[Vor. 5 
284 ANNALS OF THE MISSOURI BOTANICAL GARDEN 
infected and germinated as already noted. The jars for both 
lots contained sufficiently moist filter paper to supply the nec- 
essary amount of water for the seeds during the usual 24-hour 
period of etherization, and during this time stood at room 
temperature of 28-32? C. At the end of the period both the 
etherized and control seeds were quickly dehydrated by im- 
mersion in 50 per cent aleohol for 5 minutes, for 10 minutes 
in each of two changes of 95 per cent aleohol, and then for 
10-minute periods in acetone, 95 per cent alcohol and acetone 
in the order named. They were then air-dried before an 
eleetrie fan at high summer temperature. When thoroughly 
dry both lots were twice ground into coarse meal in a food 
grinder and finally made into a fine flour by grinding twice in 
a mill, after which they were stored in desiccators. _ 
The substrates used were a 1 per cent starch solution, made 
as outlined in the previous work, sucrose (Merck), and 
maltose (Merck), each in 2 per cent solution. Fifty cc. of 
each substrate with 1 per cent toluol were used in each flask, 
and to them were added 5 grams of the respective lots of 
barley flour. The tests were run at room temperature of 30? 
C. A preliminary test for amylases was made to determine 
the length of time desirable to run the series. By the use of 
the iodine spot-plate test for starch it was found that a little 
more than 34 hours elapsed before the action of 5 gms. of 
either flour acting on 50 cc. of starch solution failed to give 
positive results. The flasks were therefore allowed to stand 
for 13 hours after adding the flour in the case of the starch, 
and for 2 hours in the case of sucrose and maltose. 
Two flasks of each substrate with flour from etherized 
barley and a like number with control flour were employed. 
Fifty се. of each solution served as a check upon the sub- 
strate itself. At the end of the run one flask with etherized 
and one with control flour were at once heated to 15 pounds 
pressure in the autoclave, cooled, the mixture centrifuged, and 
the liquid analyzed for reducing sugars by the Shaffer method. 
The other pair of flasks was similarly treated, except that the 
contents were first centrifuged, and the liquid alone then auto- 
claved. It was believed that a comparison of the values from 
