The detail information about the media used for Bio-fertilizers, Bio-pesticide and Bio-agents production. ELE PATH-243 Bio-fertilizers, Bio-control and Bio-agents and Bio-pesticide.

3. 1) Media for Rhizobium Studies :
i) Yeast extract mannitol medium: g/l
K2HPO4 : 0.5 g
MgSO4.7H2O : 0.2 g
NaCl : 0.1 g
Mannitol : 10.0
Yeast extract : 1.0
Distilled water : 1000 ml
Agar : 20.0 g
Congo red 1% solution : 2.5 ml (only for solid medium during isolation)

5. 1) Media for Rhizobium Studies :
ii) Peptone - glucose agar : g/l
Glucose : 5.0
Peptone : 10.0
Agar : 15-10
Distilled water : 1000 ml
iii) Carbol Fuchsin stain :
Basic fuchsin : 1.0 g
Ethyl alcohol : 10.0 ml
5% phenol solution : 100.0 ml
Peptone – glucose
agar

6. 2) Nitrogen-free media for growing seedlings to test root
nodulation of legumes :
i) Jensen's medium(1942) : g/l
CaHPO4 : 1.0
K2HPO4 : 0.2
MgSO4.7H2O : 0.2
NaCl : 0.2
FeCl3 : 0.1
Water : 1000 ml
ii) Thomton's medium(1930) : g/l
Ca3(PO4)2 : 2.0
K2HPO4 : 0.5
MgSO4.7H2O : 0.2
NaCl : 0.1
FePO4 : 1.0
FeCl3 : 0.01 Water : 1000 ml

7. Note :
 Add agar 15 g/l when solid media are needed.
 Prepare a stock solution of trace elements
containing Bo 0.05%, Mn 0.05 %; Zn 0.005 %, Mo
0.005 %, Cu 0.002 % (Gibson, 1963).
 Add 1 ml of the stock solution for every 1000 ml of
the medium.
 Adjust pH to 6.5 - 7.0

8. 2) Media for Blue Green Algae :
Pringsheim's medium(Pringsheim, 1964)
KNO3 : 0.02 %
MgSO4.7H2O : 0.001 %
(NH4).HPO3 : 0.002 %
CaCl2.6H2O : 0.0005 %
FeCl3 : 0.00005 %
Chu's mediumNo. 10 (Chu. 1942)
Ca(NO3) : 0.004 %
MgSO4.7H2O : 0.0025 %
K2HPO4 : 0.0005 to 0.001 %2 %
Na2CO3 : 0.002 %
Na2SiO3 : 0.0025 %
FeCl3 : 0.0008 %

9. 2) Media for Blue Green Algae :
Cron's plant nutrient medium g/l
KCl or KNO3 : 0.75
CaSO4.2H2O : 0.50
MgSO4.7H2O : 0.50
Ca3(PO4).5H2O : 0.25
Fe3(PO4).5H2O : 0.25
Distilled water : 1 litre

10. 2) Media for Blue Green Algae :
Trace element solution g/l
LiSO4 : 0.055
CuSO4.2H2O : 0.055
ZnSO4 : 0.055
Al2SO4 : 0.055
NiSO4.7H2O : 0.055
H3BO3 : 0.62
SnCl2 : 0.03
MnCl2 : 0.40
CoCl2.6H2O : 0.055
TiO2 : 0.055
KI : 0.035
KBr : 0.035
Na2S2O3 : 0.43
Na2MO4.2H2O : 0.03
KMnO : 0.40

11. 3)Media for Azoto-bactor :
Ashby's medium g/l
Mannitol : 20.0
K2HPO4 : 0.2
MgSO4.7H2O : 0.2
NaCl : 0.2
K2SO4 : 0.1
CaCO3 : 5.0
Agar : 15.0
Distilled water : 1000.0 ml

12. 3)Media for Azoto-bactor :
Jensen's medium g/ml
Sucrose : 20.0
K2HPO4 : 1.0
MgSO4.7H2O : 1.0
NaCl : 0.5
FeSO4 : 0.1
CaCO3 : 2.0
Agar : 15.0
Distilled water : 1000 ml

13. 3)Media for Azoto-bactor :
Beijerinckia Medium (Becking, 1959)
Sucrose : 20.0
KH2PO4 : 0.8
K2HPO4 : 0.2
MgSO4.7H2O : 0.5
FeCl3 : 0.1
Na2MoO4 : 0.005
Agar : 15.0
Distilled water : 1000 ml
pH : 6.5

14. 4)Medium for Azo-spirillum :
Okon et al. (1977) mediumas modified by Lakshmi Kumari et al. (1980)
a) K2HPO4 : 6.0
KH2PO4 : 4.0
Distilled water : 500 ml
b) MgSO4 : 0.2
NaCl : 0.1
CaCl2 : 0.02
NH4Cl : 1.0
Malic acid : 5.0
NaOH : 5.0
Yeast extract : 3.0
Na2MoO4 : 0.002

15. 4)Medium for Azo-spirillum :
MnSO4 : 0.001
H3BO3 : 0.0014
Cu(NO3)2 : 0.0004
ZnSO4 : 0.0021
FeCl3 : 0.002
Distilled water : 500 ml
Bromothymol blue : 2 ml
(0.5% alcoholic soln.)
The phosphate buffer portion of the medium was made in half
of the total volume required and also contained enough agar
(1.5 to 2.0 %) for solidification.
Part (a) and (b) were sterilized separately, mixed while hot,
poured into plates and allowed to set.

16. 5) Medium for Gluconacetobacter diazotrophicus :
:Composition of semisolid LGIP medium
(Cavalcante and Dobereiner, 1988)
Dipotassium hydrogen phosphate : 0.200 g
Potassium dihydrogen phosphate : 0.600 g
Magnesium sulphate : 0.200 g
Calcium chloride : 0.020 g
Sodium molybdate : 0.002 g
Ferric chloride : 0.010 g
Bromothymol blue (0.5%) solution : 5 ml
Cane-sugar : 100 g
Yeast extract : 0.02g
Agar : 18 g
Distilled water : 1000 ml
Final pH : 6.0
Composition of semisolid
acetic LGIP medium
Semisolid LGIP medium was
acidified with acetic acid to pH
4.5 and agar concentration
was increased to 2.2 g 1-1
according to Cavalcante and
Dobereiner (1988).

17. 6) Media for Determining Phosphate Solubilization :
Katzelson andBose medium(1959) :
Soil extract : 100 ml
(Autoclave soil suspension, filter and use clear filtrate)
Glucose : 1.0 g
Agar : 2.0 g
Sterilize in 100 ml lots, cool and add 5 ml of 10 % KH2PO4 and 10
ml of 10 % CaCl2 to each flask.
 Adjust pH to 7.0 with sterile N/10 NaOH. Pour plates
immediately and allow to solidify.

18. 6) Media for Determining Phosphate Solubilization :
Pikovskaya's medium (Modifiedby Sundara Rao and Sinha, 1963)
Glucose : 10.0
Ca3(PO4)2 : 5.0
(NH4)2SO4 : 0.5
KCl : 0.2
MgSO4.7H2O : 0.1
MnSO4 : trace
FeSO4 : trace
Yeast extract : 0.5
Agar : 15.0
Distilled water : 1000ml

19. 6) Media for Determining Phosphate Solubilization :
Barton's reagent :
a) Dissolve 25 g ammonium molybdate in 400 ml of H2O.
b) Dissolve 1.25 g ammonium metavanadate in 300 ml of
boiling water, cool and then add 250 ml conc. HNO3.
After wards, mix A and B solutions and make up to a
litre.

20. 7) Media for fungal bio-agents ( Trichoderma, Verticillium,
Beauveria, Metarhizium, Nomuraea, Paecilomyces) :
PDA Media Composition :
Potato : 200 g
Dextrose : 20 g
Agar-agar : 20 g
Distilled water : 1000 ml