Cylindrocladiella

Boesew., Canad. J. Bot. 60: 2289. 1982. Fig. 4.

Fig. 4. Cylindrocladiella spp. A–D. Conidiophores of Cylindrocladiella spp. on infected alfalfa seeds A–B. Cylindrocladiella spp. sporulating on the seed coat of Medicago sativa. C. Root rot of M. sativa seedling. D. Cylindrocladiella spp. on carnation leaf. E–AA. Asexual morph. E–I. Penicillate conidiophores. E. Cylindrocladiella longistipitata (ex-type CBS 116075). F. Cylindrocladiella kurandica (ex-type CBS 129577). G. Cylindrocladiella lanceolata (ex-type CBS 129566). H. Cylindrocladiella pseudoparva (ex-type CBS 129560). I. Cylindrocladiella nederlandica (ex-type CBS 152.91). J–N. Penicillate conidiogenous apparatus. J. Cylindrocladiella hawaiiensis (ex-type CBS 129569). K. Cylindrocladiella australiensis (ex-type CBS 129567). L. Cylindrocladiella natalensis (ex-type CBS 114943). M. Cylindrocladiella cymbiformis (ex-type CBS 129553). N. Cylindrocladiella ellipsoidea (ex-type CBS 129573). O–S. Subverticillate conidiophores. O. Cylindrocladiella australiensis (ex-type CBS 129567). P. Cylindrocladiella longiphialidica (ex-type CBS 129557). Q. Cylindrocladiella pseudohawaiiensis (ex-type CBS 210.94). R, S. Cylindrocladiella natalensis (ex-type CBS 114943). T–Y. Terminal vesicles of stipe extensions. T. Cylindrocladiella hawaiiensis (ex-type CBS 129569). U. Cylindrocladiella stellenboschensis (ex-type CBS 110668). V. Cylindrocladiella cymbiformis (ex-type CBS 129553). W. Cylindrocladiella variabilis (ex-type CBS 129561). X. Cylindrocladiella lanceolata (ex-type CBS 129566). Y. Cylindrocladiella kurandica (ex-type CBS 129577). Z, AA. Conidia. Z. Cylindrocladiella natalensis (ex-type CBS 114943). AA. Cylindrocladiella brevistipitata (ex-type CBS 142783). Scale bars: E–I, O = 50 μm; J–N, P–AA = 10 μm.

Synonym: Nectricladiella Crous & C.L. Schoch, Stud. Mycol. 45: 54. 2000.

Classification: Sordariomycetes, Hypocreomycetidae, Hypocreales, Nectriaceae.

Type species: Cylindrocladiella parva (P.J. Anderson) Boesew., basionym: Cylindrocladium parvum P.J. Anderson. Representative strain: CBS 114524 = ATCC 28272.

DNA barcodes (genus): LSU, ITS.

DNA barcodes (species): his3, tef1, tub2. Table 3. Fig. 5.

Table 3. DNA barcodes of accepted Cylindrocladiella spp.

Species Isolates1 GenBank accession numbers2 References
ITS LSU his3 tef1 tub2
Cylindrocladiella addiensis CBS 143794T MH111383 MH111393 MH111388 Present study
CBS 143793 MH111385 MH111395 MH111390 Present study
CBS 143795 MH111384 MH111394 MH111389 Present study
C. australiensis CBS 129567T JN100624 JN099222 JN098932 JN099060 JN098747 Lombard et al. (2012)
C. brevistipitata CBS 142786T MF444940 MF444926 Lombard et al. (2017)
C. camelliae IMI 346845 AF220952 JN099249 AY793509 JN099087 AY793471 Van Coller et al., 2005, Lombard et al., 2012
C. clavata CBS 129564T JN099095 JN099135 JN098858 JN098974 JN098752 Lombard et al. (2012)
C. cymbiformis CBS 129553T JN099103 JN099143 JN098866 JN098988 JN098753 Lombard et al. (2012)
C. elegans CBS 338.92T AY793444 JN099201 AY793512 JN099039 AY793474 van Coller et al. (2005), Lombard et al. (2012)
C. ellipsoidea CBS 129573T JN099094 JN099134 JN098857 JN098973 JN098757 Lombard et al. (2012)
C. hahajimaensis MAFF 238172T JN687561 JN687562 Inderbitzin et al. (2012)
C. hawaiiensis CBS 129569T JN100621 JN099219 JN098929 JN099057 JN098761 Lombard et al. (2012)
C. horticola CBS 142784T MF444911 MF444938 MF444924 Lombard et al. (2017)
C. humicola CBS 142779T MF444906 MF444933 MF444919 Lombard et al. (2017)
C. infestans CBS 111795T AF220955 JN099199 AY793513 JN099037 AF320190 Schoch et al., 2000, Crous et al., 2001, van Coller et al. (2005), Lombard et al. (2012)
C. kurandica CBS 129577T JN100646 JN099245 JN098953 JN099083 JN098765 Lombard et al. (2012)
C. lageniformis CBS 340.92T AF220959 JN099165 AY793520 JN099003 AY793481 Schoch et al. (2000), van Coller et al. (2005), Lombard et al. (2012)
C. lanceolata CBS 129566T JN099099 JN099139 JN098862 JN098978 JN098789 Lombard et al. (2012)
C. lateralis CBS 142788T MF444914 MF444942 MF444928 Lombard et al. (2017)
C. longiphialidica CBS 129557T JN100585 JN099264 JN098851 JN098966 JN098790 Lombard et al. (2012)
C. longistipitata CBS 116075T AF220958 JN099155 AY793546 JN098993 AY793506 Schoch et al. (2000), van Coller et al. (2005), Lombard et al. (2012)
C. microcylindrica CBS 111794T AY793452 JN099203 AY793523 JN099041 AY793483 van Coller et al. (2005), Lombard et al. (2012)
C. natalensis CBS 114943T JN100588 JN099178 JN098895 JN099016 JN098794 Lombard et al. (2012)
C. nederlandica CBS 152.91T JN100603 JN099195 JN098910 JN099033 JN098800 Lombard et al. (2012)
C. novazelandica CBS 486.77T AF220963 JN099212 AY793525 JN099050 AY793485 Schoch et al. (2000), van Coller et al. (2005), Lombard et al. (2012)
C. nauliensis CBS 143792T MH111387 MH111397 MH111392 Present study
CBS 143791 MH111386 MH111396 MH111391 Present study
C. parva CBS 114524 AF220964 JN099171 AY793526 JN099009 AY793486 Schoch et al. (2000), van Coller et al. (2005), Lombard et al. (2012)
C. peruviana IMUR 1843T AF220966 JN099266 AY793540 JN098968 AY793500 Schoch et al. (2000), van Coller et al. (2005), Lombard et al. (2012)
C. pseudocamelliae CBS 129555T JN100577 JN099256 JN098843 JN098958 JN098814 Lombard et al. (2012)
C. pseudohawaiiensis CBS 210.94T JN099128 JN099174 JN098890 JN099012 JN098819 Lombard et al. (2012)
C. pseudoinfestans CBS 114531T AF220957 JN099166 AY793548 JN099004 AY793508 Schoch et al. (2000), van Coller et al. (2005), Lombard et al. (2012)
C. pseudoparva CBS129560T JN100620 JN099218 JN098927 JN099056 JN098824 Lombard et al. (2012)
C. queenslandica CBS 129574T JN099098 JN099098 JN098861 JN098977 JN098826 Lombard et al. (2012)
C. reginae CBS 142782T MF444909 MF444936 MF444922 Lombard et al. (2017)
C. stellenboschensis CBS 110668T JN100615 JN099213 JN098922 JN099051 JN098829 Lombard et al. (2012)
C. terrestris CBS 142789T MF444915 MF444943 MF444929 Lombard et al. (2017)
C. thailandica CBS 129571T JN100582 JN099261 JN098848 JN098963 JN098834 Lombard et al. (2012)
C. variabilis CBS 129561T JN100643 JN099242 JN098950 JN099080 JN098719 Lombard et al. (2012)
C. viticola CBS 112897T AY793468 JN099226 AY793544 JN099064 AY793504 van Coller et al. (2005), Lombard et al. (2012)
C. vitis CBS 142517T KY979751 KY979806 KY979891 KY979918 Crous et al. (2017a)
1

CBS: Westerdijk Fungal Biodiversity Institute, Utrecht, the Netherlands; IMI: International Mycological Institute, CABI-Bioscience, Egham, Bakeham Lane, U.K; IMUR: Institute of Mycology, University of Recife, Recife, Brazil; MAFF: Genetic Resources Centre, National Agriculture and Food Research Organization (NARO), NARO Genebank, Ibaraki, Japan. T indicates ex-type strains.

2

ITS: internal transcribed spacers and intervening 5.8S nrDNA; LSU: 28S large subunit nrDNA; his3: partial histone H3 gene; tef1: partial translation elongation factor 1-alpha gene; tub2: partial β-tubulin gene.

Fig. 5. The Maximum Likelihood (ML) consensus tree of Cylindrocladiella spp. inferred from the combined ITS (547 bp), tef1 (527 bp) and tub2 (502 bp) sequence alignment. Thickened lines indicate branches present in the ML, Maximum parsimony (MP) and Bayesian consensus trees. Branches with ML-bootstrap (BS) & MP-BS = 100 % and posterior probabilities (PP) = 1.00 are in blue. Branches with ML-BS & MP-BS ≥ 75 % and PP ≥ 0.95 are in green. The scale bar indicates 0.02 expected changes per site. The tree is rooted to Gliocladiopsis sagariensis CBS 199.55. Ex-type strains are indicated in bold. GenBank accession numbers are indicated in Table 3 and in Lombard et al., 2012, Lombard et al., 2017. TreeBASE: S22340.

Ascomata perithecial, superficial, solitary, basal stroma absent, globose to obpyriform, collapsing laterally when dry, smooth, with minute, brown setae arising from ascomatal wall surface, red, changing colour in KOH, ostiole consisting of clavate cells, lined with inconspicuous periphyses. Asci unitunicate, 8-spored, cylindrical, sessile, thin-walled, with a flattened apex, and a refractive apical apparatus. Ascospores uniseriate, overlapping, hyaline, ellipsoid to fusoid with obtuse ends, smooth, 1-septate. Conidiophores monomorphic, penicillate, or dimorphic (penicillate and subverticillate), mononematous, hyaline. Penicillate conidiophores consist of a stipe, a penicillate arrangement of fertile branches, a stipe extension, and a terminal vesicle. Subverticillate conidiophores consist of a stipe, and one or two series of phialides. Stipe septate, hyaline, smooth. Stipe extensions aseptate, straight, thick-walled, with one basal septum, terminating in a thin-walled vesicle of characteristic shape. Conidiogenous apparatus with primary branches 0–1-septate, secondary branches aseptate, terminating in 2–4 phialides. Phialides cylindrical, straight or doliiform to reniform to cymbiform, hyaline, aseptate, apex with minute periclinal thickening and collarette. Conidia cylindrical, rounded at both ends, straight, (0–)1(–3)-septate, frequently slightly flattened at base, held in asymmetrical clusters by hyaline slime. Chlamydospores brown, thick-walled, more frequently arranged in chains than clusters (adapted from Boesewinkel 1982 and Lombard et al. 2012).

Culture characteristics: Colonies on MEA white to pale brick when young, becoming pale brick to dark sepia when mature, fluffy, cottony, effuse to convex with papillate surface, margin entire, undulate, lobate, or fimbriate, sometimes with abundant chlamydospores forming microsclerotia within medium.

Optimal media and cultivation conditions: CLA to induce sporulation of the asexual morph at 25 °C, while for the sexual morph sterile toothpicks placed on SNA is used at 20 °C.

Distribution: Worldwide.

Hosts: Soil-borne, weak pathogen of forestry, agricultural and horticultural crops (Crous, 2002, Lombard et al., 2012).

Disease symptoms: Leaf spots, cutting rot, stem cankers, damping-off and root rot.

Notes: Boesewinkel (1982) established the asexual genus Cylindrocladiella, based on C. parva, to accommodate several cylindrocladium-like species characterised by small (<20 μm long), 1-septate, cylindrical conidia and aseptate stipe extensions. Initially, Cylindrocladiella spp. were linked to the sexual genus Nectricladiella (Crous and Wingfield, 1993, Schoch et al., 2000). Following the implementation of the International Code of Nomenclature for algae, fungi and plants (ICN; McNeill et al. 2012), Rossman et al. (2013) proposed that the generic name Cylindrocladiella be retained over Nectricladiella. Recently, Lombard et al. (2015) showed that the genus Cylindrocladiella formed a monophyletic group in the Nectriaceae, closely related to the genera Aquanectria and Gliocladiopsis. To date, 36 species of Cylindrocladiella have been recognised (Crous and Wingfield, 1993, Van Coller et al., 2005, Inderbitzin et al., 2012, Lombard et al., 2012, Lombard et al., 2015, Lombard et al., 2017, Crous et al., 2017a), of which two are linked to nectricladiella-like sexual morphs (Schoch et al., 2000, Crous, 2002, Lombard et al., 2012, Lombard et al., 2015). These fungi are generally soil-borne and regarded as saprobes or weak pathogens of numerous plant hosts (Crous, 2002, Van Coller et al., 2005, Scattolin and Montecchio, 2007, Lombard et al., 2012). Disease symptoms associated with Cylindrocladiella infection include leaf spots and root, stem and cutting rots (Crous et al., 1991b, Peerally, 1991, Crous and Wingfield, 1993, Crous, 2002, Van Coller et al., 2005, Scattolin and Montecchio, 2007, Lombard et al., 2012).

References: Boesewinkel 1982 (morphology and pathogenicity); Crous & Wingfield 1993 (morphology); Schoch et al., 2000, Lombard et al., 2012, Lombard et al., 2017 (morphology and phylogeny); Crous 2002 (morphology, pathogenicity and monograph); van Coller et al. 2005 (morphology, pathogenicity and phylogeny).


Cylindrocladiella addiensis

L. Lombard & Crous, sp. nov. MycoBank MB824497.

Etymology: Name refers to Addis Ababa, Ethiopia, from where this fungus was collected.

Cultures sterile. Cylindrocladiella addiensis differs from its closest phylogenetic neighbours, C. elegans and C. noveazelandica, by unique fixed alleles in three loci based on the alignments deposited in TreeBASE (S22340): ITS position 92(T); tef1 positions 32(T), 80(C), 84(C), 154(indel), 155(A), 156(G), 157(indel), 199(A), 244(G), 261(G), 368(T), 398(T), 458(G/C), 466(G), 467(C), 475(T), 478(C), 483(T), 485(T), 487(T), 490(T) and 492(G); tub2 position 174(indel).

Culture characteristics: Colonies convex, cottony, with smooth margins, cream; reverse pale luteous; chlamydospores not seen; reaching 45–65 mm after 1 wk on MEA at 24 °C in ambient light.

Materials examined: Ethiopia, Addis Ababa, from soil, 2010, coll. P.W. Crous, isol. L. Lombard [holotype CBS 143794 (maintained as metabolically inactive culture), isotype cultures CBS 143793, CBS 143795 (also maintained as metabolically inactive cultures).

Note: None of the three isolates of C. addiensis could be induced to sporulate on MEA, PDA, OA, SNA or SNA amended with carnation leaf pieces.

Cylindrocladiella nauliensis L. Lombard & Crous, sp. nov. MycoBank MB824500. Fig. 6.

Fig. 6. Cylindrocladiella nauliensis (ex-type CBS 143792). A–D. Penicillate conidiophores. E–H. Penicillate conidiogenous apparatus. I–L. Terminal vesicles of stipe extensions. M, N. Subverticillate conidiophores. O. Conidia. Scale bars: A–D = 50 μm; E–O = 10 μm.

Etymology: Name refers to the area Aek Nauli, Indonesia, from where this fungus was collected.

Conidiophores 35–55 × 3–6 μm, dimorphic, penicillate and subverticillate, mononematous and hyaline, comprising a stipe, a penicillate arrangement of fertile branches, a stipe extension and a terminal vesicle; stipe septate, hyaline, smooth; stipe extension 100–135 μm long, 4–5 μm wide, aseptate, straight, thick-walled with one basal septum, terminating in thin-walled, broadly clavate to ellipsoidal vesicles sometimes with papillate apex. Penicillate conidiogenous apparatus 10–22 × 2–4 μm, with primary branches aseptate, secondary branches 8–15 × 2–4 μm, aseptate, each terminal branch producing 2–4 phialides; phialides 8–13 × 2–3 μm, elongate doliiform to reniform to cymbiform, hyaline, aseptate, apex with minute periclinal thickening and collarette. Subverticillate conidiophores abundant, comprised of a septate stipe and rarely primary branches terminating in 2–4 phialides; primary branches 25–45 × 2–4 μm, straight, hyaline, 0–1-septate; phialides 12–32 × 2–3 μm, cymbiform to cylindrical, hyaline, aseptate, apex with minute periclinal thickening and collarette. Conidia (10–)11–13(–14) × 2–3 μm (av. = 12 × 2 μm), cylindrical, rounded at both ends, straight, 1-septate, frequently slightly flattened at base, held in asymmetrical clusters by hyaline slime. Sexual morph unknown.

Culture characteristics: Colonies convex, cottony, with smooth margins, cream with pale luteous to brick centre; reverse pale luteous to honey with sepia centre; chlamydospores moderate throughout medium arranged in chains; reaching 60–70 mm after 1 wk on MEA at 24 °C in ambient light.

Materials examined: Indonesia, Sumatra Utara, Aek Nauli, from soil, May 2005, coll. M.J. Wingfield, isol. L. Lombard (holotype CBS H-23400, culture ex-type CBS 143792), isotype culture CBS 143791 (metabolically inactive).

Notes: Cylindrocladiella nauliensis is closely related to C. longistipitata. The stipe extensions of C. nauliensis (up to 135 μm long) are shorter than those of C. longistipitata (up to 216 μm long; Lombard et al. 2012). Additionally, the conidia of C. nauliensis are smaller [(10–)11–13(–14) × 2–3 μm (av. = 12 × 2 μm)] than those of C. longistipitata [(12–)14–16(–17) × 2–4 μm (av. = 15 × 3 μm); Lombard et al. 2012].

Authors: L. Lombard & P.W. Crous