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 2 - 



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 

 5 N 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, (b) total nitrogen, and (c) ammonia and alpha-XH 2 

 nitrogen. 



Analyses on the alcohol-ether insoluble fraction included the 

 following: (a) reducing sugars after acid hydrolysis, (b) 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 (c) weight 

 of ash. The "crude fiber" would largely be cellulose plus protein 

 which was yet undissolved by acid hydrolysis, hence the results 



