Chapter M: J. Berkeley (6)

M. de Seynes made the _Hymenomycetes_ an especial object of study,[A] but he can give us no information on the germination and growth of the spore. Hitherto almost nothing is positively known. As to the form of the spore, it is always at first spherical, which it retains for a long time, while attached to the basidia, and in some species, but rarely, this form is final, as in _Ag. terreus_, &c. The most usual form is either ovoid or regularly elliptic. All the _Coprini_ have the spores oval, ovoid, more or less elongated or attenuated from the hilum, which is more translucent than the rest of the spore. This last form is rather general amongst the Leucospores, in _Amanita_, _Lepiota_, &c. At other times the spores are fusiform, with regularly attenuated extremities, as in _Ag. ermineus_, Fr., or with obtuse extremities, as in _Ag. rutilans_, Sch. In _Hygrophorus_ they are rather irregular, reniform, or compressed in the centre all round. Hoffmann[B] has given a figure taken from _Ag. chlorophanus_, and Seynes verified it upon _Ag. ceraceus_, Sow. (See figures on page 121.)

The exospore is sometimes roughened, with more or less projecting warts, as may be seen in _Russula_, which much resembles _Lactarius_ in this as in some other particulars. The spores of the _Dermini_ and the _Hyporhodii_ often differ much from the sphærical form. In _Ag. pluteus_, Fr., and _Ag. phaiocephalus_, Bull, there is already a commencement of the polygonal form, but the angles are much rounded. It is in _Ag. sericeus_, _Ag. rubellus_, &c., that the polygonal form becomes most distinct. In _Dermini_ the angles are more or less pronounced, and become rather acute in _Ag. murinus_, Sow., and _Ag. ramosus_, Bull. The passage from one to the other may be seen in the stellate form of the conidia of _Nyctalis_.

It is almost always the external membrane that is coloured, which is subject to as much variation as the form. The more fine and more delicate shades are of rose, yellow-dun or yellow, violet, ashy-grey, clear fawn colour, yellow-orange, olive-green, brick-red, cinnamon-brown, reddish-brown, up to sepia-black and other combinations. It is only by the microscope and transparency that one can make sure of these tints; upon a sufficient quantity of agglomerated spores the colour may be distinguished by the naked eye. Colour, which has only a slight importance when considered in connection with other organs, acquires much in the spores, as a basis of classification.

With the growth of Agarics from the mycelium, or spawn, we are not deficient in information, but what are the conditions necessary to cause the spores themselves to germinate before our eyes and produce this mycelium is but too obscure. In the cultivated species we proceed on the assumption that the spores have passed a period of probation in the intestines of the horse, and by this process have acquired a germinating power, so that when expelled we have only to collect them, and the excrement in which they are concealed, and we shall secure a crop.[C] As to other species, we know that hitherto all attempts to solve the mystery of germination and cultivation has failed. There are several species which it would be most desirable to cultivate if the conditions could be discovered which are essential to germination.[D] In the same manner the _Boleti_ and _Hydnei_--in fact, all other hymenomycetal fungi, with the exception of the _Tremellini_--still require to be interrogated by persevering experiment and close inquiry as to their mode of germination, but more especially as to the essential conditions under which alone a fruitful mycelium is produced.

The germination of the spore has been observed in some of the _Tremellini_. Tulasne described it in _Tremella violacea_.[E] These spores are white, unilocular, and filled with a plastic matter of homogeneous appearance. From some portion of their surface an elongated germ filament is produced, into which the contents of the reproductive cell pass until quite exhausted. Other spores, perhaps more abundant, have a very different kind of vegetation. From their convex side, more rarely from the outer edge, these particular spores emit a conical process, generally shorter than themselves, and directed perpendicularly to the axis of their figure. This appendage becomes filled with protoplasm at the expense of the spore, and its free and pointed extremity finally dilated into a sac, at first globose and empty. This afterwards admits into its cavity the plastic matter contained in its support, and, increasing, takes exactly the form of a new spore, without, however, quite equalling in size the primary or mother spore. The spore of the new formation long retains its pedicel, and the mother spore which produced it, but these latter organs are then entirely empty and extremely transparent. Sometimes two secondary spores are thus engendered from the same spore, and their pedicels may be implanted on the same or on different sides, so as to be parallel in the former case, and growing in opposite directions in the latter. The fate of these secondary spores was not determined.

In _Dacrymyces deliquescens_ are found mingled amongst the spores immense numbers of small round or ovoid unilocular bodies, without appendages of any kind, which long puzzled mycologists. Tulasne ascertained that they are derived from the spores of this fungus when they have become free, and rest on the surface of the hymenium. Each of the cells of the spore emits exteriorly one or several of these corpuscles, supported on very short slender pedicels, which remain after the corpuscles are detached from them. This latter circumstance evidences that new corpuscles succeed the firstborn one on each pedicel as long as there remains any plastic matter within the spore. The latter, in fact, in consequence of this labour of production, becomes gradually emptied, and yet preserves the generative pedicels of the corpuscles, even when it no longer contains any solid or coloured matter. These pedicels are not all in the same plane, as may be ascertained by turning the spore on its longitudinal axis; but it often seems to be so when they are looked at in profile, on account of the very slight distance which then separates them one from another. It will also be remarked that they are in this case often implanted all on the same side of the reproductive body, and most often on its convex side. Their fecundity is exhausted with the plastic contents of the spore. The corpuscles, when placed in the most favourable conditions, have never given the least sign of vegetation; they have also remained for a long time in water without experiencing any appreciable alteration.

All the individuals of _Dacrymyces deliquescens_ do not produce these corpuscles in the same abundance; those which bear the most are recognizable by the pale tint of the reproductive dust with which they are covered; in others, where this dust preserves its golden appearance, only a few corpuscles are found. The spores which produce corpuscles do not appear at all apt to germinate. On the other hand, multitudes of spores will germinate which had not produced any corpuscles. Tulasne remarks on this, that these observations would authorize us to think that all spores, though perfectly identical to our eyes, have not, without distinction, the same fate, nor doubtless the same nature; and, in the second place, that these two kinds of bodies, if they are not always isolated, yet are most frequently met with on distinct individuals. This author claims for the corpuscles in question that they are spermatia, and thinks that their origin is only so far unusual in that they proceed from veritable spores.

The whole of the _Gasteromycetes_ have as yet to be challenged as to the mode and conditions of germination and development. It is probable that these will not materially differ from those which prevail in _Hymenomycetes_.

The germination in _Æcidium_ has been followed out by Tulasne,[F] either by placing the pseudospores in a drop of water, or confining them in a moist atmosphere, or by placing the leaves on which the _Æcidium_ flourishes upon water. The pseudospores plunged in water germinated more readily than the others. If the conditions were favourable, germination would take place in a few hours. _Æcidium Ranunculacearum_, D. C., on leaves of figwort, gives rarely more than one germinating filament, which soon attains three times the length of the diameter of the pseudospore. This filament generally remains simple, sometimes torulose, and distorted in a long spire. Sometimes it has been seen divided into two branches, nearly equal to each other. The spore in germinating empties itself of its plastic contents, contracts, and diminishes in size. The pseudospores of _Æcidium crassum_, P., emit three long filaments, which describe spirals, imitating the twistings of the stem of a bean or bindweed. In _Æcidium Violæ_, Schum, one filament is produced, which frequently rolls up its anterior extremity into a spire, but more often this same extremity rises in a large ovoid, irregular vesicle, which continues the axis of the filament, or makes with it a more or less decided angle. In whatever manner placed, this vesicle attracts to it all the orange protoplasm, and hardly does this become settled and complete before the vesicle becomes the starting point of a new development, for it begins to produce at its apex a filament, more slender than the previous one, stiff, and unbranched.

According to M. Tulasne, the germination of the pseudospores of _Æcidium Euphorbiæ_ on _Euphorbia sylvatica_ differ in some respects from the preceding. When dropped upon water these spores very soon emit a short tube, which ordinarily curves in an arch or circle, almost from its origin, attaining a length of from three to six times the diameter of the spore; then this tube gives rise to four spicules, each of which produces a small obovate or reniform sporule; the generation of these sporules absorbs all the plastic matter contained in the germ-tube, which permits of the observation that it was divided into four cells corresponding with the number of spicules. These sporules germinate very rapidly from an indefinite point of their surface, emitting a filiform process, which is flexuous and very delicate, not extending more in length than three times that of the long axis of the sporule, often less, reproducing at its summit a new sporule, differing in form and size from that which preceded it. This sporule of the second formation becomes at its apex a vital centre, and sprouts one or more linear buds, of which the elongation is occasionally interrupted by the formation of vesicular swellings. As Tulasne observes, the pseudospores of the _Æcidium_ and the greater number of Uredines are easily wetted with water before arriving at maturity; but when they are ripe, on the contrary, they appear to be clothed with a greasy matter which protects them from the liquid, forcing them almost all to rest on the surface.

The pseudospores of _Roestelia_ are produced in strings or chaplets, as in _Æcidium_, with this difference, that instead of being contiguous they are separated by narrow isthmuses. The ripe pseudospores are enveloped in a thick tegument, of a dark brown colour. They germinate readily on water, producing a filament fifteen times as long as the diameter of the spore. This filament is sometimes rolled or curved. Towards its extremity it exhibits protuberances which resemble the rudiments of ramuli, or they terminate in a vesicle which gives rise to a slender filament. The tegument of these pseudospores, above all in those which have germinated, and have consequently become more transparent, it is easy to see has many pores, or round ostioles.

In _Peridermium_ the pseudospores, when dropped upon water, germinate at any point of their surface. Sometimes two unequal filaments issue from the same spore. After forty-eight hours of vegetation in the air, the greater part had already emitted a multitude of thick little branches, themselves either simple or branched, giving to the filaments a peculiar aspect. Tulasne did not on any occasion observe the formation of secondary spores.

In the Uredines proper the germination seems to be somewhat similar, or at least not offering sufficient differences to warrant special reference in _Uredo_, _Trichobasis_, _Lecythea_, &c. In _Coleosporium_ there are two kinds of spores, one kind consisting of pulverulent single cells, and the other of elongated septate cells, which break up into obovate joints. Soon after the maturity of the pulverulent spores, each begins to emit a long tube, which is habitually simple, and produces at its summit a reproductive cellule, or reniform sporule. The orange protoplasm passes along the colourless tubes to the terminal sporule at the end of its vegetation. The two forms of spores in this genus are constantly found on the same leaf, and in the same pulvinule, but generally the pulverulent spores abound at the commencement of the summer. The reniform sporules begin to germinate in a great number as soon as they are free; some few extend a filament which remains simple and uniform, but more commonly it forms at its extremity a second sporule. If this does not become isolated, to play an independent life, the filament is continued, and new vesicles are repeated many times.

In _Melampsora_ the summer spores are of the _Lecythea_ type, and were included in that genus till their relation with _Melampsora_ was clearly made out. The winter spores are in solid pulvinules, and their fructification takes place towards the end of winter or in the spring. This phenomenon consists in the production of cylindrical tubes, which start from the upper extremity of the wedge-shaped spores, or more rarely from the base. These tubes are straight or twisted, simple or bifurcated, and each of them very soon emits four monosporous spicules, at the same time that they become septate. The sporules are in this instance globose.

In _Uromyces_ germination follows precisely the same type as that of the upper cell of _Puccinia_; in fact, Tulasne states that it is very difficult to say in what they differ from the _Pucciniæ_ which are accidentally unilocular.

In _Cystopus_ a more complex method prevails, which will be examined more closely hereafter.

In _Puccinia_, as already observed when describing their structure, the pseudospores are two-celled. From the pores of each cell, which are near the central septum, springs a clavate tube, which attains two or three times the total length of the fruit, and of which the very obtuse extremity curves more or less in the manner of a crozier.[G] This tube, making a perfectly uncoloured transparent membrane, is filled with a granular and very pale plastic matter at the expense of the generative cell, which is soon rendered vacant; then it gives rise to four spicules, usually on the same side, and at the summit of these produces a reniform cellule. The four sporules so engendered exhaust all the protoplasm at first contained in the generative cell, so that their united capacity proves to be evidently much insufficient to contain it, the more so as it leads to the belief that this matter undergoes as it condenses an elaboration which diminishes its size. In all cases the spicule originates before the sporule which it carries, and also attains its full length when the sporule appears. The form of the latter is at first globular, then ellipsoid, and more or less curved. All these phases of vegetation are accomplished in less than twelve hours, and if the spore is mature and ready for germination, it is sufficient to provoke it by keeping the pseudospores in a humid atmosphere. During this process the two cells do not separate, nor does one commence germination before the other, but both simultaneously. When the sporules are produced, the protospore, somewhat analogous to a prothallus, has performed its functions and decays. Towards the time of the falling of the sporules they are nearly all divided into four unequal cells by transverse and parallel septa. These sporules in time produce, from any point on their surface, a filament, which reproduces a new sporule, resembling the first, but generally smaller. This sporule of the second generation ordinarily detaches itself from its support before germinating.

The pseudospores of _Triphragmium ulmariæ_ have been seen in April germinating on old leaves of the meadowsweet which survived the winter, whilst at the same time new tufts of the spores were being developed on the leaves of the year. These fruits of the spring vegetation would not germinate the same year. Each cell in germination emits a long cylindrical filament, containing a brownish protoplasm, on which four spicules, bearing as many sporules, are generated.

The germination of the black fruits of _Phragmidium_ only appears to take place in the spring. It greatly resembles that in _Puccinia_, except that the filament is shorter, and the sporules are spherical and orange-coloured, instead of being kidney-shaped and pale. In the species found on the leaves of the common bramble, the filament emitted by each cell attains three or four times the length of the fruit. The granular orange protoplasm which fills it passes ere long into the sporules, which are engendered at the extremity of pointed spicules. After the long warty fruits are emptied of their contents they still seem as dark as before, but the pores which are pierced in the sides, through which the germinating filaments have proceeded, are more distinctly visible.

It will be observed that throughout all these allied genera of _Uromyces_, _Puccinia_, _Triphragmium_, and _Phragmidium_ the same type of germination prevails, which confirms the accuracy of their classification together, and renders still less probable the supposed affinity of _Phragmidium_ with _Sporidesmium_, which was at one time held by very astute mycologists, but which is now abandoned. This study of germination leads also to a very definite conclusion with regard to the genus _Uromyces_--that it is much more closely related to _Puccinia_ and its immediate allies than to other unicellular Uredines.

The germination of the pseudospores of the gelatinous Uredines of the genus _Podisoma_ was studied by Tulasne.[H] These pretended spores, he writes, are formed of two large conical cells, opposed by their base and easily separating. They vary in length. The membrane of which they are formed is thin and completely colourless in most of them, though much thicker and coloured brown in others. It is principally the spores with thin membranes that emit from near the middle very obtuse tubes, into which by degrees, as they elongate, the contents of the parent utricles pass. Each of the two cells of the supposed spore may originate near its base four of these tubes, opposed to each other at their point of origin, and their subsequent direction; but it is rather rare for eight tubes, two by two, to decussate from the same spore or basidium. Usually there are only two or three which are completely developed, and these tend together towards the surface of the fungus, which they pass, and expand at liberty in the air. The tubes generally become thicker by degrees as they elongate, some only slightly exceeding the length of the protospores. Others attain three or four times that length, according to the greater or less distance between the protospore and the surface of the plant. In the longest tubes it is easy to observe how the colouring matter passes to their outer extremity, leaving the portion nearest to the parent cell colourless and lifeless. When nearly attaining their ultimate dimensions, all the tubes are divided towards their outer extremity by transverse septa into unequal cells; then simple and solitary processes, of variable length and form, but attenuated upwards, proceed from each segment of the initial tube, and produce at their extremity an oval spore (teleutospore, Tul.), which is slightly curved and unilocular. These spores absorb all the orange endochrome from the original tubes. They appear in immense numbers on the surface of the fungus, and when detached from their spicules fall upon the ground or on any object which may be beneath them. So freely are they deposited that they may be collected on paper, or a slip of glass, like a fine gold-coloured powder. Again, these secondary spores (teleutospores) are capable of germination, and many of them will be found to have germinated on the surface of the _Podisoma_ whence they originated. The germ filament which they produce springs habitually from the side, at a short distance from the hilum, which indicates the point of attachment to the original spicule. These filaments will attain to from fifteen to twenty times the diameter of the spore in length before branching, and are in themselves exceedingly delicate. The tubes which issue from the primary spores (protospores, Tul.) are not always simple, but sometimes forked; and the cells which are ultimately formed at their extremities, though producing filiform processes, do not always generate secondary spores (teleutospores) at their apices. This mode of germination, it will be seen, resembles greatly that which takes place in _Puccinia_.

The germination of the Ustilagines was in part examined by Tulasne, but since has received accessions through the labours of Dr. A. Fischer von Waldheim.[I] Nothing, however, of any importance is added to our knowledge of the germination of _Tilletia_, which was made known as early as 1847.[J] After some days a little obtuse tube is protruded through the epispore, bearing at its apex long fusiform bodies, which are the sporules of the first generation. These conjugate by means of short transverse tubes, after the manner of the threads of _Zygnema_. Afterwards long elliptical sporules of the second generation are produced on short pedicels by the conjugated fusiform bodies of the first generation. (Fig. 89, _ss._) Ultimately these sporules of the second generation germinate, and generate, on short spicules, similar sporules of a third generation. (Fig. 89, _st._)

In _Ustilago (flosculorum)_ germination takes place readily in warm weather. The germ tube is rather smaller at its base than further on. In from fifteen to eighteen hours the contents become coarsely granular; at the same time little projections appear on the tube which are narrowed at the base, into which some of the protoplasm passes. These ultimately mature into sporules. At the same time a terminal sporule generally appears on the threads. Secondary sporules frequently grow from the primary, which are rather smaller, and these occasionally give rise to a third generation.

In _Urocystis (pompholygodes)_ the germinating tubes spring exclusively from the darker central cells of the clusters. From these are developed at their extremity three or four linear bodies, as in _Tilletia_, but after this no further development has as yet been traced. It may be remarked here that Waldheim observed similar conjugation of the sporules in some species of _Ustilago_ as have been remarked in the sporules of the first generation in _Tilletia_.

Returning to _Cystopus_, as the last of the Uredines, we must briefly recapitulate the observations made by Professor de Bary,[K] who, by the bye, claims for them an affinity with _Peronospora_ (Mucedines but too well known in connection with the potato disease), and _not_ with the Uredines and their allies. In this genus there are two kinds of reproductive organs, those produced on the surface of the plant bursting through the cuticle in white pustules, and which De Bary terms _conidia_, which are generated in chains, and certain globose bodies termed _oogonia_, which are developed on the mycelium in the internal tissues of the foster plant. When the conidia are sown on water they rapidly absorb the moisture, and swell; the centre of one of the extremities soon becomes a large obtuse papilla resembling the neck of a bottle. This is filled with a granular protoplasm, in which vacuoles are formed. Soon, however, these vacuoles disappear, and very fine lines of demarcation separate the protoplasm into from five to eight polyhedric portions, each presenting a little faintly-coloured vacuole in the centre (_a_). Soon after this division the papilla at the extremity swells, opens itself, and at the same time the five to eight bodies which had formed in the interior are expelled one by one (_b_). These are zoospores, which at first take a lenticular form, and group themselves before the mouth of the parent cell in a globose mass (_c._) Very soon, however, they begin to move, and then vibratile cilia show themselves (_d_), and by means of these appendages the entire globule moves in an oscillating manner as one by one the zoospores disengage themselves, each becoming isolated and swimming freely in the surrounding fluid. The movement is precisely that of the zoospores of Algæ.

The generation of the zoospores commences within from an hour and a half to three hours after the sowing of the conidia on water. From the oogonia, or resting spores, similar zoospores, but in greater number, are generated in the same manner, and their conduct after becoming free is identical. Their movements in the water usually last from two to three hours, then they abate, the cilia disappear, and the spore becomes immovable, takes a globose form, and covers itself with a membrane of cellulose. Afterwards the spore emits, from any point whatever of its surface, a thin, straight or flexuous tube, which attains a length of from two to ten times the diameter of the spore. The extremity becomes clavate or swollen, after the manner of a vesicle, which receives by degrees the whole of the protoplasm.

De Bary then proceeds to describe experiments which he had performed by watering growing plants with these zoospores, the result being that the germinating tubes did not penetrate the epidermis, but entered by the stomates, and there put forth an abundant mycelium which traversed the intercellular passages. Altogether the germination of these conidia or zoospores offers so many differences from the ordinary germination of the Uredines, and is so like that which prevails in _Peronospora_, in addition to the fact of both genera producing winter spores or oogonia, that we cannot feel surprised that the learned mycologist who made these observations should claim for _Cystopus_ an affinity with _Peronospora_ rather than with the plants so long associated with it amongst the _Coniomycetes_.

In passing from these to the _Mucedines_, therefore, we cannot do so more naturally than by means of that genus of white moulds to which we have just alluded. The erect branched threads bear at the tip of their branchlets spores, or conidia, which conduct themselves in a like manner to the organs so named in _Cystopus_, and oogonia or resting spores developed on the mycelium within the tissues of the foster plant also give origin to similar zoospores.

The conidia are borne upon erect, elongated filaments, originating from the creeping mycelium. These threads are hollow, and rarely septate; the upper portion divided into numerous branches, and these again are subdivided, the ultimate ramuli each terminated by a single conidium. This body when mature is oval or elliptical, filled with protoplasm, but there is a diversity in their mode of germination. In the greater part, of which _P. effusa_ may be taken as an example, the conidia have the function of simple spores. Placed in favourable conditions, each of them puts forth a germ-tube, the formation of which does not differ in any essential point from what is known of the spores of the greater part of fungi.

The short oval conidia of _P. gangliformis_ have little obtuse papillæ at their apex, and it is at this point that germination commences.

The conidia of _P. densa_ are similar, but the germination is different. When placed in a drop of water, under favourable circumstances, the following changes may be observed in from four to six hours. The protoplasm, at first uniformly distributed in all the conidia, appears strewn with semi-lenticular, and nearly equidistant vacuoles, of which the plane face is immediately in contact with the periphery of the protoplasm. These vacuoles number from sixteen to eighteen in _P. macrocarpa_, but are less numerous in _P. densa_. A short time after the appearance of the vacuoles the entire conidium extends itself so that the papilla disappears. Suddenly it reappears, elongates itself, its attenuated membrane vanishes, and the protoplasm is expelled by the narrow opening that remains in place of the papilla. In normal cases the protoplasm remains united in a single mass that shows a clear but very delicate outline. When it has reached the front of the opening in the conidium, which is thus emptied, the mass remains immovable. In _P. densa_ it is at first of a very irregular form, but assumes by degrees a regular globose shape. This is deprived of a distinct membrane, the vacuoles that disappeared in the expulsion again become visible, but soon disappear for a second time. The globule becomes surrounded with a membrane of cellulose, and soon puts out from the point opposite to the opening of the conidium a thick tube which grows in the same manner as the germ-tube of the conidia in other species. Sometimes the expulsion of the protoplasm is not completely accomplished; a portion of it remaining in the membrane of the conidium detaches itself from the expelled portion, and while this is undergoing changes takes the form of a vesicle, which is destroyed with the membrane. It is very rare that the protoplasm is not evacuated, and that the conidia give out terminal or lateral tubes in the manner that is normal to other species without papillæ. The germination just described does not take place unless the conidia are entirely surrounded by water; it is not sufficient that they repose upon its surface. Besides, there is another condition which, without being indispensable, has a sensible influence on the germination of _P. macrocarpa_, and that is the exclusion of light. To ascertain if the light or the darkness had any influence, two equal sowings were placed side by side, the one under a clear glass bell, the other under a blackened glass bell. Repeated many times, these experiments always gave the same result--germination in from four to six hours in the conidia under the blackened glass; no change in those under the clear glass up to the evening. In the morning germination was completed.

The conidia of _P. umbelliferarum_ and _P. infestans_[L] show an analogous structure. These bodies, if their development be normal, become zoosporangia. When they are sown upon water, one sees at the end of some hours the protoplasm divided by very fine lines, and each of the parts furnished with a small central vacuole. Then the papilla of the conidium disappears. In its place appears a rounded opening, by which the parts of the protoplasm are expelled rapidly, one after the other. Each of these, when free, immediately takes the form of a perfect zoospore, and commences to agitate itself. In a few moments the sporangium is empty and the spores disappear from the field of the microscope.

The zoospores are oval or semi-oval, and in _P. infestans_ the two cilia spring from the same point on the inferior border of the vacuole. Their number in a sporangium are from six to sixteen in _P. infestans_, and from six to fourteen in _P. umbelliferarum_. The movement of the zoospores ceases at the end of from fifteen to thirty minutes. They become motionless, cover themselves with a membrane of cellulose, and push out slender bent germ-tubes which are rarely branched. It is but seldom that two tubes proceed from the same spore. The same development of the zoospores in _P. infestans_ is favoured by the exclusion of the light. Placed in a position moderately lighted or protected by a blackened bell, the conidia very readily produced zoospores.

A second form of germination of the conidia in _P. infestans_, when sown upon a humid body or on the surface of a drop of water, consists in the conidium emitting from its summit a simple tube, the extremity of which swells itself into the form of an oval vesicle, drawing to itself, little by little, all the protoplasm contained in the conidium. Then it isolates itself from the germ-tube by a septum, and takes all the essential characteristics of the parent conidium. This secondary conidium can sometimes engender a third cellule by a similar process. These secondary and tertiary productions have equally the character of sporangia. When they are plunged into water, the ordinary production of zoospores takes place.

Lastly, there is a third mode of germination which the conidia of _P. infestans_ manifest, and which consists in the conidium emitting from its summit a simple or branched germ-tube. This grows in a similar manner to the conidia first named as of such species as _P. effusa_. The conditions which control this form of germination cannot be indicated, since some conidia which germinate after this manner will sometimes be found mixed with others, the majority of which furnish zoospores. It may be that the conidia themselves are in some sort of abnormal condition.

In all the species examined the conidia possess the power of germination from the moment of their maturity. The younger they are the more freely they germinate. They can retain this power for some days or weeks, provided they are not entirely dried. Dessication in an ordinary temperature seemed sufficient to destroy the faculty of germinating in twenty-four hours, when the conidia had been removed from the leaves on which they were produced. They none of them retained the faculty during a few months, hence they cannot preserve it during the winter.

The germs of _Peronospora_ enter the foster plant if the spores are sown upon a part suitable for the development of the parasite. It is easy to convince one's self that the mycelium, springing from the penetrating germs, soon takes all the characters that are found in the adult state. Besides, when cultivated for some time, conidiiphorous branches can be seen growing, identical with those to which it owes its origin. Such cultivation is so readily accomplished that it can be made upon cut leaves preserved fresh in a moist atmosphere.

In the species of _Peronospora_ that inhabit perennial plants, or annual plants that last through the winter, the mycelium hidden in the tissues of the foster-plant lasts with it. In the spring it recommences vegetation, and emits its branches into the newly-formed organs of its host, there to fructify. The _Peronospora_ of the potato is thus perennial by means of its mycelium contained in the browned tissue of the diseased tubers. When in the spring a diseased potato begins to grow, the mycelium rises in the stalk, and soon betrays itself by blackish spots. The parasites can fructify abundantly on these little stalks, and in consequence propagate themselves in the new season by the conidia coming from the vivacious mycelium.

The diseased tubers of the potato always contain the mycelium of _P. infestans_, which never fructifies there as long as the skin of the tuber is intact. But when, in cutting the tuber, the parenchyma occupied by the mycelium is exposed to the contact of the air, it covers itself with conidia-bearing branches at the end of from twenty-four to forty-eight hours. Analogous results are obtained with the stalks of the potato. It is evident that in these experiments nothing is changed except the contact of the air; the specific conditions particularly remain the same. It appears, therefore, that it is this contact alone which determines generally the production of the conidiiferous branches.[M]

The mode of germination and development in the Mucors has been studied by several observers, but most recently by Van Tieghem and Le Monnier.[N] In one of the common forms, the _Mucor phycomyces_ of some authors, and the _Phycomyces nitens_ of others, the process is given in detail. In this species germination will not take place in ordinary water, but it readily takes place in orange juice and other media. The spore loses colour, swells, and absorbs fluid around it until double its original size and ovoid. Then a thick thread is emitted from one or both extremities, which elongates and becomes branched in a pinnate manner. Sometimes the exospore is ruptured and detached loosely from the germinating spore. After about forty-eight hours from the first sowing, the mycelium will send branches into the air, which again become abundantly branched; other short submerged branches will also remain simple, or have tuft-like ramifications, each terminating in a point, so as to bristle with spiny hairs. In two or three days abruptly swollen branches, of a club shape, will make their appearance on the threads both in the air and in the fluid. Sometimes these branches are prolonged into an equal number of sporangia-bearing threads, but most frequently they divide first at their swollen summits into numerous branches, of which usually one, sometimes two or three, develop into sporangia-bearing threads, while the rest are short, pointed, and form a tuft of rootlets. Sometimes these rootlets reduce themselves to one or more rounded protuberances towards the base of the sporangia-bearing threads.

There are often also a certain number of the branches which had acquired a clavate shape, and do not erect themselves above the surface, instead of producing a fertile thread, which would seem to have been their first intention, become abruptly attenuated, and are merely prolonged into a mycelial filament. Although in other species chlamydospores are formed in such places on the mycelium, nothing of the kind has been traced in this species, more than here indicated. Occasionally, when germination is arrested prematurely, certain portions of the hyphæ, in which the protoplasm maintains its vitality, become partitioned off. This may be interpreted as a tendency towards the formation of chlamydospores, but there is no condensation of protoplasm, or investiture with a special membrane. Later on this isolated protoplasm is gradually altered, separating into somewhat regular ovoid or fusiform granules, which have, to a certain extent, the appearance of spores in an ascus, but they seem to be incapable of germination.

Another method of reproduction, not uncommon in _Mucorini_, is described by Van Tieghem in this species. Conjugating threads on the substratum by degrees elaborate zygospores, but these, contrary to the mode in other species, are surrounded by curious branched processes which emanate from the arcuate cells on either side of the newly-developed zygospore. This system of reproduction is again noticed more in detail in the chapter on polymorphism.

M. de Seynes has given the details of his examination of the sporidia of _Morchella esculenta_ during germination.[O] A number of these sporidia, placed in water in the morning, presented, at nine o'clock of the same evening, a sprout from one of the extremities, measuring half the length of the spore. In the morning of the next day this sprout had augmented, and become a filament three or four times as long. The next day these elongated filaments exhibited some transverse divisions and some ramifications. On the third day, the germination being more advanced, many more of the sporidia were as completely changed, and presented, in consequence of the elongation, the appearance of a cylindrical ruffle, the cellular prolongations arising from the germination having a tendency towards one of the extremities of the longer axis of the sporidium, and more often to the two opposed extremities, either simultaneously or successively. Out of many hundreds of sporidia examined during germination, he had only seen a very few exceptions to this rule, among which he had encountered the centrifugal tendency to vegetate by two opposed filaments, proving that if it bears a second by the side of the primal filament situated at one of the poles, a second would also be seen from the side of the filament coming from the opposite pole.

Before being submitted to the action of water, the contents of the sporidia seemed formed of two distinct parts, one big drop of yellow oil of the same form as the sporidium, with the space between it and the cell wall occupied by a clear liquid, more fluid and less refractive, nearly colourless, or at times slightly roseate. As the membrane absorbed the water by which it was surrounded, the quantity of this clear liquid was augmented, and the rosy tint could be more easily distinguished. All the contents of the spore, which up to this time remained divided into two parts, presented altogether one aspect, only containing numerous granulations, nearly of equal size, completely filling it, and reaching the inner face of the sporic membrane.

After this time the sporidium augments in size very rapidly, becoming at times irregular, and sometimes even as much as from two to three times its original dimensions, then there appears at the surface, usually at one of the poles of the ellipse, a small prominence, with an extremely fine membrane, which does not appear to separate itself from that which surrounds the sporidium, and it is difficult to say whether it is a prolongation of the internal membrane going across the outside, or simply a prolongation caused by a continuation of tissue of an unique membrane. Sometimes there may be seen at the point where the primal filament issues from the sporidium a circular mark, which appears to indicate the rupture of the external membrane. From this time another change comes over the contents. We again find the yellow oily liquid, now occupying the external position, with some drops of colourless or roseate liquid in the centre, so that the oily liquid and the more limpid fluid interchange the positions which they occupied previous to the commencement of germination. Whether these two fluids have undergone any change in their constitution is difficult to determine, at all events the oily liquid appears to be less refractive and more granular, and it may be that it is a product of new formation, containing some of the elements of the primitive oily drop. Having regard to the delicate character of the membrane of the germinating filaments, De Seynes supposed that it might offer greater facility for the entrance of water by endosmose, and account for the rapid enlargement of the sporidia. By a series of experiments he became satisfied that this was the case to a considerable extent, but he adds:--"I cannot help supposing that a greater absorption of greasy matter in the cell which is the first product of germination raises an objection to an aqueous endosmose. One can also see in this experience a proof of the existence of two special membranes, and so suppose that the germinative cell is the continuation of the internal membrane, the external membrane alone being susceptible of absorbing the liquids, at least with a certain rapidity."

In other _Discomycetes_ germination takes place in a similar manner. Boudier[P] narrates that in _Ascobolus_, when once the spore reaches a favourable place, if the circumstances are good, _i.e._, if the temperature is sufficiently high and the moisture sufficient, it will germinate. The time necessary for this purpose is variable, some hours sufficing for some species; those of _A. viridis_, for example, germinate in eight or ten hours, doubtless because, being terrestrial, it has in consequence less heat. The spore slightly augments in size, then opens, generally at one or other extremity, sometimes at two, or at any point on its surface, in order to pass the mycelium tubes. At first simple, without septa, and granular in the interior, above all at the extremity, these tubes, the rudiment of the mycelium, are not long in elongating, in branching, and later in having partitions. These filaments are always colourless, only the spore may be coloured, or not. Coemans has described them as giving rise to two kinds of conidia,[Q] the one having the form of _Torula_, when they give rise to continuous filaments, the other in the form of _Penicillium_, when they give birth to partitioned filaments. De Seynes could never obtain this result. Many times he had seen the _Penicillium glaucum_ invade his sowings, but he feels confident that it had nothing to do with the _Ascobolus_. M. Woronin[R] has detailed some observations on the sexual phenomena which he has observed in _Ascobolus_ and _Peziza_, and so far as the scolecite is concerned these have been confirmed by M. Boudier.

There is no reason for doubt that in other of the _Discomycetes_ the germination of the sporidia is very similar to that already seen and described, whilst in the _Pyrenomycetes_, as far as we are aware, although the production of germinating tubes is by no means difficult, development has not been traced beyond this stage.[S]

[A] Seynes, J. de, "Essai d'une Flore Mycologique de la Montpellier,"
&c. (1863), p. 30.

[B] Hoffman, "Icones Analyticæ Fungorum."

[C] The spores of Agarics which are devoured by flies, however,
though returned in their dung in an apparently perfect state,
are quite effete. It is, we believe, principally by the
_Syrphidæ_, which devour pollen, that fungus spores are
consumed.

[D] All attempts at Chiswick failed with some of the more esculent
species, and Mr. Ingram at Belvoir, and the late Mr. Henderson
at Milton, were unsuccessful with native and imported spawn.

[E] Tulasne, "On the Organization of the Tremellini," "Ann. des. Sci.
Nat." 3^me sér. xix. (1853), p. 193.

[F] Tulasne, "Mémoire sur les Urédinées."

[G] Tulasne, in his "Memoirs on the Uredines."

[H] Mr. Berkeley has lately published a species under the name of _P.
Ellisii_, in which the gelatinous element is scarcely
discernible till the plant is moistened. There are two septa in
this species, and another species or form has lately been
received from Mr. Ellis which has much shorter pedicels, and
resembles more closely _Puccinia_, from which it is chiefly
distinguished by its revivescent character.

[I] Von Waldheim, on the "Development of the Ustilagineæ," in
"Pringsheim's Jahrbucher," vol. vii. (1869); translated in
"Transactions of N. Y. State Agricultural Society for 1870."

[J] Berkeley, on the "Propagation of Bunt," in "Trans. Hort. Soc.
London," ii. (1847), p. 113; Tulasne, second memoir, in "Ann.
des. Sci. Nat." ii. (4^me sér.), p. 77; Cooke, in "Journ.
Quekett Micro. Club," i. p. 170.

[K] De Bary, "Recherches," &c. in "Annales des Sciences Naturelles"
(4^me sér.), xx. p. 5; Cooke in "Pop. Sci. Rev." iii. (1864),
p. 459.

[L] This is the mould which produces the potato murrain.

[M] De Bary, "Champignons parasitiques," in "Annales des Sci. Nat."
(4^me sér.), xx. p. 5; Cooke, "Microscopic Fungi," cap. xi. p.
138; "Popular Science Review," iii. 193 (1864).

[N] Van Tieghem and Le Monnier, "Researches on Mucorini," in "Ann.
des Sci. Nat." (1873), xvii. p. 261; Summary in "Quart. Journ.
Micro. Science" (2nd ser.), xiv. p. 49.

[O] Seynes, "Essai d'une Flore Mycologique."

[P] Boudier, "Mémoire sur l'Ascoboles," pt. i. iv. f. 13-15.

[Q] Coemans, "Spicilége Mycologique," i. p. 6.

[R] Woronin, "Abhandlungen der Senchenbergischen Naturfor.
Gesellschaft" (1865), p. 333.

[S] In the very important observations made by Dr. Cunningham at
Calcutta, on substances floating in the atmosphere, it appeared
that the sporidia of many _Sphæriæ_ actually germinated after
being taken up by the air. The multitude of fungus spores which
were observed in every case was quite extraordinary.

VIII.

SEXUAL REPRODUCTION.

The existence of some sort of sexual reproduction in Fungi has long been suspected, although in earlier instances upon insufficient grounds; but of late years observations have multiplied and facts accumulated which leave no doubt of its existence. If the _Saprolegniæ_ are left out of the question as disputed Fungi, there still remain a number of well authenticated instances of the phenomena of copulation, and many other facts which indicate some sort of sexual relationship. The precise manner in which those minute bodies, so common amongst the _Sphæronemei_, which we prefer to call stylospores, perform their functions is still to a great extent a mystery; yet it is no longer doubted that certain species of _Aposphæria_, _Phoma_, _Septoria_, &c., are only conditions of some species of _Sphæria_, often developed and matured in close proximity to them on the same host. In _Æcidium_, _Roestelia_, &c., spermogonia are produced plentifully on or near the same spots on which the fructification appears, either simultaneously or at a later period.[A] The relation of _Cytispora_ to _Valsa_ was suspected by Fries very many years ago, and, as since demonstrated, with very good reason. All attempts, however, to establish anything like sexual reproduction in the higher forms of _Hymenomycetes_ have at present been unsuccessful; and the same may be said of the _Gasteromycetes_; but in _Ascomycetes_ and _Physomycetes_ instances abound.

We know not whether any importance is to be attached to the views of M. A. S. Oersted,[B] which have not since been confirmed, but which have been cited with some approval by Professor de Bary, as to a trace of sexual organs in _Hymenomycetes_. He is supposed to have seen in _Agaricus variabilis_, P., oocysts or elongated reniform cells, which spring up like rudimentary branches of the filaments of the mycelium, and enclose an abundant protoplasm, if not even a nucleus. At the base of these oocysts appear the presumed antheridia, that is to say, one or two slender filaments, which generally turn their extremities towards the oocysts, and which more rarely are applied to them. Then, without ulteriorily undergoing any appreciable modifications, the fertile cell or oocyst becomes enveloped in a network of filaments of mycelium which proceed from the one which bears it, and this tissue forms the rudiments of the cap. The reality of some kind of fecundation in this circumstance, and the mode of the phenomena, if there is one, are for the present equally uncertain. If M. Oersted's opinion is confirmed, naturally the whole of the cap will be the product of fecundation. Probably Karsten (Bonplandia, 1862, p. 62) saw something similar in _Agaricus campestris_, but his account is obscure.

In _Phycomyces_ the organs of reproduction have been subjected to close examination by Van Tieghem,[C] and although he failed to discover chlamydospores in this, he describes them in other Mucors. In this species, besides the regular sexual development, by means of sporangia, there is a so-called sexual reproduction by means of zygospores, which takes place in this wise. The threads which conjugate to form the zygospores are slender and erect on the surface of the substratum. Two of these threads come into close contact through a considerable length, and clasp each other by alternate protuberances and depressions. Some of the protuberances are prolonged into slender tubes. At the same time the free extremities of the threads dilate, and arch over one towards the other until their tops touch like a vice, each limb of which rapidly increases in size. Each of these arcuate, clavate cells has now a portion of its extremity isolated by a partition, by means of which a new hemispherical cell is formed at the end of each thread at its point of junction with the opposed thread. These cells become afterwards cylindrical by pressure, the protoplasm is aggregated into a mass, the double membrane at the point of first contact is absorbed, and the two confluent masses of protoplasm form a zygospore invested with a tubercular coat and enveloped by the primary wall of the two conjugating cells. During this formation of the zygospore, the two arched cells whence the zygospore originated develop a series of dichotomous processes in close proximity to the walls which separate them from the zygospore. These processes appear at first on one of the arcuate cells in successive order. The first makes its appearance above upon the convex side; the succeeding ones to the right and left in descending order; the last is in the concavity beneath. It is only after the development of this that the first process appears on the opposite cell, which is followed by others in the same order. These dichotomous processes are nothing more than branches developed from the arcuate, or mother cells. During all these changes, while the zygospore enlarges, the wall of the arcuate cells becomes coloured brown. This colouring is more marked on the convex side, and it shows itself first in the cell on which the dichotomous branches are first produced, and which retains the darker tint longer than the other. The zone from whence the processes issue, and also the processes themselves, have their walls blackened deeply, while the walls of the conjugated cells, which continue to clothe the zygospore during the whole of its development, are bluish-black. By pressure, the thin brittle coat which envelopes the zygospore is ruptured, and the coat of the zygospore exposed, formed of a thick cartilaginous membrane, studded with large irregular warts.

The germination of the zygospores in this species has not as yet been observed, but it is probably the same or very similar to that observed in other species of _Mucor_. In these the rough tuberculate epispore splits on one side, and its internal coat elongates itself and protrudes as a tube filled with protoplasm and oil globules, terminating in an ordinary sporangium. Usually the amount of nutriment contained in the zygospore is exhausted by the formation of the terminal sporangium, according to Brefeld;[D] but Van Tieghem and Le Monnier remark that in their examinations they have often seen a partition formed at about a third of the length of the principal filament from the base, below which a strong branch is given off, and this is also terminated by a large sporangium.