1916] Brandt: Notes on Trillium 41 



their spores in the early part of their growing season before or shortly 

 after the young shoots appear above ground. Miss Alden (1912) gives 

 a review of the rather meager literature on this general subject in 

 connection with her pertinent study of stages in the seasonal develop- 

 ment of Uvidaria sessilifolia. Howard (1915) has made a series of 

 interesting experiments in an effort to break the summer rest period 

 of bulbous plants. 



With the object, then, of finding out how nearly the behavior of 

 our forms of Trillium in the resting season agreed with that of the 

 eastern forms, collections and observations were begun in the autumn 

 of 1913. Collecting was begun while the plants were in the pollen 

 mother-cell stage, late in September, and was discontinued in April, 

 when most of the plants were going out of flower. Collecting was 

 resumed in August, 1914, and continued until November 10, 1915. 



i\Iost of the material was collected in the hills near Berkeley, on 

 the eastern side of San Francisco Bay, but from the latter part of 

 May to the early part of August, 1915, T. sessile var. giganteum H. 



6 A. was collected at Point Arena, Mendocino County. From time to 

 time T. ovatum Pursh was collected for purposes of comparison, at 

 Mill Valley, Marin County, and at Gualala, Mendocino County. 



Up to the autumn of 1915 all specimens were trimmed and killed 

 in the field to avoid wilting. In these last collections, made for the 

 express purpose of securing a large supply of material showing chro- 

 mosome reduction in the pollen mother-cells, the material was brought 

 into the laboratory and examined before killing, suitable precautions 

 having been taken to keep it fresh. The following killing solutions 

 were used: Bouin's solution, Zenker's mixture, Flemming's fluid 

 (strong), and Farmer's absolute alcohol-acetic acid solution. The last 

 two gave the best results. 



All material was imbedded in paraffin. Anthers were sectioned 



7 to lOfx thick, ovaries 7 to 25fx, according to the stage of development 

 of the ovule, and young stems just forming lateral organs were sec- 

 tioned 10 or 12/x thick. The stains used were aqueous safranin and 

 Delafield 's haematoxylon in combination or alone ; safranin and ' ' light 

 green"; the safranin, gentian-violet, orange G. combination; and 

 Haidenhain's iron alum haematoxylon. The last stain was particu- 

 larly useful in studies of chromosomes, while the most generally suc- 

 cessful was the safranin ' ' light green ' ' combination. 



