0 g Potatoes were sliced and kept in 500ml of boiling water for 30 minutes. These boiled potatoes were then strained through cheese cloth. The volume was adjusted to one litre with distilled water and to this; 20 g glucose and 15 g agar were added. 8 Trebouxia medium Soil water Protease peptone Glucose Bristol s solution 140 ml 10g 20g 860ml Bristol s solution To 940 ml of distilled water the following contents were added. ml Stock Solution g/400ml 10 NaNO CaCl 2 2 H 2 O MgSO 4 7 H 2 O 3.0

6 K 2 HPO KH 2 PO NaCl Citric Acid PIV Metal Solution 1.3 PIV Metal solution To 1000ml, of disodium salt of was added and to which the following contents were added. EDTA 0.750g FeCl 3. 6H 2 O MnCl 2. 4 H 2 O ZnCl 2 CaCl 2. 6 H 2 O Na 2 Mo O 4. 2 H 2 O 97 mg 41 mg 5mg 2 mg 4 mg Soil water preparation 20 g of loamy soil was air dried and dissolved in 1000ml of distilled water along with a pinch of CaCO 3 was added to soil water prior to sterilization. The above contents were mix well and kept in a steaming condition for 3 hours for two successive days. Collect the extract by filtering through Whatman No.1 filter paper and sterilize the extract under 15 psi at 121 C minutes in an autoclave for further use.

7 161 9 Bold s Basal Medium (BBM) Component Stock Solution Quantity Used (g L-1 dh2o) Macronutrients NaNO ml CaCl 2 2H 2 O ml MgSO 4 7H 2 O ml K 2 HPO ml KH 2 PO ml NaCl ml Alkaline EDTA Solution 1 ml EDTA KOH Acidified Iron Solution 1 ml FeSO 4 7H 2 O 4.98 H 2 SO 4 Boron Solution 1 ml 1 ml H 3 BO

8 162 Trace Metals Solution 1 ml ZnSO 7HO 8.82 MnCl 2 4H 2 O 1.44 MoO CuSO 4 5H 2 O 1.57 Co(NO3) 2 6H 2 O Dy-III Medium Component Stock Solution Quantity Used (g L-1 dh2o) MES buffer 200 mg CaCl 2 2H 2 O ml Na 2 SiO 3 9H 2 O ml MgSO 4 7H 2 O ml NH 4 NO ml NaNO ml KCl ml Na 2 β-glycerophosphate ml H 3 BO ml Trace metals solution 1 ml Vitamins solution 1 ml

9 163 Trace Metals Solution Component 1 Stock Solution Quantity used (g L-1 dh2o) Na 2 EDTA (anhydrous) g Fe (iron wire) g MnCl 2 4H 2 O g ZnSO 4 7H 2 O g Na 2 MoO 4 6H 2 O 50 1 ml CoCl 2 6H 2 O 29 1 ml Vitamins Solution Component 1 Stock Solution Quantity used (g L-1 dh2o) Thiamine HCl 200 mg Biotin ml Cyanocobalamin ml 11 Aaronson Medium Composition NH 4 Cl KH 2 PO 4 MgSO 4.7H 2 O 0.5 g 0.3 g 1.0 g

10 164 MgCO 3 C 6 H 9 NO 6 (Nitrilotriacetic acid) CaCO 3 C 5 H 9 NO 4 (Glutamic acid) Glucose Arginine Hystidine monochloride Biotin (Vitamin H) Thiamine HCl (Vitamin B1) Microelement stock solution 0.4 g 0.2 g 0.05 g 3.0 g 10.0 g 0.4 g 0.4 g g g 10 ml Microelement Stock Solution Fe(NH4) 2 (SO4) 2.6H 2 O H 3 BO 3 MnSO 4.H 2 O CoSO 4.7H 2 O CuSO4.5H 2 O ZnSO 4.H 2 O (NH4) 6 Mo 7 O 24.4H 2 O Na 3 VO 4.16H 2 O 0.5 g g g g g 0.25 g g g

11 Cramer and Myers Medium (NH4) 2 HPO 4 KH 2 PO 4 MgSO 4.7H 2 O CaCl 2 Na 3 C 6 H 5 O 7.2H 2 O Fe 2 (SO4) 3. nh 2 O MnCl 2.4H 2 O CoSO 4.7H 2 O ZnSO 4.7H 2 O Na 2 MoO 4.2H 2 O CuSO 4.5H 2 O H 3 BO 3 Thiamine HCl Cyanocobalamin Glucose 1 g 1 g 0.2 g 0.02 g 0.8 g 3mg 1.8 mg 1.5 mg 0.4 mg 0.2 mg 0.02 mg 2.48 mg 0.5 mg 0.02 mg 10 g 13. Beijerinck Medium Stock I Stock II 100 ml 40 ml

12 166 Stock Micronutrients III 60 ml 1 ml Stock I NH 4 NO 3 K 2 HPO 4 MgSO 4 7H 2 O CaCl 2.2H 2 O 1.5 g 0.2 g 0.2 g 0.1 g Stock II KH 2 PO g Stock III K 2 HPO g Micronutrients H 3 BO 3 MnCl 2.4H 2 O EDTA CuSO 4.5H 2 O ZnSO 4.H 2 O CoCl 2.6H 2 O FeSO 4.7H 2 O (NH4) 6 Mo 7 O 24.4H 2 O 10 g 5g 50 g 1.5 g 22 g 1.5 g 5g 1g