A

Euphorbia peplus

1881.

Roots placed in sol of 2 to 1000 of C. of amm at 2° 30 P.m, examined 8° am on 17th — near tips much brown matter, as usual, even extending to all the cells except the 2 extreme terminal ones.

In these old plants, the brown matter does not always run up so far; some rows of cells being faint brown with no granular matter —

The cells with the gran. matter generally alternate with the transparent ones, but sometimes 2 rows of intermediate transparent cells. —

(Acetic a. during 2 hr. did not dissolve the granular matter — nor did alcohol during 24 hr — but rendered them paler.

Irrigation with S. Ether no quick effect.)

Cells with brown granular matter are after ammonia are have been cast off from the as roots. grow)

In upper & thick part of root the matter can hardly

(B

Euphorbia

be called granular, but consist of variously shaped, always rounded, brown hyaline matter more or less aggregated — within some of the smaller masses, minute transparent speres can be seen. I cd see no movement in these bodies — This matter lies in cells, not in tubes — The cells in the upper parts of roots with granular matter, are almost continuous within the outer layer of cells or epidermis — These succeeded by 2 or 3 rows of parenchymatous, & then come a few of inner cells with agg. matter, which lies close to the control fibrous tissue.

(Stem of the plants, the roots of which had been in C. of ammonia, as above described.─ No aggregation — Epidermic cells with purple fluid or fine granular matter; a solution

C

Euphorbia

of 7 to 1000 produced no effect after 1 hr on these slices; nor after 21 hr. except that the purple in epidermic cells somewhat granular or dotted— no inner circle of granular matter.

(Longitudinal & transverse slice of thick upper part of root of fresh plant, which had not been exposed to C. of am. were examined no aggregation, but after about 5' after irrigating with the ammonia, (7 to 1000) all the external & interior normal cells opake with brown granular matter — Oct 19th so it was next morning with no change in the granular matter.)

(Oct 19th, examined 4 uninjured & some injured roots from fresh plant after having immersed for 3' or 4' in Boiling water no aggregation —

Then placed in sol. of C. of amm 7 to 1000 for about 1/2 hr no true granular aggregation, but I am not sure that brown matter not collected on walls — I shd preexamine a root & irrigate

(A stem, which had rested for 48 hr in C. of am 2 to 1000 was sliced obliquely, but no evidence of absorption— fluid contents of epidermis cells purple — no aggregation.

(D

Euphorbia peplus

Oct 28th

Youngish plant placed roots for 3 or 4° in water which had just boiled water — no aggregation, but was rendered rather opake, & in many cells (in one fine sub-root, but only a few another root) transparent spheres, which seem to be the primordial contracted utricle enclosing coagulated matter. These spheres have no relation to granular aggregated matter, for they occurred in 4 or 5 adjoining rows of cells: sometimes, I believe, 2 or 3 spheres in same cell. — Irrigated copiously with sol. of 7 C. of amm to 1000; but no irrigation aggregation ensued after 25'. Added Acetic. a. which under the roots so clear that I cd just see central spiral vessel, but still no granular aggregated matter in 2 uninjured fibrate carefully examined.

This case important for shows that aggregation either a vital phenomenon, or that C. of A does not act on contents of cell after exposure to temp of nearly 212° F. 29th looked next morning at

[Dv]

these same rootlets on Acetic a. no change & looked at some other roots nowhere any granular aggregation.

E

Euphorbia peplus

Oct 29th placed a thin root under glass & examined & fd no aggregation, partially dried glass & added drop of C. of am. (7 to 1000) & in 20 seconds cd just distinguish fine granular matter — appears first as slight brownish cloud. — In course of 1 or 2 minutes whole length of rootlet with granular matter, but is perhaps absorbed by walls — Old experiments show that chiefly absorbed by tip — A Two pieces of a root, cut off at both ends became granular.

In 2 instances I found 3 adjoining rows of cells with granular matter, the middle one row, however, after a few cells in longest became quite transparent — I traced one row of transparent cells continuously for a great distance. —

In this specimen there never came never came the dark

F

Eup. Peplus

brown masses a little above tip.— Here & there a single cell was quite filled with brown granular matter so as to be opake.

The primordial utricle sometimes contracts & draws the granular matter from square end-wall, so that the brown matter ends in curved line. Hence probably the cause of square masses becoming after a time spherical.

I can see no difference in size or shape of the cells with granular matter & the empty ones. — Looked again no difference

I must say that if many scores of rootlets are looked at— alternation is certainly the general rule, but not rarely 2 with granular, or 2 with no contents run side by side. —

G

Euphorbia peplus

Oct. 30th.

Stem, transverse & oblique slices left in C. of A (7 to 1000) for 20 hr.— no aggregation; but on the walls of in the elongated purple epidermic cells, the some granular matter is attached.—)

Roots in Phosphate of Ammonia (4 to 1000) for 21 1/2 hr. —

The generally alternate rows of cells with granular matter, as distinct as & in some places coarser than from C. of A, but never so brown — In some roots the alternate rows differ only by the one set being of a very pale brown, but granular, no doubt present, yet so small as to be (a) invisible — In one rootlet, just above tip, spherical aggregation of brown matter. (I suspect acted on by dissolved by Glycerine??)

Nitrate of A. 2 to 1000 for 22 hr. In one rootlet with brownish spheres above tip, including brown granular matter; in upper part of some rootlet alternate rows differing only being slightly browner, but with no granular matter — In 2d rootlet cut off at both ends & so higher up root, a good deal of fine granular matter in the alternate rows, but not so much as from C. of Ammonia.— In a 3d rootlet — no sphere near tip, & in most part rows of brownish cells with no granular matter, but in some of the cells plain granules (B)

[Gv]

(a) Rexamined fresh rootlets. Nov. I 11° a.m in the Phosphate of A 4 to 1000 since 11° a.m on Oct 29th & now splendid brown spherical masses near tip & high up the rootlet in the usually alternate rows of cells more & plainer granules, than I have seen with C. of A. — Irrigated with glycerine & water & examined Nov. 2d 8° a.m — the brown masses not affected near tip in one rootlet very brownish, the granules in alternate rows as plain as ever; could this rootlet be dead? (Nov. 3d. 8° 30' same result. In one rootlet the inner vessels in endoderm seemed to me continuous, with much aggregated matter.

(B) Rexamined fresh rootlets at 11° 15' Nov. I, which had been immersed in the Nitrate (2 to 1000) since 11° on Oct 29th — Now much granular matter in the alternate rows, sometimes heaped into balls into in middle of cell. — Brown masses near tips not conspicuous.

H

Euphorbia peplus 2 1/2 to 437 [x] 2 [=] [illeg]

Oct 30

Carbonate of Soda (2 gr. to 2. oz) during 24 hrs —Three rootlets examined, square very brown masses in cells near tip, apparently but little granular─ Higher up rootlets, some rows of cells coarsely granular; other rows only brownish & no granules visible. — Does acid hold matter in solution? Phosphate & Nitrate opposed. —

In one place 4 adjoining rows contained granular matter

Oct 31st Some of These rootlets kept in acetic A. for about 20 hr.

The brown masses above apex quite distinct, but more transparent, but the alternate rows of cells, with granules no longer distinguishable — gran either dissolved or colour to water or somehow lost.—

(Some of same spec. left in glycerine & water for 20 hr— brown masses above tip still plain — some in root-cap; this looks as if excretions. No granular matter left but the rows of brownish C cells still brown — I saw some rows of such brownish cells [illeg] divide into 2 rows of brownish cells & into 1 white & 1 brown —

Also row of colourless cells divide into brown & colourless rows.— A second rootlet with alternate rows no longer distinguishable except by brownish tint, ie no granules visible.─

I

Euphorbia peplus

Oct 30th

Rootlets exposed for 24° 40' to C. of Am. 1 to 10,000

Rootlet (1) no brown masses near tip, alternate rows with brownish granular matter higher up.

(2) a few rather brown & coarsely granular masses near tip; alternate rows higher up with granular matter unmistakable

(3) no brown masses near tip. — higher up alternate rows conspicuously

(4) cut off both ends — very obscure evidence of alternate rows

(5) no clear brown masses near tip, but wall apparently have & the kind with brown matter — higher up plainly alternate rows with granular matter.

Oct 31'. Three rootlets left for about 20 hr in glycerine & water under cover glass & the alternate rows no longer distinguishable, for in all the P.m. contracted into one or 2 or 3 regular or irregular spheres or ovals or into irregular masses, & I suppose grain then all collected into heaps.

K

Euphorbia peplus

Oct 31

I made sections of high up of thin roots, after immersion for about 20 hrs. in sol. of 4 to 1000— I cd see here & there epidermic cells bearing hairs, within which were the cells with aggregated granules — I certainly feel sure that think these in epidermis — corresponding with root-cap in Frank's drawing — Sachs speaks of r epidermis to root —

In the higher part of roots almost all the rows of cells, just within epidermis, contain the granular matter, & the alternations cease. —

Nov. 1 These slices were well irrigated with glycerine & water & left till this morning ie for about 21 hr & every single granule has disappeared, either dissolved or aggregation has ceased — It shd be remembered that it was slices which were thus exposed. —

L

Euphorbia peplus

Nov I

Root 5 mm above highest lateral rootlet, surface brown, but very near where it become green, & probably probably beneath ground—

Much plainly marked aggregated granular matter — also in dark-coloured elongated cells of epidermis — transverse slices examined

These slices after remaining in water for about 20 hrs exhibited agg. grain plainly— may be dead. Left in glycerine & water for 24 hr. grain still left & orange. Nov. 3d 8° 20' irrigated same spec. with acetic acid.— Nov. 4th 9° a.m. In parts granular matter quite distinct in part only fine brownish matter distinguishable in which no grain present.

Nov. 2d) Roots were placed in sol 4 to 1000 on Nov Oct. 30th at 12° & were today on Nov 1st examined at 4° 30' P.m placed in glycerine & water — were examined on Nov 2d 11° 45'.— In 3 of rootlets granular matter in the alternate rows as plain or plainer & browner than in fresh specimens after ammonia — In 2 rootlets which were very pale rootlets no aggregated matter. — As these rootlets were so long submerged in the sol. they may have been killed.

M

Euphorbia peplus

Nov. 3d

The roots of a fresh plant were put in sol. of 4 to 1000 at 8° 7' a.m & were put in water at 10° 45 A.m. so exposed for only about 2° 40' & in all five rootlets (6 were examined) the alternate rows contained abundance of granular matter; but not much above root-cap. At 11° a.m added glycerine & water: by 4° P.m. the granules not dissolved. Nov. 4th 8° a.m by next morning granules in same state.

At 11°, 3 of the above rootlets were irrigated with acetic a. & at 8° a.m next morning 4th, examined— granular matter still present, but I think reduced in quantity — roots rendered very brown.

Roots of same plant, exposed for about 2° 40; to the sol, were transversely sliced just a little beneath uppermost lateral rootlet & there was plenty of granular matter in the exterior cells. At 11° 50' added glycerine & water to these slices

Nov 4th 8° 30' a.m the granular matter certainly seems broken down into the finest brown granules, too small to be seen.

N

Euphorbia peplus

Nov. 5

Some roots which had been in C. of A. (4 to 1000) since 2°. 15' Oct. 30 were placed in alcohol for 18° no aggregation, was visible— were then irrigated with C. of ammonia (7 to 1000) & after 28° no trace of aggregation caused, & the roots were so transparent that I could easily see spiral vessels. —I infer from this case & boiling water that aggregation a vital process.—

Some oldish roots which had undergone aggregation from C. of Ammonia were irrigated with caustic potash, & all granules have disappeared in the exterior cells & in endoderm. — But I must repeat this observation

(Roots in C. of A (on 7th) (7 to 1000) from 8° 10' a.m to 10° 20' a.m with good aggregation then placed in weak Caustic potash & examined this morning (8th) at 8° 30' — In most places no trace of granules left, but at cut off end of one rootlet numerous transparent granules in tubes on both sides of spiral vessels— Therefore it appears that cold Caustic Potash does not efficiently remove the aggregated matter.

O

Eu. peplus

Nov 9th

I was so astounded at agg. in radicles of E. myrsinites without any ammonia that I have looked at fresh root of E. peplus — examined a straight one with care — no grains in exterior cells or in endoderm. — Irrigated with C. of A. (7 to 1000) & in 3' grain in the alternate rows & in the endoderm. In 10' after the appearance of the granules irrigated with weak Caustic Potash & left for 24°, & now not a single grain left in the exterior cells or in those round spiral vessels— the whole as transparent as glass

P

Eu. peplus

Nov. 10th

A germinating seed with thin radicle 1/2 inch in length examined — no agg: grains of any kind but the roots were not torn open.─ irrigated with C. of A. (7 to 1000) & in 45" the brown masses formed near tip conspicuously— higher up rows of brown cells with no grains distinguishable — higher up some cells with grains — Too opake to see spiral vessel. — The brown masses above tip did not altering shapes. — The brown tubes with fine matter extended some height up radicle— I also saw, after compression some very thin tubes, (thinner than the cells with brown matter) with agg. matter, & which I suppose are incipient milk-tubes. These thin tubes were very near surface — Do they give rise to the cells with agg. matter?

(Nov 11' radicle only 7 mm. long, kept for 20° in sol. of C. of A. (4 to 1000) was heated for a few minutes in a moderately strong Caustic potash & water, & then torn open:

Q

Euphorbia peplus

Nov 11th

& it is certain that undivided milk tube containing the usual hyaline, almost solid, aggregated granules extended down close to tip, beneath root-cap terminating in blunt points — They run very near exterior & round spiral vessels — many of them — not excessively thin. The squarish brown masses (caused by the ammonia) just above the tip & the fine brown cells in rows & scattered, in which granules were not visible, lie exteriorly to the tubes & have no connection with them.

This last specimen had been kept in the same ammonia for the same time as the one, which had been heated in caustic Potash. —

I conclude that the brown cells in rows have no sort of connection morphologically with milk-tubes, but they seem may functionally. —

R

Euphorbia peplus

Nov 14th

Upper part of stem — good oblique long slices —

No agg. structure — irrigated well at 11° 45' on 13th with C. of A. 7 to 1000 & examined 8°- 9° A.m. 14th —

Now groups of cells in epidermis & long continued rows with granular matter exactly as in roots!!

Also in one place a vessel or a row of cells above the spiral vessels in same state ie with brown brown granular matter —(Before irrigation Ch. grains remarkably perfect & distinct — after the C. of A. in part granular, in part lie in heaps, & occasionally distinctly confluent— but only a few cases of such confluence.

(The sole strange fact that remains is the tendency to alternation in the cells of epidermis or above the epidermis of root. —

S

E. peplus

Nov 14'

I have looked at radicle of young seedling in C. of A. (4 to 1000) since Nov. 10th — well defined defined brown masses above tip— & rarely here & there an exterior cell with very fine granular matter. — Milk-tube splendidly clear with agg. matter which is either hyaline hyaline & white, or brown brown, but clearer & coarser than the matter granular matter in the few exterior cells —

Added Caustic Potash 11° 30' to see of brown cells affected, while not so the milk tubes — Nov 15th The brown squares heaps near tip & few granular cells have all disappeared whereas the milk tubes as plain or plainer than ever—

There must be chemical difference between the content of surface cells & milk-tubes.

Nov. 15th looked at 2 more young radicles with Cots in seed-coats immersed in 7 to 1000 for about 20°.— brown masses above tip plain, but the alternating rows of cells very obscure — slightly browner with here & there fine granular matter. —

T

E. peplus

Nov 15.—

Roots of moderately old plant, left in C. of A. (4 to 1000) for 3 days), tips of fine rootlets brown, & then twice heated for about 5' in caustic Potash, & frayed out & split, so that I could see whole contents & spiral vessels & not a trace I could see of any milk vessel (& I think De Bary says same thing) — We know positively that Caustic Potash does not cause disappearance of milk-vessels with contents, but makes them plainer— Of course all the aggregated granular matter was removed from tips & alternate rows of cells — Now in E. myrsinites in which, the alternate rows of cells with granular matter are not present, there are abundant milk vessels in lateral rootlets. — (I have been looking again at lateral rootlet of E. myrsinites after C. of A. & the milk-tubes, though not

(It is case of partial replacement, but with some chemical difference in contents. —

U

E. peplus

immersion are beautifully plain — There are a good many exterior cells; not epidermic with brown granular matter — sometimes several in a row & in one place these alternated with colourless cells in rows — but the phenomenon not distinguishable without careful examination.— Nothing like so many milk-tubes as in primary radicle. —

Added Caustic Potash (15' 10° 30') to see if brown granular matter will be dissolved. —

(15th I have made transverse (generally rather oblique) section across roots about 1/2 way up — I cannot make out whether the very brown endoderm cells which surround vascular tissue are tubes or cells— I certainly think there are exterior cells ie epidermic without the brown matter. — Added at 11° Caustic Pot. to see if the brown endoderm matter will disappear. Nov. 16th 8° 30' a.m it has disappeared no grain left & only here & there a brown stain. This shows that the endoderm cells contain matter like →

[Uv]

that in the exterior cells— In thin section up the somewhat thick roots 1/2 way up them, I distinctly saw, now that they were closed with potash a few milk-tubes with hyaline globules not dissolved by the Potash.—

(I have looked at another root 1/2 way up after being long in C. of A. & not in Potash— tessellated with square brown exterior cells— long narrow cells, no round with very dark brown contents near spiral vessels & a few milk-tubes.

The root was split & frayed.—

X

Euphorbia peplus. —Roots

Dec. 6'

Roots put into old sol. of aniline, strong 1 to 10, 000 at 8° 10' a m — taken out & washed at 11° 30'— stained pink, especially —root-cap—

(This bears on action of C. of A. on root-cap)—

Some aggregation certainly caused — Watts says is formed from Ammonia by [2 words illeg] C. & H. for the H in Ammonia — so that Nitrogen is present.

Roots placed Dec 5 2° 30' P.m in sol of 4 to 1000 examined from 9° to 12° on 6th. — F. made thin longitudinal slice of moderately thick roots, & on the several slices, I counted 62 root-hairs arising from the empty cells & not one from the granular cells. In these thicker roots the epidermic cells (in roughly alternate rows) & the cells close to vascular bundle, & here & there single isolated layers, more or less superficial in the parenchyma contained the usual brown granular matter. Besides these there were the latex-ducts with the hyaline, oddly shaped, more or

Y

Eup. Peplus

confluent grains. The parenchymatous cells were always single & often occurred in 2 layers between the epidermic & central cells.

In one place all the rows of cells contained the brown granular matter (& have no hair-roots) but these rows were distinctly darker & lighter brown.— new & good case. —

(Dec 6th Roots placed in osmic A. (1 per cent, 2 drops added to watch glass of water), left in for 2° 10' & then added C. of A. 7 to 1000 — As soon as I looked at all mottled (like Drosera) with black aggregation — Examined this morning splendid aggregation, in exterior epidermic cells — in the thicker roots in the parenchyma (which often stand in chequer arrangement) & on epidermis cells close to vascular bundle— Here in latter agg. matter very coarse grained, coarser than the grain in milk-tubes.

(I then tried the same sol. of osmic A. on root out of water & there was the same darker agg, in 1° as before observed after the C. of A. had been added; but roots perhaps not so transparent. æ

[Yv]

Dec. 7'

Some roots which had long been in C. of A 4 to 1000 were left in same sol. of osmic A. & now the aggregation was less distinct— higher up the roots many of the cells being filled with dark orange pulpy matter instead of granular. — Yet granules very distinct in other cells — Appearance of roots wonderful— The same long row of cells when followed up changed into empty ones & vice versâ empty ones turned into granular ones. —