28

Sept 1 [1880]

1)

Trypsin on leaves (together with Thymol)

10° 40' a.m. Ivy — Lime & Hazel leaves with chain of dots of trypsin sol — on wet litmus paper under moistened Bell glass —

(Sept 3d 8° 10 a.m. put in fresh drops)

(1st temp of room 11°15 66°7 — 10°30 P. 2 65°)

2d 6° 50 a.m 64° — 2° 45' 66° — 10 67°)

(Sept 3d 8° 15' a.m 66°

11° 5' put tips of leaves, under Bell-glass to soak in Trypsin & in distilled water — of Lime, Hazel, Ivy, sage, Hornbeam, & Elm.

Sep 2d 7° 46' a.m. Lime immersed part pale, veins I think pinkish

Hazel — rinsed parts pale with here & there green cells — bathed margin brown. — veins I think pinkish

Hornbeam. Margin on one side brown — (other leaves unaffected part in [bottom] Hornbeam — leaf with base cut off.

Sept. 2. 2° P.m added young Lime leaf to Trypsin & to water.

(Sept 3d 8° a.m. Lime, vein certainly reddened — parts of leaf paler & parts brownish — The young leaf — part paler — what is cause?

Hazel part very brown — large parts pale green — I doubt about veins pinking — Hornbeam parts very dark .) Water no action

29

Sep 3

2)

Young lime from trypsin in [solution] taken out of trypsin abt 12 noon & put on a cover glass in glycerine & water —

The chief effect is due to infiltration as was proved by comparing it with a healthy bit of the leaf boiled in glycerine & water. The contents of the cells in the trypsin piece is yellower or browner than the boiled specimen — It is almost impossible to make out the chlorophyll grains in the trypsin specn. The cells look as if filled up with a nearly homogeneous brown green mass. In the boiled one the contents have a granulated look from the chlorophyll grains being more distinct — The boiled one looked at again 2.40 pm has become about as yellow and as homogeneous as the trypsin one

30

Sept 4th

3)

7° 30 a.m. — Lime — much reddened even part not immersed vein brown — fluid dreadfully putrid & turbid

Hazel part not immersed much blackened

Vein of upper blackened part more discoloured than immersed part.

Hornbeam parts blackened in upper part — last Hornbeam a little affected n one side of immersed part.

Elm square patches blackened in upper part

Sage — upper part much blackened —

Ivy alone not affected

The blackening of upper part seem to show that access of air somehow important (see p. 5 from Frank diameter of Elm leaf

Leaves in plain water not affected.

(No action from the drops on the [base] on damp paper under the Bell-glass.)

Sept 5th 8° a.m Lime leaf & sage tips affected & discoloured by plain water —

Sept 6th 8° a.m only a trifle more affected)

30v

Sept 5th

4)

8° a.m Ivy-leaf — When I (yesterday?) passed with pin's head & placed drop of Trypsin now brown marks. Now passed in another transfer lime parallel to last & in those lime I smoked through fine tube & then placed drops. On opposite side rubbed lime with bluntend of ivory needle & placed some trypsin on old Hazel & Lime leaf —

(also 8.15' on very old ivy leaf)

(a worm near base did not pass head)

31

Sept 5th

8° 20 put starch in not diluted trypsin not dissolved.

8° 25 — drops of trypsin on disc & on edge of Hazel & Lime, twigs standing in water under Bell-glass.)

Sept 7th 8° a.m The Lime with edge in fluid considerably discoloured, not that with fluid on disc

7th at 3° P.m — I think a close difference; but then the trypsin had gone bad & cd not act.

31

Sep 4

5)

Trypsin on Sages

A piece of an old lime leaf taken out of trypsin & put in methyl spirit last night. It was cut so that it consisted of an uninjured and all injured strip side by side. It was put in a solution of Iodine & Iodine of potassium & the under surface examined & it was found that in the injured strip which had been in trypsin there was no starch and hardly any debris even in the guard-cells of the stomata while the other half which had not been in trypsin had starch in it.

Lime

Bits of the lower side of leaf which had been pulled out of a worm's burrow were put in Iodine solution & it was found that the guard cells were almost emptied of their contents & contained no starch, whereas those near the base of the lead has starch in plenty.

(The unimmersed brown parts of the elm

33

leaf are more opaque than the green parts this is a good deal due to little veins down to smallest ramifications being reddish brown — many of the chlorophyll & grains are stained the same colour, & in other places they are of a dark dull green