Having examined a leaf in water, and found the contents of the cells homogeneous, I placed it in a few drops of a solution of one part of the carbonate to 437 of water, and attended to the cells immediately beneath the glands, but did not use a very high power. No aggregation was visible in 3 m.; but after 15 m. small spheres of protoplasm were formed, more especially beneath the long-headed marginal glands; the process, however, in this case took place with unusual slowness. In 25 m. conspicuous spherical masses were present in the cells of the pedicels for a length about equal to that of the glands; and in 3 hrs. to that of a third or half of the whole tentacle.

If tentacles with cells containing only very pale pink fluid, and apparently but little protoplasm, are placed in a few drops of a weak solution of one part of the carbonate to 4375 of water (1 gr. to 10 oz.), and the highly transparent cells beneath the glands are carefully observed under a high power, these may be seen first to become slightly cloudy from the formation of numberless, only just perceptible, granules, which rapidly grow larger either from coalescence or from attracting more protoplasm from the surrounding fluid. On one occasion I chose a singularly pale leaf, and gave it, whilst under the microscope, a single drop of a stronger solution of one part to 437 of water; in this case the contents of the cells did not become cloudy, but after 10 m. minute irregular granules of protoplasm could be detected, which soon increased into irregular masses and globules of a greenish or very pale purple tint; but these never formed perfect spheres, though incessantly changing their shapes and positions.

With moderately red leaves the first effect of a solution of the carbonate generally is the formation of two or three, or of several, extremely minute purple spheres which rapidly increase in size. To give an idea of the rate at which such spheres increase in size, I may mention that a rather pale purple leaf placed under a slip of glass was given a drop of a solution of one part to 292 of water, and in 13 m. a few minute spheres of protoplasm were formed; one of these, after 2 hrs. 30 m., was about two-thirds of the diameter of the cell. After 4 hrs. 25 m. [page 46] it nearly equalled the cell in diameter; and a second sphere about half as large as the first, together with a few other minute ones, were formed. After 6 hrs. the fluid in which these spheres floated was almost colourless. After 8 hrs. 35 m. (always reckoning from the time when the solution was first added) four new minute spheres had appeared. Next morning, after 22 hrs., there were, besides the two large spheres, seven smaller ones, floating in absolutely colourless fluid, in which some flocculent greenish matter was suspended.

At the commencement of the process of aggregation, more especially in dark red leaves, the contents of the cells often present a different appearance, as if the layer of protoplasm (primordial utricle) which lines the cells had separated itself and shrunk from the walls; an irregularly shaped purple bag being thus formed. Other fluids, besides a solution of the carbonate, for instance an infusion of raw meat, produce this same effect. But the appearance of the primordial utricle shrinking from the walls is certainly false;* for before giving the solution, I saw on several occasions that the walls were lined with colourless flowing protoplasm, and after the bag-like masses were formed, the protoplasm was still flowing along the walls in a conspicuous manner, even more so than before. It appeared indeed as if the stream of protoplasm was strengthened by the action of the carbonate, but it was impossible to ascertain whether this was really the case. The bag-like masses, when once formed, soon begin to glide slowly round the cells, sometimes sending out projections which separate into little spheres; other spheres appear in the fluid surrounding the bags, and these travel much more quickly. That the small spheres are separate is often shown by sometimes one and then another travelling in advance, and sometimes they revolve round each other. I have occasionally seen spheres of this kind proceeding up and down the same side of a cell, instead of round it. The bag-like masses after a time generally divide into two rounded or oval masses, and these undergo the changes shown in figs. 7 and 8. At other times spheres appear within the bags; and these coalesce and separate in an endless cycle of change.

After leaves have been left for several hours in a solution of the carbonate, and complete aggregation has been effected, the

* With other plants I have often seen what appears to be a true shrinking of the primordial utricle from the walls of the cells, caused by a solution of carbonate of ammonia, as likewise follows from mechanical injuries. [page 47]

stream of protoplasm on the walls of the cells ceases to be visible; I observed this fact repeatedly, but will give only one instance. A pale purple leaf was placed in a few drops of a solution of one part to 292 of water, and in 2 hrs. some fine purple spheres were formed in the upper cells of the pedicels, the stream of protoplasm round their walls being still quite distinct; but after an additional 4 hrs., during which time many more spheres were formed, the stream was no longer distinguishable on the most careful examination; and this no doubt was due to the contained granules having become united with the spheres, so that nothing was left by which the movement of the limpid protoplasm could be perceived. But minute free spheres still travelled up and down the cells, showing that there was still a current. So it was next morning, after 22 hrs., by which time some new minute spheres had been formed; these oscillated from side to side and changed their positions, proving that the current had not ceased, though no stream of protoplasm was visible. On another occasion, however, a stream was seen flowing round the cell-walls of a vigorous, dark-coloured leaf, after it had been left for 24 hrs. in a rather stronger solution, namely, of one part of the carbonate to 218 of water. This leaf, therefore, was not much or at all injured by an immersion for this length of time in the above solution of two grains to the ounce; and on being afterwards left for 24 hrs. in water, the aggregated masses in many of the cells were re-dissolved, in the same manner as occurs with leaves in a state of nature when they re-expand after having caught insects.

In a leaf which had been left for 22 hrs. in a solution of one part of the carbonate to 292 of water, some spheres of protoplasm (formed by the self-division of a bag-like mass) were gently pressed beneath a covering glass, and then examined under a high power. They were now distinctly divided by well-defined radiating fissures, or were broken up into separate fragments with sharp edges; and they were solid to the centre. In the larger broken spheres the central part was more opaque, darker-coloured, and less brittle than the exterior; the latter alone being in some cases penetrated by the fissures. In many of the spheres the line of separation between the outer and inner parts was tolerably well defined. The outer parts were of exactly the same very pale purple tint, as that of the last formed smaller spheres; and these latter did not include any darker central core.

From these several facts we may conclude that when vigorous dark-coloured leaves are subjected to the action of carbonate of [page 48] ammonia, the fluid within the cells of the tentacles often aggregates exteriorly into coherent viscid matter, forming a kind of bag. Small spheres sometimes appear within this bag, and the whole generally soon divides into two or more spheres, which repeatedly coalesce and redivide. After a longer or shorter time the granules in the colourless layer of protoplasm, which flows round the walls, are drawn to and unite with the larger spheres, or form small independent spheres; these latter being of a much paler colour, and more brittle than the first aggregated masses. After the granules of protoplasm have been thus attracted, the layer of flowing protoplasm can no longer be distinguished, though a current of limpid fluid still flows round the walls.

If a leaf is immersed in a very strong, almost concentrated, solution of carbonate of ammonia, the glands are instantly blackened, and they secrete copiously; but no movement of the tentacles ensues. Two leaves thus treated became after 1 hr. flaccid, and seemed killed; all the cells in their tentacles contained spheres of protoplasm, but these were small and discoloured. Two other leaves were placed in a solution not quite so strong, and there was well-marked aggregation in 30 m. After 24 hrs. the spherical or more commonly oblong masses of protoplasm became opaque and granular, instead of being as usual translucent; and in the lower cells there were only innumerable minute spherical granules. It was evident that the strength of the solution had interfered with the completion of the process, as we shall see likewise follows from too great heat.

All the foregoing observations relate to the exterior tentacles, which are of a purple colour; but the green pedicels of the short central tentacles are acted on by the carbonate, and by an infusion of raw meat, in exactly the same manner, with the sole difference that the aggregated masses are of a greenish colour; so that the process is in no way dependent on the colour of the fluid within the cells.

Finally, the most remarkable fact with respect to this salt is the extraordinary small amount which suffices to cause aggregation. Full details will be given in the seventh chapter, and here it will be enough to say that with a sensitive leaf the absorption by a gland of 1/134400 of a grain (.000482 mgr.) is enough to cause in the course of one hour well-marked aggregation in the cells immediately beneath the gland.

The Effects of certain other Salts and Fluids.—Two leaves were placed in a solution of one part of acetate of ammonia to about [page 49] 146 of water, and were acted on quite as energetically, but perhaps not quite so quickly, as by the carbonate. After 10 m. the glands were black, and in the cells beneath them there were traces of aggregation, which after 15 m. was well marked, extending down the tentacles for a length equal to that of the glands. After 2 hrs. the contents of almost all the cells in all the tentacles were broken up into masses of protoplasm. A leaf was immersed in a solution of one part of oxalate of ammonia to 146 of water; and after 24 m. some, but not a conspicuous, change could be seen within the cells beneath the glands. After 47 m. plenty of spherical masses of protoplasm were formed, and these extended down the tentacles for about the length of the glands. This salt, therefore, does not act so quickly as the carbonate. With respect to the citrate of ammonia, a leaf was placed in a little solution of the above strength, and there was not even a trace of aggregation in the cells beneath the glands, until 56 m. had elapsed; but it was well marked after 2 hrs. 20 m. On another occasion a leaf was placed in a stronger solution, of one part of the citrate to 109 of water (4 grs. to 1 oz.), and at the same time another leaf in a solution of the carbonate of the same strength. The glands of the latter were blackened in less than 2 m., and after 1 hr. 45 m. the aggregated masses, which were spherical and very dark-coloured, extended down all the tentacles, for between half and two-thirds of their lengths; whereas in the leaf immersed in the citrate the glands, after 30 m., were of a dark red, and the aggregated masses in the cells beneath them pink and elongated. After 1 hr. 45 m. these masses extended down for only about one-fifth or one-fourth of the length of the tentacles.

Two leaves were placed, each in ten minims of a solution of one part of nitrate of ammonia to 5250 of water (1 gr. to 12 oz.), so that each leaf received 1/576 of a grain (.1124 mgr.). This quantity caused all the tentacles to be inflected, but after 24 hrs. there was only a trace of aggregation. One of these same leaves was then placed in a weak solution of the carbonate, and after 1 hr. 45 m. the tentacles for half their lengths showed an astonishing degree of aggregation. Two other leaves were then placed in a much stronger solution of one part of the nitrate to 146 of water (3 grs. to 1 oz.); in one of these there was no marked change after 3 hrs.; but in the other there was a trace of aggregation after 52 m., and this was plainly marked after 1 hr. 22 m., but even after 2 hrs. 12 m. there was certainly not more aggregation than would have fol- [page 50] lowed from an immersion of from 5 m. to 10 m. in an equally strong solution of the carbonate.

Lastly, a leaf was placed in thirty minims of a solution of one part of phosphate of ammonia to 43,750 of water (1 gr. to 100 oz.), so that it received 1/1600 of a grain (.04079 mgr.); this soon caused the tentacles to be strongly inflected; and after 24 hrs. the contents of the cells were aggregated into oval and irregularly globular masses, with a conspicuous current of protoplasm flowing round the walls. But after so long an interval aggregation would have ensued, whatever had caused inflection.

Only a few other salts, besides those of ammonia, were tried in relation to the process of aggregation. A leaf was placed in a solution of one part of chloride of sodium to 218 of water, and after 1 hr. the contents of the cells were aggregated into small, irregularly globular, brownish masses; these after 2 hrs. were almost disintegrated and pulpy. It was evident that the protoplasm had been injuriously affected; and soon afterwards some of the cells appeared quite empty. These effects differ altogether from those produced by the several salts of ammonia, as well as by various organic fluids, and by inorganic particles placed on the glands. A solution of the same strength of carbonate of soda and carbonate of potash acted in nearly the same manner as the chloride; and here again, after 2 hrs. 30 m., the outer cells of some of the glands had emptied themselves of their brown pulpy contents. We shall see in the eighth chapter that solutions of several salts of soda of half the above strength cause inflection, but do not injure the leaves. Weak solutions of sulphate of quinine, of nicotine, camphor, poison of the cobra, &c., soon induce well-marked aggregation; whereas certain other substances (for instance, a solution of curare) have no such tendency.

Many acids, though much diluted, are poisonous; and though, as will be shown in the eighth chapter, they cause the tentacles to bend, they do not excite true aggregation. Thus leaves were placed in a solution of one part of benzoic acid to 437 of water; and in 15 m. the purple fluid within the cells had shrunk a little from the walls, yet when carefully examined after 1 hr. 20 m., there was no true aggregation; and after 24 hrs. the leaf was evidently dead. Other leaves in iodic acid, diluted to the same degree, showed after 2 hrs. 15 m. the same shrunken appearance of the purple fluid within the cells; and these, after 6 hrs. 15 m., were seen under a high power to be filled with excessively minute spheres of dull reddish protoplasm, [page 51] which by the next morning, after 24 hrs., had almost disappeared, the leaf being evidently dead. Nor was there any true aggregation in leaves immersed in propionic acid of the same strength; but in this case the protoplasm was collected in irregular masses towards the bases of the lower cells of the tentacles.

A filtered infusion of raw meat induces strong aggregation, but not very quickly. In one leaf thus immersed there was a little aggregation after 1 hr. 20 m., and in another after 1 hr. 50 m. With other leaves a considerably longer time was required: for instance, one immersed for 5 hrs. showed no aggregation, but was plainly acted on in 5 m.; when placed in a few drops of a solution of one part of carbonate of ammonia to 146 of water. Some leaves were left in the infusion for 24 hrs., and these became aggregated to a wonderful degree, so that the inflected tentacles presented to the naked eye a plainly mottled appearance. The little masses of purple protoplasm were generally oval or beaded, and not nearly so often spherical as in the case of leaves subjected to carbonate of ammonia. They underwent incessant changes of form; and the current of colourless protoplasm round the walls was conspicuously plain after an immersion of 25 hrs. Raw meat is too powerful a stimulant, and even small bits generally injure, and sometimes kill, the leaves to which they are given: the aggregated masses of protoplasm become dingy or almost colourless, and present an unusual granular appearance, as is likewise the case with leaves which have been immersed in a very strong solution of carbonate of ammonia. A leaf placed in milk had the contents of its cells somewhat aggregated in 1 hr. Two other leaves, one immersed in human saliva for 2 hrs. 30 m., and another in unboiled white of egg for 1 hr. 30 m., were not action on in this manner; though they undoubtedly would have been so, had more time been allowed. These same two leaves, on being afterwards placed in a solution of carbonate of ammonia (3 grs. to 1 oz.), had their cells aggregated, the one in 10 m. and the other in 5 m.

Several leaves were left for 4 hrs. 30 m. in a solution of one part of white sugar to 146 of water, and no aggregation ensued; on being placed in a solution of this same strength of carbonate of ammonia, they were acted on in 5 m.; as was likewise a leaf which had been left for 1 hr. 45 m. in a moderately thick solution of gum arabic. Several other leaves were immersed for some hours in denser solutions of sugar, gum, and starch, and they had the contents of their cells greatly aggregated. This [page 52] effect may be attributed to exosmose; for the leaves in the syrup became quite flaccid, and those in the gum and starch somewhat flaccid, with their tentacles twisted about in the most irregular manner, the longer ones like corkscrews. We shall hereafter see that solutions of these substances, when placed on the discs of leaves, do not incite inflection. Particles of soft sugar were added to the secretion round several glands and were soon dissolved, causing a great increase of the secretion, no doubt by exosmose; and after 24 hrs. the cells showed a certain amount of aggregation, though the tentacles were not inflected. Glycerine causes in a few minutes well-pronounced aggregation, commencing as usual within the glands and then travelling down the tentacles; and this I presume may be attributed to the strong attraction of this substance for water. Immersion for several hours in water causes some degree of aggregation. Twenty leaves were first carefully examined, and re-examined after having been left immersed in distilled water for various periods, with the following results. It is rare to find even a trace of aggregation until 4 or 5 and generally not until several more hours have elapsed. When however a leaf becomes quickly inflected in water, as sometimes happens, especially during very warm weather, aggregation may occur in little over 1 hr. In all cases leaves left in water for more than 24 hrs. have their glands blackened, which shows that their contents are aggregated; and in the specimens which were carefully examined, there was fairly well-marked aggregation in the upper cells of the pedicels. These trials were made with cut off-leaves, and it occurred to me that this circumstance might influence the result, as the footstalks would not perhaps absorb water quickly enough to supply the glands as they continued to secrete. But this view was proved erroneous, for a plant with uninjured roots, bearing four leaves, was submerged in distilled water for 47 hrs., and the glands were blackened, though the tentacles were very little inflected. In one of these leaves there was only a slight degree of aggregation in the tentacles; in the second rather more, the purple contents of the cells being a little separated from the walls; in the third and fourth, which were pale leaves, the aggregation in the upper parts of the pedicels was well marked. In these leaves the little masses of protoplasm, many of which were oval, slowly changed their forms and positions; so that a submergence for 47 hrs. had not killed the protoplasm. In a previous trial with a submerged plant, the tentacles were not in the least inflected. [page 53]

Heat induces aggregation. A leaf, with the cells of the tentacles containing only homogeneous fluid, was waved about for 1 m. in water at 130o Fahr. (54o.4 Cent.) and was then examined under the microscope as quickly as possible, that is in 2 m. or 3 m.; and by this time the contents of the cells had undergone some degree of aggregation. A second leaf was waved for 2 m. in water at 125o (51o.6 Cent.) and quickly examined as before; the tentacles were well inflected; the purple fluid in all the cells had shrunk a little from the walls, and contained many oval and elongated masses of protoplasm, with a few minute spheres. A third leaf was left in water at 125o, until it cooled, and when examined after 1 hr. 45 m., the inflected tentacles showed some aggregation, which became after 3 hrs. more strongly marked, but did not subsequently increase. Lastly, a leaf was waved for 1 m. in water at 120o (48o.8 Cent.) and then left for 1 hr. 26 m. in cold water; the tentacles were but little inflected, and there was only here and there a trace of aggregation. In all these and other trials with warm water the protoplasm showed much less tendency to aggregate into spherical masses than when excited by carbonate of ammonia.

Redissolution of the Aggregated Masses of Protoplasm.—As soon as tentacles which have clasped an insect or any inorganic object, or have been in any way excited, have fully re-expanded, the aggregated masses of protoplasm are redissolved and disappear; the cells being now refilled with homogeneous purple fluid as they were before the tentacles were inflected. The process of redissolution in all cases commences at the bases of the tentacles, and proceeds up them towards the glands. In old leaves, however, especially in those which have been several times in action, the protoplasm in the uppermost cells of the pedicels remains in a permanently more or less aggregated condition. In order to observe the process of redissolution, the following observations were made: a leaf was left for 24 hrs. in a little solution of one part of carbonate of ammonia to 218 of water, and the protoplasm was as usual aggregated into numberless purple spheres, which were incessantly changing their forms. The leaf was then washed and placed in distilled water, and after 3 hrs. 15 m. some few of the spheres began to show by their less clearly defined edges signs of redissolution. After 9 hrs. many of them had become elongated, and the surrounding fluid in the cells was slightly more coloured, showing plainly that redissolution had commenced. After 24 hrs., though many cells still contained spheres, here and there one [page 54] could be seen filled with purple fluid, without a vestige of aggregated protoplasm; the whole having been redissolved. A leaf with aggregated masses, caused by its having been waved for 2 m. in water at the temperature of 125o Fahr., was left in cold water, and after 11 hrs. the protoplasm showed traces of incipient redissolution. When again examined three days after its immersion in the warm water, there was a conspicuous difference, though the protoplasm was still somewhat aggregated. Another leaf, with the contents of all the cells strongly aggregated from the action of a weak solution of phosphate of ammonia, was left for between three and four days in a mixture (known to be innocuous) of one drachm of alcohol to eight drachms of water, and when re-examined every trace of aggregation had disappeared, the cells being now filled with homogeneous fluid.

We have seen that leaves immersed for some hours in dense solutions of sugar, gum, and starch, have the contents of their cells greatly aggregated, and are rendered more or less flaccid, with the tentacles irregularly contorted. These leaves, after being left for four days in distilled water, became less flaccid, with their tentacles partially re-expanded, and the aggregated masses of protoplasm were partially redissolved. A leaf with its tentacles closely clasped over a fly, and with the contents of the cells strongly aggregated, was placed in a little sherry wine; after 2 hrs. several of the tentacles had re-expanded, and the others could by a mere touch be pushed back into their properly expanded positions, and now all traces of aggregation had disappeared, the cells being filled with perfectly homogeneous pink fluid. The redissolution in these cases may, I presume, be attributed to endosmose.]

On the Proximate Causes of the Process of Aggregation.

As most of the stimulants which cause the inflection of the tentacles likewise induce aggregation in the contents of their cells, this latter process might be thought to be the direct result of inflection; but this is not the case. If leaves are placed in rather strong solutions of carbonate of ammonia, for instance of three or four, and even sometimes of only two grains to the ounce of water (i.e. one part to 109, or 146, or [page 55] 218, of water), the tentacles are paralysed, and do not become inflected, yet they soon exhibit strongly marked aggregation. Moreover, the short central tentacles of a leaf which has been immersed in a weak solution of any salt of ammonia, or in any nitrogenous organic fluid, do not become in the least inflected; nevertheless they exhibit all the phenomena of aggregation. On the other hand, several acids cause strongly pronounced inflection, but no aggregation.

It is an important fact that when an organic or inorganic object is placed on the glands of the disc, and the exterior tentacles are thus caused to bend inwards, not only is the secretion from the glands of the latter increased in quantity and rendered acid, but the contents of the cells of their pedicels become aggregated. The process always commences in the glands, although these have not as yet touched any object. Some force or influence must, therefore, be transmitted from the central glands to the exterior tentacles, first to near their bases causing this part to bend, and next to the glands causing them to secrete more copiously. After a short time the glands, thus indirectly excited, transmit or reflect some influence down their own pedicels, inducing aggregation in cell beneath cell to their bases.

It seems at first sight a probable view that aggregation is due to the glands being excited to secrete more copiously, so that sufficient fluid is not left in their cells, and in the cells of the pedicels, to hold the protoplasm in solution. In favour of this view is the fact that aggregation follows the inflection of the tentacles, and during the movement the glands generally, or, as I believe, always, secrete more copiously than they did before. Again, during the re-expansion [page 56] of the tentacles, the glands secrete less freely, or quite cease to secrete, and the aggregated masses of protoplasm are then redissolved. Moreover, when leaves are immersed in dense vegetable solutions, or in glycerine, the fluid within the gland-cells passes outwards, and there is aggregation; and when the leaves are afterwards immersed in water, or in an innocuous fluid of less specific gravity than water, the protoplasm is redissolved, and this, no doubt, is due to endosmose.

Opposed to this view, that aggregation is caused by the outward passage of fluid from the cells, are the following facts. There seems no close relation between the degree of increased secretion and that of aggregation. Thus a particle of sugar added to the secretion round a gland causes a much greater increase of secretion, and much less aggregation, than does a particle of carbonate of ammonia given in the same manner. It does not appear probable that pure water would cause much exosmose, and yet aggregation often follows from an immersion in water of between 16 hrs. and 24 hrs., and always after from 24 hrs. to 48 hrs. Still less probable is it that water at a temperature of from 125o to 130o Fahr. (51o.6 to 54o.4 Cent.) should cause fluid to pass, not only from the glands, but from all the cells of the tentacles down to their bases, so quickly that aggregation is induced within 2 m. or 3 m. Another strong argument against this view is, that, after complete aggregation, the spheres and oval masses of protoplasm float about in an abundant supply of thin colourless fluid; so that at least the latter stages of the process cannot be due to the want of fluid to hold the protoplasm in solution. There is still stronger evidence that aggregation is independent of secretion; for the papillae, described in the first chapter, with which the [page 57] leaves are studded are not glandular, and do not secrete, yet they rapidly absorb carbonate of ammonia or an infusion of raw meat, and their contents then quickly undergo aggregation, which afterwards spreads into the cells of the surrounding tissues. We shall hereafter see that the purple fluid within the sensitive filaments of Dionaea, which do not secrete, likewise undergoes aggregation from the action of a weak solution of carbonate of ammonia.

The process of aggregation is a vital one; by which I mean that the contents of the cells must be alive and uninjured to be thus affected, and they must be in an oxygenated condition for the transmission of the process at the proper rate. Some tentacles in a drop of water were strongly pressed beneath a slip of glass; many of the cells were ruptured, and pulpy matter of a purple colour, with granules of all sizes and shapes, exuded, but hardly any of the cells were completely emptied. I then added a minute drop of a solution of one part of carbonate of ammonia to 109 of water, and after 1 hr. examined the specimens. Here and there a few cells, both in the glands and in the pedicels, had escaped being ruptured, and their contents were well aggregated into spheres which were constantly changing their forms and positions, and a current could still be seen flowing along the walls; so that the protoplasm was alive. On the other hand, the exuded matter, which was now almost colourless instead of being purple, did not exhibit a trace of aggregation. Nor was there a trace in the many cells which were ruptured, but which had not been completely emptied of their contents. Though I looked carefully, no signs of a current could be seen within these ruptured cells. They had evidently been killed by the pressure; and the matter which they [page 58] still contained did not undergo aggregation any more than that which had exuded. In these specimens, as I may add, the individuality of the life of each cell was well illustrated.

A full account will be given in the next chapter of the effects of heat on the leaves, and I need here only state that leaves immersed for a short time in water at a temperature of 120oFahr. (48o.8 Cent.), which, as we have seen, does not immediately induce aggregation, were then placed in a few drops of a strong solution of one part of carbonate of ammonia to 109 of water, and became finely aggregated. On the other hand, leaves, after an immersion in water at 150o (65o.5 Cent.), on being placed in the same strong solution, did not undergo aggregation, the cells becoming filled with brownish, pulpy, or muddy matter. With leaves subjected to temperatures between these two extremes of 120o and 150o Fahr. (48o.8 and 65o.5 Cent.), there were gradations in the completeness of the process; the former temperature not preventing aggregation from the subsequent action of carbonate of ammonia, the latter quite stopping it. Thus, leaves immersed in water, heated to 130o (54o.4 Cent.), and then in the solution, formed perfectly defined spheres, but these were decidedly smaller than in ordinary cases. With other leaves heated to 140o (60o Cent.), the spheres were extremely small, yet well defined, but many of the cells contained, in addition, some brownish pulpy matter. In two cases of leaves heated to 145o (62o.7 Cent.), a few tentacles could be found with some of their cells containing a few minute spheres; whilst the other cells and other whole tentacles included only the brownish, disintegrated or pulpy matter.

The fluid within the cells of the tentacles must be in an oxygenated condition, in order that the force or [page 59] influence which induces aggregation should be transmitted at the proper rate from cell to cell. A plant, with its roots in water, was left for 45 m. in a vessel containing 122 oz. of carbonic acid. A leaf from this plant, and, for comparison, one from a fresh plant, were both immersed for 1 hr. in a rather strong solution of carbonate of ammonia. They were then compared, and certainly there was much less aggregation in the leaf which had been subjected to the carbonic acid than in the other. Another plant was exposed in the same vessel for 2 hrs. to carbonic acid, and one of its leaves was then placed in a solution of one part of the carbonate to 437 of water; the glands were instantly blackened, showing that they had absorbed, and that their contents were aggregated; but in the cells close beneath the glands there was no aggregation even after an interval of 3 hrs. After 4 hrs. 15 m. a few minute spheres of protoplasm were formed in these cells, but even after 5 hrs. 30 m. the aggregation did not extend down the pedicels for a length equal to that of the glands. After numberless trials with fresh leaves immersed in a solution of this strength, I have never seen the aggregating action transmitted at nearly so slow a rate. Another plant was left for 2 hrs. in carbonic acid, but was then exposed for 20 m. to the open air, during which time the leaves, being of a red colour, would have absorbed some oxygen. One of them, as well as a fresh leaf for comparison, were now immersed in the same solution as before. The former were looked at repeatedly, and after an interval of 65 m. a few spheres of protoplasm were first observed in the cells close beneath the glands, but only in two or three of the longer tentacles. After 3 hrs. the aggregation had travelled down the pedicels of a few of the tentacles [page 60] for a length equal to that of the glands. On the other hand, in the fresh leaf similarly treated, aggregation was plain in many of the tentacles after 15 m.; after 65 m. it had extended down the pedicels for four, five, or more times the lengths of the glands; and after 3 hrs. the cells of all the tentacles were affected for one-third or one-half of their entire lengths. Hence there can be no doubt that the exposure of leaves to carbonic acid either stops for a time the process of aggregation, or checks the transmission of the proper influence when the glands are subsequently excited by carbonate of ammonia; and this substance acts more promptly and energetically than any other. It is known that the protoplasm of plants exhibits its spontaneous movements only as long as it is in an oxygenated condition; and so it is with the white corpuscles of the blood, only as long as they receive oxygen from the red corpuscles;* but the cases above given are somewhat different, as they relate to the delay in the generation or aggregation of the masses of protoplasm by the exclusion of oxygen.

Summary and Concluding Remarks.—The process of aggregation is independent of the inflection of the tentacles and of increased secretion from the glands. It commences within the glands, whether these have been directly excited, or indirectly by a stimulus received from other glands. In both cases the process is transmitted from cell to cell down the whole length of the tentacles, being arrested for a short time at each transverse partition. With pale-coloured leaves the first change which is perceptible, but only

* With respect to plants, Sachs, 'Trait de Bot.' 3rd edit., 1874, p. 864. On blood corpuscles, see 'Quarterly Journal of Microscopical Science,' April 1874, p. 185.' [page 61]

under a high power, is the appearance of the finest granules in the fluid within the cells, making it slightly cloudy. These granules soon aggregate into small globular masses. I have seen a cloud of this kind appear in 10 s. after a drop of a solution of carbonate of ammonia had been given to a gland. With dark red leaves the first visible change often is the conversion of the outer layer of the fluid within the cells into bag-like masses. The aggregated masses, however they may have been developed, incessantly change their forms and positions. They are not filled with fluid, but are solid to their centres. Ultimately the colourless granules in the protoplasm which flows round the walls coalesce with the central spheres or masses; but there is still a current of limpid fluid flowing within the cells. As soon as the tentacles fully re-expand, the aggregated masses are redissolved, and the cells become filled with homogeneous purple fluid, as they were at first. The process of redissolution commences at the bases of the tentacles, thence proceeding upwards to the glands; and, therefore, in a reversed direction to that of aggregation.

Aggregation is excited by the most diversified causes,—by the glands being several times touched,—by the pressure of particles of any kind, and as these are supported by the dense secretion, they can hardly press on the glands with the weight of a millionth of a grain,*—by the tentacles being cut off close beneath

* According to Hofmeister (as quoted by Sachs, 'Trait de Bot.' 1874, p. 958), very slight pressure on the cell-membrane arrests immediately the movements of the protoplasm, and even determines its separation from the walls. But the process of aggregation is a different phenomenon, as it relates to the contents of the cells, and only secondarily to the layer of protoplasm which flows along the walls; though no doubt the effects of pressure or of a touch on the outside must be transmitted through this layer. [page 62]

the glands,—by the glands absorbing various fluids or matter dissolved out of certain bodies,—by exosmose,—and by a certain degree of heat. On the other hand, a temperature of about 150o Fahr. (65o.5 Cent.) does not excite aggregation; nor does the sudden crushing of a gland. If a cell is ruptured, neither the exuded matter nor that which still remains within the cell undergoes aggregation when carbonate of ammonia is added. A very strong solution of this salt and rather large bits of raw meat prevent the aggregated masses being well developed. From these facts we may conclude that the protoplasmic fluid within a cell does not become aggregated unless it be in a living state, and only imperfectly if the cell has been injured. We have also seen that the fluid must be in an oxygenated state, in order that the process of aggregation should travel from cell to cell at the proper rate.

Various nitrogenous organic fluids and salts of ammonia induce aggregation, but in different degrees and at very different rates. Carbonate of ammonia is the most powerful of all known substances; the absorption of 1/134400 of a grain (.000482 mg.) by a gland suffices to cause all the cells of the same tentacle to become aggregated. The first effect of the carbonate and of certain other salts of ammonia, as well as of some other fluids, is the darkening or blackening of the glands. This follows even from long immersion in cold distilled water. It apparently depends in chief part on the strong aggregation of their cell-contents, which thus become opaque, and do not reflect light. Some other fluids render the glands of a brighter red; whilst certain acids, though much diluted, the poison of the cobra-snake, &c., make the glands perfectly white and opaque; and this seems to depend on the coagulation of their contents without [page 63] any aggregation. Nevertheless, before being thus affected, they are able, at least in some cases, to excite aggregation in their own tentacles.

That the central glands, if irritated, send centrifugally some influence to the exterior glands, causing them to send back a centripetal influence inducing aggregation, is perhaps the most interesting fact given in this chapter. But the whole process of aggregation is in itself a striking phenomenon. Whenever the peripheral extremity of a nerve is touched or pressed, and a sensation is felt, it is believed that an invisible molecular change is sent from one end of the nerve to the other; but when a gland of Drosera is repeatedly touched or gently pressed, we can actually see a molecular change proceeding from the gland down the tentacle; though this change is probably of a very different nature from that in a nerve. Finally, as so many and such widely different causes excite aggregation, it would appear that the living matter within the gland-cells is in so unstable a condition that almost any disturbance suffices to change its molecular nature, as in the case of certain chemical compounds. And this change in the glands, whether excited directly, or indirectly by a stimulus received from other glands, is transmitted from cell to cell, causing granules of protoplasm either to be actually generated in the previously limpid fluid or to coalesce and thus to become visible.

Supplementary Observations on the Process of Aggregation in the Roots of Plants.

It will hereafter be seen that a weak solution of the carbonate of ammonia induces aggregation in the cells of the roots of Drosera; and this led me to make a few trials on the roots of other plants. I dug up in the latter part of October the first weed which I met with, viz. Euphorbia peplus, being care- [page 64] ful not to injure the roots; these were washed and placed in a little solution of one part of carbonate of ammonia to 146 of water. In less than one minute I saw a cloud travelling from cell to cell up the roots, with wonderful rapidity. After from 8 m. to 9 m. the fine granules, which caused this cloudy appearance, became aggregated towards the extremities of the roots into quadrangular masses of brown matter; and some of these soon changed their forms and became spherical. Some of the cells, however, remained unaffected. I repeated the experiment with another plant of the same species, but before I could get the specimen into focus under the microscope, clouds of granules and quadrangular masses of reddish and brown matter were formed, and had run far up all the roots. A fresh root was now left for 18 hrs. in a drachm of a solution of one part of the carbonate to 437 of water, so that it received 1/8 of a grain, or 2.024 mg. When examined, the cells of all the roots throughout their whole length contained aggregated masses of reddish and brown matter. Before making these experiments, several roots were closely examined, and not a trace of the cloudy appearance or of the granular masses could be seen in any of them. Roots were also immersed for 35 m. in a solution of one part of carbonate of potash to 218 of water; but this salt produced no effect.

I may here add that thin slices of the stem of the Euphorbia were placed in the same solution, and the cells which were green instantly became cloudy, whilst others which were before colourless were clouded with brown, owing to the formation of numerous granules of this tint. I have also seen with various kinds of leaves, left for some time in a solution of carbonate of ammonia, that the grains of chlorophyll ran together and partially coalesced; and this seems to be a form of aggregation.

Plants of duck-weed (Lemna) were left for between 30 m. and 45 m. in a solution of one part of this same salt to 146 of water, and three of their roots were then examined. In two of them, all the cells which had previously contained only limpid fluid now included little green spheres. After from 1 1/2 hr. to 2 hrs. similar spheres appeared in the cells on the borders of the leaves; but whether the ammonia had travelled up the roots or had been directly absorbed by the leaves, I cannot say. As one species, Lemna arrhiza, produces no roots, the latter alternative is perhaps the most probable. After about 2 1/2 hrs. some of the little green spheres in the roots were broken up into small granules which exhibited Brownian movements. Some duck-weed was also left for 1 hr. 30 m. in a solution of one part of [page 65] carbonate of potash to 218 of water, and no decided change could be perceived in the cells of the roots; but when these same roots were placed for 25 m. in a solution of carbonate of ammonia of the same strength, little green spheres were formed.

A green marine alga was left for some time in this same solution, but was very doubtfully affected. On the other hand, a red marine alga, with finely pinnated fronds, was strongly affected. The contents of the cells aggregated themselves into broken rings, still of a red colour, which very slowly and slightly changed their shapes, and the central spaces within these rings became cloudy with red granular matter. The facts here given (whether they are new, I know not) indicate that interesting results would perhaps be gained by observing the action of various saline solutions and other fluids on the roots of plants. [page 66]



CHAPTER IV.

THE EFFECTS OF HEAT ON THE LEAVES.

Nature of the experiments—Effects of boiling water—Warm water causes rapid inflection— Water at a higher temperature does not cause immediate inflection, but does not kill the leaves, as shown by their subsequent re-expansion and by the aggregation of the protoplasm— A still higher temperature kills the leaves and coagulates the albuminous contents of the glands.

IN my observations on Drosera rotundifolia, the leaves seemed to be more quickly inflected over animal substances, and to remain inflected for a longer period during very warm than during cold weather. I wished, therefore, to ascertain whether heat alone would induce inflection, and what temperature was the most efficient. Another interesting point presented itself, namely, at what degree life was extinguished; for Drosera offers unusual facilities in this respect, not in the loss of the power of inflection, but in that of subsequent re-expansion, and more especially in the failure of the protoplasm to become aggregated, when the leaves after being heated are immersed in a solution of carbonate of ammonia.*

* When my experiments on the effects of heat were made, I was not aware that the subject had been carefully investigated by several observers. For instance, Sachs is convinced ('Trait de Botanique,' 1874, pp. 772, 854) that the most different kinds of plants all perish if kept for 10 m. in water at 45o to 46o Cent., or 113o to 115o Fahr.; and he concludes that the protoplasm within their cells always coagulates, if in a damp condition, at a temperature of between 50oand 60o Cent., or 122o to 140o Fahr. Max Schultze and Khne (as quoted by Dr. Bastian in 'Contemp. Review,' 1874, p. 528) "found that the protoplasm of plant-cells, with which they experimented, was always killed and [[page 67]] altered by a very brief exposure to a temperature of 118 1/2o Fahr. as a maximum." As my results are deduced from special phenomena, namely, the subsequent aggregation of the protoplasm and the re-expansion of the tentacles, they seem to me worth giving. We shall find that Drosera resists heat somewhat better than most other plants. That there should be considerable differences in this respect is not surprising, considering that some low vegetable organisms grow in hot springs—cases of which have been collected by Prof. Wyman ('American Journal of Science,' vol. xliv. 1867). Thus, Dr. Hooker found Confervae in water at 168o Fahr.; Humboldt, at 185o Fahr.; and Descloizeaux, at 208o Fahr.) [page 67]

[My experiments were tried in the following manner. Leaves were cut off, and this does not in the least interfere with their powers; for instance, three cut off leaves, with bits of meat placed on them, were kept in a damp atmosphere, and after 23 hrs. closely embraced the meat both with their tentacles and blades; and the protoplasm within their cells was well aggregated. Three ounces of doubly distilled water was heated in a porcelain vessel, with a delicate thermometer having a long bulb obliquely suspended in it. The water was gradually raised to the required temperature by a spirit-lamp moved about under the vessel; and in all cases the leaves were continually waved for some minutes close to the bulb. They were then placed in cold water, or in a solution of carbonate of ammonia. In other cases they were left in the water, which had been raised to a certain temperature, until it cooled. Again in other cases the leaves were suddenly plunged into water of a certain temperature, and kept there for a specified time. Considering that the tentacles are extremely delicate, and that their coats are very thin, it seems scarcely possible that the fluid contents of their cells should not have been heated to within a degree or two of the temperature of the surrounding water. Any further precautions would, I think, have been superfluous, as the leaves from age or constitutional causes differ slightly in their sensitiveness to heat.

It will be convenient first briefly to describe the effects of immersion for thirty seconds in boiling water. The leaves are rendered flaccid, with their tentacles bowed backwards, which, as we shall see in a future chapter, is probably due to their outer surfaces retaining their elasticity for a longer period than their inner surfaces retain the power of contraction. The purple fluid within the cells of the pedicels is rendered finely granular, but there is no true aggregation; nor does this follow [page 68] when the leaves are subsequently placed in a solution of carbonate of ammonia. But the most remarkable change is that the glands become opaque and uniformly white; and this may be attributed to the coagulation of their albuminous contents.

My first and preliminary experiment consisted in putting seven leaves in the same vessel of water, and warming it slowly up to the temperature of 110o Fahr. (43o.3 Cent.); a leaf being taken out as soon as the temperature rose to 80o (26o.6 Cent.), another at 85o, another at 90o, and so on. Each leaf, when taken out, was placed in water at the temperature of my room, and the tentacles of all soon became slightly, though irregularly, inflected. They were now removed from the cold water and kept in damp air, with bits of meat placed on their discs. The leaf which had been exposed to the temperature of 110o became in 15 m. greatly inflected; and in 2 hrs. every single tentacle closely embraced the meat. So it was, but after rather longer intervals, with the six other leaves. It appears, therefore, that the warm bath had increased their sensitiveness when excited by meat.

I next observed the degree of inflection which leaves underwent within stated periods, whilst still immersed in warm water, kept as nearly as possible at the same temperature; but I will here and elsewhere give only a few of the many trials made. A leaf was left for 10 m. in water at 100o (37o.7 Cent.), but no inflection occurred. A second leaf, however, treated in the same manner, had a few of its exterior tentacles very slightly inflected in 6 m., and several irregularly but not closely inflected in 10 m. A third leaf, kept in water at 105o to 106o (40o.5 to 41o.1 Cent.), was very moderately inflected in 6 m. A fourth leaf, in water at 110o (43o.3 Cent.), was somewhat inflected in 4 m., and considerably so in from 6 to 7 m.

Three leaves were placed in water which was heated rather quickly, and by the time the temperature rose to 115o-116o (46o.1 to 46o.06 Cent.), all three were inflected. I then removed the lamp, and in a few minutes every single tentacle was closely inflected. The protoplasm within the cells was not killed, for it was seen to be in distinct movement; and the leaves, having been left in cold water for 20 hrs., re-expanded. Another leaf was immersed in water at 100o (37.7o Cent.), which was raised to 120o (48o.8 Cent.); and all the tentacles, except the extreme marginal ones, soon became closely inflected. The leaf was now placed in cold water, and in 7 hrs. 30 m. it had partly, and in 10 hrs. fully, re-expanded. On the following morning it was immersed in a weak solution of carbonate of [page 69] ammonia, and the glands quickly became black, with strongly marked aggregation in the tentacles, showing that the protoplasm was alive, and that the glands had not lost their power of absorption. Another leaf was placed in water at 110o (43o.3 Cent.) which was raised to 120o (48o.8 Cent.); and every tentacle, excepting one, was quickly and closely inflected. This leaf was now immersed in a few drops of a strong solution of carbonate of ammonia (one part to 109 of water); in 10 m. all the glands became intensely black, and in 2 hrs. the protoplasm in the cells of the pedicels was well aggregated. Another leaf was suddenly plunged, and as usual waved about, in water at 120o, and the tentacles became inflected in from 2 m. to 3 m., but only so as to stand at right angles to the disc. The leaf was now placed in the same solution (viz. one part of carbonate of ammonia to 109 of water, or 4 grs. to 1 oz., which I will for the future designate as the strong solution), and when I looked at it again after the interval of an hour, the glands were blackened, and there was well-marked aggregation. After an additional interval of 4 hrs. the tentacles had become much more inflected. It deserves notice that a solution as strong as this never causes inflection in ordinary cases. Lastly a leaf was suddenly placed in water at 125o (51o.6 Cent.), and was left in it until the water cooled; the tentacles were rendered of a bright red and soon became inflected. The contents of the cells underwent some degree of aggregation, which in the course of three hours increased; but the masses of protoplasm did not become spherical, as almost always occurs with leaves immersed in a solution of carbonate of ammonia.]

We learn from these cases that a temperature of from 120o to 125o (48o.8 to 51o.6 Cent.) excites the tentacles into quick movement, but does not kill the leaves, as shown either by their subsequent re-expansion or by the aggregation of the protoplasm. We shall now see that a temperature of 130o (54o.4 Cent.) is too high to cause immediate inflection, yet does not kill the leaves.

[Experiment 1.—A leaf was plunged, and as in all cases waved about for a few minutes, in water at 130o (54o.4 Cent.), but there was no trace of inflection; it was then placed in cold water, and after an interval of 15 m. very slow movement was [page 70] distinctly seen in a small mass of protoplasm in one of the cells of a tentacle.* After a few hours all the tentacles and the blade became inflected.

Experiment 2.—Another leaf was plunged into water at 130o to 131o, and as before there was no inflection. After being kept in cold water for an hour, it was placed in the strong solution of ammonia, and in the course of 55 m. the tentacles were considerably inflected. The glands, which before had been rendered of a brighter red, were now blackened. The protoplasm in the cells of the tentacles was distinctly aggregated; but the spheres were much smaller than those generated in unheated leaves when subjected to carbonate of ammonia. After an additional 2 hrs. all the tentacles, excepting six or seven, were closely inflected.

Experiment 3.—A similar experiment to the last, with exactly the same results.

Experiment 4.—A fine leaf was placed in water at 100o (37o.7 Cent.), which was then raised to 145o (62o.7 Cent.). Soon after immersion, there was, as might have been expected, strong inflection. The leaf was now removed and left in cold water; but from having been exposed to so high a temperature, it never re-expanded.

Experiment 5.—Leaf immersed at 130o (54o.4 Cent.), and the water raised to 145o (62o.7 Cent.), there was no immediate inflection; it was then placed in cold water, and after 1 hr. 20 m. some of the tentacles on one side became inflected. This leaf was now placed in the strong solution, and in 40 m. all the submarginal tentacles were well inflected, and the glands blackened. After an additional interval of 2 hrs. 45 m. all the tentacles, except eight or ten, were closely inflected, with their cells exhibiting a slight degree of aggregation; but the spheres of protoplasm were very small, and the cells of the exterior tentacles contained some pulpy or disintegrated brownish matter.

Experiments 6 and 7.—Two leaves were plunged in water at 135o (57o.2 Cent.) which was raised to 145o (62o.7 Cent.); neither became inflected. One of these, however, after having been left for 31 m. in cold water, exhibited some slight inflection, which increased after an additional interval of 1 hr. 45 m., until

* Sachs states ('Trait de Botanique,' 1874, p. 855) that the movements of the protoplasm in the hairs of a Cucurbita ceased after they were exposed for 1 m. in water to a temperature of 47o to 48o Cent., or 117o to 119o Fahr. [page 71]

all the tentacles, except sixteen or seventeen, were more or less inflected; but the leaf was so much injured that it never re-expanded. The other leaf, after having been left for half an hour in cold water, was put into the strong solution, but no inflection ensued; the glands, however, were blackened, and in some cells there was a little aggregation, the spheres of protoplasm being extremely small; in other cells, especially in the exterior tentacles, there was much greenish-brown pulpy matter.

Experiment 8.—A leaf was plunged and waved about for a few minutes in water at 140o (60oCent.), and was then left for half an hour in cold water, but there was no inflection. It was now placed in the strong solution, and after 2 hrs. 30 m. the inner submarginal tentacles were well inflected, with their glands blackened, and some imperfect aggregation in the cells of the pedicels. Three or four of the glands were spotted with the white porcelain-like structure, like that produced by boiling water. I have seen this result in no other instance after an immersion of only a few minutes in water at so low a temperature as 140o, and in only one leaf out of four, after a similar immersion at a temperature of 145o Fahr. On the other hand, with two leaves, one placed in water at 145o (62o.7 Cent.), and the other in water at 140o (60oCent.), both being left therein until the water cooled, the glands of both became white and porcelain-like. So that the duration of the immersion is an important element in the result.

Experiment 9.—A leaf was placed in water at 140o (60o Cent.), which was raised to 150o(65o.5 Cent.); there was no inflection; on the contrary, the outer tentacles were somewhat bowed backwards. The glands became like porcelain, but some of them were a little mottled with purple. The bases of the glands were often more affected than their summits. This leaf having been left in the strong solution did not undergo any inflection or aggregation.

Experiment 10.—A leaf was plunged in water at 150o to 150 1/2o (65o.5 Cent.); it became somewhat flaccid, with the outer tentacles slightly reflexed, and the inner ones a little bent inwards, but only towards their tips; and this latter fact shows that the movement was not one of true inflection, as the basal part alone normally bends. The tentacles were as usual rendered of a very bright red, with the glands almost white like porcelain, yet tinged with pink. The leaf having been placed in the strong solution, the cell-contents of the tentacles became of a muddy-brown, with no trace of aggregation. [page 72]

Experiment 11.—A leaf was immersed in water at 145o (62o.7 Cent.), which was raised to 156o (68o.8 Cent.). The tentacles became bright red and somewhat reflexed, with almost all the glands like porcelain; those on the disc being still pinkish, those near the margin quite white. The leaf being placed as usual first in cold water and then in the strong solution, the cells in the tentacles became of a muddy greenish brown, with the protoplasm not aggregated. Nevertheless, four of the glands escaped being rendered like porcelain, and the pedicels of these glands were spirally curled, like a French horn, towards their upper ends; but this can by no means be considered as a case of true inflection. The protoplasm within the cells of the twisted portions was aggregated into distinct though excessively minute purple spheres. This case shows clearly that the protoplasm, after having been exposed to a high temperature for a few minutes, is capable of aggregation when afterwards subjected to the action of carbonate of ammonia, unless the heat has been sufficient to cause coagulation.]

Concluding Remarks.—As the hair-like tentacles are extremely thin and have delicate walls, and as the leaves were waved about for some minutes close to the bulb of the thermometer, it seems scarcely possible that they should not have been raised very nearly to the temperature which the instrument indicated. From the eleven last observations we see that a temperature of 130o (54o.4 Cent.) never causes the immediate inflection of the tentacles, though a temperature from 120o to 125o (48o.8 to 51o.6 Cent.) quickly produces this effect. But the leaves are paralysed only for a time by a temperature of 130o, as afterwards, whether left in simple water or in a solution of carbonate of ammonia, they become inflected and their protoplasm undergoes aggregation. This great difference in the effects of a higher and lower temperature may be compared with that from immersion in strong and weak solutions of the salts of ammonia; for the former do not excite movement, whereas the latter act energetically. A temporary suspension of the [page 73] power of movement due to heat is called by Sachs* heat-rigidity; and this in the case of the sensitive-plant (Mimosa) is induced by its exposure for a few minutes to humid air, raised to 120o-122o Fahr., or 49o to 50o Cent. It deserves notice that the leaves of Drosera, after being immersed in water at 130o Fahr., are excited into movement by a solution of the carbonate so strong that it would paralyse ordinary leaves and cause no inflection.

The exposure of the leaves for a few minutes even to a temperature of 145o Fahr. (62o.7 Cent.) does not always kill them; as when afterwards left in cold water, or in a strong solution of carbonate of ammonia, they generally, though not always, become inflected; and the protoplasm within their cells undergoes aggregation, though the spheres thus formed are extremely small, with many of the cells partly filled with brownish muddy matter. In two instances, when leaves were immersed in water, at a lower temperature than 130o (54o.4 Cent.), which was then raised to 145o (62o.7 Cent.), they became during the earlier period of immersion inflected, but on being afterwards left in cold water were incapable of re-expansion. Exposure for a few minutes to a temperature of 145o sometimes causes some few of the more sensitive glands to be speckled with the porcelain-like appearance; and on one occasion this occurred at a temperature of 140o (60o Cent.). On another occasion, when a leaf was placed in water at this temperature of only 140o, and left therein till the water cooled, every gland became like porcelain. Exposure for a few minutes to a temperature of 150o (65o.5 Cent.) generally produces this effect, yet many glands retain a

* 'Trait de Bot.' 1874, p. 1034. [page 74]

pinkish colour, and many present a speckled appearance. This high temperature never causes true inflection; on the contrary, the tentacles commonly become reflexed, though to a less degree than when immersed in boiling water; and this apparently is due to their passive power of elasticity. After exposure to a temperature of 150o Fahr., the protoplasm, if subsequently subjected to carbonate of ammonia, instead of undergoing aggregation, is converted into disintegrated or pulpy discoloured matter. In short, the leaves are generally killed by this degree of heat; but owing to differences of age or constitution, they vary somewhat in this respect. In one anomalous case, four out of the many glands on a leaf, which had been immersed in water raised to 156o (68o.8 Cent.), escaped being rendered porcellanous;* and the protoplasm in the cells close beneath these glands underwent some slight, though imperfect, degree of aggregation.

Finally, it is a remarkable fact that the leaves of Drosera rotundifolia, which flourishes on bleak upland moors throughout Great Britain, and exists (Hooker) within the Arctic Circle, should be able to withstand for even a short time immersion in water heated to a temperature of 145o.

It may be worth adding that immersion in cold

* As the opacity and porcelain-like appearance of the glands is probably due to the coagulation of the albumen, I may add, on the authority of Dr. Burdon Sanderson, that albumen coagulates at about 155o, but, in presence of acids, the temperature of coagulation is lower. The leaves of Drosera contain an acid, and perhaps a difference in the amount contained may account for the slight differences in the results above recorded.

It appears that cold-blooded animals are, as might have been expected, far more sensitive to an increase of temperature than is Drosera. Thus, as I hear from Dr. Burdon Sanderson, a frog begins to be distressed in water at a temperature of only 85o Fahr. At 95o the muscles become rigid, and the animal dies in a stiffened condition. [page 75]

water does not cause any inflection: I suddenly placed four leaves, taken from plants which had been kept for several days at a high temperature, generally about 75o Fahr. (23o.8 Cent.), in water at 45o (7o.2 Cent.), but they were hardly at all affected; not so much as some other leaves from the same plants, which were at the same time immersed in water at 75o; for these became in a slight degree inflected. [page 76]



CHAPTER V.

THE EFFECTS OF NON-NITROGENOUS AND NITROGENOUS ORGANIC FLUIDS ON THE LEAVES.

Non-nitrogenous fluids—Solutions of gum arabic—Sugar—Starch—Diluted alcohol—Olive oil— Infusion and decoction of tea—Nitrogenous fluids—Milk—Urine—Liquid albumen—Infusion of raw meat—Impure mucus—Saliva—Solution of isinglass—Difference in the action of these two sets of fluids—Decoction of green peas—Decoction and infusion of cabbage—Decoction of grass leaves.

WHEN, in 1860, I first observed Drosera, and was led to believe that the leaves absorbed nutritious matter from the insects which they captured, it seemed to me a good plan to make some preliminary trials with a few common fluids, containing and not containing nitrogenous matter; and the results are worth giving.

In all the following cases a drop was allowed to fall from the same pointed instrument on the centre of the leaf; and by repeated trials one of these drops was ascertained to be on an average very nearly half a minim, or 1/960 of a fluid ounce, or .0295 ml. But these measurements obviously do not pretend to any strict accuracy; moreover, the drops of the viscid fluids were plainly larger than those of water. Only one leaf on the same plant was tried, and the plants were collected from two distant localities. The experiments were made during August and September. In judging of the effects, one caution is necessary: if a drop of any adhesive fluid is placed on an old or feeble leaf, the glands of which have ceased to secrete copiously, the drop sometimes dries up, especially if the plant [page 77] is kept in a room, and some of the central and submarginal tentacles are thus drawn together, giving to them the false appearance of having become inflected. This sometimes occurs with water, as it is rendered adhesive by mingling with the viscid secretion. Hence the only safe criterion, and to this alone I have trusted, is the bending inwards of the exterior tentacles, which have not been touched by the fluid, or at most only at their bases. In this case the movement is wholly due to the central glands having been stimulated by the fluid, and transmitting a motor impulse to the exterior tentacles. The blade of the leaf likewise often curves inwards, in the same manner as when an insect or bit of meat is placed on the disc. This latter movement is never caused, as far as I have seen, by the mere drying up of an adhesive fluid and the consequent drawing together of the tentacles.

First for the non-nitrogenous fluids. As a preliminary trial, drops of distilled water were placed on between thirty and forty leaves, and no effect whatever was produced; nevertheless, in some other and rare cases, a few tentacles became for a short time inflected; but this may have been caused by the glands having been accidentally touched in getting the leaves into a proper position. That water should produce no effect might have been anticipated, as otherwise the leaves would have been excited into movement by every shower of rain.

[Gum arabic.—Solutions of four degrees of strength were made; one of six grains to the ounce of water (one part to 73); a second rather stronger, yet very thin; a third moderately thick, and a fourth so thick that it would only just drop from a pointed instrument. These were tried on fourteen leaves; the drops being left on the discs from 24 hrs. to 44 hrs.; generally about [page 78] 30 hrs. Inflection was never thus caused. It is necessary to try pure gum arabic, for a friend tried a solution bought ready prepared, and this caused the tentacles to bend; but he afterwards ascertained that it contained much animal matter, probably glue.

Sugar.—Drops of a solution of white sugar of three strengths (the weakest containing one part of sugar to 73 of water) were left on fourteen leaves from 32 hrs. to 48 hrs.; but no effect was produced.

Starch.—A mixture about as thick as cream was dropped on six leaves and left on them for 30 hrs., no effect being produced. I am surprised at this fact, as I believe that the starch of commerce generally contains a trace of gluten, and this nitrogenous substance causes inflection, as we shall see in the next chapter.

Alcohol, Diluted.—One part of alcohol was added to seven of water, and the usual drops were placed on the discs of three leaves. No inflection ensued in the course of 48 hrs. To ascertain whether these leaves had been at all injured, bits of meat were placed on them, and after 24 hrs. they were closely inflected. I also put drops of sherry-wine on three other leaves; no inflection was caused, though two of them seemed somewhat injured. We shall hereafter see that cut off leaves immersed in diluted alcohol of the above strength do not become inflected.

Olive Oil.—drops were placed on the discs of eleven leaves, and no effect was produced in from 24 hrs. to 48 hrs. Four of these leaves were then tested by bits of meat on their discs, and three of them were found after 24 hrs. with all their tentacles and blades closely inflected, whilst the fourth had only a few tentacles inflected. It will, however, be shown in a future place, that cut off leaves immersed in olive oil are powerfully affected.

Infusion and Decoction of Tea.—Drops of a strong infusion and decoction, as well as of a rather weak decoction, of tea were placed on ten leaves, none of which became inflected. I afterwards tested three of them by adding bits of meat to the drops which still remained on their discs, and when I examined them after 24 hrs. they were closely inflected. The chemical principle of tea, namely theine, was subsequently tried and produced no effect. The albuminous matter which the leaves must originally have contained, no doubt, had been rendered insoluble by their having been completely dried.]

We thus see that, excluding the experiments with water, sixty-one leaves were tried with drops of the [page 79] above-named non-nitrogenous fluids; and the tentacles were not in a single case inflected.

[With respect to nitrogenous fluids, the first which came to hand were tried. The experiments were made at the same time and in exactly the same manner as the foregoing. As it was immediately evident that these fluids produced a great effect, I neglected in most cases to record how soon the tentacles became inflected. But this always occurred in less than 24 hrs.; whilst the drops of non-nitrogenous fluids which produced no effect were observed in every case during a considerably longer period.

Milk.—Drops were placed on sixteen leaves, and the tentacles of all, as well as the blades of several, soon became greatly inflected. The periods were recorded in only three cases, namely, with leaves on which unusually small drops had been placed. Their tentacles were somewhat inflected in 45 m.; and after 7 hrs. 45 m. the blades of two were so much curved inwards that they formed little cups enclosing the drops. These leaves re-expanded on the third day. On another occasion the blade of a leaf was much inflected in 5 hrs. after a drop of milk had been placed on it.

Human Urine.—Drops were placed on twelve leaves, and the tentacles of all, with a single exception, became greatly inflected. Owing, I presume, to differences in the chemical nature of the urine on different occasions, the time required for the movements of the tentacles varied much, but was always effected in under 24 hrs. In two instances I recorded that all the exterior tentacles were completely inflected in 17 hrs., but not the blade of the leaf. In another case the edges of a leaf, after 25 hrs. 30 m., became so strongly inflected that it was converted into a cup. The power of urine does not lie in the urea, which, as we shall hereafter see, is inoperative.

Albumen (fresh from a hen's egg), placed on seven leaves, caused the tentacles of six of them to be well inflected. In one case the edge of the leaf itself became much curled in after 20 hrs. The one leaf which was unaffected remained so for 26 hrs., and was then treated with a drop of milk, and this caused the tentacles to bend inwards in 12 hrs.

Cold Filtered Infusion of Raw Meat.—This was tried only on a single leaf, which had most of its outer tentacles and the blade inflected in 19 hrs. During subsequent years, I repeatedly used this infusion to test leaves which had been experimented [page 80] on with other substances, and it was found to act most energetically, but as no exact account of these trials was kept, they are not here introduced.

Mucus.—Thick and thin mucus from the bronchial tubes, placed on three leaves, caused inflection. A leaf with thin mucus had its marginal tentacles and blade somewhat curved inward in 5 hrs. 30 m., and greatly so in 20 hrs. The action of this fluid no doubt is due either to the saliva or to some albuminous matter* mingled with it, and not, as we shall see in the next chapter, to mucin or the chemical principle of mucus.

Saliva.—Human saliva, when evaporated, yields from 1.14 to 1.19 per cent. of residue; and this yields 0.25 per cent. of ashes, so that the proportion of nitrogenous matter which saliva contains must be small. Nevertheless, drops placed on the discs of eight leaves acted on them all. In one case all the exterior tentacles, excepting nine, were inflected in 19 hrs. 30 m.; in another case a few became so in 2 hrs., and after 7 hrs. 30 m. all those situated near where the drop lay, as well as the blade, were acted on. Since making these trials, I have many scores of times just touched glands with the handle of my scalpel wetted with saliva, to ascertain whether a leaf was in an active condition; for this was shown in the course of a few minutes by the bending inwards of the tentacles. The edible nest of the Chinese swallow is formed of matter secreted by the salivary glands; two grains were added to one ounce of distilled water (one part to 218), which was boiled for several minutes, but did not dissolve the whole. The usual-sized drops were placed on three leaves, and these in 1 hr. 30 m. were well, and in 2 hrs. 15 m. closely, inflected.

Isinglass.—Drops of a solution about as thick as milk, and of a still thicker solution, were placed on eight leaves, and the tentacles of all became inflected. In one case the exterior tentacles were well curved in after 6 hrs. 30 m., and the blade of the leaf to a partial extent after 24 hrs. As saliva acted so efficiently, and yet contains so small a proportion of nitrogenous matter, I tried how small a quantity of isinglass would act. One part was dissolved in 218 parts of distilled water, and drops were placed on four leaves. After 5 hrs. two of these were considerably and two moderately inflected; after 22 hrs. the former were greatly and the latter much more inflected. In the course of 48 hrs.

* Mucus from the air-passages is said in Marshall, 'Outlines of Physiology,' vol. ii. 1867, p. 364, to contain some albumen.

Mller's 'Elements of Physiology,' Eng. Trans. vol. i., p. 514. [page 81]

from the time when the drops were placed on the leaves, all four had almost re-expanded. They were then given little bits of meat, and these acted more powerfully than the solution. One part of isinglass was next dissolved in 437 of water; the fluid thus formed was so thin that it could not be distinguished from pure water. The usual-sized drops were placed on seven leaves, each of which thus received 1/960 of a grain (.0295 mg.). Three of them were observed for 41 hrs., but were in no way affected; the fourth and fifth had two or three of their exterior tentacles inflected after 18 hrs.; the sixth had a few more; and the seventh had in addition the edge of the leaf just perceptibly curved inwards. The tentacles of the four latter leaves began to re-expand after an additional interval of only 8 hrs. Hence the 1/960 of a grain of isinglass is sufficient to affect very slightly the more sensitive or active leaves. On one of the leaves, which had not been acted on by the weak solution, and on another, which had only two of its tentacles inflected, drops of the solution as thick as milk were placed; and next morning, after an interval of 16 hrs., both were found with all their tentacles strongly inflected.]

Altogether I experimented on sixty-four leaves with the above nitrogenous fluids, the five leaves tried only with the extremely weak solution of isinglass not being included, nor the numerous trials subsequently made, of which no exact account was kept. Of these sixty-four leaves, sixty-three had their tentacles and often their blades well inflected. The one which failed was probably too old and torpid. But to obtain so large a proportion of successful cases, care must be taken to select young and active leaves. Leaves in this condition were chosen with equal care for the sixty-one trials with non-nitrogenous fluids (water not included); and we have seen that not one of these was in the least affected. We may therefore safely conclude that in the sixty-four experiments with nitrogenous fluids the inflection of the exterior tentacles was due to the absorption of [page 82] nitrogenous matter by the glands of the tentacles on the disc.

Some of the leaves which were not affected by the non-nitrogenous fluids were, as above stated, immediately afterwards tested with bits of meat, and were thus proved to be in an active condition. But in addition to these trials, twenty-three of the leaves, with drops of gum, syrup, or starch, still lying on their discs, which had produced no effect in the course of between 24 hrs. and 48 hrs., were then tested with drops of milk, urine, or albumen. Of the twenty-three leaves thus treated, seventeen had their tentacles, and in some cases their blades, well inflected; but their powers were somewhat impaired, for the rate of movement was decidedly slower than when fresh leaves were treated with these same nitrogenous fluids. This impairment, as well as the insensibility of six of the leaves, may be attributed to injury from exosmose, caused by the density of the fluids placed on their discs.

[The results of a few other experiments with nitrogenous fluids may be here conveniently given. Decoctions of some vegetables, known to be rich in nitrogen, were made, and these acted like animal fluids. Thus, a few green peas were boiled for some time in distilled water, and the moderately thick decoction thus made was allowed to settle. Drops of the superincumbent fluid were placed on four leaves, and when these were looked at after 16 hrs., the tentacles and blades of all were found strongly inflected. I infer from a remark by Gerhardt* that legumin is present in peas "in combination with an alkali, forming an incoagulable solution," and this would mingle with boiling water. I may mention, in relation to the above and following experiments, that according to Schiff certain forms of albumen

* Watts' 'Dictionary of Chemistry,' vol. iii., p. 568.

'Leons sur la Phys. de la Digestion,' tom. i, p. 379; tom. ii. pp. 154, 166, on legumin. [page 83]

exist which are not coagulated by boiling water, but are converted into soluble peptones.

On three occasions chopped cabbage-leaves* were boiled in distilled water for 1 hr. or for 1 1/4 hr.; and by decanting the decoction after it had been allowed to rest, a pale dirty green fluid was obtained. The usual-sized drops were placed on thirteen leaves. Their tentacles and blades were inflected after 4 hrs. to a quite extraordinary degree. Next day the protoplasm within the cells of the tentacles was found aggregated in the most strongly marked manner. I also touched the viscid secretion round the glands of several tentacles with minute drops of the decoction on the head of a small pin, and they became well inflected in a few minutes. The fluid proving so powerful, one part was diluted with three of water, and drops were placed on the discs of five leaves; and these next morning were so much acted on that their blades were completely doubled over. We thus see that a decoction of cabbage-leaves is nearly or quite as potent as an infusion of raw meat.

About the same quantity of chopped cabbage-leaves and of distilled water, as in the last experiment, were kept in a vessel for 20 hrs. in a hot closet, but not heated to near the boiling-point. Drops of this infusion were placed on four leaves. One of these, after 23 hrs., was much inflected; a second slightly; a third had only the submarginal tentacles inflected; and the fourth was not at all affected. The power of this infusion is therefore very much less than that of the decoction; and it is clear that the immersion of cabbage-leaves for an hour in water at the boiling temperature is much more efficient in extracting matter which excites Drosera than immersion during many hours in warm water. Perhaps the contents of the cells are protected (as Schiff remarks with respect to legumin) by the walls being formed of cellulose, and that until these are ruptured by boiling-water, but little of the contained albuminous matter is dissolved. We know from the strong odour of cooked cabbage-leaves that boiling water produces some chemical change in them, and that they are thus rendered far more digestible and nutritious to man. It is therefore an interesting

* The leaves of young plants, before the heart is formed, such as were used by me, contain 2.1 per cent. of albuminous matter, and the outer leaves of mature plants 1.6 per cent. Watts' 'Dictionary of Chemistry,' vol. i. p. 653. [page 84]

fact that water at this temperature extracts matter from them which excites Drosera to an extraordinary degree.

Grasses contain far less nitrogenous matter than do peas or cabbages. The leaves and stalks of three common kinds were chopped and boiled for some time in distilled water. Drops of this decoction (after having stood for 24 hrs.) were placed on six leaves, and acted in a rather peculiar manner, of which other instances will be given in the seventh chapter on the salts of ammonia. After 2 hrs. 30 m. four of the leaves had their blades greatly inflected, but not their exterior tentacles; and so it was with all six leaves after 24 hrs. Two days afterwards the blades, as well as the few submarginal tentacles which had been inflected, all re-expanded; and much of the fluid on their discs was by this time absorbed. It appears that the decoction strongly excites the glands on the disc, causing the blade to be quickly and greatly inflected; but that the stimulus, differently from what occurs in ordinary cases, does not spread, or only in a feeble degree, to the exterior tentacles.

I may here add that one part of the extract of belladonna (procured from a druggist) was dissolved in 437 of water, and drops were placed on six leaves. Next day all six were somewhat inflected, and after 48 hrs. were completely re-expanded. It was not the included atropine which produced this effect, for I subsequently ascertained that it is quite powerless. I also procured some extract of hyoscyamus from three shops, and made infusions of the same strength as before. Of these three infusions, only one acted on some of the leaves, which were tried. Though druggists believe that all the albumen is precipitated in the preparation of these drugs, I cannot doubt that some is occasionally retained; and a trace would be sufficient to excite the more sensitive leaves of Drosera. [page 85]



CHAPTER VI.

THE DIGESTIVE POWER OF THE SECRETION OF DROSERA.

The secretion rendered acid by the direct and indirect excitement of the glands—Nature of the acid—Digestible substances—Albumen, its digestion arrested by alkalies, recommences by the addition of an acid—Meat—Fibrin—Syntonin—Areolar tissue—Cartilage—Fibro-cartilage— Bone—Enamel and dentine—Phosphate of lime—Fibrous basis of bone—Gelatine—Chondrin— Milk, casein and cheese—Gluten—Legumin—Pollen—Globulin—Haematin—Indigestible substances—Epidermic productions—Fibro-elastic tissue—Mucin—Pepsin—Urea—Chitine— Cellulose—Gun-cotton—Chlorophyll—Fat and oil—Starch—Action of the secretion on living seeds—Summary and concluding remarks.

AS we have seen that nitrogenous fluids act very differently on the leaves of Drosera from non-nitrogenous fluids, and as the leaves remain clasped for a much longer time over various organic bodies than over inorganic bodies, such as bits of glass, cinder, wood, &c., it becomes an interesting inquiry, whether they can only absorb matter already in solution, or render it soluble,—that is, have the power of digestion. We shall immediately see that they certainly have this power, and that they act on albuminous compounds in exactly the same manner as does the gastric juice of mammals; the digested matter being afterwards absorbed. This fact, which will be clearly proved, is a wonderful one in the physiology of plants. I must here state that I have been aided throughout all my later experiments by many valuable suggestions and assistance given me with the greatest kindness by Dr. Burdon Sanderson. [page 86]

It may be well to premise for the sake of any reader who knows nothing about the digestion of albuminous compounds by animals that this is effected by means of a ferment, pepsin, together with weak hydrochloric acid, though almost any acid will serve. Yet neither pepsin nor an acid by itself has any such power.* We have seen that when the glands of the disc are excited by the contact of any object, especially of one containing nitrogenous matter, the outer tentacles and often the blade become inflected; the leaf being thus converted into a temporary cup or stomach. At the same time the discal glands secrete more copiously, and the secretion becomes acid. Moreover, they transmit some influence to the glands of the exterior tentacles, causing them to pour forth a more copious secretion, which also becomes acid or more acid than it was before.

As this result is an important one, I will give the evidence. The secretion of many glands on thirty leaves, which had not been in any way excited, was tested with litmus paper; and the secretion of twenty-two of these leaves did not in the least affect the colour, whereas that of eight caused an exceedingly feeble and sometimes doubtful tinge of red. Two other old leaves, however, which appeared to have been inflected several times, acted much more decidedly on the paper. Particles of clean glass were then placed on five of the leaves, cubes of albumen on six, and bits of raw meat on three, on none of which was the secretion at this time in the least acid. After an interval of 24 hrs., when almost all the tentacles on