100 KANSAS ACADEMY OF SCIENCE. 
above classes of soil, with the result that in both cases tubercles were formed in 
the same relative proportion to the above. This shows that Kansas soil, being 
once inoculated, can be used to inoculate other soils. 
INOCULATING WITH TUBERCULOUS Roots.— After remaining in loose soil about 
a:month, some of the roots which had previously produced tubercles were taken 
to inoculate a pot of yellow soy-beans, The plants grew well and ranked among 
the best in the greenhouse. On washing out the roots, large and numerous 
tubercles were discovered, which were by far the best of any produced in the 
greenhouse during this experiment. Likewise, washed roots that had been air- 
dried in diffused light for about the same time were placed in another pot;, 
tubercles were formed, but neither the growth of the plant nor the tubercles 
were“equal to the above. In the former case the roots had more or less soil ad- 
hering to their surface, but in the latter there was practically none. 
EFFeEctT oF INOCULATING OTHER LEGUMES wiTH MassacuHusetts Sort.—Four 
pots each of adzuki beans ( Phaseolus radiatus), cow-peas, Canada field peas, 
alfalfa, and red clover were planted, half of these being inoculated with Massa- 
chusetts soil and the other half not treated. On the roots of the adzuki beans 
and the cow-peas no nodules were apparent in any of the pots; the alfalfashowed 
several; and on the clover and Canada field peas they were very numerous; but 
no difference could be detected on any of them that was due to the Massachu- 
setts soil. Evidently these plants were attacked by a different kind of organism 
from that attacking the soy-bean. 
ROOT TUBERCLES UNDER THE MICROSCOPE. 
PREPARATION OF StipEs.—This phase of the subject was taken up with the 
hope of observing the way micro-organisms behave within the tissues of the root. 
Tubercles were cut from the roots of plants seventy-two and ninety-nine days old, 
respectively, which had been grown in the greenhouse under rather unfavorable 
circumstances. These were placed in one per cent. chromic acid for eighteen 
hours, after which they were washed out and placed in fifteen per cent. alcohol 
for seventeen hours, then in thirty per cent. for nine hours, then fifty, sixty, 
eighty and ninety per cent., and absolute alcoho! for six hours each, more or less, 
at convenience. They were then transferred to one-half alcohol and one-half tur- 
peutine for seven hours previous to placing them in pure turpentine. Following 
this treatment paraffine was added sufficient to make a saturated solution. This 
was placed on a radiator for twelve or fifteen hours to keep the paraffine melted 
and thus to more thoroughly saturate the tubercles, when they were removed to 
a water-bath and kept in paraffine at a temperature of fifty-eight degrees C. for 
two or three hours. The tubercles with the melted paraffine were then poured 
into a paper box, which was floated on the surface of water until the paraffine 
formed a scum on its upper surface, after which the whole was rapidly cooled by 
immersing it. From this solid paraffine pieces containing tubercles were cut out 
and mounted for the microtome. 
When the sections were cut, they were placed on a glass slide previously 
covered with a thin coat of. albumen solution to make them stick. This was 
then held over an alcohol lamp until the paraffine was all melted. After being 
allowed to cool, the paraffine was dissolved off with turpentine, and the specimen 
carried back through the various strengths of alcohol until it could be placed 
in water. It was then put into asolution of hematoxylon for twenty minutes to 
stain it and, after being brought up through alcohol to turpentine, was mounted 
in Canada balsam. The apparent infecting mycelium in the tubercle absorbed 
the stain more readily than the cell tissue, and could be seen with a Zeiss micro- 
scope fitted with 1-12 (2mm.) homogeneous objective anda No. 4 eyepiece. This 
