Phylogenetic evaluation
The ML analyses primarily based on the rpb2 dataset (Fig. S1) confirmed the phylogenetic placement of strains CBS 149229, CBS 148468, CBS 148467 and CBS 149228 inside Trichocomaceae. Detailed statistics and the mannequin chosen for the ML evaluation is given in Desk 2. The phylogram was rooted with Paecilomyces brunneolus CBS 370.70 T. Briefly, the ML evaluation grouped studied strains into 15 lineages, together with outgroup. These lineages are in settlement with knowledge printed in research on Trichocomaceae13,14,25. Primarily based on these knowledge, CBS 149229 and CBS 148468 will be categorised in Rasamsonia, and CBS 148468 and CBS 148467 in Talaromyces sect. Bacillispori.
The Rasamsonia dataset contained sequences from 17 strains, together with the outgroup Trichocoma paradoxa (CBS 103.73 T). The mixed dataset (ITS, BenA, and CaM) contained 1664 websites, together with alignment gaps. Of those, 1007 had been conserved and 435 parsimony-informative. Detailed outcomes for every single gene dataset together with corresponding fashions are given in Desk 2. Strains CBS 148468 and CBS 149229 type a lineage inside Rasamsonia (Fig. 1). This lineage is sister to a clade containing R. brevistipitata, R. columbiensis, R. frigidotolerans, R. oblata, R. pulvericola and R. sabulosa. Single locus timber of particular person loci (ITS, BenA, CaM, rpb2) are proven in Fig. S2.
Most chance tree generated from the mixed evaluation of ITS, BenA, and CaM sequence knowledge. BS/pp values are given on the nodes. The tree was rooted to Trichocoma paradoxa (CBS 103.73 T). The brand new species Rasamsonia chlamydospora Spetik & Houbraken sp. nov. is highlighted in daring. Tex-type pressure.
The Talaromyces sect. Bacillispori dataset consisted of sequences from 10 strains, together with the outgroups Talaromyces subinflatus (CBS 652.95 T) and Talaromyces flavus (CBS 310.38 T). The mixed dataset (ITS, BenA, CaM and rpb2) contained 2512 websites, together with alignment gaps. Of those, 1632 had been conserved, 541 parsimony-informative and 817 distinctive. Detailed outcomes for every single gene dataset together with corresponding fashions are displayed in Desk 2. The multilocus evaluation resolved CBS 148467 and CBS 149228 in a totally supported clade, sister to T. bacillisporus CBS 296.48 T (84% BS, 1.00 pp) (Fig. 2). Moreover, a single locus ITS ML/BI evaluation was carried out (Fig. S3), displaying the distant place of T. hachijoensis to the newly proposed T. clematidis.
Most chance tree generated from the mixed evaluation of ITS, BenA, CaM and rpb2 sequence knowledge. BS/pp values are given on the nodes. The tree was rooted to Talaromyces subinflatus (CBS 652.95 T) and Talaromyces flavus (CBS 310.38 T). The brand new species Talaromyces clematidis Spetik & Houbraken sp. nov. is highlighted in daring. Tex-type pressure.
Taxonomy
Primarily based on the outcomes of the phylogenetic evaluation and the morphological examination (see beneath below Taxonomy and Dialogue), we suggest the names Rasamsonia chlamydospora for CBS 148468 and CBS 149229 and Talaromyces clematidis for CBS 148467 and CBS 149228.
Rasamsonia chlamydospora Spetik & Houbraken sp. nov. (Fig. 3).
Rasamsonia chlamydospora (CBS 149229 T). Colonies 7 d, 37 °C from left to proper (prime row) CYA, DG18, MEA and OA; (backside row) CYA reverse, DG18 reverse, YES and CREA. (A) In situ element of the colony on MEA; (B–E) Conidiophores; (F, G) Chlamydospores; (H) Conidia. Scale bars: 500 μm (A); 10 μm (B–H); 5 μm (G).
Mycobank quantity: MB 845365.
Etymology:—refers to chlamydospores, the globose to subglobose swollen cells produced by this species.
Sort:—CZECH REPUBLIC, Břeclav: Lednice, college backyard (48° 47′ 33.4″ N 16° 47′ 55.7″ E), remoted from the basis of Clematis ´Snow Queen´ (Ranunculaceae), 2021, M. Spetik, Holotype: CBS H-25023, ex-type dwelling tradition CBS 149229 = DTO 473-E5 = MEND-F-0752.
Barcodes: ITS ON863770; LSU ON863795; BenA ON873765; CaM ON938198; rpb2 ON938202.
Colony diameter (7 d, in mm): 25 °C: CYA 10–13; CYAS No progress; DG18 2; MEA 25–28; OA 12–21; YES 10–12; CREA No progress. 37 °C: CYA 31–35; CYAS No progress; DG18 36–38; MEA > 65; OA > 65; YES 46–52; CREA 18–25.
Colony diameter at totally different temperatures (7 d in mm). On CYA: 15–21 °C No progress; 24 °C 5–7; 27 °C 16–18; 30 °C 20–23; 33 °C 27–30; 36 °C 28–33; 37 °C 31–35; 40 °C 26–29; 45 °C 15–16; 48 °C 7–8; > 50 °C No progress. On MEA: 15–18 °C No progress; 21 °C 9–11; 24 °C 25–27; 27 °C 45–46; 30 °C 56–58; 33 °C 66–71; 36 °C 72–76; 37 °C 70–74; 40 °C 75; 45 °C 39–45; 48 °C 21–25; 50 °C 12–13; 52 °C 5; 55 °C No progress. Cardinal progress temperatures: Minimal, on CYA between 21 and 24 °C and between 18 and 21 °C on MEA, optimum round 36 °C and most on CYA 48 °C, on MEA 52 °C. Colonies grown on CYA and MEA at numerous temperatures (15–45 °C) after 7 d are proven on Fig. S4.
Colony characters (37 °C, 7 d): CYA: Colonies low, aircraft; margin low, total; mycelium mild brown; texture floccose; sporulation reasonably dense; soluble pigments absent; exudates absent; reverse white to mild brown. MEA: Colonies reasonably deep, radially sulcate; margin low, total; mycelium olivaceous with white spots close to the centre; texture velvety; sporulation dense; soluble pigments absent; exudates absent; reverse brown. DG18: Colonies raised, sulcate; margin low, total; mycelium white to olivaceous; texture velvety to floccose; sporulation dense; soluble pigments absent; exudates absent; reverse brown. YES: Colonies flat, raised in centre; margin low, total; mycelium olivaceous with white spots; texture velvety to floccose; sporulation dense; soluble pigments absent; exudates absent; reverse white. OA: Colonies flat; margin irregular; mycelium yellow to olivaceous; texture floccose; sporulation dense; soluble pigments absent; exudates absent; reverse brown. CYAS: No progress. CREA: Average progress, acid manufacturing absent.
Micromorphology: Mycelium 2–3.5 μm diam, distinctly rough-walled. Conidiophores monoverticillate, typically with subterminal branches. Stipes rough-walled, 15–70 × 2–3 μm. Phialides 3–6 per stipe, {smooth}, acerose to flask-shaped, 10–15 × 2–3 μm. Conidia {smooth}, cylindrical, 3.5–4.5 × 2.5–3 μm. Swollen cells resembling chlamydospores current, {smooth}, globose to subglobose, 6–8 μm. Ascomata not noticed.
Notes: Most Rasamsonia species are thermotolerant or thermophilic, together with R. chlamydospora. Thermophilic fungi are these with a most progress temperature of fifty °C or increased, and a minimal progress temperature of 20 °C or above26. The minimal progress temperature of R. chlamydospora is determined by the expansion medium, and the minimal progress temperature on MEA is between 18 and 21 °C. Comparability of microscopic options between all Rasamsonia species is given in Desk 3. Rasamsonia chlamydospora shares the manufacturing of olive-brown conidia and cylindrical phialides that regularly taper in the direction of the apices with different species however will be distinguished from different Rasamsonia species by the manufacturing of swollen cells that resemble chlamydospores. Evaluating sequences obtained from the kind pressure with these of R. brevistipitata, the phylogenetically closest species, confirmed a pairwise nucleotide distinction of 27 bp in ITS, 63 bp in BenA, 79 bp in CaM and 88 bp in rpb2.
Extra specimens examined:—CZECH REPUBLIC, Breclav: Lednice, college backyard (48° 47′ 33.3″ N 16° 47′ 55.6″ E), remoted from the basis of Clematis ´Snow Queen´ (Ranunculaceae), 2021, M. Spetik, dwelling tradition CBS 148468 = DTO 743-E4 = MEND-F-0751.
Talaromyces clematidis Spetik & Houbraken sp. nov. (Fig. 4).
Talaromyces clematidis (CBS 149229 T). Colonies 7 d, 25 °C from left to proper (prime row), MEA, DG18, YES and OA; (backside row) MEA reverse, DG18 reverse, YES reverse; MEA (14 d 25 °C). (A) Ascomata in situ on OA; (B, C) Conidiophores; (D) Conidia; (E) Asci and conidia; (F) Ascospores. Scale bars: 1000 μm (A), 10 μm (all others).
Mycobank quantity: MB 845200.
Etymology:—refers back to the host, Clematis.
Sort:—CZECH REPUBLIC, Břeclav: Lednice, college backyard (48°47′ 33.5″ N 16° 47′ 55.8″ E), remoted from the basis of Clematis ´Snow Queen´ (Ranunculaceae), 2021, M. Spetik, Holotype: CBS H-25024, ex-type dwelling tradition CBS 149228 = DTO 473-E3 = MEND-F-0750.
Barcodes: ITS ON863768; LSU ON863793; BenA ON873763; CaM ON938196; rpb2 ON938200.
Colony diameter (25 °C, in mm), 7 d: CYA 2; CYAS No progress; DG18 3–5; MEA 8–11; OA 6–9; YES 4–6; CREA 25 °C No progress. 14 d: CYA 4–5; CYAS No progress; DG18 10–13; MEA 21–26; OA 15–18; YES 8–9; CREA No progress. Colonies grown on CYA, MEA, DG18, YES, OA at 25 °C after 7 d and 14 d are displayed in (Fig. S5).
Colony diameter at totally different temperatures (mm). On CYA, 7 d: 21 °C No progress; 24 °C 2; 27 °C 3–4; 2; 33 °C 2; 36 °C No progress. On MEA, 7 d: 15 °C 3–4; 18 °C 3–6; 21 °C 7; 24 °C 3–4; 27 °C 10–12; 30 °C 11–12; 33 °C 8; 36 °C 2; 37 °C No progress. On CYA 14 d: 15 °C 2; 18 °C 3–4; 21 °C 3; 24 °C 4; 27 °C 5; 30 °C 4–5; 33 °C 4–5; CYA 36 °C No progress. On MEA, 14 d: 15 °C 8; 18 °C 12–14; 21 °C 18–21; 24 °C 22–25; 27 °C 25–30; 30 °C 26–28; 33 °C 18–20; 36 °C 7; 37 °C No progress. Cardinal progress temperatures: Minimal 15 °C; optimum between 27 and 30 °C and most at 36 °C. Colonies grown on CYA and MEA at numerous temperatures (15–36 °C) after 14 d are displayed on (Fig. S6).
Colony characters (25 °C, 7 d): CYA: Progress restricted; colonies flat; margin aircraft, irregular; mycelium white; texture floccose; sporulation absent; soluble pigments absent; exudates absent; reverse white. MEA: Colonies raised in centre; margin low, total (2 mm); mycelium white; texture floccose; sporulation dense; soluble pigments absent; exudates absent; reverse white. DG18: Colonies low; margin total; mycelium white; sporulation absent; soluble pigments absent; exudates absent; reverse pale yellow. YES: Progress restricted; colonies medium raised; margin total, round; mycelium white; texture floccose; sporulation absent; soluble pigments absent; exudates absent; reverse white. OA: Colonies flat; margin total; mycelium white at margin, olivaceous in centre; texture floccose; sporulation dense; soluble pigments absent; exudates absent; reverse white. CYAS: No progress. CREA: No progress.
Micromorphology: Conidiophores monoverticillate, typically with subtermal branches. Stipes smooth-walled, 15–30 × 2 μm. Phialides normally in teams of three–6 per stipe, typically solitary, lateral of terminal on vegetative hyphae, {smooth}, acerose, 7–15 × 2–2.5 μm. Conidia {smooth}, ellipsoidal 3.5–5 × 2–3 μm. Ascomata yellow, subglobose to ovoidal, 100–600 μm; asci globose to subglobose 11–14 μm; ascospores {smooth}, ellipsoid, 6–7.5 × 4.5–5.5 μm.
Notes: Talaromyces clematidis varieties a well-supported sister clade to T. bacillisporus. There are numerous characters (e.g., colony diameters on CYA at 25 °C and 37 °C, and ascospore ornamentation) that can be utilized to differentiate each species, and a few of them are summarized in Table4. The pairwise nucleotide variations between the kind strains of each species are 45 nucleotides in ITS, 73 bp in BenA, 94 bp in CaM and 112 bp in rpb2.
Extra specimens examined:—CZECH REPUBLIC, Břeclav: Lednice, college backyard (48° 47′ 33.4″ N 16° 47′ 55.7″ E), remoted from the basis of Clematis ´Snow Queen´ (Ranunculaceae), 2021, M. Spetik, dwelling tradition CBS 148467 = DTO 743-E2 = MEND-F-0749.
