174 CEOWN-GALL OF PLANTS. 



crown-galls of the sugar beet, as compared with the unaffected parts 

 of the same roots, slightly more water, considerably less cane sugar, 

 the presence of invert sugar, double the quantity of ash, and in all 

 but one instance more than double the amount of raw protein. Six 

 analyses were made and the calculations are expressed in per cents 

 of fresh substance. They all agree, except that two blanks occur in 

 the invert-sugar line, and tv/o in the raw-protein line. 



The same facts respecting cane sugar, invert sugar, pure ash, and 

 raw protein are shown still more strikingl}' in a table where the 

 amounts are calculated in 100 parts of the dr}^ substance. No 

 invert sugar was found in the normal parts of the roots, but 0.91 

 to 1.52 per cent in the galls. An average of the six analyses shows 

 that the dry substance of the galls contained 50 per cent cane sugar 

 as against 61 per cent in the normal parts of the roots. The average 

 per cent of ash in the roots examined was 2.78 and in the galls 6.05. 

 The average per cent of raw protein in the roots was 4,09 and in the 

 galls 9.80. 



ANALYSIS OF FLASK CULTURES OF BACTEEIUIVI TUMEFACIENS. 



Anal3"ses of flask cul tinges of the daisy oi-ganism after some months' 

 growth in 750 cc. filtered river water containing 35 grams c. p. 

 calcium carbonate, 14 grams Witte's white peptonum siccum, and 35 

 grams Merck's c. p. dextrose were made for us by Dr. Carl Ij. Alsberg, 

 with the following results: 



Received July 26, from Doctor Smith, five flasks of the culture, labeled, "Daisy 

 (newest strain)." The reaction of the culture medium [inoculated March 29, 1910] 

 was distinctly alkaline; the bottom of the flask contained much calcium carbonate, 

 which was filtered off. The filtrate was alkaline. A small portion, when acidified 

 with acetic acid and treated with ammonium oxalate, gave a heavy precipitate of 

 calcium oxalate, showing that a considerable amount of the calcium carbonate had 

 been dissolved. The solution reduced Fehling's solution powerfully, showing the 

 presence either of aldehyde or of sugar. Subsequent investigations showed the 

 absence of aldehyde, so that this reduction must be attributed to sugar. Other flasks 

 of the same lot, the analysis of which was taken one or two months later, still showed 

 a large quantity of sugar present. The filtrate, which was alkaline, was preserved and 

 examined. It did not reduce ammonium silver nitrate solution, and therefore can 

 not have contained any aldehyde. It gave a powerful reaction with potassium iodide, 

 resulting in the formation of considerable iodoform. Hence, the main constituent of 

 the distillate was ethyl alcohol. The residue in the distilling flask was now acidified 

 with sidphuric acid, and the distillation re)>ea(ed. The distillate proved to be very 

 acid, and had an odor resembling acetic acid. It was made ammoniacal and con- 

 centrated to a small bulk. The neutral solution resulting was treated with silver 

 nitrate, yielding a crj^stalline precipitate. This was recrystallized in hot water, 

 yielding large white needles ; 0.3969 gi'am of this silver salt yields 0.2559 gram of silver, 

 or 64.46 per cent of silver. Sil,ver acetate contains theoretically 64.67 per cent of 

 silver; hence the volatile acid can not be anything else than acetic acid. 



The results obtained with this single culture flask were exactly duplicated with 

 two other flasks. 

 213 



