Pinguicula vulgaris--Structure of leaves--Number of insects and other objects caught-- Movement of the margins of the leaves--Uses of this movement--Secretion, digestion, and absorption--Action of the secretion on various animal and vegetable substances--The effects of substances not containing soluble nitrogenous matter on the glands--Pinguicula grandiflora--Pinguicula lusitanica, catches insects--Movement of the leaves, secretion and digestion.
PINGUICULA VULGARIS.--This plant grows in moist places, generally on mountains. It bears on an average eight, rather thick, oblong, light green leaves, having scarcely any footstalk. A full-sized leaf is about 1 1/2 inch in length and 3/4 inch in breadth. The young central leaves are deeply concave, and project upwards; the older ones towards the outside are flat or convex, and lie close to the ground, forming a rosette from 3 to 4 inches in diameter. The margins of the leaves are incurved. Their upper surfaces are thickly covered with two sets of glandular hairs, differing in the size of the glands and in the length of their pedicels. The larger glands have a circular outline as seen from above, and are of moderate thickness; they are divided by radiating partitions into sixteen cells, containing light-green, homogeneous fluid. They are supported on elongated, unicellular pedicels (containing a nucleus with a nucleolus) which rest on slight prominences. The small glands differ only in being formed of about half the number of cells, containing much paler fluid, and supported on much shorter pedicels. Near the midrib, towards the base of the leaf, the pedicels are multicellular, are longer than elsewhere, and bear smaller glands. All the glands secrete a colourless fluid, which is so viscid that I have seen a fine thread drawn out to a length of 18 inches; but the fluid in this case was secreted by a gland which had been excited. The edge of the leaf is translucent, and does not bear any glands; and here the spiral vessels, proceeding from the midrib, terminate in cells marked by a spiral line, somewhat like those within the glands of Drosera.
The roots are short. Three plants were dug up in North Wales on June 20, and carefully washed; each bore five or six unbranched roots, the longest of which was only 1.2 of an inch. Two rather young plants were examined on September 28; these had a greater number of roots, namely eight and eighteen, all under 1 inch in length, and very little branched.
I was led to investigate the habits of this plant by being told by Mr. W. Marshall that on the mountains of Cumberland many insects adhere to the leaves.
[A friend sent me on June 23 thirty-nine leaves from North Wales, which were selected owing to objects of some kind adhering to them. Of these leaves, thirty-two had caught 142 insects, or on an average 4.4 per leaf, minute fragments of insects not being included. Besides the insects, small leaves belonging to four different kinds of plants, those of Erica tetralix being much the commonest, and three minute seedling plants, blown by the wind, adhered to nineteen of the leaves. One had caught as many as ten leaves of the Erica. Seeds or fruits, commonly of Carex and one of Juncus, besides bits of moss and other rubbish, likewise adhered to six of the thirty-nine leaves. The same friend, on June 27, collected nine plants bearing seventy-four leaves, and all of these, with the exception of three young leaves, had caught insects; thirty insects were counted on one leaf, eighteen on a second, and sixteen on a third. Another friend examined on August 22 some plants in Donegal, Ireland, and found insects on 70 out of 157 leaves; fifteen of these leaves were sent me, each having caught on an average 2.4 insects. To nine of them, leaves (mostly of Erica tetralix) adhered; but they had been specially selected on this latter account. I may add that early in August my son found leaves of this same Erica and the fruits of a Carex on the leaves of a Pinguicula in Switzerland, probably Pinguicula alpina; some insects, but no great number, also adhered to the leaves of this plant, which had much better developed roots than those of Pinguicula vulgaris. In Cumberland, Mr. Marshall, on September 3, carefully examined for me ten plants bearing eighty leaves; and on sixty-three of these (i.e. on 79 per cent.) he found insects, 143 in number; so that each leaf had on an average 2.27 insects. A few days later he sent me some plants with sixteen seeds or fruits adhering to fourteen leaves. There was a seed on three leaves on the same plant. The sixteen seeds belonged to nine different kinds, which could not be recognised, excepting one of Ranunculus, and several belonging to three or four distinct species of Carex. It appears that fewer insects are caught late in the year than earlier; thus in Cumberland from twenty to twenty-four insects were observed in the middle of July on several leaves, whereas in the beginning of September the average number was only 2.27. Most of the insects, in all the foregoing cases, were Diptera, but with many minute Hymenoptera, including some ants, a few small Coleoptera, larvae, spiders, and even small moths.]
We thus see that numerous insects and other objects are caught by the viscid leaves; but we have no right to infer from this fact that the habit is beneficial to the plant, any more than in the before given case of the Mirabilis, or of the horse-chestnut. But it will presently be seen that dead insects and other nitrogenous bodies excite the glands to increased secretion; and that the secretion then becomes acid and has the power of digesting animal substances, such as albumen, fibrin, &c. Moreover, the dissolved nitrogenous matter is absorbed by the glands, as shown by their limpid contents being aggregated into slowly moving granular masses of protoplasm. The same results follow when insects are naturally captured, and as the plant lives in poor soil and has small roots, there can be no doubt that it profits by its power of digesting and absorbing matter from the prey which it habitually captures in such large numbers. It will, however, be convenient first to describe the movements of the leaves.
Movements of the Leaves.--That such thick, large leaves as those of Pinguicula vulgarisshould have the power of curving inwards when excited has never even been suspected. It is necessary to select for experiment leaves with their glands secreting freely, and which have been prevented from capturing many insects; as old leaves, at least those growing in a state of nature, have their margins already curled so much inwards that they exhibit little power of movement, or move very slowly. I will first give in detail the more important experiments which were tried, and then make some concluding remarks.
[Experiment 1.--A young and almost upright leaf was selected, with its two lateral edges equally and very slightly incurved. A row of small flies was placed along one margin. When looked at next day, after 15 hrs., this margin, but not the other, was found folded inwards, like the helix of the human ear, to the breadth of 1/10 of an inch, so as to lie partly over the row of flies (fig. 15). The glands on which the flies rested, as well as those on the over-lapping margin which had been brought into contact with the flies, were all secreting copiously.
FIG. 15. (Pinguicula vulgaris.) Outline of leaf with left margin inflected over a row of small flies.
Experiment 2.--A row of flies was placed on one margin of a rather old leaf, which lay flat on the ground; and in this case the margin, after the same interval as before, namely 15 hrs., had only just begun to curl inwards; but so much secretion had been poured forth that the spoon-shaped tip of the leaf was filled with it.
Experiment 3.--Fragments of a large fly were placed close to the apex of a vigorous leaf, as well as along half one margin. After 4 hrs. 20 m. there was decided incurvation, which increased a little during the afternoon, but was in the same state on the following morning. Near the apex both margins were inwardly curved. I have never seen a case of the apex itself being in the least curved towards the base of the leaf. After 48 hrs. (always reckoning from the time when the flies were placed on the leaf) the margin had everywhere begun to unfold.
Experiment 4.--A large fragment of a fly was placed on a leaf, in a medial line, a little beneath the apex. Both lateral margins were perceptibly incurved in 3 hrs., and after 4 hrs. 20 m. to such a degree that the fragment was clasped by both margins. After 24 hrs. the two infolded edges near the apex (for the lower part of the leaf was not at all affected) were measured and found to be .11 of an inch (2.795 mm.) apart. The fly was now removed, and a stream of water poured over the leaf so as to wash the surface; and after 24 hrs. the margins were .25 of an inch (6.349 mm.) apart, so that they were largely unfolded. After an additional 24 hrs. they were completely unfolded. Another fly was now put on the same spot to see whether this leaf, on which the first fly had been left 24 hrs., would move again; after 10 hrs. there was a trace of incurvation, but this did not increase during the next 24 hrs. A bit of meat was also placed on the margin of a leaf, which four days previously had become strongly incurved over a fragment of a fly and had afterwards re-expanded; but the meat did not cause even a trace of incurvation. On the contrary, the margin became somewhat reflexed, as if injured, and so remained for the three following days, as long as it was observed.
Experiment 5.--A large fragment of a fly was placed halfway between the apex and base of a leaf and halfway between the midrib and one margin. A short space of this margin, opposite the fly, showed a trace of incurvation after 3 hrs., and this became strongly pronounced in 7 hrs. After 24 hrs. the infolded edge was only .16 of an inch (4.064 mm.) from the midrib. The margin now began to unfold, though the fly was left on the leaf; so that by the next morning (i.e. 48 hrs. from the time when the fly was first put on) the infolded edge had almost completely recovered its original position, being now .3 of an inch (7.62 mm.), instead of .16 of an inch, from the midrib. A trace of flexure was, however, still visible.
Experiment 6.--A young and concave leaf was selected with its margins slightly and naturally incurved. Two rather large, oblong, rectangular pieces of roast meat were placed with their ends touching the infolded edge, and .46 of an inch (11.68 mm.) apart from one another. After 24 hrs. the margin was greatly and equally incurved (see fig. 16) throughout this space, and for a length of .12 or .13 of an inch (3.048 or 3.302 mm.) above and below each bit; so that the margin had been affected over a greater length between the two bits, owing to their conjoint action, than beyond them. The bits of meat were too large to be clasped by the margin, but they were tilted up, one of them so as to stand almost vertically. After 48 hrs. the margin was almost unfolded, and the bits had sunk down. When again examined after two days, the margin was quite unfolded, with the exception of the naturally inflected edge; and one of the bits of meat, the end of which had at first touched the edge, was now .067 of an inch (1.70 mm.) distant from it; so that this bit had been pushed thus far across the blade of the leaf.
FIG. 16. (Pinguicula vulgaris.) Outline of leaf, with right margin inflected against two square bits of meat.
Experiment 7.--A bit of meat was placed close to the incurved edge of a rather young leaf, and after it had re-expanded, the bit was left lying .11 of an inch (2.795 mm.) from the edge. The distance from the edge to the midrib of the fully expanded leaf was .35 of an inch (8.89 mm.); so that the bit had been pushed inwards and across nearly one-third of its semi-diameter.
Experiment 8.--Cubes of sponge, soaked in a strong infusion of raw meat, were placed in close contact with the incurved edges of two leaves,--an older and younger one. The distance from the edges to the midribs was carefully measured. After 1 hr. 17 m. there appeared to be a trace of incurvation. After 2 hrs. 17 m. both leaves were plainly inflected; the distance between the edges and midribs being now only half what it was at first. The incurvation increased slightly during the next 4 1/2 hrs., but remained nearly the same for the next 17 hrs. 30 m. In 35 hrs. from the time when the sponges were placed on the leaves, the margins were a little unfolded--to a greater degree in the younger than in the older leaf. The latter was not quite unfolded until the third day, and now both bits of sponge were left at the distance of .1 of an inch (2.54 mm.) from the edges; or about a quarter of the distance between the edge and midrib. A third bit of sponge adhered to the edge, and, as the margin unfolded, was dragged backwards, into its original position.
Experiment 9.--A chain of fibres of roast meat, as thin as bristles and moistened with saliva, were placed down one whole side, close to the narrow, naturally incurved edge of a leaf. In 3 hrs. this side was greatly incurved along its whole length, and after 8 hrs. formed a cylinder, about 1/20 of an inch (1.27 mm) in diameter, quite concealing the meat. This cylinder remained closed for 32 hrs., but after 48 hrs. was half unfolded, and in 72 hrs. was as open as the opposite margin where no meat had been placed. As the thin fibres of meat were completely overlapped by the margin, they were not pushed at all inwards, across the blade.
Experiment 10.--Six cabbage seeds, soaked for a night in water, were placed in a row close to the narrow incurved edge of a leaf. We shall hereafter see that these seeds yield soluble matter to the glands. In 2 hrs. 25 m. the margin was decidedly inflected; in 4 hrs. it extended over the seeds for about half their breadth, and in 7 hrs. over three-fourths of their breadth, forming a cylinder not quite closed along the inner side, and about .7 of an inch (1.778 mm.) in diameter. After 24 hrs. the inflection had not increased, perhaps had decreased. The glands which had been brought into contact with the upper surfaces of the seeds were now secreting freely. In 36 hrs. from the time when the seeds were put on the leaf the margin had greatly, and after 48 hrs. had completely, re-expanded. As the seeds were no longer held by the inflected margin, and as the secretion was beginning to fail, they rolled some way down the marginal channel.
Experiment 11.--Fragments of glass were placed on the margins of two fine young leaves. After 2 hrs. 30 m. the margin of one certainly became slightly incurved; but the inflection never increased, and disappeared in 16 hrs. 30 m. from the time when the fragments were first applied. With the second leaf there was a trace of incurvation in 2 hrs. 15 m., which became decided in 4 hrs. 30 m., and still more strongly pronounced in 7 hrs., but after 19 hrs. 30 m. had plainly decreased. The fragments excited at most a slight and doubtful increase of the secretion; and in two other trials, no increase could be perceived. Bits of coal-cinders, placed on a leaf, produced no effect, either owing to their lightness or to the leaf being torpid.
Experiment 12.--We now turn to fluids. A row of drops of a strong infusion of raw meat were placed along the margins of two leaves; squares of sponge soaked in the same infusion being placed on the opposite margins. My object was to ascertain whether a fluid would act as energetically as a substance yielding the same soluble matter to the glands. No distinct difference was perceptible; certainly none in the degree of incurvation; but the incurvation round the bits of sponge lasted rather longer, as might perhaps have been expected from the sponge remaining damp and supplying nitrogenous matter for a longer time. The margins, with the drops, became plainly incurved in 2 hrs. 17 m. The incurvation subsequently increased somewhat, but after 24 hrs. had greatly decreased.
Experiment 13.--Drops of the same strong infusion of raw meat were placed along the midrib of a young and rather deeply concave leaf. The distance across the broadest part of the leaf, between the naturally incurved edges, was .55 of an inch (13.97 mm.). In 3 hrs. 27 m. this distance was a trace less; in 6 hrs. 27 m. it was exactly .45 of an inch (11.43 mm.), and had therefore decreased by .1 of an inch (2.54 mm.). After only 10 hrs. 37 m. the margin began to re-expand, for the distance from edge to edge was now a trace wider, and after 24 hrs. 20 m. was as great, within a hair's breadth, as when the drops were first placed on the leaf. From this experiment we learn that the motor impulse can be transmitted to a distance of .22 of an inch (5.590 mm.) in a transverse direction from the midrib to both margins; but it would be safer to say .2 of an inch (5.08 mm.) as the drops spread a little beyond the midrib. The incurvation thus caused lasted for an unusually short time.
Experiment 14.--Three drops of a solution of one part of carbonate of ammonia to 218 of water (2 grs. to 1 oz.) were placed on the margin of a leaf. These excited so much secretion that in 1 h. 22 m. all three drops ran together; but although the leaf was observed for 24 hrs., there was no trace of inflection. We know that a rather strong solution of this salt, though it does not injure the leaves of Drosera, paralyses their power of movement, and I have no doubt, from the following case, that this holds good with Pinguicula.
Experiment 15.--A row of drops of a solution of one part of carbonate of ammonia to 875 of water (1 gr. to 2 oz.) was placed on the margin of a leaf. In 1 hr. there was apparently some slight incurvation, and this was well-marked in 3 hrs. 30 m. After 24 hrs. the margin was almost completely re-expanded.
Experiment 16.--A row of large drops of a solution of one part of phosphate of ammonia to 4375 of water (1 gr. to 10 oz.) was placed along the margin of a leaf. No effect was produced, and after 8 hrs. fresh drops were added along the same margin without the least effect. We know that a solution of this strength acts powerfully on Drosera, and it is just possible that the solution was too strong. I regret that I did not try a weaker solution.
Experiment 17.--As the pressure from bits of glass causes incurvation, I scratched the margins of two leaves for some minutes with a blunt needle, but no effect was produced. The surface of a leaf beneath a drop of a strong infusion of raw meat was also rubbed for 10. m. with the end of a bristle, so as to imitate the struggles of a captured insect; but this part of the margin did not bend sooner than the other parts with undisturbed drops of the infusion.]
We learn from the foregoing experiments that the margins of the leaves curl inwards when excited by the mere pressure of objects not yielding any soluble matter, by objects yielding such matter, and by some fluids--namely an infusion of raw meat and a week solution of carbonate of ammonia. A stronger solution of two grains of this salt to an ounce of water, though exciting copious secretion, paralyses the leaf. Drops of water and of a solution of sugar or gum did not cause any movement. Scratching the surface of the leaf for some minutes produced no effect. Therefore, as far as we at present know, only two causes--namely slight continued pressure and the absorption of nitrogenous matter--excite movement. It is only the margins of the leaf which bend, for the apex never curves towards the base. The pedicels of the glandular hairs have no power of movement. I observed on several occasions that the surface of the leaf became slightly concave where bits of meat or large flies had long lain, but this may have been due to injury from over-stimulation.
The shortest time in which plainly marked movement was observed was 2 hrs. 17 m., and this occurred when either nitrogenous substances or fluids were placed on the leaves; but I believe that in some cases there was a trace of movement in 1 hr. or 1 hr. 30 m. The pressure from fragments of glass excites movement almost as quickly as the absorption of nitrogenous matter, but the degree of incurvation thus caused is much less. After a leaf has become well incurved and has again expanded, it will not soon answer to a fresh stimulus. The margin was affected longitudinally, upwards or downwards, for a distance of .13 of an inch (3.302 mm.) from an excited point, but for a distance of .46 of an inch between two excited points, and transversely for a distance of .2 of an inch (5.08 mm.). The motor impulse is not accompanied, as in the case of Drosera, by any influence causing increased secretion; for when a single gland was strongly stimulated and secreted copiously, the surrounding glands were not in the least affected. The incurvation of the margin is independent of increased secretion, for fragments of glass cause little or no secretion, and yet excite movement; whereas a strong solution of carbonate of ammonia quickly excites copious secretion, but no movement.
One of the most curious facts with respect to the movement of the leaves is the short time during which they remain incurved, although the exciting object is left on them. In the majority of cases there was well-marked re-expansion within 24 hrs. from the time when even large pieces of meat, &c., were placed on the leaves, and in all cases within 48 hrs. In one instance the margin of a leaf remained for 32 hrs. closely inflected round thin fibres of meat; in another instance, when a bit of sponge, soaked in a strong infusion of raw meat, had been applied to a leaf, the margin began to unfold in 35 hrs. Fragments of glass keep the margin incurved for a shorter time than do nitrogenous bodies; for in the former case there was complete re-expansion in 16 hrs. 30 m. Nitrogenous fluids act for a shorter time than nitrogenous substances; thus, when drops of an infusion of raw meat were placed on the midrib of a leaf, the incurved margins began to unfold in only 10 hrs. 37 m., and this was the quickest act of re-expansion observed by me; but it may have been partly due to the distance of the margins from the midrib where the drops lay.
We are naturally led to inquire what is the use of this movement which lasts for so short a time? If very small objects, such as fibres of meat, or moderately small objects, such as little flies or cabbage-seeds, are placed close to the margin, they are either completely or partially embraced by it. The glands of the overlapping margin are thus brought into contact with such objects and pour forth their secretion, afterwards absorbing the digested matter. But as the incurvation lasts for so short a time, any such benefit can be of only slight importance, yet perhaps greater than at first appears. The plant lives in humid districts, and the insects which adhere to all parts of the leaf are washed by every heavy shower of rain into the narrow channel formed by the naturally incurved edges. For instance, my friend in North Wales placed several insects on some leaves, and two days afterwards (there having been heavy rain in the interval) found some of them quite washed away, and many others safely tucked under the now closely inflected margins, the glands of which all round the insects were no doubt secreting. We can thus, also, understand how it is that so many insects, and fragments of insects, are generally found lying within the incurved margins of the leaves.
The incurvation of the margin, due to the presence of an exciting object, must be serviceable in another and probably more important way. We have seen that when large bits of meat, or of sponge soaked in the juice of meat, were placed on a leaf, the margin was not able to embrace them, but, as it became incurved, pushed them very slowly towards the middle of the leaf, to a distance from the outside of fully .1 of an inch (2.54 mm.), that is, across between one-third and one-fourth of the space between the edge and midrib. Any object, such as a moderately sized insect, would thus be brought slowly into contact with a far larger number of glands, inducing much more secretion and absorption, than would otherwise have been the case. That this would be highly serviceable to the plant, we may infer from the fact that Drosera has acquired highly developed powers of movement, merely for the sake of bringing all its glands into contact with captured insects. So again, after a leaf of Dionaea has caught an insect, the slow pressing together of the two lobes serves merely to bring the glands on both sides into contact with it, causing also the secretion charged with animal matter to spread by capillary attraction over the whole surface. In the case of Pinguicula, as soon as an insect has been pushed for some little distance towards the midrib, immediate re-expansion would be beneficial, as the margins could not capture fresh prey until they were unfolded. The service rendered by this pushing action, as well as that from the marginal glands being brought into contact for a short time with the upper surfaces of minute captured insects, may perhaps account for the peculiar movements of the leaves; otherwise, we must look at these movements as a remnant of a more highly developed power formerly possessed by the progenitors of the genus.
In the four British species, and, as I hear from Prof. Dyer, in most or all the species of the genus, the edges of the leaves are in some degree naturally and permanently incurved. This incurvation serves, as already shown, to prevent insects from being washed away by the rain; but it likewise serves for another end. When a number of glands have been powerfully excited by bits of meat, insects, or any other stimulus, the secretion often trickles down the leaf, and is caught by the incurved edges, instead of rolling off and being lost. As it runs down the channel, fresh glands are able to absorb the animal matter held in solution. Moreover, the secretion often collects in little pools within the channel, or in the spoon-like tips of the leaves; and I ascertained that bits of albumen, fibrin, and gluten, are here dissolved more quickly and completely than on the surface of the leaf, where the secretion cannot accumulate; and so it would be with naturally caught insects. The secretion was repeatedly seen thus to collect on the leaves of plants protected from the rain; and with exposed plants there would be still greater need of some provision to prevent, as far as possible, the secretion, with its dissolved animal matter, being wholly lost.
It has already been remarked that plants growing in a state of nature have the margins of their leaves much more strongly incurved than those grown in pots and prevented from catching many insects. We have seen that insects washed down by the rain from all parts of the leaf often lodge within the margins, which are thus excited to curl farther inwards; and we may suspect that this action, many times repeated during the life of the plant, leads to their permanent and well-marked incurvation. I regret that this view did not occur to me in time to test its truth.
It may here be added, though not immediately bearing on our subject, that when a plant is pulled up, the leaves immediately curl downwards so as almost to conceal the roots,--a fact which has been noticed by many persons. I suppose that this is due to the same tendency which causes the outer and older leaves to lie flat on the ground. It further appears that the flower-stalks are to a certain extent irritable, for Dr. Johnson states that they "bend backwards if rudely handled."*
Secretion, Absorption, and Digestion.--I will first give my observations and experiments, and then a summary of the results.
[The Effects of Objects containing Soluble Nitrogenous Matter.
(1) Flies were placed on many leaves, and excited the glands to secrete copiously; the secretion always becoming acid, though not so before. After a time these insects were rendered so tender that their limbs and bodies could be separated by a mere touch, owing no doubt to the digestion and disintegration of their muscles. The glands in contact with a small fly continued to secrete for four days, and then became almost dry. A narrow strip of this leaf was cut off, and the glands of the longer and shorter hairs, which had lain in contact for the four days with the fly, and those which had not touched it, were compared under the microscope and presented a wonderful contrast. Those which had been in contact were filled with brownish granular matter, the others with homogeneous fluid. There could therefore be no doubt that the former had absorbed matter from the fly.
(2) Small bits of roast meat, placed on a leaf, always caused much acid secretion in the course of a few hours--in one case within 40 m. When thin fibres of meat were laid along the margin of a leaf which stood almost upright, the secretion ran down to the ground. Angular bits of meat, placed in little pools of the secretion near the margin, were in the course of
* 'English Botany,' by Sir J.E. Smith; with coloured figures by J. Sowerby; edit. of 1832, tab. 24, 25, 26.
two or three days much reduced in size, rounded, rendered more or less colourless and transparent, and so much softened that they fell to pieces on the slightest touch. In only one instance was a very minute particle completely dissolved, and this occurred within 48 hrs. When only a small amount of secretion was excited, this was generally absorbed in from 24 hrs. to 48 hrs.; the glands being left dry. But when the supply of secretion was copious, round either a single rather large bit of meat, or round several small bits, the glands did not become dry until six or seven days had elapsed. The most rapid case of absorption observed by me was when a small drop of an infusion of raw meat was placed on a leaf, for the glands here became almost dry in 3 hrs. 20 m. Glands excited by small particles of meat, and which have quickly absorbed their own secretion, begin to secrete again in the course of seven or eight days from the time when the meat was given them.
(3) Three minute cubes of tough cartilage from the leg-bone of a sheep were laid on a leaf. After 10 hrs. 30 m. some acid secretion was excited, but the cartilage appeared little or not at all affected. After 24 hrs. the cubes were rounded and much reduced in size; after 32 hrs. they were softened to the centre, and one was quite liquefied; after 35 hrs. mere traces of solid cartilage were left; and after 48 hrs. a trace could still be seen through a lens in only one of the three. After 82 hrs. not only were all three cubes completely liquefied, but all the secretion was absorbed and the glands left dry.
(4) Small cubes of albumen were placed on a leaf; in 8 hrs. feebly acid secretion extended to a distance of nearly 1/10 of an inch round them, and the angles of one cube were rounded. After 24 hrs. the angles of all the cubes were rounded, and they were rendered throughout very tender; after 30 hrs. the secretion began to decrease, and after 48 hrs. the glands were left dry; but very minute bits of albumen were still left undissolved.
(5) Smaller cubes of albumen (about 1/50 or 1/60 of an inch, .508 or .423 mm.) were placed on four glands; after 18 hrs. one cube was completely dissolved, the others being much reduced in size, softened, and transparent. After 24 hrs. two of the cubes were completely dissolved, and already the secretion on these glands was almost wholly absorbed. After 42 hrs. the two other cubes were completely dissolved. These four glands began to secrete again after eight or nine days.
(6) Two large cubes of albumen (fully 1/20 of an inch, 1.27 mm.) were placed, one near the midrib and the other near the margin of a leaf; in 6 hrs. there was much secretion, which after 48 hrs. accumulated in a little pool round the cube near the margin. This cube was much more dissolved than that on the blade of the leaf; so that after three days it was greatly reduced in size, with all the angles rounded, but it was too large to be wholly dissolved. The secretion was partially absorbed after four days. The cube on the blade was much less reduced, and the glands on which it rested began to dry after only two days.
(7) Fibrin excites less secretion than does meat or albumen. Several trials were made, but I will give only three of them. Two minute shreds were placed on some glands, and in 3 hrs. 45 m. their secretion was plainly increased. The smaller shred of the two was completely liquefied in 6 hrs. 15 m., and the other in 24 hrs.; but even after 48 hrs. a few granules of fibrin could still be seen through a lens floating in both drops of secretion. After 56 hrs. 30 m. these granules were completely dissolved. A third shred was placed in a little pool of secretion, within the margin of a leaf where a seed had been lying, and this was completely dissolved in the course of 15 hrs. 30 m.
(8) Five very small bits of gluten were placed on a leaf, and they excited so much secretion that one of the bits glided down into the marginal furrow. After a day all five bits seemed much reduced in size, but none were wholly dissolved. On the third day I pushed two of them, which had begun to dry, on to fresh glands. On the fourth day undissolved traces of three out of the five bits could still be detected, the other two having quite disappeared; but I am doubtful whether they had really been completely dissolved. Two fresh bits were now placed, one near the middle and the other near the margin of another leaf; both excited an extraordinary amount of secretion; that near the margin had a little pool formed round it, and was much more reduced in size than that on the blade, but after four days was not completely dissolved. Gluten, therefore, excites the glands greatly, but is dissolved with much difficulty, exactly as in the case of Drosera. I regret that I did not try this substance after having been immersed in weak hydrochloric acid, as it would then probably have been quickly dissolved.
(9) A small square thin piece of pure gelatine, moistened with water, was placed on a leaf, and excited very little secretion in 5 hrs. 30 m., but later in the day a greater amount. After 24 hrs. the whole square was completely liquefied; and this would not have occurred had it been left in water. The liquid was acid.
(10) Small particles of chemically prepared casein excited acid secretion, but were not quite dissolved after two days; and the glands then began to dry. Nor could their complete dissolution have been expected from what we have seen with Drosera.
(11) Minute drops of skimmed milk were placed on a leaf, and these caused the glands to secrete freely. After 3 hrs. the milk was found curdled, and after 23 hrs. the curds were dissolved. On placing the now clear drops under the microscope, nothing could be detected except some oil-globules. The secretion, therefore, dissolves fresh casein.
(12) Two fragments of a leaf were immersed for 17 hrs., each in a drachm of a solution of carbonate of ammonia, of two strengths, namely of one part to 437 and 218 of water. The glands of the longer and shorter hairs were then examined, and their contents found aggregated into granular matter of a brownish-green colour. These granular masses were seen by my son slowly to change their forms, and no doubt consisted of protoplasm. The aggregation was more strongly pronounced, and the movements of the protoplasm more rapid, within the glands subjected to the stronger solution than in the others. The experiment was repeated with the same result; and on this occasion I observed that the protoplasm had shrunk a little from the walls of the single elongated cells forming the pedicels. In order to observe the process of aggregation, a narrow strip of leaf was laid edgeways under the microscope, and the glands were seen to be quite transparent; a little of the stronger solution (viz. one part to 218 of water) was now added under the covering glass; after an hour or two the glands contained very fine granular matter, which slowly became coarsely granular and slightly opaque; but even after 5 hrs. not as yet of a brownish tint. By this time a few rather large, transparent, globular masses appeared within the upper ends of the pedicels, and the protoplasm lining their walls had shrunk a little. It is thus evident that the glands of Pinguicula absorb carbonate of ammonia; but they do not absorb it, or are not acted on by it, nearly so quickly as those of Drosera.
(13) Little masses of the orange-coloured pollen of the common pea, placed on several leaves, excited the glands to secrete freely. Even a very few grains which accidentally fell on a single gland caused the drop surrounding it to increase so much in size, in 23 hrs., as to be manifestly larger than the drops on the adjoining glands. Grains subjected to the secretion for 48 hrs. did not emit their tubes; they were quite discoloured, and seemed to contain less matter than before; that which was left being of a dirty colour, including globules of oil. They thus differed in appearance from other grains kept in water for the same length of time. The glands in contact with the pollen-grains had evidently absorbed matter from them; for they had lost their natural pale-green tint, and contained aggregated globular masses of protoplasm.
(14) Square bits of the leaves of spinach, cabbage, and a saxifrage, and the entire leaves of Erica tetralix, all excited the glands to increased secretion. The spinach was the most effective, for it caused the secretion evidently to increase in 1 hr. 40 m., and ultimately to run some way down the leaf; but the glands soon began to dry, viz. after 35 hrs. The leaves of Erica tetralix began to act in 7 hrs. 30 m., but never caused much secretion; nor did the bits of leaf of the saxifrage, though in this case the glands continued to secrete for seven days. Some leaves of Pinguicula were sent me from North Wales, to which leaves of Erica tetralixand of an unknown plant adhered; and the glands in contact with them had their contents plainly aggregated, as if they had been in contact with insects; whilst the other glands on the same leaves contained only clear homogeneous fluid.
(15) Seeds.--A considerable number of seeds or fruits selected by hazard, some fresh and some a year old, some soaked for a short time in water and some not soaked, were tried. The ten following kinds, namely cabbage, radish, Anemone nemorosa, Rumex acetosa, Carex sylvatica, mustard, turnip, cress, Ranunculus acris, and Avena pubescens, all excited much secretion, which was in several cases tested and found always acid. The five first-named seeds excited the glands more than the others. The secretion was seldom copious until about 24 hrs. had elapsed, no doubt owing to the coats of the seeds not being easily permeable. Nevertheless, cabbage seeds excited some secretion in 4 hrs. 30 m.; and this increased so much in 18 hrs. as to run down the leaves. The seeds or properly the fruits of Carex are much oftener found adhering to leaves in a state of nature than those of any other genus; and the fruits of Carex sylvatica excited so much secretion that in 15 hrs. it ran into the incurved edges; but the glands ceased to secrete after 40 hrs. On the other hand, the glands on which the seeds of the Rumex and Avena rested continued to secrete for nine days.
The nine following kinds of seeds excited only a slight amount of secretion, namely, celery, parsnip, caraway, Linum grandiflorum, Cassia, Trifolium pannonicum, Plantago, onion, and Bromus. Most of these seeds did not excite any secretion until 48 hrs. had elapsed, and in the case of the Trifolium only one seed acted, and this not until the third day. Although the seeds of the Plantago excited very little secretion, the glands continued to secrete for six days. Lastly, the five following kinds excited no secretion, though left on the leaves for two or three days, namely lettuce, Erica tetralix, Atriplex hortensis, Phalaris canariensis, and wheat. Nevertheless, when the seeds of the lettuce, wheat, and Atriplex were split open and applied to leaves, secretion was excited in considerable quantity in 10 hrs., and I believe that some was excited in six hours. In the case of the Atriplex the secretion ran down to the margin, and after 24 hrs. I speak of it in my notes "as immense in quantity and acid." The split seeds also of the Trifolium and celery acted powerfully and quickly, though the whole seeds caused, as we have seen, very little secretion, and only after a long interval of time. A slice of the common pea, which however was not tried whole, caused secretion in 2 hrs. From these facts we may conclude that the great difference in the degree and rate at which various kinds of seeds excite secretion, is chiefly or wholly due to the different permeability of their coats.
Some thin slices of the common pea, which had been previously soaked for 1 hr. in water, were placed on a leaf, and quickly excited much acid secretion. After 24 hrs. these slices were compared under a high power with others left in water for the same time; the latter contained so many fine granules of legumin that the slide was rendered muddy; whereas the slices which had been subjected to the secretion were much cleaner and more transparent, the granules of legumin apparently having been dissolved. A cabbage seed which had lain for two days on a leaf and had excited much acid secretion, was cut into slices, and these were compared with those of a seed which had been left for the same time in water. Those subjected to the secretion were of a paler colour; their coats presenting the greatest differences, for they were of a pale dirty tint instead of chestnut-brown. The glands on which the cabbage seeds had rested, as well as those bathed by the surrounding secretion, differed greatly in appearance from the other glands on the same leaf, for they all contained brownish granular matter, proving that they had absorbed matter from the seeds.
That the secretion acts on the seeds was also shown by some of them being killed, or by the seedlings being injured. Fourteen cabbage seeds were left for three days on leaves and excited much secretion; they were then placed on damp sand under conditions known to be favourable for germination. Three never germinated, and this was a far larger proportion of deaths than occurred with seeds of the same lot, which had not been subjected to the secretion, but were otherwise treated in the same manner. Of the eleven seedlings raised, three had the edges of their cotyledons slightly browned, as if scorched; and the cotyledons of one grew into a curious indented shape. Two mustard seeds germinated; but their cotyledons were marked with brown patches and their radicles deformed. Of two radish seeds, neither germinated; whereas of many seeds of the same lot not subjected to the secretion, all, excepting one, germinated. Of the two Rumex seeds, one died and the other germinated; but its radicle was brown and soon withered. Both seeds of the Avena germinated, one grew well, the other had its radicle brown and withered. Of six seeds of the Erica none germinated, and when cut open after having been left for five months on damp sand, one alone seemed alive. Twenty-two seeds of various kinds were found adhering to the leaves of plants growing in a state of nature; and of these, though kept for five months on damp sand, none germinated, some being then evidently dead.
The Effects of Objects not containing Soluble Nitrogenous Matter.
(16) It has already been shown that bits of glass, placed on leaves, excite little or no secretion. The small amount which lay beneath the fragments was tested and found not acid. A bit of wood excited no secretion; nor did the several kinds of seeds of which the coats are not permeable to the secretion, and which, therefore, acted like inorganic bodies. Cubes of fat, left for two days on a leaf, produced no effect.
(17) A particle of white sugar, placed on a leaf, formed in 1 hr. 10 m. a large drop of fluid, which in the course of 2 additional hours ran down into the naturally inflected margin. This fluid was not in the least acid, and began to dry up, or more probably was absorbed, in 5 hrs. 30 m. The experiment was repeated; particles being placed on a leaf, and others of the same size on a slip of glass in a moistened state; both being covered by a bell-glass. This was done to see whether the increased amount of fluid on the leaves could be due to mere deliquescence; but this was proved not to be the case. The particle on the leaf caused so much secretion that in the course of 4 hrs. it ran down across two-thirds of the leaf. After 8 hrs. the leaf, which was concave, was actually filled with very viscid fluid; and it particularly deserves notice that this, as on the former occasion, was not in the least acid. This great amount of secretion may be attributed to exosmose. The glands which had been covered for 24 hrs. by this fluid did not differ, when examined under the microscope, from others on the same leaf, which had not come into contact with it. This is an interesting fact in contrast with the invariably aggregated condition of glands which have been bathed by the secretion, when holding animal matter in solution.
(18) Two particles of gum arabic were placed on a leaf, and they certainly caused in 1 hr. 20 m. a slight increase of secretion. This continued to increase for the next 5 hrs., that is for as long a time as the leaf was observed.
(19) Six small particles of dry starch of commerce were placed on a leaf, and one of these caused some secretion in 1 hr. 15 m., and the others in from 8 hrs. to 9 hrs. The glands which had thus been excited to secrete soon became dry, and did not begin to secrete again until the sixth day. A larger bit of starch was then placed on a leaf, and no secretion was excited in 5 hrs. 30 m.; but after 8 hrs. there was a considerable supply, which increased so much in 24 hrs. as to run down the leaf to the distance of 3/4 of an inch. This secretion, though so abundant, was not in the least acid. As it was so copiously excited, and as seeds not rarely adhere to the leaves of naturally growing plants, it occurred to me that the glands might perhaps have the power of secreting a ferment, like ptyaline, capable of dissolving starch; so I carefully observed the above six small particles during several days, but they did not seem in the least reduced in bulk. A particle was also left for two days in a little pool of secretion, which had run down from a piece of spinach leaf; but although the particle was so minute no diminution was perceptible. We may therefore conclude that the secretion cannot dissolve starch. The increase caused by this substance may, I presume, be attributed to exosmose. But I am surprised that starch acted so quickly and powerfully as it did, though in a less degree than sugar. Colloids are known to possess some slight power of dialysis; and on placing the leaves of a Primula in water, and others in syrup and diffused starch, those in the starch became flaccid, but to a less degree and at a much slower rate than the leaves in the syrup; those in water remaining all the time crisp.]
From the foregoing experiments and observations we see that objects not containing soluble matter have little or no power of exciting the glands to secrete. Non-nitrogenous fluids, if dense, cause the glands to pour forth a large supply of viscid fluid, but this is not in the least acid. On the other hand, the secretion from glands excited by contact with nitrogenous solids or liquids is invariably acid, and is so copious that it often runs down the leaves and collects within the naturally incurved margins. The secretion in this state has the power of quickly dissolving, that is of digesting, the muscles of insects, meat, cartilage, albumen, fibrin, gelatine, and casein as it exists in the curds of milk. The glands are strongly excited by chemically prepared casein and gluten; but these substances (the latter not having been soaked in weak hydrochloric acid) are only partially dissolved, as was likewise the case with Drosera. The secretion, when containing animal matter in solution, whether derived from solids or from liquids, such as an infusion of raw meat, milk, or a weak solution of carbonate of ammonia, is quickly absorbed; and the glands, which were before limpid and of a greenish colour, become brownish and contain masses of aggregated granular matter. This matter, from its spontaneous movements, no doubt consists of protoplasm. No such effect is produced by the action of non-nitrogenous fluids. After the glands have been excited to secrete freely, they cease for a time to secrete, but begin again in the course of a few days.
Glands in contact with pollen, the leaves of other plants, and various kinds of seeds, pour forth much acid secretion, and afterwards absorb matter probably of an albuminous nature from them. Nor can the benefit thus derived be insignificant, for a considerable amount of pollen must be blown from the many wind-fertilised carices, grasses, &c., growing where Pinguicula lives, on to the leaves thickly covered with viscid glands and forming large rosettes. Even a few grains of pollen on a single gland causes it to secrete copiously. We have also seen how frequently the small leaves of Erica tetralix and of other plants, as well as various kinds of seeds and fruits, especially of Carex, adhere to the leaves. One leaf of the Pinguicula had caught ten of the little leaves of the Erica; and three leaves on the same plant had each caught a seed. Seeds subjected to the action of the secretion are sometimes killed, or the seedlings injured. We may, therefore, conclude that Pinguicula vulgaris, with its small roots, is not only supported to a large extent by the extraordinary number of insects which it habitually captures, but likewise draws some nourishment from the pollen, leaves, and seeds of other plants which often adhere to its leaves. It is therefore partly a vegetable as well as an animal feeder.
This species is so closely allied to the last that it is ranked by Dr. Hooker as a sub-species. It differs chiefly in the larger size of its leaves, and in the glandular hairs near the basal part of the midrib being longer. But it likewise differs in constitution; I hear from Mr. Ralfs, who was so kind as to send me plants from Cornwall, that it grows in rather different sites; and Dr. Moore, of the Glasnevin Botanic Gardens, informs me that it is much more manageable under culture, growing freely and flowering annually; whilst Pinguicula vulgaris has to be renewed every year. Mr. Ralfs found numerous insects and fragments of insects adhering to almost all the leaves. These consisted chiefly of Diptera, with some Hymenoptera, Homoptera, Coleoptera, and a moth. On one leaf there were nine dead insects, besides a few still alive. He also observed a few fruits of Carex pulicaris, as well as the seeds of this same Pinguicula, adhering to the leaves. I tried only two experiments with this species; firstly, a fly was placed near the margin of a leaf, and after 16 hrs. this was found well inflected. Secondly, several small flies were placed in a row along one margin of another leaf, and by the next morning this whole margin was curled inwards, exactly as in the case of Pinguicula vulgaris.
This species, of which living specimens were sent me by Mr. Ralfs from Cornwall, is very distinct from the two foregoing ones. The leaves are rather smaller, much more transparent, and are marked with purple branching veins. The margins of the leaves are much more involuted; those of the older ones extending over a third of the space between the midrib and the outside. As in the two other species, the glandular hairs consist of longer and shorter ones, and have the same structure; but the glands differ in being purple, and in often containing granular matter before they have been excited. In the lower part of the leaf, almost half the space on each side between the midrib and margin is destitute of glands; these being replaced by long, rather stiff, multicellular hairs, which intercross over the midrib. These hairs perhaps serve to prevent insects from settling on this part of the leaf, where there are no viscid glands by which they could be caught; but it is hardly probable that they were developed for this purpose. The spiral vessels proceeding from the midrib terminate at the extreme margin of the leaf in spiral cells; but these are not so well developed as in the two preceding species. The flower-peduncles, sepals, and petals, are studded with glandular hairs, like those on the leaves.
The leaves catch many small insects, which are found chiefly beneath the involuted margins, probably washed there by the rain. The colour of the glands on which insects have long lain is changed, being either brownish or pale purple, with their contents coarsely granular; so that they evidently absorb matter from their prey. Leaves of the Erica tetralix, flowers of a Galium, scales of grasses, &c. likewise adhered to some of the leaves. Several of the experiments which were tried on Pinguicula vulgaris were repeated on Pinguicula lusitanica, and these will now be given.
[(1) A moderately sized and angular bit of albumen was placed on one side of a leaf, halfway between the midrib and the naturally involuted margin. In 2 hrs. 15 m. the glands poured forth much secretion, and this side became more infolded than the opposite one. The inflection increased, and in 3 hrs. 30 m. extended up almost to the apex. After 24 hrs. the margin was rolled into a cylinder, the outer surface of which touched the blade of the leaf and reached to within the 1/20 of an inch of the midrib. After 48 hrs. it began to unfold, and in 72 hrs. was completely unfolded. The cube was rounded and greatly reduced in size; the remainder being in a semi-liquefied state.
(2) A moderately sized bit of albumen was placed near the apex of a leaf, under the naturally incurved margin. In 2 hrs. 30 m. much secretion was excited, and next morning the margin on this side was more incurved than the opposite one, but not to so great a degree as in the last case. The margin unfolded at the same rate as before. A large proportion of the albumen was dissolved, a remnant being still left.
(3) Large bits of albumen were laid in a row on the midribs of two leaves, but produced in the course of 24 hrs. no effect; nor could this have been expected, for even had glands existed here, the long bristles would have prevented the albumen from coming in contact with them. On both leaves the bits were now pushed close to one margin, and in 3 hrs. 30 m. this became so greatly inflected that the outer surface touched the blade; the opposite margin not being in the least affected. After three days the margins of both leaves with the albumen were still as much inflected as ever, and the glands were still secreting copiously. With Pinguicula vulgaris I have never seen inflection lasting so long.
(4) Two cabbage seeds, after being soaked for an hour in water, were placed near the margin of a leaf, and caused in 3 hrs. 20 m. increased secretion and incurvation. After 24 hrs. the leaf was partially unfolded, but the glands were still secreting freely. These began to dry in 48 hrs., and after 72 hrs. were almost dry. The two seeds were then placed on damp sand under favourable conditions for growth; but they never germinated, and after a time were found rotten. They had no doubt been killed by the secretion.
(5) Small bits of a spinach leaf caused in 1 hr. 20 m. increased secretion; and after 3 hrs. 20 m. plain incurvation of the margin. The margin was well inflected after 9 hrs. 15 m., but after 24 hrs. was almost fully re-expanded. The glands in contact with the spinach became dry in 72 hrs. Bits of albumen had been placed the day before on the opposite margin of this same leaf, as well as on that of a leaf with cabbage seeds, and these margins remained closely inflected for 72 hrs., showing how much more enduring is the effect of albumen than of spinach leaves or cabbage seeds .
(6) A row of small fragments of glass was laid along one margin of a leaf; no effect was produced in 2 hrs. 10 m., but after 3 hrs. 25 m. there seemed to be a trace of inflection, and this was distinct, though not strongly marked, after 6 hrs. The glands in contact with the fragments now secreted more freely than before; so that they appear to be more easily excited by the pressure of inorganic objects than are the glands of Pinguicula vulgaris. The above slight inflection of the margin had not increased after 24 hrs., and the glands were now beginning to dry. The surface of a leaf, near the midrib and towards the base, was rubbed and scratched for some time, but no movement ensued. The long hairs which are situated here were treated in the same manner, with no effect. This latter trial was made because I thought that the hairs might perhaps be sensitive to a touch, like the filaments of Dionaea.
(7) The flower-peduncles, sepals and petals, bear glands in general appearance like those on the leaves. A piece of a flower-peduncle was therefore left for 1 hr. in a solution of one part of carbonate of ammonia to 437 of water, and this caused the glands to change from bright pink to a dull purple colour; but their contents exhibited no distinct aggregation. After 8 hrs. 30 m. they became colourless. Two minute cubes of albumen were placed on the glands of a flower-peduncle, and another cube on the glands of a sepal; but they were not excited to increased secretion, and the albumen after two days was not in the least softened. Hence these glands apparently differ greatly in function from those on the leaves.]
From the foregoing observations on Pinguicula lusitanica we see that the naturally much incurved margins of the leaves are excited to curve still farther inwards by contact with organic and inorganic bodies; that albumen, cabbage seeds, bits of spinach leaves, and fragments of glass, cause the glands to secrete more freely;--that albumen is dissolved by the secretion, and cabbage seeds killed by it;--and lastly that matter is absorbed by the glands from the insects which are caught in large numbers by the viscid secretion. The glands on the flower-peduncles seem to have no such power. This species differs from Pinguicula vulgarisand grandiflora in the margins of the leaves, when excited by organic bodies, being inflected to a greater degree, and in the inflection lasting for a longer time. The glands, also, seem to be more easily excited to increased secretion by bodies not yielding soluble nitrogenous matter. In other respects, as far as my observations serve, all three species agree in their functional powers.