Insectivorous Plants. Darwin Charles
reflected light under the microscope, fine streaks of white opaque matter could be seen in the one, and traces of similar streaks in the other. The drops were replaced on the leaves, which re-expanded after 10 days; and now nothing was left except a very little transparent acid fluid.
Experiment 5. – This experiment was slightly varied, so that the albumen might be more quickly exposed to the action of the secretion. Two cubes, each of about 1/40 of an inch (.635 mm.), were placed on the same leaf, and two similar cubes on another leaf. These were examined after 21 hrs. 30 m., and all four were found rounded. After 46 hrs. the two cubes on the one leaf were completely liquefied, the fluid being perfectly transparent; on the other leaf some opaque white streaks could still be seen in the midst of the fluid. After 72 hrs. these streaks disappeared, but there was still a little viscid fluid left on the disc; whereas it was almost all absorbed on the first leaf. Both leaves were now beginning to re-expand.]
The best and almost sole test of the presence of some ferment analogous to pepsin in the secretion appeared to be to neutralise the acid of the secretion with an alkali, and to observe whether the process of digestion ceased; and then to add a little acid and observe whether the process recommenced. This was done, and, as we shall see, with success, but it was necessary first to try two control experiments; namely, whether the addition of minute drops of water of the same size as those of the dissolved alkalies to be used would stop the process of digestion; and, secondly, whether minute drops of weak hydrochloric acid, of the same strength and size as those to be used, would injure the leaves. The two following experiments were therefore tried: —
Experiment 6. – Small cubes of albumen were put on three leaves, and minute drops of distilled water on the head of a pin were added two or three times daily. These did not in the least delay the process; for, after 48 hrs., the cubes were completely dissolved on all three leaves. On the third day the leaves began to re-expand, and on the fourth day all the fluid was absorbed.
Experiment 7. – Small cubes of albumen were put on two leaves, and minute drops of hydrochloric acid, of the strength of one part to 437 of water, were added two or three times. This did not in the least delay, but seemed rather to hasten, the process of digestion; for every trace of the albumen disappeared in 24 hrs. 30 m. After three days the leaves partially re-expanded, and by this time almost all the viscid fluid on their discs was absorbed. It is almost superfluous to state that cubes of albumen of the same size as those above used, left for seven days in a little hydrochloric acid of the above strength, retained all their angles as perfect as ever.
Experiment 8. – Cubes of albumen (of 1/20 of an inch, or 2.54 mm.) were placed on five leaves, and minute drops of a solution of one part of carbonate of soda to 437 of water were added at intervals to three of them, and drops of carbonate of potash of the same strength to the other two. The drops were given on the head of a rather large pin, and I ascertained that each was equal to about 1/10 of a minim (.0059 ml.), so that each contained only 1/4800 of a grain (.0135 mg.) of the alkali. This was not sufficient, for after 46 hrs. all five cubes were dissolved.
Experiment 9. – The last experiment was repeated on four leaves, with this difference, that drops of the same solution of carbonate of soda were added rather oftener, as often as the secretion became acid, so that it was much more effectually neutralised. And now after 24 hrs. the angles of three of the cubes were not in the least rounded, those of the fourth being so in a very slight degree. Drops of extremely weak hydrochloric acid (viz. one part to 847 of water) were then added, just enough to neutralise the alkali which was still present; and now digestion immediately recommenced, so that after 23 hrs. 30 m. three of the cubes were completely dissolved, whilst the fourth was converted into a minute sphere, surrounded by transparent fluid; and this sphere next day disappeared.
Experiment 10. – Stronger solutions of carbonate of soda and of potash were next used, viz. one part to 109 of water; and as the same-sized drops were given as before, each drop contained 1/1200 of a grain (.0539 mg.) of either salt. Two cubes of albumen (each about 1/40 of an inch, or .635 mm.) were placed on the same leaf, and two on another. Each leaf received, as soon as the secretion became slightly acid (and this occurred four times within 24 hrs.), drops either of the soda or potash, and the acid was thus effectually neutralised. The experiment now succeeded perfectly, for after 22 hrs. the angles of the cubes were as sharp as they were at first, and we know from experiment 5 that such small cubes would have been completely rounded within this time by the secretion in its natural state. Some of the fluid was now removed with blotting-paper from the discs of the leaves, and minute drops of hydrochloric acid of the strength of the one part to 200 of water was added. Acid of this greater strength was used as the solutions of the alkalies were stronger. The process of digestion now commenced, so that within 48 hrs. from the time when the acid was given the four cubes were not only completely dissolved, but much of the liquefied albumen was absorbed.
Experiment 11. – Two cubes of albumen (1/40 of an inch, or .635 mm.) were placed on two leaves, and were treated with alkalies as in the last experiment, and with the same result; for after 22 hrs. they had their angles perfectly sharp, showing that the digestive process had been completely arrested. I then wished to ascertain what would be the effect of using stronger hydrochloric acid; so I added minute drops of the strength of 1 per cent. This proved rather too strong, for after 48 hrs. from the time when the acid was added one cube was still almost perfect, and the other only very slightly rounded, and both were stained slightly pink. This latter fact shows that the leaves were injured,20 for during the normal process of digestion the albumen is not thus coloured, and we can thus understand why the cubes were not dissolved.]
From these experiments we clearly see that the secretion has the power of dissolving albumen, and we further see that if an alkali is added, the process of digestion is stopped, but immediately recommences as soon as the alkali is neutralised by weak hydrochloric acid. Even if I had tried no other experiments than these, they would have almost sufficed to prove that the glands of Drosera secrete some ferment analogous to pepsin, which in presence of an acid gives to the secretion its power of dissolving albuminous compounds.
Splinters of clean glass were scattered on a large number of leaves, and these became moderately inflected. They were cut off and divided into three lots; two of them, after being left for some time in a little distilled water, were strained, and some discoloured, viscid, slightly acid fluid was thus obtained. The third lot was well soaked in a few drops of glycerine, which is well known to dissolve pepsin. Cubes of albumen (1/20 of an inch) were now placed in the three fluids in watch-glasses, some of which were kept for several days at about 90o Fahr. (32o.2 Cent.), and others at the temperature of my room; but none of the cubes were dissolved, the angles remaining as sharp as ever. This fact probably indicates that the ferment is not secreted until the glands are excited by the absorption of a minute quantity of already soluble animal matter, – a conclusion which is supported by what we shall hereafter see with respect to Dionaea. Dr. Hooker likewise found that, although the fluid within the pitchers of Nepenthes possesses extraordinary power of digestion, yet when removed from the pitchers before they have been excited and placed in a vessel, it has no such power, although it is already acid; and we can account for this fact only on the supposition that the proper ferment is not secreted until some exciting matter is absorbed.
On three other occasions eight leaves were strongly excited with albumen moistened with saliva; they were then cut off, and allowed to soak for several hours or for a whole day in a few drops of glycerine. Some of this extract was added to a little hydrochloric acid of various strengths (generally one to 400 of water), and minute cubes of albumen were placed in the mixture.21 In two of these trials the cubes were not in the least acted on; but in the third the experiment was successful. For in a vessel containing two cubes, both were reduced in size in 3 hrs.; and after 24 hrs. mere streaks of undissolved albumen were left. In a second vessel, containing two minute ragged bits of albumen, both were likewise reduced in size in 3 hrs., and after 24 hrs. completely disappeared. I then added a little weak hydrochloric acid to both vessels, and placed fresh cubes of albumen in them; but these were not acted on. This latter fact is intelligible according to the high authority of Schiff,22 who has demonstrated, as he believes, in opposition to the view held by some physiologists, that a certain small amount of pepsin
Sachs remarks ('Trait de Bot.' 1874, p. 774), that cells which are killed by freezing, by too great heat, or by chemical agents, allow all their colouring matter to escape into the surrounding water.
As a control experiment bits of albumen were placed in the same glycerine with hydrochloric acid of the same strength; and the albumen, as might have been expected, was not in the least affected after two days.
'Leons phys. de la Digestion,' 1867, tom. ii. pp. 114-126.