200 BOTANICAL GAZETTE [SEPTEMBER 
preserving, the tissue was heated in a water bath for one hour at 
70° C. and set aside for at least one week before proceeding with 
the analysis. Later the epicotyls were cut with scissors into 
2-5 mm. lengths, transferred to ashless filter paper extraction cups, 
and the preserving liquid filtered through the cups. Two extrac- 
tions followed, one with hot 95 per cent alcohol for 4 hours and the 
other with hot ether for 2 hours. Then the tissue was powdered 
in a mortar and finally extracted for 12 hours more with hot 95 per 
cent alcohol. This procedure separated the sample into alcohol- 
ether soluble and insoluble portions. The dry weight of each was 
determined by methods which are described below. That of the. 
insoluble fraction was found simply by- drying to constant weight 
in an oven at 104°C. But the soluble fraction was concentrated 
upon a water bath to about 450 cc., transferred to a 500 cc. volu- 
metric flask, and made up to the mark. Then an aliquot part 
(usually 100 cc.) was taken for dry weight determination, and later 
for ashing. The final drying was carried out in a vacuum desic- 
cator over CaCl. 
Analysis of the alcohol-ether soluble fraction was carried out 
on the remaining 400 cc. This was evaporated to small volume to 
free it from alcohol, taken up with water, and transferred to a 500 Cc. 
volumetric flask. Fats and lipoids were precipitated from solu- 
tion by the addition of 3-5 cc. of chloroform and 10-15 cc. of 
5N HC. After shaking, the solution was made up to volume and 
set aside in an ice box for 24 hours to settle. The clear supernatant 
liquid was then decanted and filtered. No determinations were 
made upon this fat and lipoid residue. The water solution was 
divided into portions for the following determinations: (a) car- 
bohydrates, (6) total cde and (c) ammonia and’ alpha-NH; 
nitrogen. 
Analyses on the alcohol-ether insoluble fraction included the 
following: (a) reducing sugars after acid hydrolysis, (6) total nitro- 
gen of the fraction, (c) total nitrogen of the portion rendered soluble 
by acid hydrolysis, (d) “crude fiber” (dry weight only), which was 
that portion remaining insoluble after acid hydrolysis, and (e) weight 
of ash. The “crude fiber” would largely be cellulose plus protein 
which was yet undissolved by acid hydrolysis, hence the results 
