Feb. 5, 1917 
Leafspot-Rot of Pond Lilies 
223 
tainers were left covered for five days, but after eight days no signs of infection had 
appeared. 
A species of Altemaria obtained from a leafspot of lotus was tested upon four 
leaves of N. odoraia. No infection had resulted after eight days. 
Laboratory, August 8, 1914.—Isolations 217 and 220 were inoculated on four 
leaves each of N. odoraia. After four days small infection spots had started on all the 
inoculated leaves and at the majority of points of inoculation. After eight days the 
infections had developed into the typical irregular spots. The four control leaves 
were sound. 
Laboratory, October 16, 1914. Isolations 205 and 225.—Inoculations were 
made with isolation 205 on four leaves of N. odoraia, and one of N. daubeniana, and 
with isolation 225 on three leaves of N. odoraia and one of N. zanzihariensis. These 
inoculations were from 45-day-old com-meal-agar cultures of each isolation. In 5 to 6 
days all inoculations with 205 tad taken. Two out of three leaves inoculated with 
225 showed infection but the third, together with the leaf of N. zanzihariensis , remained 
healthy. Five leaves of N. odorata and one each of N. daubeniana and N. zanzibarien- 
sis held through the test as controls remained sound. In this case the fungus retained 
its infective power in culture after being held 45 days at laboratory temperature 
(20°-25° C.). 
Laboratory, October 24, 1914. Isolations 205 and 225.—Six leaves of N. 
odorata and three leaves of one of the night-blooming species of Nymphaea were inocu¬ 
lated on the upper surface with isolation 205, and two leaves of N. odorata with isolation 
225. Four leaves of each species were floated upside down in the glass containers and 
inoculated with isolation 225. 
After three days small reddish infection spots were starting at most of the inoculation 
points in all the leaves inoculated on the upper surface. After six days all these 
inoculations had taken, giving the typical spots, which were now up to 5 or 6 mm. in 
diameter. Most of the spots were surrounded by a halo of pale yellowish green tissue 
grading off to the normal green of the leaf. No fungus mycelium could be found in 
this outer halo. It seems thus evident that a substance or substances produced by 
the fungus may diffuse into the living leaf tissues and cause the death of the cells 
ahead of the actual fungus invasion. 
After three days no infections were apparent on the inverted leaves. After five 
days infection areas were starting at inoculation points at the water surface or where 
the leaf epidermis was covered only by a very thin film of water. Inoculated portions 
covered by an appreciable depth of water (for example, several millimeters) still 
showed no signs of infection although in many cases the spore masses were still resting 
directly on the leaf epidermis. Infections thus took place where the lower epidermis 
was not submerged, but much more slowly than where inoculations were made on the 
upper epidermis. 
The four control leaves run with these two sets of inoculations were and remained 
sound throughout. 
On October 30, reisolation of the fungus was made from leaves inoculated October 
24 with isolation 225. This reisolation was designated as isolation 249. Subsequent 
inoculations with this isolation gave typical infections. 
Greenhouse, November 9, 1914. Isolations 205, 225, and 249, using 2- to 4- 
weeks-old cultures. —Inoculations were made with isolation 205 on two upper leaf 
surfaces each of N. omarana , N. rubra , N. dentata , N. odorata; with isolation 225 on 
N. rubra , N. zanzihariensis , N. omarana , and N. odorata; and with isolation 249 on N. 
daubeniana and N. capensis. Two tubs containing N. caerulea and two containing N. 
tuberosa were retained as controls. All inoculated and control leaves were covered by 
shallow bell jars for four days after inoculation and in each species two to four leaves 
were inoculated. After three days evidences of infection were observed on many of 
the inoculated leaves. After five days every inoculated leaf showed the typical spots, 
