Calvatia craniiformis

The species was first described as Bovista craniiformis by Lewis David de Schweinitz in 1832.[2] Elias Fries transferred it to the then newly circumscribed genus Calvatia in 1849,[3] setting Calvatia craniiformis as the type and only species.[lower-alpha 2] Scott Bates and colleagues suggest that the name is synonymous with Lycoperdon delicatum published by Miles Joseph Berkeley and Moses Ashley Curtis in 1873 (not the L. delicatum published by Berkeley in 1854), as is Lycoperdon missouriense published by William Trelease in 1891.[5] The form C. craniiformis f. gardneri, published by Yosio Kobayasi in 1932 (originally Lycoperdon gardneri Berk. 1875),[6] since been elevated to the distinct species Calvatia gardneri.[7]

In their 1962 monograph on North American Calvatia, mycologists Sanford Myron Zeller and Alexander H. Smith set C. craniiformis as the type species of the stirps (a grouping of related species) Craniiformis, containing species with a large sterile base and a persistent cottony gleba. Other species they included in this stirps were C. umbrina, C. diguetti, C. lycoperdoides, C. rubroflava, C. ochrogleba, C. excipuliformis (since transferred by some authorities to Handkea), and C. elata.[8]

Calvatia craniiformis is commonly known as the "brain-shaped puffball"[9] or the "skull-shaped puffball".[10] The specific epithet craniiformis derives from the Ancient Greek words cranion, meaning "brain", and forma, "a form".[11]

Cluster of fruit bodies found in New Zealand

The fruit bodies of Calvatia craniiformis grow to dimensions of 6–20 cm (2.4–7.9 in) tall by 8–20 cm (3.1–7.9 in) wide, and have a form ranging from pear-shaped,[12] to flattened-spherical, to obovate (roughly egg-shaped), to tunicate (like an inverted cone). At the bottom is a thick, often crumpled base attached to a cord-like rhizomorph,[9] which is often encrusted with surrounding soil.[5] The rhizomorphs are well developed, and when cut into longitudinal section, reveal three distinct tissues: an outer cortex, a subcortical layer, and a central core.[13] The thin and fragile exoperidium (the outer layer of "skin") is whitish-gray to gray, and initially smooth before becoming areolate (divided by cracks into discrete areas). The base extends up one-third to one-half way into the puffball (where it becomes the columnella) tapering to a point.[9] The gleba is initially whitish, and then yellow-green, and finally brownish-green in older specimens with mature spores.[12]

Spores are spherical, hyaline (translucent),[12] and measure 2.5–3.4 μm in diameter.[14] They are thick-walled with a short pedicel (a tubelike extension),[12] and are ornamented with tiny spines (verrucae) that are roughly equidistant from each other.[14] Capillitial threads are long, hyaline, and branched, measuring 2.4–4 μm thick.[9] They are septate and occasionally have pits on their walls.[12] The exoperidium comprises thick-walled, inflated hyphae mixed with sphaerocysts (spherical cells), while the endoperidium is made of tightly interwoven, thick-walled hyphae.[5] In the rhizomorphs, the hyphae in the central core are several times as thick as those in the surrounding subcortex.[13]

Spores and capillitial threads

Using light microscopy, the spores of Calvatia craniiformis are generally indistinguishable from those of C. rubroflava and C. gigantea; electron microscopy reveals that each has distinctive spore ornamentation. C. craniiformis features small, well-separated verrucae (wartlike projections) up to 0.2 μm tall with rounded tips. In comparison, C. gigantea has larger verrucae (up to 0.4 μm tall) that are more irregularly arranged.[15]

Calvatia craniiformis is edible when the gleba is firm and white, unlike the pictured specimen.

Calvatia craniiformis is an edible species.[18] Young puffballs with a firm, white gleba have a mild odor and pleasant taste.[12] Early 20th-century mycologist Charles McIlvaine noted over a century ago that "the slightest change to yellow makes it bitter."[19] Versatile in cooking, the puffball absorbs flavors well.[20]

In the United States, the Ojibwe used the powdery gleba as a hemostatic agent to staunch the flow of nosebleeds:[21] the spore powder was inhaled through the nostrils. It is now known that this practice can lead to the pulmonary disease lycoperdonosis, which causes symptoms similar to pneumonia.[22] It is also used as a hemostatic agent in Chinese and Japanese folk medicines.[23]

Although Calvatia craniiformis is generally considered a saprobic species, in controlled laboratory conditions, an ectomycorrhizae between the fungus and American sweetgum (Liquidambar styraciflua) was reported in a 1966 publication. A Chinese study showed that C. craniifromis would readily form mycorrhiza with poplar seedlings on unsterilized, but not on sterilized soil.[24] Later research was unable to establish any similar association between C. craniiformis and Pinus ponderosa.[25] Brain puffballs grow singly or in groups in fields and open woods, hardwood forests, and wet areas.[9][12] In Asia, it has been recorded from China,[26] India,[27] Indonesia,[28] Japan,[5] Malaysia,[29] and South Korea.[30] The brain puffball has been recorded from Australia.[31] In North America, its range includes the eastern and southern United States,[12] and Mexico.[32] In Michigan, it is one of the few macrofungi found regularly in black locust plantations.[20] Fruit bodies serve as a food source for several species of flies.[33]

Extracts of the puffball have strong antitumor activity in mouse models attributable to protein-bound polysaccharides, the compounds calvatan, craniformin, and a tautomer of rubroflavin. Calvatan is thought to act by stimulating the immune response, rather than by killing cells.[25] Craniformin, originally reported in 1997, is an azoformamide compound. Three sterol compounds have been identified from the fungus: ergosta-4,6,8(14), 22-tetraene-3-one, ergosta-7,22-diene-3-ol, and ergosterol peroxide.[23]

Three azo- and azoxyformamides compounds have been isolated and identified from the puffball and tested for their ability to inhibit the growth of plants: 4-methoxybenzene-1-ONN-azoxyformamide, 4-methoxybenzene-1-azoformamide, and 4-hydroxybenzene-1-azoformamide. Only the first compound was significantly inhibitory, suggesting that the presence of the azoxy moiety is required for growth inhibition.[34]