84 CROWN-GALL OF PLANTS. 



REINELT S EXPERIMENT. 



The experiment performed in Reinelt's manner gave the following 

 result at the end of 5 days: 



Second dilution from tube containing the cube — plate 4, nothing; 

 plate 5, nothing; plate 6, 2 small white colonies, nature doubtful; 

 plate 7, nothing. 



The dilutions were made as follows: After the 4-millimeter cube, 

 which was cut with a cold knife, had remained in the tube of bouillon 

 about 10 minutes, one 3-miUimeter loop of this fluid was transferred 

 to tube 2, which was shaken; then two 3-millimeter loops from this 

 tube were transferred to tube 3, which was shaken. The plates were 

 then poured from tube 3, the following amounts of fluid being put 

 into each: Plate 4 received three 3-millimeter loops; plate 5 received 

 two 3-millimeter loops; plate 6 received one 3-millimeter loop ; plate 

 7 received one 3-millimeter loop. 



After the cube had stood in tube 1 for 2 hours it was mashed in 

 the bouillon with a sterile scalpel as well as it could be (but not 

 nearly as effectually as the remainder of the tumor which was ground 

 with sand), and after standing an hour longer (to diffuse) 4 addi- 

 tional plates were poured, inoculating as follows directly from the 

 tube containing the crushed beet : Plate 68 received three 3-millimeter 

 loops; plate 69 received two 3-millimeter loops; plate 70 received one 

 3-millimeter loop; plate 71 received one 1-millimeter loop. 



At the end of the fifth day all were free from gall colonies. The 

 same was true at the end of 15 days. 



OTHER ATTEMPTS AT ISOLATION. 



At the same time as the above, Miss Brown nuT,de attempts to cul- 

 tivate out the organism believed to be the cause of the sugar-beet 

 tumor from galled sugar beets from Arlington, Va. ; Blissfield, Mich. ; 

 and Faii-field, Wash. On her numerous poured plates she obtained a 

 small sprinkling of colonies which appeared to be the right thing, and 

 with subcultures from 30 of these made inoculations upon sugar beets 

 in one of our houses, and also with the more hopeful a few inocula- 

 tions upon daisies, tomatoes, etc. Of the whole lot inoculated, 

 greatly to our surprise, not a single one has produced galls on sugar 

 beet. The only results obtained up to the time this bulletin goes to 

 the press are tiny beginnings of overgrowth in needle pricks on a 

 few daisies and oleanders, and 2 somewhat larger, but small hyper- 

 plasias, on 2 tomato stems. There can be no doubt about the 

 growths being tumor growths and due to what was inserted, l)ut 

 5 colonies only of the 30 have yielded these results. The remainder 

 have failed. 



