Lecanicillium gracile (Cordycipitaceae), a new species isolated from mineral building materials Author Ponizovskaya, Valeria B. 0000-0001-5437-1016 Department of Mycology and Algology, Faculty of Biology, Lomonosov Moscow State University, 1 - 12 Leninskie Gory, 119234 Moscow, Russia. v.ponizovskaya@gmail.com Author Grum-Grzhimaylo, Alexey A. 0000-0001-5512-6223 Department of Microbial Ecology, Netherlands Institute of Ecology (NIOO-KNAW), 6708 PB Wageningen, The Netherlands. a.grgr@nioo.knaw.nl Author Georgieva, Marina L. 0000-0002-7657-6474 Department of Mycology and Algology, Faculty of Biology, Lomonosov Moscow State University, 1 - 12 Leninskie Gory, 119234 Moscow, Russia. & Gause Institute of New Antibiotics, 11 - 1 Bolshaya Pirogovskaya, 119021 Moscow, Russia. i-marina@yandex.ru Author Kokaeva, Lyudmila Y. 0000-0003-1384-3552 Department of Mycology and Algology, Faculty of Biology, Lomonosov Moscow State University, 1 - 12 Leninskie Gory, 119234 Moscow, Russia. & All-Russian Lorh Research Institute of Potato Farming, 140051 Moscow, Russia. kokaeval@gmail.com Author Bilanenko, Elena N. 0000-0001-6323-2237 Department of Mycology and Algology, Faculty of Biology, Lomonosov Moscow State University, 1 - 12 Leninskie Gory, 119234 Moscow, Russia. e_bilanenko@mail.ru text Phytotaxa 2020 2020-05-20 443 3 265 278 http://dx.doi.org/10.11646/phytotaxa.443.3.3 journal article 10.11646/phytotaxa.443.3.3 1179-3163 13878083 Lecanicillium gracile V.B. Ponizovskaya, A.A. Grum-Grzhim., Georgieva & Bilanenko , sp. nov. MycoBank MB830764 Figs. 2–3 Etymology:— Latin, gracile = slender or thin; referring to slim phialides and thin mycelium. Type:— RUSSIA . Vladimir region : Kideksha village, Church of Boris and Gleb (XII century), wall area with high moisture content and precipitation of salts at the surface, loose limestone, 13/June/2013 , collected by V.B. Ponizovskaya, N.L. Rebrikova, isolated by V.B. Ponizovskaya ( holotype CBS H-23932), ex-type culture Kd1 (CBS 142821). TABLE 2 . Taxa used in the phylogenetic analyses with their GenBank accession numbers. Newly isolated strains and generated accessions are marked in bold.
Species Isolation source Voucher ITS TEF1a
Akanthomyces attenuatus Carpocapsa pomonella larva CBS 170.76, type EF679164 -
Akanthomyces lecanii Coccus viridis CBS 102067, IMI 331550R, type EF513009 KM283818
Akanthomyces muscarius Trialeurodes vaporariorum CBS 143.62, IMI 068689, epitype AJ292388 KR064305
Lecanicillium cauligalbarum Lepidopteran larva GZUIFRZHJ01, type MH730663 MH801920
Lecanicillium acerosum Crinipellis perniciosa in Theobroma cacao CBS 418.81, ex-type EF641893 KM283810
Lecanicillium aphanocladii Agaricus bisporus CBS 101286 AJ292430 -
Lecanicillium aphanocladii Agaricus bitorquis CBS 376.77 AJ292431 -
Lecanicillium aphanocladii Agaricus bisporus CBS 797.84 KM283811 -
Lecanicillium antillanum Agaricales CBS 350.85, ex-type AJ292392 DQ522350
Lecanicillium aranearum Araneida CBS 726.73a, epitype AJ292464 EF468781
Lecanicillium araneicola Araneae BTCC F35, NBRC 105407, ex- type AB378506 -
Lecanicillium coprophilum Faece of Marmota monax CGMCC 3.18988 MH177614 MH184585
Lecanicillium coprophilum Faece of Marmota monax CGMCC 3.18987 MH177615 MH184586
Lecanicillium coprophilum Faece of Marmota monax CGMCC 3.18986, ex-type MH177616 MH184587
Lecanicillium dimorphum Agaricus bisporus CBS 363.86, JCM 23128, ex-type AJ292429 EF468784
Lecanicillium flavidum Abies alba CBS 342.80, ex-type EF641878 -
Lecanicillium fungicola var. aleophilum Agaricus bisporus CBS 357.80, ex-type AF324876 KM283815
Lecanicillium fungicola var. fungicola Agaricus bisporus CBS 992.69, epitype EF641889 KM283816
Lecanicillium fusisporum Coltricia perennis CBS 164.70, ex-type AJ292428 EF468783
Lecanicillium gracile Plaster from interior wall CBS 142816 LT598647 MN602797
Lecanicillium gracile Plaster from interior wall VKPM F -1421 MF682449 MN602799
Lecanicillium gracile Limestone from interior wall CBS 142821, ex-type MF682448 MN602801
Lecanicillium kalimantanense Coleoptera (Staphylinoidea?) in suspended soil NBRC 105406, BTCC F23, ex- type AB360356 -
Lecanicillium longisporum Icerya purchasi CBS 126.27, IMI 021167, type AJ292385 -
Lecanicillium nodulosum Scale insect ( Coccidae ) IMI 338014R, type EF513012 -
......continued on the next page TABLE 2. (Continued)
Species Isolation source Voucher ITS TEF1a
Lecanicillium pissodis Pissodes strobi CBS 118231 - KM283822
Lecanicillium primulinum Soil under unidentified plant JCM 18525, FKI–6172, ex-type AB712266 -
Lecanicillium restrictum Surface of the wooden barrel CCF 5252, ex-type LT548279 LT626943
Lecanicillium saksenae Forest soil CBS 532.81, IMI 179841, ex-type AJ292432 EF469067
Lecanicillium sp. Diaphorina citri cadavers ZJLSP07 KM013699 -
Lecanicillium sp. Plaster from interior wall CBS 142817 LT598645 MN602798
Lecanicillium sp. Plaster from interior wall CBS 142818 LT598646 MN602800
Lecanicillium sabanense Pulvinaria caballeroramosae ANDES-F 1024, holotype KC633232 KC633266
Lecanicillium subprimulinum dead wood KUMCC 17-0148, ex-type MG585314 MG585317
Lecanicillium tenuipes Spider CBS 309.85 AJ292391 DQ522341
Lecanicillium testudineum Carapace of Trachemys scripta elegans CCF 5201, CBS 141096, ex-type LT548278 LT626942
Lecanicillium testudineum Carapace of Trachemys scripta elegans CCF 5545 LT992874 LT992868
Lecanicillium testudineum Carapace of Trachemys scripta elegans CCF 5546 LT992871 LT992868
Lecanicillium uredinophilum Insect KUN 101466 MG948305 MG948315
Lecanicillium uredinophilum Pucciniastrum agrimoniae on Agrimonia pilosa KACC 44082, ex-type - KM283806
Lecanicillium wallacei Lepidopteran larva CBS 101237, IMI 331549, holotype EF641891 EF469073
Parengyodontium album Fresco CBS 368.72 LC092891 -
Parengyodontium album Human ear secretions IHEM 4198 LC092887 -
Parengyodontium album Human brain abscess CBS 504.83, ex-type LC092880 -
Parengyodontium album Limestone from interior wall CBS 142824 LT549076 MN602802
Simplicillium lanosoniveum Hemileia vastatrix CBS 704.86 AJ292396 DQ522358
Simplicillium obclavatum Air above sugarcane field CBS 311.74, IMI 185383, ex-type AJ292394 EF468798
Verticillium indonesiacum Araneae BTCC-F36, ex-type AB378516 -
FIGURE 1 . Two-gene-based phylogeny (ITS rDNA+TEF1a) of Lecanicillium and closely related species. Our new isolates are marked in bold. Bayesian topology is displayed with the PP/ML support values over each node. Thickened branches indicated strong combined support (PP> 0.94, ML> 90). T – type/ex-type strains. Culture characteristics: Colonies on PDA reaching 14–30 mm diam. in 10 days and 20–39 mm diam. in 14 days. Colonies white, cottony, centrally raised, colony surface or at least marginal parts more or less radially wrinkled, margin entire, no exudate and soluble pigments, reverse yellowish to creamish. Colonies on PCA reaching 19–24 mm diam. in 10 days and 26–31 mm diam. in 14 days. Colonies white, floccose, sometimes waxy near margin, more or less centrally raised, margin entire, submerged, no exudate and soluble pigments, reverse creamish. Colonies on MYA reaching 21–26 mm diam. in 10 days and 27–33 mm diam. in 14 days. Colonies white, cottony, more or less centrally raised, colony surface or marginal parts more or less radially wrinkled, margin entire, no exudate and soluble pigments, reverse creamish to brownish. Phialides (6–)10–23(–33) μm long (mean ± standard deviation 15.7±5.3), basal part (0.5–)0.8–1.5 (1±0.2) μm wide, tip 0.2–0.9 μm wide (0.5±0.1), slender, tapering toward the tip, produced solitary or less often in whorls of 2–3 (rarely 4–5) on prostrate conidiophores; sometimes also produced on short erect conidiophores or secondarily formed on the preceeding phialides. Conidia formed in heads in the apex of the phialides, dimorphic, macroconidia mostly with pointed ends, fusiform to slightly falcate, smooth-walled, 1-celled, 3.6–8.0 × 1.7–0.8 μm (5.4±0.9 × 1.2±0.2), microconidia ovate, obovate, ellipsoidal, sometimes fusiform and slightly curved, smooth-walled, 1-celled, 1.1–3.3 × 0.7–1.4 μm (2.3±0.45 × 1.1±0.2). No microscopic crystals observed. FIGURE 2 . Lecanicillium gracile (CBS 142821, ex-type). a–c 14 days-old (25 °C, dark regime) colonies on PDA, PCA and MYA in 9 cm Petri plates, respectively. d–f Reverses of 14 days-old (25 °C, dark regime) colonies on PDA, PCA and MYA in 9 cm Petri plates, respectively. g–h Phialides and conidia (SEM). i–j Conidia. Scale bars: g–j = 10 μm. FIGURE 3 . Lecanicillium gracile (CBS 142821, ex-type). a–c Prostrate conidiophores with solitary phialides or phialides produced in whorls (LM). d Short erect conidiophore with phialides (LM). e–f Prostrate conidiophores with phialides produced in whorls (LM). g–h Phialides secondarily born on the preceeding phialides (LM). i Conidia (LM). j A drawing of prostrate conidiophores with phialides. k A drawing of conidiophores with phialides and conidia. Scale bars: a = 50 μm; b–k = 10 μm. FIGURE 4 . Temperature-dependent growth profiles of Lecanicillium gracile isolates. Colony diam. after 14 d on PDA (n=3, mean±SD). Sexual stage not observed. Temperature optimum for growth of Kd1 (CBS 142821) is 20–23 °C ( 19–33 mm diam. in 14 d), reduced growth at 28 °C ( 5–7 mm diam. in 14 d), no growth occurring at 30 °C ( Fig. 4 ). Temperature optimum for growth of 57-9-2 (CBS 142816) is 23–25 °C ( 30–38 mm diam. in 14 d), reduced growth at 33 °C ( 3–7 mm diam. in 14 d), no growth occurring at 35 °C ( Fig. 4 ). Temperature optimum for growth of G4-5 (VKPM F-1421) is 23–25 °C ( 26–39 mm diam. in 14 d), reduced growth at 33 °C ( 2–3 mm diam. in 14 d), no growth occurring at 35 °C ( Fig. 4 ). ITS differences with the closest taxa Lecanicillium gracile (MF682448, Kd1, type ) differs from L. restrictum (LT548279, CCF 5252, type ) by 22 SNPs, two 1-bp deletions, three 1-bp insertions in a total of 1034 character alignment (97.4 % similarity). Lecanicillium gracile (MF682448, Kd1, type ) differs from L. testudineum (LT548278, CCF 5201, type ) by 24 SNPs, one 1-bp deletion, three 1-bp insertions in a total of 1033 character alignment (97.3 % similarity). Lecanicillium gracile (MF682448, Kd1, type ) differs from L. coprophilum (MH177616, CGMCC 3.18986, type ) by 22 SNPs, three 1-bp deletions, one 1-bp insertion in a total of 541 character alignment (95.2 % similarity). Lecanicillium gracile (MF682448, Kd1, type ) differs from P. album (LC092880, CBS 504.83, type ) by 15 SNPs, seven deletions (five 1-bp deletions and two 4-bp deletions), one 1-bp insertion in a total of 548 character alignment (94.7 % similarity). TEF1a differences with the closest taxa Lecanicillium gracile (MN602801, Kd1, type ) differs from L. restrictum (LT626943, CCF 5252, type ) by 33 SNPs in a total of 459 character alignment (92.8 % similarity). Lecanicillium gracile (MN602801, Kd1, type ) differs from L. testudineum (LT626942, CCF 5201, type ) by 27 SNPs in a total of 459 character alignment (94.1 % similarity). Lecanicillium gracile (MN602801, Kd1, type ) differs from L. coprophilum (MH184587, CGMCC 3.18986, type ) by 26 SNPs in a total of 412 character alignment (93.7 % similarity).