420 Persoonia – Volume 42, 2019 Kazachstania molopis Fungal Planet description sheets 421 Fungal Planet 926 – 19 July 2019 Kazachstania molopis Gouliamova, R.A. Dimitrov, sp. nov. Etymology. mo­lo­pis, referring to the host beetle Molops piceus (Cara­ Notes — In our previous article we determined the lower bidae) from which two new strains were isolated. and upper bounds for the range of species discrimination in Classification — Saccharomycetaceae, Saccharomycetales, the Kazachstania clade based on sequence identity value Saccharomycetes. (SI) and distance between physiological profiles (DPP): SI (98.5–83.7 %) and DPP (8–18) (Dimitrov & Gouliamova 2019). After 7 d at 25 °C in 5 % glucose broth, the cells are ovoid to el- A phylogenetic analysis of combined ITS and LSU sequences lipsoidal, 2–4 × 4–7 μm, occurring singly or in clusters. Asexual placed the new strain IMB 4R on a separate branch between K. viti­ reproduction occurs by multilateral budding. Poorly developed cola and K. kunashiriensis. Pairwise analysis of sequences pseudohyphae can be present. After 7 d at 25 °C on YPGA in a multiple alignment showed that the new strains show (yeast extract, pepton, glucose agar) the colony is cream, bu- 87.95 % identity (847 identical nt., 90 nt subst., 123 gaps) with tyrous, glistening, convex and with an entire margin. Dalmau K. kunashiriensis and 85.49 % identity (884 identical nt., 137 plate culture after 10 d on morphology agar did not show pseu- subst., 138 gaps) with K. viticola. The new strains can be dif- dohyphae or true hyphae. Sexual reproduction was detected ferentiated from both K. kunashiriensis and K. viticola based on yeast extract, malt extract, peptone, glucose (YM) and on 14 common physiological characteristics. The new species McClary acetate agar. Conjugation between independent cells can assimilate L-sorbose, D-ribose, sucrose, maltose, α-methyl- was observed. Asci contained one to four globose ascospores. D-glucoside, cellobiose, salicin, arbutin, melezitose, soluble Fermentation — Glucose and galactose are fermented. starch, ribitol, D-glucitol, 2-keto-D-gluconate and quinic acid. Sucrose, maltose, lactose and raffinose are not fermented. It cannot grow in the presence of 10 % NaCl. In addition the Carbon assimilation — D-glucose, D-galactose, L-sorbose, new species can be differentiated from K. viticola based on its D-ribose, sucrose, maltose, α,α-trehalose, α-methyl-D-gluco- ability to assimilate α,α-trehalose and its inability to assimilate side, cellobiose (delayed), salicin, arbutin (delayed), melezitose, D-gluconate and growth in the presence of 15 % NaCl. The new soluble starch, glycerol, ribitol, D-glucitol, D-mannitol, D-gluco- species differ from K. kunashiriensis based on its inability to no 1,5-lactone, 2-keto-D-gluconate (delayed), ethanol, quinic assimilate L-lysine. The obtained SI and DPP data for the new acid are assimilated. D-xylose, D-arabinose, D-glucosamine, strain IMB 4R fall within the limits for species discrimination of L-arabinose, L-rhamnose, melibiose, raffinose, lactose, inulin, the Kazachstania clade. Thus, based on our results we propose meso-erythritol, myo-inositol, xylitol, D-gluconate, D-glucuro- a new yeast species, Kazachstania molopis, to accommodate nate, D-galacturonate, succinate, citrate, DL-lactate, methanol, Bulgarian yeast strains IMB 4R and IMB 4 (100 % SI in both propane 1,2 diol, butane 2,3 diol, galactonic acid, galactitol, ITS and LSU sequences). So far, only three species of Kazach­ galactonic acid and saccharate are not assimilated. stania were isolated from insects. A strain of K. spencerorum Nitrogen assimilation — Nitrate, nitrite, ethylamine, creatine, was isolated from larva of a Psychidae moth (Lepidoptera) creatinine, L-lysine, cadaverine and imidazole are not assimi- collected from an acacia tree (South Africa) (CBS database). lated. Three strains of K. intestinalis were isolated from the gut of the Other tests — Starch formation test is negative. Growth in passalid beetle, O. disjunctus, collected from rotten oak tree 10 % is negative. Growth in 0.01 % is negative. Growth in 50 % (Virginia, USA) (Suh & Zhou 2011). Recently two strains of glucose is negative. Urea hydrolysis and DBB reaction tests are K. chrysolinae were isolated from the guts of Chrysolinae polita negative. Growth without all vitamins test is negative. Growth in Bulgaria (Gouliamova & Dimitrov unpubl. data). at 25 °C is positive. Growth at 30 °C is negative. Typus. BULGARIA, Nature park Zlatni Pyasatsi from the gut of beetle Molops T 78 Kazachstania spencerorum DBVPG 6746 D89898_U84227 piceus (Carabidae, Coleoptera) collected in oak forest under fallen tree trunk, 95 T Kluyveromyces hubeiensis AS2 1536 AY325966_AY325967 23–24 Apr. 2009, D. Gouliamova (holotype IMB 4R preserved in metaboli- T 100 Kazachstania lodderae CBS 2757 AY046160_U68551 cally inactive state, ex-type cultures NBIMCC 9029 and CBS 12448; ITS T 100 Kazachstania piceae CBS 7738 AY046159_AF399767 65 T and D1/D2 LSU sequences GenBank KC118123 and KC878454, MycoBank Kazachstania rosinii NRRLY 17919 AY046158_U84232 MB802456) Kazachstania hellenica CBS 107045T EF620862_EF620861 100 70 T 86 Kazachstania zonata NBRC 100504 EU075199_AB198187 ULGARIA Additional material examined. B , same details as type, IMB4 = T Kazachstania gamospora NBRC 11056 EU075198_AB0873 NBIMCC 9028 = CBS 12566, ITS and D1/D2 LSU sequences GenBank T Kazachstania africana NRRLY 8276 AY046155-AY048159 70 92 HM627145 and HM627092. T 56 Kazachstania martiniae NRRLY 409 AY046157_AY881651 Kazachstania viticola NRRLY 27206T AY046162_AF398482 98 Kazachstania molopis IMB4RT KC118123 _KC878454 T 91 Kazachstania kunashirensis CBS7662 KY103649_KY107927 Kazachstania psyhrophila CBS 12679T JX656699_JX564243 Kazachstania intestinalis ATCC MYA 4658T HQ260334_HQ260337 Kazachstania humilis NRRLY 17074T AY046174_AF399778 Kazachstania pseudohumilis CBS 11404T FJ888526 0.005 Phylogenetic tree obtained by the analysis of combined ITS and LSU rDNA Colour illustrations. Krushuna Waterfalls, Bulgaria. Molops piceus (Photo sequences of Kazachstania molopis IMB 4RT and related species using a credit: Ruslan Panin, http://carabidae.org); bottom to top: morphology of neighbour-joining method (Kimura two-parameter model; MEGA v. 7; 100 cells of Kazachstania molopis IMB4RT in 5 % glucose broth after 1 wk; asci bootstrap replicates). Kazachstania humilis and K. pseudohumilis represent with ascospores in YM agar. Scale bars = 5 μm (cell morphology), 10 μm an outgroup species. GenBank accession numbers of ITS and LSU rDNA (ascospores). sequences are presented on the tree. Dilnora E. Gouliamova, The Stephan Angeloff Institute of Microbiology, Bulgarian Academy of Sciences, 26 Acad. Georgi Bonchev, Sofia 1113, Bulgaria; e-mail: [email protected] Roumen A. Dimitrov, National Center of Infectious and Parasitic Diseases, 26 Yanko Sakazov blvd, Sofia 1504, Bulgaria; e-mail: [email protected] © 2019 Naturalis Biodiversity Center & Westerdijk Fungal Biodiversity Institute.