Two new families of the Pezizales: Karstenellaceae and Pseudor hizinaceae Harr i Harmaja Department of Botany, University of Helsinki, SF-00170 Helsinki, Finland HARMAJA H . 1974: Two new families of the Pezizales: Karstenellaceae and Pseu­ dorhizinaceae. - Karstenia 14: 109- 112. The author considers especially the sporal, anatomical and cytological characters of the genera Karstenella Harmaja and Pseudorhizina J ach. to warrant the establishment of a new mono typic family for each : Karstenelloceae Harmaja and Pseudorhizinaceae Harmaja. Certain characters relevant to the family level taxonomy have been observed by the author in both genera. Features apparently diagnostic of the family Karstenellaceae are the p re­ sence of two nuclei in the spores, the lack of a cyanophilic perispore in all stages. of spore development, the simple structure of the excipulum which is exclusively composed of textura intricata, and the subicular characters. The genera of the Pezizales with tetranucleate spores are considered to form three different families on the basis of both sporal and anatomical differences: H elvellaceae Dum., Pseudorhizinaceae and Rhizinaceae Bon. The lack of a cyanophilic perispore in the mature spores and the simple structure and thick-walled hyphae of the excipulum are important distinguishing features of the family Pseudorhizinaceae. Comparisons are given between Pseudorhizinaceae and the two other families. I. Karstenellaceae Harmaja The description of the genus Karstenella valuable publication of 1972 the new tribe Harmaja was based on the new species K. Karstenelleae Korf was established in the vernalis Harmaja described in the same paper subfamily Pyronematoideae of the family (HARMAJA 1969b). Even then I noted that Pyronemataceae Corda. In that paper he the genus possessed such characters that it writes that Karstenella is »clearly a most un­ was impossible to place it in any known usual member of the order, and a monotypic family of the Pezizales. Prof. Dr. Richard P. family to accommodate it would not be Korf, on borrowing the type of K . vernalis, unreasonable». Considering my original found that the spores contained two nuclei opinion, and as I also regard Prof. Korf's each, which is a unique number in the order concept of the family Pyronemataceae too Pezizales. He therefore agreed that either a wide, e.g. in view of EcKBLAD's study ( 1968 ) new tribe or even a new family should be the excipular anatomy appears to deserve established for this genm (KoRF 1972 and in more emphasis, my own conclusion regarding litt.). In one of my letters to Prof. Korf I Karstenella agrees with this alternative suggested that the genus might possibly be discussed by him. Accordingly I propose a related to Pyronema Carus or Octospora new monotypic family Karstenellaceae, for Gray. KoRF ( 1972 and in litt.) thought that this genus, in the suborder Pezizineae. P·yronema was not a bad guess, and in his In the following those characters of Kars- 109 tenellaceae will be cited which appear to have mounts. 10 ) The asci and the spores within most taxonomic significance at the family a single ascocarp are at different stages of level · for a detailed description of the development. 11) The asci have inamyloid features of Karstenella the original descri­ walls. 12 ) The contents of the paraphyses ption (HARMAJA 1969b ) should be consulted. show no green reaction with Melzer's reagent. 1) The ascocarp is flat, fairly small, very 13) The excipulum is composed throughout thin as compared with its diameter and of textura intricata with narrow and thin­ appressed to the substrate throughout. 2) A walled hyphae. 14) Specialized hairs are subiculum of whitish hyphae is present. 3) lacking. 15 ) The subicular hyphae have thick, Red pigment is present in the ascocarp (in papillose, and strongly cyanophilic walls. the contents of the paraphyses) . 4) The spo­ res are binucleate. According to present Karstenellaceae Harmaja, n. famil. - Fa­ knowledge (cf. KoRF 1972) this is the only milia ex ordine Pezizales, subordine Pezizin­ case in the Pezizales. 5) The spores lack a eae. Ascocarpus tenuzsszmus, applanatus, cyanophilic perispore-periplasm during their colore aurantiaco, ad subiculum e hyphis whole development, i.e. belong to the crassitunicatis, papillatis et cyanophilibus: Morchella type (HARMAJA 1974b). 6) The Excipulum omnino e textura intricata. Asci spores are completely smooth during their parietibus ad apicibus haud manifeste in­ whole development, also with an oil immer­ crassatis, iodo haud caerulescentes. S parae sion lens. 7) The spores are hyaline. 8) The hyalinae, satis tenuitunicatae, eguttulatae, spore contents are ± homogeneous, lacking sine vacuolis gaseosis, cum duobus nucleis. oil drops. 9) De Bary bubbles are lacking in Paraphyses intus iodo haud virides. - Type: the spores at all stages in heated cotton blue Karstenella Harmaja. II. Pseudorhizinaceae Harmaja The genus Pseudorhizina Jachevsky (H el­ dies (HARMAJA 1969a, 1973 ) I had failed vellella Imai, Ochromitra Vel., Gyromitrodes to observe the presence of this layer during Vas.) , if recognized at all, was long regarded the immature stages of the spores. The cyano­ as mono typic with H elvella sphaerospora philic layer is very thin, ca. 0.2- 0.3 f!m Peck as its only species. However, in a recent when thickest, smooth or slightly uneven, its paper (HARMAJA 1973 ) it was demonstrated formation beginning evenly all over the spore that the genus is valid and natural, and it wall. In young spores in this genus (in P. was amended to include two species: H elvella sphaerospora) the perispore is very occasion­ californica Phill. and the type species ally seen to be partly detached from the mentioned above. The paper gave both the spore wall proper, so that its existence differences between the genera Pseudorhizina cannot be in doubt. and Gyromitra and those between the two Secondly, the spores of both species of Pseudorhizina species. Two additional chara­ Pseudorhizina were found to be tetranucleate, cters common to both species will be reported which confirms the affinity of this genus to here together with the observations justifying H elvella and Gyromitra. The nuclei are the establishment of a separate family to carminophobic in mature spores, but in the accommodate Pseudorhizin·a. immature ones they ~tain to some extent, so Firstly, I observed that the spores of both that the presence of four nuclei in the middle P. sphaerospora and P. californica (part o.f of the spore could be demonstrated. The the holotype at Kew, K, studied) belong to median position of the nuclei was exactly the Otidea type (HARMAJA 1974b), i.e., they as in the young spores of the certainly possess a strongly cyanophilic perispore­ tetranucleate species H elvella acetabulum, periplasin complex when immature ( exclu­ studied for control. The large central oil drop ding the very youngest stages when only a developing later in that species forces the cyanophobic true wall and moderately cyano­ nuclei to both ends of the spore, pressing philic contents are observed), but lack such them against the spore wall. Staining the a coating when mature. In my earlier stu- Pseudorhizina spores by the usual acetocar- 110 mine + boiling method gave poor results, cyanophobic). 3) The nuclei of ripe spores but preceding treatment with KOH (see from dried specimens are carminophobic in HARMAJA 1974c) considerably improved the Pseudorhizina (even after repeated boiling), absorption of the dye in this genus, too (how­ while in the H elvellaceae they are carmino­ ever, the nuclei in the mature spores philic. 4) A certain proportion of mature continued to be carminophobic). spores of Pseudorhizina (though it is very Details about the characters of the species small in P. californica) contain in heated cot­ of Pseudorhizina as well as references to ton blue one de Bary bubble each, while further literature may be found in HARMAJA these are completely lacking in the H elvella-­ 1973, so in the following I will restrict the ceae. 5 ) The hyphae of the excipulum (those discussion to the characters which I consider of the subhymenium excluded) are thick­ diagnostic at the family level, and will give walled in Pseudorhizina, the thickness varying the features which separate the proposed between ca. 0.7 and 2.0 flm. This probably monotypic family Pseudorhizinaceae from the accounts for the poor reviving capacity of two other families _( in my sense) of the; the hyphae of dried specimens, at least in Pezizales with tetranucleate spores, viz. H el­ heated cotton blue, where they remain col­ vellaceae Dum. (with Helvella only; see lapsed even after repeated boiling. The HARMAJA 1974a) and Rhizinaceae Bon. hyphae of the H elvellaceae have distinctly (with Gyromitra and Rhizina; see HARMAJA thinner walls and revive easily when prepared 1969a and 1973) . Some of these differences for a microscopic mount. 6) As is already have not been reported before. known, the construction of the excipuli dif­ A. Differences from H elvellaceae. 1) The fers, that of Pseudorhizina being composed stipe very often stains red (see HARMAJA of textura intricata throughout, while that 1973) in Pseudorhizina but never does so in of the H elvellaceae has an ental part of t. the H elvellaceae. 2) The mature spores of intricata and a distinct ectal layer of t. Pseudorhizina are devoid of a cyanophilic prismatica to t. angularis. perispore-periplasm complex, as already re­ ported (HARMAJA 1969a), while such a B. Differences from Rhizinaceae ( cf. also complex appears to be present in the mature HARMAJA 1973 ) . 1) The same as the first spores of Helvellaceae, though not all the difference fro IT). the H elvellaceae (see species have yet been studied by me. In the above) . 2) Exactly the same as the second H elvellaceae (and elsewhere) it seems to be difference from the H elvellaceae ( cf. also generally believed that a perisporial sheath HARMAJA 1969a). The Rhizinaceae spores (often regarded as »exosporium») is present also belong to the P eziza type, or the only when the spore is seen to be ornament­ persistently perisporous type (HARMAJA ed, but in many genera (HARMAJA 1969a, 1974b).