A manual of rice seed health testing - IRRI books - International Rice ...
A manual of rice seed health testing - IRRI books - International Rice ...
A manual of rice seed health testing - IRRI books - International Rice ...
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After autoclaving the medium, add the following<br />
filter-sterilized antibiotic solutions<br />
(millipore filter 0.4 m pore diameter):<br />
Ampicillin sodium<br />
10 mg (1 ml<br />
<strong>of</strong> a 10 mg/ml solution<br />
in 75% ethanol)<br />
Pheneticillin potassium 50 mg (5 ml<br />
<strong>of</strong> a 10 mg/ml aqueous<br />
solution)<br />
Starch medium (Lelliot and Stead 1987)<br />
Soluble (potato) starch 2.0 g<br />
Nutrient agar<br />
28.0 g<br />
Distilled water<br />
1.0 liter<br />
Dissolve the nutrient agar powder in 500 ml<br />
distilled water and heat into solution.<br />
Dissolve the starch in 10 ml distilled water<br />
by gently heating and add to the nutrient<br />
agar solution. Make up to 1 liter volume,<br />
Sterilize by autoclaving. Pour the medium<br />
into individual petri dishes.<br />
Tryptic soy agar (TSA) (commercially available<br />
as powder preparation)<br />
Bacto-dextrose<br />
2.5 g<br />
K 2<br />
HPO 4<br />
2.5 g<br />
Bacto-soytone<br />
3.0 g<br />
NaCl<br />
5.0 g<br />
Bacto-tryptone<br />
17.0 g<br />
Distilled water<br />
1.0 liter<br />
II. Stains<br />
The following solutions are used for gram<br />
staining.<br />
Acetone-alcohol decolorizer<br />
Acetone<br />
Ethanol<br />
Crystal violet<br />
Ammonium oxalate<br />
Crystal violet<br />
Ethanol 95%<br />
Deionized water<br />
lodine solution<br />
Iodine<br />
Potassium iodide<br />
Deionized water<br />
Safranin counterstain<br />
Safranin 0<br />
Ethanol 95%<br />
Deionized water<br />
50.0 ml<br />
50.0 ml<br />
0.8 g<br />
2.0 g<br />
20.0 mI<br />
80.0 ml<br />
0.33 g<br />
0.66 g<br />
100.00 ml<br />
0.25 g<br />
10.00 ml<br />
90.00 ml<br />
III. Buffer<br />
Phosphate-buffered saline (PBS)<br />
Na 2<br />
HPO 4<br />
1.15 g<br />
KCI<br />
0.20 g<br />
K 2<br />
HPO 4<br />
0.20 g<br />
NaCl<br />
8.00 g<br />
IV. Others<br />
Oxidase test<br />
Tetramethylphenylene diamine<br />
dihydrochloride or<br />
Kovacs’reagent<br />
1% solution<br />
110 <strong>Rice</strong> <strong>seed</strong> <strong>health</strong> <strong>testing</strong> <strong>manual</strong>